JPS63501221A - Method for producing aloe products, products obtained by the method and compositions thereof - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Method for producing aloe products, products obtained by the method and compositions thereof Related applications This application is serial number 810.025 filed on December 11, 1985. No. 810,025 was filed on July 12, 1985. This is a partial continuation of the application with serial number 754,859, which was filed under serial number 754.859. Part of the application with serial number @750,321 filed on June 28, 1985 750.321 is a serial number filed on September 12, 1984. No. 649,967 is a partial continuation of the application filed in May 1982. This is a partial continuation of the application with serial number 375,720 filed on May 7th. Before Serial number 810,025 is a method for producing aloe products and a method for producing aloe products. The resulting product is entitled: Said Syria, number #754,859, No. 750,321, 649.967@; and No. 375,720 are aloe vera (Aloe vera) product manufacturing method.

and pl (7) January 1. Field of invention This invention processes an aloe plant and extracts parts of the plant for topical and internal use. The present invention relates to techniques for processing into dosage compositions as well as compositions containing parts of this aloe.

2. Description of the prior art and other information Approximately 325 types of aloe are known. , many of which are native to Africa. Aloe barbadensis s　is　s) is native to northern Africa and was introduced to the island of Barbados around 1630. It was. A type of Aloe barbadensis [Aloe chinensis Baker] chinensis Baker) is William Anderson (WiIam AnderSOn) from China in 1817. It was introduced in Curacao. This was during the 19th century when industry began to decline. It was cultivated in Barbados for its laxative properties until about Kura power or aloe often Also known as Barbados aloe, it comes from the Dutch islands of Aruba and Bonar. ing. The laxative aloe market will continue to grow as better and safer laxatives are developed. disappeared.

This plant contains two different juice substances.

One is made from a transparent cell gel, the other is a yellow juice and has an external appearance. Contained in the inner sheath cells of vascular bundles at the junction between the skin (skin) and the inner fiber bundles. (In Figures 1 and 2, 1 is a transparent cell gel and 2 is an anthraquinone-containing gel. 3 is the rind).

For centuries, this yellow juice has been dried and used as a laxative.

For example, in the Dutch islands of Aruba and Bonnard, leaves are cut in March and April. Place the cut pieces in a V-shaped pot tilted so that the latex is introduced into the cooking mold. The residual limb is placed facing downward. Barrow E. Tyler, Pharmacog Nosy, pp. 60-63, Law and Fuebiger, Philadelphia 198. 1 (Varro E, Tyler, PHARt4ACOGt40SY pp , Bo ~ 63 (Lea and Febiger, Ph1ladelphi a, 1981)). Aloe barbadensis mira()li　I　Ier)ma The color of the dried latex of aloe or other aloe leaves is reddish-black or brownish-black. It varies in color from dark brown to dark brown. Each taste of dried latex makes me nauseous It's bitter because it's what makes it happen. Moreover, the odor is characteristic and unpleasant. this is Contains many anthraquinone glycosides, one of the major ones being barbaroin It is called (aloe emotin anthrone C-10 glucoside). This dry la Tex has been sold as a laxative for centuries and is commonly referred to as aloe. It will be revealed.

(±) Barbaroin This active laxative component varies both quantitatively and qualitatively depending on the species and growing environment conditions. do. For example, Kura Chikara Aloe is compared to Cape Aloe (cape Al0e). It contains 2.5 times more aloe emo gain than other types of aloe. Free chrysophanic acid, which does not exist in Roe, ) in appreciable amounts (Tyler, supra).

Many companies sell aloe products that contain large amounts of yellow sap and claim to be efficacious. There is. The two types of juice are juice extraction, which is used by many manufacturers. mixed with each other by the process.

The following types of aloe are dried for their yellow sap, which is used as a laxative. has been used commercially.

Arlie Otzle et al., The United States Dispensary Agency Physicians Flumacology (J, B, Lippencott, Philharmonic) Ladelphia.

1980, pages 42-43) (ArthUr oso+ et al, Ding HEUNI ED 5 TATES DISPENSATION AND PHY SICIANS’PHARHACOLOGY, (J, B, LippenCott CO,, Ph1ladelphia.

1980) DD, 42-43) ゛1. Aloe Perry Baker (Benigumi Asahi ■汀 Kumijo Ω.

This true SOCOtrine aloe is a perennial plant, High altitudes from 0 feet to 3,000 feet above sea level, especially in limestone areas of the island of Ra. It grows abundantly throughout the country and is also found in East Africa and Arabia.

It has a trunk one foot tall, topped with brown-tipped thorns. It has densely rosette-shaped pale green or pale red fleshy, apical leaves with margins. ing. ″ ゛2. Aloe Barbadensis Mil-(A, Vera at L tt.

A. Vulgaris Lamarck). This species is the source of Kuraiki Oaroko and is very short It has a tree-like stem and is a light blue-green, tip-shaped, hugging sweet with stiff, pale thorns. I'm wearing it. It has bright yellow flowers arranged in spikes. A, Ba rubadensis is native to southeastern Europe, northern Africa, and Madagascar.

It is grown in Italy, Sicilo, Malta and especially in Western India . ″ “3. Aloe ferox mirror ) is one of three South African tree-like species that produce Cape aloe; It is one of the tallest species of this genus. C As per the original text] This is 5 to 15 feet long It has branched stems 4 to 6 inches in diameter, with a 1-long stem with barbs at the top. ．． Bearing dense rosettes of 30 to 50 5 to 2 foot long tip-shaped leaves. Several triangular species with a pure tall trunk, the top of which is edged with large horn-shaped teeth. It has square or rectangular pale blue-green leaves. It is native to the Cape Colony. Ru. ′ Pa5. Aloe Spicata Baker (A, El Valcornuta Berger) (Aloe 5 picata Baker.

(A, Eruvar, cornuta Berger)) is a tropical South African It is a tall branched aloe that grows naturally in mosquitoes.

It has pale glossy succulent leaves with white blotches and bell-shaped yellow flowers. It has panicles. ``Recent status of the yellow sap part of aloe as a laxative is in the text of Goodman and GiIman. It is best summarized as:

``This yellow sap has not been tested in controlled clinical comparisons with other anthraquinones. Although it has never been done, it is said to be the most stimulating of these laxatives. It is. This causes considerable abdominal pain and pelvic congestion, and if administered in excess, nephritis may occur. may occur. This is stated in the US patent solely for pharmaceutical reasons. ing. Both aloe and aloin, which are mixtures of active glycosides, should be discarded. be. ″ Gutsudman and Gilman, eds., The Pharmacological Basis of Sera. Beautics, 984 (Macmillan Publishing, New York, 1975) ( Goodman and Gilman, Eds, THE PHARHAC OLOGICAL BASIS 0FTHERAPEυTlC5, at 984 ．． (Mac old 11an Publishing Co.

Inc., New York 1975)).

Aloe is characterized by its spear-shaped leaves with jagged edges and sharp tips. It is a tropical or subtropical plant that is cultivated. This plant has been healthy for centuries It is often thought that it has therapeutic effects, and the basics of such properties are It has been used without a clear understanding or scientific analysis of its meaning. Cobble ( No. 3,892,853 to Cobble and Coats No. 4,178,372. These two patents are for aloe vera. Stabilized by extracting mucilage from the leaves and adding a mild oxidizing agent (Machi 02) teaches a method for producing aloe juice that is said to be bacteriologically stable (i.e., bacteriologically stable) are doing.

These patents include removing yellow sap, or due to the presence of yellow sap. No mention is made of any untoward properties in aloe juice. Actually゛8 Both the 53 and °372 patents show that the unfavorable properties before treatment are ” ('853 patent, column 3, lines 41-43, ° 372 patent, column 3, lines 42-47). Also, extracts and derivatives of the aloe plant Attempts to manufacture various commercial products have met with varying degrees of success and failure. I've been doing it.

The problem to which the invention is directed This lack of knowledge about the aloe plant and its characteristics has led to this Previous methods used to treat plants and their components are those of Gutdman and Due to the presence of a yellow sap known by Gilman to have laxative properties For a method that produces a final product that does not consistently give the desired results, use a bifurcated aloe infusion. Grind the whole leaf (pressure roller) or grind it (e.g. Thompson aloe leaf slitter) Aloe Vera Juice Make an Aloe vera juice and then filter this juice. It includes various steps of stabilization. Next, add the obtained solution to other solutions. or mixed with reagents, for example in cosmetics, health food drinks, or topical cartilage. The company was creating the desired final product that could be used.

The main drawback of such traditional methods is that they conflict with the intended use of the final product. In addition to Not realizing that the various components of aloe leaves have sexual properties. , and not taking this into account.

For example, research conducted in conjunction with the process of the present invention has shown that certain components of aloe vera is cytotoxic, U.S. Pat. No. 3,892,853, while other components are cytotoxic. Aloe leaf ingredients are used advantageously in the manufacture of beverages to stimulate the growth of It has been found that it can actually be harmful when used in construction. winters Winters et al. The cytotoxic effects of these fractions on normal human cells and cultured tumor cells are The formulation may have altered levels of lectin-like activity introduced during the commercial process; Contains substances that can significantly disrupt in vitro adsorption and growth of human cells. We conclude that this suggests that Id　page 95. Winters et al. To the Applicant, he disclosed that Coat as a commercially stabilized aloe preparation. It is reported that materials obtained from s) were used.

Winters et al. Although beneficial for cell growth, traditional commercial aloe preparations are actually toxic. In contrast, the stabilized Aloe vera glu fraction was The researchers found that the drug was equally cytotoxic to normal cells and tumor cells. did. [W, D., Winters et al. Aloe extract has been shown to be effective against human normal cells in vitro. ``Effects ofal'' oe Extract on Human Normal and Tumor Ce1ls InVitrO”, ECO, BO DING ANY) 35 (1) 89-95 (1981)).

This is obtained from virtually all leaves (or at least most of them) of the Aloe Vera plant. Juice containing an impure mixture or compound of various characteristic parts of leaves or Products obtained by using conventional processes for producing extracts necessarily , which is useful for one end use but actually pollutes for another end use. It contained the constituent ingredients. If such contamination exists, does it constitute a hazardous substance? If not, at least the resulting juice is desired for some other end use. It constitutes a diluted mixture of ingredients rather than a concentrate with the properties of It becomes.

More recent research has shown that the yellow sap is also toxic to human skin cells and Ipan E, Danhoff et al., “Stabilized Aloe Vera: Effects on Human Skin Cells” Hibiki “Drug and Cosmetic Industry (IVanE, Dan hof et at,,”5 tabilized Aloe Vera: E ffect on Humann 5kin Ce1ls”, DRUG AN D CO3ME ICIND) 52-54, 105-106 (August 1983 month) (not recognized as prior art to the present invention), and that it has been shown to be minimized in topical products that come into contact with Ru. Idiosyncratic in processed aloe vera gel consisting of a diverse mixture of yellow sap Hypersensitivity was demonstrated. Dipid, M. 70-M, D., Ra' Hypersensitivity to Aloe ", ARCH Dermatol (DaVid) 1. MOrrOW, I (, D ,, etal, “Hypersensitivity to Aloe”, ARCH, DERMATOL, ) 116, 1064-1065 (1980 (September). The adverse effects of yellow sap aloin on human tissue cultures are 40. This is consistent with the inflammatory reaction to the intestinal mucosa reported more than a year ago. Melvin W. Green “Stimulating effect of aloin, preliminary low note, American fur Mathematical Association (Melvin W, Green, “Th e Irritant Effect of Aloin Preliminar y Note", AMER, PHAR,, ASSOC,) 30186-1 87 (1941). The problem is the yellow sap (aloin) content of each plant. is made all the more difficult by the fact that It will be terrible.

E, R, Jansz et al. “Aloin content of local allografts” (E, R, Jansz et al, “The Aloin Content of Local A loeSpecies''', J, NATN, SC1, C0UN, (S ri Lanka)] 9(1) 107-109 (1981). Jans et al. Aloin (yellow sap) was "commonly used as a laxative." is reported.

A second problem with treating aloe vera plants is the effect of darkness on the acid content of the leaves. This is due to the influence of In the field, leaves are collected and stacked while awaiting processing. bottom The leaves of the A0 plant on the side are kept in a dark state. Items with leaves are processed before being processed. It is kept in the dark for several days to several weeks. In a matter of time the acid content can rise several times . F, R, Parcha, prolonged darkness is acid metabolism in aloe veraline leaves. F.R.Bharucha, “Effect Of: pr o+onged Darkness on Ac1d) Ietabalism in the Leaves of Aloe Vera Linn”, SC 1, AND C0LT, ) 22(7) 389-390. this is aloeve (Too much acid may cause burning or irritation to the skin.) (can cause irritation).

Cora's "372 patent uses aloe plants to minimize changes in the gel material. It teaches that plants should be grown under controlled conditions. but controlled Even under such conditions these materials can change by more than 100%. It has been processed The variable mineral content, i.e. the variable salt content, in aloe juice is Lack of consistency in consistency (e.g. viscosity) and health reasons (@cotton products, e.g. Stinging sensation due to large substantial variations in salt in ashampoo compositions In many applications, it is harmful to the production of the final product due to the presence of heat and irritation). Made of aloe The chemical stability of the product also changes the mineral content of the extracted aloe juice. influenced by. Too much salt in the extracted aloe juice may cause the topical final Layer separation can occur in aloe formulations.

This third problem is that the mineral content of the raw aloe plant varies depending on the season. Consistency in aloe juice compositions on the basis of varying Swast Institute for Natural Sauces Southwest In5 posture For Natural 5our ces) became the National Aloe Rhinoceros on or about November 11, 1982. The National Aloe 5science Council This has been announced through a publication report submitted to the Council. This report is incorporated herein by reference in its entirety and this report is not controlled. Even in the O Grande Valley (Rio Grande Va. Varying mineral concentrations found in aloe leaves collected from It shows. Sodium and chloride ion concentrations vary in the hundreds depending on the time of collection. The percentage also changes. Aloe is a succulent plant that absorbs minerals from the soil. . During the rainy season, the plant is hydrated by rain that has low mineral concentrations. .

However, during the dry season, the field is flooded with water containing much higher concentrations of minerals. be irrigated. Fertilizer also replenishes minerals to the soil and affects the mineral content. obtain. The present invention provides that the salt is present in excess concentration to produce a certain final product. In some cases this problem was overcome by dialysis.

The fourth problem: the strength of products made from extracted aloe juice. Undesirable changes in (tonicity) are caused by the first three problems mentioned above. It is something that will come. That is, (1) Aloe vera sweets vary in amount and content. (2) the acid content within the leaves, i.e. Malay The acid content is determined by the amount of time the leaves are exposed to light and kept in the dark after being cut. and (3) the mineral content varies depending on rainfall and treatment. The difference is that it changes depending on the fertilizer. These variables, like the mucilage content of aloe, Eel affects all ingredients of aloe vera gel. The rate of decomposition of this mucus is content, and this change further alters the size of the mucus polymer during processing. This will cause the situation to change. All these variables are determined by the extracted aloe vera juice. Affects osmotic pressure. Strength is the degree to which a product transfers moisture to and from the skin. Determine. Osmotic pressure affects the strength of aloe juice F. Guianon, Review of Medical Physiology) (Lang Me Dical Publications Los Altos, CA) (William F. Ganong.

REVIEW OF MEDICAL PHYSIOLOGY (Lange ) Iedical Publications, Los Altos, CA) ) 16-29,984 (1983), this may depend on the nature of the leaves used. can yield isotonic, hypertonic or hypotonic aloe products.

The fifth problem associated with aloe vera is that the active substances break down through hydrolysis. That's what it means. This decomposition is timed by the presence of water in the form of a stabilized gel. It can happen over time.

Henry H. Cobble (U.S. Pat. No. 3.89) 2,853 @) and Billy C, Coats (USA) National Patent No. 4.178.372) describes the treatment of aloe leaves with the following common characteristics: It is clear that the above problems are not addressed in the traditional process for be.

1. They added a mild oxidizing agent from the catalytic converter to fresh aloe vera gel and mixed it around 5 ℃ to about 80℃.

Cobble, column 2, lines 65-68 Cola, column 2, lines 13-16 2. The preferred oxidizing agent used for catalytic oxidation is hydrogen peroxide.

Cobble, column 3, lines 16-18 Cola, column 3, lines 43-45 3. Both prefer plants that are 4 to 5 years old.

Cobble, column 2, lines 31-33 Cola, column 2, lines 39-40 4. Suppress catalytic oxidation using non-toxic antioxidants such as tocopherols. There is.

Cobble, No. 101. Lines 8-9 Cola, column 5, lines 28-31 5. Their mild oxidation affects beta globulin proteins and possibly alpha globulin proteins. Wait a long time to completely oxidize some kind of gel substance, which appears to be Robulin protein. ing.

Cobble, column 3, lines 41-44 Cola, No. 31. Lines 43-47 6. Bill Cora designed for the treatment of orange juice in his process A commercially available machine is used.

Cone, 3rd@, lines 5-9 7. Neither Cobble's process nor Corn's process collects yellow sap; I haven't even saved this.

8. Even if the yellow sap contaminates this product, the disclosed process does not provide any means to remove this.

9. Aloe verage made by Kohn's process and Cobble's process Laboratory analysis of the yellow sap includes the amount of yellow sap, mineral content, color, taste, and gel content. Varies by hundreds of percent in consistency and osmolarity (strength) It showed stabilized aloe veraglut.

In general, both cobble and corn aloe vera processes produce a yellow sap with a slimy part. It does not provide a means to collect, store, or preserve the waste separately. subordinate Therefore, when separating the slime from the leaves, there is a greater tendency for it to be contaminated by yellow sap. Hey. Their process requires heat and oxidation, which is desirable for aloe vera gel Both components will be oxidized as well as the undesirable components. Furthermore, heat causes the gel to decompose. Facilitate. Their method cannot adjust for variable mineral content In addition, the aloe vera gel matrix molecule contains substantially acetylated mannan ( (which affects the humidifying properties of the gel) cannot be sorted and separated by size. stomach. This process is also most effective on mature 4- to 5-year-old leaves. be. This is a disadvantageous condition. This is because mature leaves are destroyed by freezing. Such freezes have occurred in the Rio Grande Valley in the last five years. This is because it happened three times. The current state of the aloe processing industry in the United States Currently, domestically grown mature leaves are virtually impossible to obtain.

Generally speaking, one problem in this technical field is the identification of effective alloys for the application. This means that the two parts or chemicals in Evera have not been completely identified.

For example, U.S. Pharmacist, August 982. John Fisher published on pages 37-45 of the issue. er) says, “In contrast to aloe latex, aloe gel usually contains Contains only small amounts of laquinone glycosides.

However, a number of substances have been identified in this gel. These substances include monosaccharides, Polysaccharides, tannins, steroid compounds, various organic acids and enzymes, saponins, vitamins such as iron, copper, calcium, magnesium, sodium, potassium Includes minerals. The contribution of these substances to the reputed activity of this plant remains unknown. ``This was recently reported in the Berkeley Medical Newsletter. (Berkeley Medical Newsletter) and so far no one has identified its uses and It was not possible to link it to any specific ingredients. what this active ingredient is knowledge, isolation of the substance, and quantitative administration of the prescribed amount of the substance. It is necessary to do so. For example, aspirin is obtained from willow bark; No one knows how much you have to chew to get a certain amount of aspirin. stomach. However, if aspirin was extracted from willow bark, a certain amount of aspirin could be ingested consistently. This form of aspirin is stable.

Aloe leaf fractionation has been the source of numerous publications in this field. do not have. Purely academic, not related to the usefulness of using fractionated parts of Aloe A paper was published by the Indian team of Handal and Das (Handal and 0aS). represented.

(1) Gaurhari Mandal and Amarend Das “Aloe Barbadensis” Structure of glucomannan isolated from Miller leaf'', Elsevilla Sciente Fifik Publishing, Amsterdam, 1980 (Gauhar i Handal and Amalendu Das, “5structure of the Glucomannan l5olated from the "Leaves of AIOe BarbadenSiS Miller", (Elsevier Scientific Publishing Compa New York, Amsterdam, 1980)) 87 pages 249-256 (2) Gaurhari Mandal and Amarend Das “Aloe Barbadensis” Structure of D-galactan isolated from mirrors”, Elsevilla Scientif Publishing Co., Ltd., Amsterdam, 1980 (Gaurhari) ) landal and Amalendu Das, “5structureo f the D-Galactan l5olated from Aloe ( 3) Gaurhari Mandal, Rina Ghosh and Amarend Das, “Aloe Barbadensis Miller's properties of polysaccharides, Part II, Structure of acidic oligosaccharides'' (I Indian Journal of Chemistry, September 1983 (Gaurh ari Handal, Rina Ghosh and AmalenduD as,Characterization of Polysaccharid es of Aloe Barbadensis Miller: Partl l-3structure ofan Ac1dic Oligosacchar ide”, (Indian Journal) (3) above is a predecessor to the present invention. It is not recognized as a process technology.

Ordinary "L5" L shellfish FIELD OF THE INVENTION The present invention relates generally to the production of purified aloe products and is described in general detail. improved the aloe plant by separating the various components of the leaves of the aloe plant. Regarding the treatment method, in detail, it is pH stable and the salt content is virtually constant. An improved version of the aloe plant that is capable of achieving a given level of strength. The present invention relates to a method for producing a mucilaginous, substantially anthraquinone-free extract.

The present invention also provides a method for adding active chemicals in aloe plants to whole leaves or said improved viscosity. A second method of physically extracting from a liquid, substantially anthraquinone-free extract. directed towards the law. According to this process, the chemicals are further freeze-dried and And if necessary, by irradiating this with gamma rays or microwear. Sterilized. This form of chemical is virtually non-degradable and can be administered in defined amounts. I can do it.

The present invention also provides for the activity in aloe plants in the form produced by the second process described above. directed towards chemicals. This active chemical is an acetylated mannose monomer is a virtually non-degradable lyophilized ordered linear polymer. It was. The mannose monomers are preferably linked to each other by β(1→4) bonds. has been done. This active chemical is produced by analytical chemistry and bioassay methods. measured, standardized and characterized.

Used for gels, fillets, extracts, etc. ``Substantially free of anthraquinone'' I mean full. My invention mucilaginous substantially anthraquinone An extract containing less than about 0.00101% by weight of anthraquinone.

``The edict for pH stabilization allows the pH to be reproduced within 0.5 pH units over a three-year period. from a given Aloe fraction and variety grown in a given region, with a pH of I mean extract. By the words ``substantially free of salt content'' , the extract (from a given Aloe fraction and variety grown in a given region) is 3. I am interested in a property that exhibits a salt content that is reproducible within ±20 ppm over a period of 20 years. I'm tasting it. By the term “given strength” we mean ±20m0 over a period of 3 years. I consider the strength of an extract from a given aloe fraction to have a reproducible osmotic pressure of 5 m Taste. The word “active chemical” refers to the wound healing and wound healing properties of aloe vera. I mean substances that are effective for sterilization and other beneficial properties. “Substantially non-degradable” According to the terms, substances whose molecular weight decreases by less than 5% over 2 years and I mean a substance that retains more than 95% of its biological activity over a period of years. “substantial The term ``mannan acetylated'' refers to partially or substantially completely acetylated mannan. I mean fully acetylated mannose monomer.

For example, topical products are compatible with the skin's natural acidic mantle (pH about 4 to about 6). It is desirable to have a pH stable product that is

In the case of damaged skin, this pH is roughly equivalent to serum or plasma (pH approximately 7.4). It should be very similar. This level varies based on the intended use. Tailored for skin care products. The total salt content is about 100 to about 500 million It should be such that it produces an osmotic pressure of Smol (m05m). For example, oil For normal skin, an isotonic to hypertonic solution (280-500 mOsm); normal skin is 180-380 m05m; dry skin is 100-280 m03m Requires solution.

In rough detail my inventive method essentially extracts anthraquinones from the leaves of the aloe plant. This method has the property of extracting aloe gel that does not contain It includes the process of

(a) Wash aloe leaves in a disinfectant solution to remove virtually all surface dirt and (b) removing at least the first end of the washed leaf; Steps to remove: (C) Drain and collect the anthraquinone-rich sap from the washed and cut leaves. and (d) removing the outer skin from the leaves to substantially collect the anthraquinone. The process of producing a gel that does not contain

``Removes dirt and bacteria from virtually all surfaces'' means ( 1) Remove dirt until the remaining dirt is less than 0.1% by weight of the leaf weight. and (2) less than 100 bacteria per gram of leaves remain on the surface. It is meant to kill surface bacteria.

A person skilled in the art will understand that instead of steps (b), (C3 and (d)) (b) crush the washed aloe leaves and (C) dialyse the crushed leaves to preferably chemically removes both components i.e. anthraquinones, minerals and acids, and It is said that the skin can be removed to create a gel that is virtually anthraquinone-free. You will recognize that. More preferably, the aloe leaves or the whole plant are can be collected from fields that are sufficiently clean that the washing process can be omitted; The aloe vera extract is extracted by crushing, squeezing and/or extrusion to obtain the desired aloe fraction. Components, e.g., anthraquinone-free components, are tested for dialysis or restriction as described herein. Extracted from whole juice using external filtration. This juice fraction contains each component separated and subsequently remixed in any desired combination, as desired, as described herein. stabilized so that

Preferably, the substantially anthraquinone-free juice is produced from the leaves of the aloe plant. My method of building includes the following steps: (a) Clean aloe leaves in a disinfectant solution to remove dirt and grime from virtually all surfaces. (b) removing the first end and the second end from the washed leaf; the step of removing the part; (C) Drain the anthraquinone-rich sap from the washed and cut leaves, The process of collecting; (d) aloege substantially free of anthraquinone by removing the outer skin from the leaves; a step of manufacturing a fillet; and (e) crushing the substantially anthraquinone-free aloe gel fillet; A process of homogenization to produce a juice substantially free of anthraquinones.

My preferred process is to add aloe juice or aloe vera 51) l) to osmotically adjust the concentration of anthraquinone or to roughly adjust the concentration of anthraquinone. Ultrafiltration is required to reduce the concentration to less than 100 ppb or even less than 100 ppb. It may include degrees.

These processes allow processors to process large leaves, small leaves, or leaves less than one year old. allows you to use This is due to the polymer size found in mature leaves. This is because they can be sorted and processed from smaller, immature leaves.

One of the advantages of this process is that the Being able to treat damaged leaves, which was previously thought to be impossible, and Undesirable contaminants can be removed by dialysis.

Most preferably my method is to roughly filter the aloe juice to remove the fibrous material. It is assumed that In this form, aloe juice can be taken internally and It can be incorporated into a huge variety of products for external use.

The process of my invention further includes the said aloe glufillet as a preferred fruit 1H. or the aloe juice or the aloe vera fraction may contain preservatives, fragrances or Any FDA-approved additive (Generally Recognized as Safe, i.e., “GRA S” (generally recognized as 5afe) additive) The composition may further contain one or more additives selected from the group consisting of:

Most preferably, the preservatives are benzoic acid, potassium sorbate, methylparaben and and propylparaben or other FDA approved for topical or internal use. additives selected from the group consisting of:

The juice of my invention can be irradiated, freeze-dried, sonicated, pasteurized or or pasteurized, thereby preserving the juice .

This ultrafiltration (dialysis) process involves membrane technology. This membrane technology Allows selection of filters with different pore sizes depending on leaf condition and any combination of the following can be achieved: (1) allo A small pore size filter (preferably approx. 100 daltons).

(2) Large pores that can separate acid from aloe vera gel if necessary size filter (preferably about 500 daltons).

(3) The yellow sap component can be separated from the aloe vera gel if necessary. A very large pore size filter (preferably around 2000 daltons).

(4) and gel matrix polymers, and divide them by molecular weight. A filter with a roughly large pore size (preferably about io, o oo~100,000 Dalton).

As an ultrafiltration device, Romicon 4-column (Romicon, 100 Cummings) Burn, Woburn, HA 01801. Model NO, HF4 SSS , Membrane Type P)l 50.30 Membrane Plan No., 11526.5-43 5-43-p (Romicon 4-column (Romicon Co ,.

100 CuCumm1n Park, Woburn, HA 01801. )todel No, HF4sss, Membrane Type PH 50, 30 Membrane Nos, H526, 5-435-43-p) is recommended.

In another preferred embodiment, the washing step of the process breaks up the fillets. substantially anthraquinone-free solution in a tumbler washer prior to The method may include a step of washing the Loegle fillet.

An effective skin wound gel can be produced by the process of my invention. , preferably from 0.1 to 100% by weight of substantially anthiraquinone-free aloe juice, O~2 heavy weight allantoin, 0~6% by weight panthenol, and Excipients or carriers may be included.

See Example 7. The concentration of allantoin and/or panthenol is F It is regulated by the DA. Twin Merman, The Essential Guide Two Nonprescription Drugs (Harbor and Row New York, 1983) (Zimmerman, THE ESSENTIAL GUI DE　TONONPRESCRIPTIONODRUGS　(Harper　a nd Row, New York.

1983)).

The second process of my invention is to extract the active chemicals in the aloe plant from the leaves of the aloe plant. This process includes the following steps: (a) Wash aloe leaves in a disinfectant solution to remove virtually all surface dirt and (b) removing at least a first end from said washed leaf; (b) removing bacteria from said washed leaf; a step of removing; (C) draining the sap containing a good amount of anthraquinone from the washed and cut leaves; storing and collecting; (d) removing the outer skin from the leaves to substantially remove the anthracite; A step of producing an aloe gel fillet that does not contain non-alcohol; (e) Intraquinone-free aloe gel fillets were crushed, homogenized, and dissolved A process for producing a substantially anthraquinone-free aloe juice comprising the substance; (f) Add a water-soluble lower aliphatic polar solvent to this aloe juice to produce active chemicals. precipitating a substance, thereby producing a heterogeneous solution; ((1) Water-soluble lower aliphatic polar solvent and dissolved substances from the heterogeneous solution removing the substance and isolating the precipitated active chemical; and (h) Drying the precipitated active chemical.

Those skilled in the art will understand that instead of steps (b), (C) and (d), (b) Crush the washed aloe leaves, and (C) dialyze the crushed leaves to separate the undesirable two parts. That is, anthraquinones, minerals and acids are chemically removed, and the outer skin is removed. to produce a substantially anthraquinone-free gel, which is then subjected to step (e). (f), ((1) and (h) can be used to extract the active chemical; You will recognize what I mean.

A person skilled in the art will also understand that steps (b), (c), (d) and (e) can be replaced by and crushed the washed aloe leaves to extract an anthraquinone-rich solution containing dissolved substances. Extrude aloe juice, then pass this aloe juice through steps (f), (0) and It is recognized that active chemicals can be extracted by Probably. All anthraquinones and all ions are water soluble and in liquid form. This process allows the active chemicals to remain in the solvent phase and not precipitate. Effectively separated from laquinones and harmful ions.

Those skilled in the art will also recognize that alternatives to steps (b), (c), (d) and (e) First, crush the whole washed aloe leaf, filter out the fibrous material, and homogenize the residue. to produce anthraquinone-rich aloe juice containing dissolved substances. We need to be aware of what we can do. This aloe juice is then added to the step (t'), ((1), l"l' and (h)) to extract the active chemical.

The active chemicals are converted into anthraquinones by this process for the reasons mentioned above. and are effectively separated from harmful ions.

A person skilled in the art will be able to extract the active chemicals in the Aloe plant from the leaves of the Aloe plant. It will be appreciated that yet another method of producing :(a) Clean aloe leaves in a disinfectant solution to remove virtually all surface dirt. Step of removing bacteria; (b) removing the outer skin from the leaves and removing the aloe grophyte; The process of manufacturing lettuce; (C) Crushing and homogenizing the aloe gel fillets to obtain an aliquot containing dissolved substances; Process of producing loe juice; (d) water-soluble lower aliphatic compounds in this aloe juice; A polar solvent is added to precipitate the active chemical, thereby forming a heterogeneous solution. process; (e) Remove the water-soluble lower aliphatic solvent and dissolved substances from this heterogeneous solution. isolating the precipitated active chemical; and (f) Drying the precipitated active chemical.

As mentioned above, all anthraquinones and all ions are water soluble. This process allows the active chemical to remain in the liquid solvent phase and not precipitate. The substances are effectively separated from anthraquinones and harmful ions.

``'Removes dirt and bacteria from virtually all surfaces'' means (1) ) removing dirt to the extent that the remaining dirt is less than 0.1% by weight of the leaf heavy equipment; and (2) less than 100 remaining surface bacteria per gram of leaf. It is meant to kill surface bacteria.

Preferably all of the above extracting the active chemicals in the aloe plant from the leaves of the aloe plant. In all processes, four times the amount of water-soluble low-grade The active chemical is precipitated by adding an aliphatic polar solvent. Preferred water-soluble lower aliphatic Polar solvents are methanol, ethanol and propatool. most preferred The solvent is ethanol.

Those skilled in the art will appreciate that the above-mentioned preferred Other water-soluble lower aliphatic polar solvents can be used in place of the white solvent. You will recognize that.

In the above extraction process, the active chemicals are mixed with a water-soluble lower aliphatic polar solvent and aloe Preferably, the juice mixture is left to settle for about 4 hours. This mixture 4. Allowing the compound to settle for 4 hours provides an optimal yield of the active chemical; It has been found that over time, the precipitated active chemicals begin to decompose. But 2 It was also found that a significant amount of active chemical was recovered after a 4 hour precipitation period. It was.

Those skilled in the art will know that the optimum precipitation time is ambient temperature, pressure and water solubility. It will be appreciated that it depends on the nature of the aliphatic polar solvent.

In the above extraction process, heat can promote hydrolysis and decomposition of active chemicals. , the precipitated active chemicals are dried by freeze drying rather than oven drying. This is desirable.

In all of the above extraction processes, any fiber present in the aloe juice (cellulose) is also precipitated by water-soluble lower aliphatic polar solvents (pre cipitated), which is precipitated early and activated by the addition of this solvent. It has a lower density than chemical substances. This fibrous material is formed after the active chemicals are allowed to settle out. , stops at the surface of the solvent. Therefore, it can be removed very easily. this One skilled in the art would know that before adding the solvent, one could alternatively filter the aloe juice and add fibers. You will realize that it is possible to remove such substances.

Preferably, in all of the above processes the aloe leaves or whole plant are , collected from fields that are sufficiently clean that the washing step can be omitted.

The dried precipitated active ingredient is irradiated with gamma rays or microwaves if desired. It can sterilize and preserve sexual chemicals.

The present invention therefore provides a new and improved method for the production of aloe vera products. It is something to do.

The present invention further provides for the prevention of undesirable combinations of distinctly characteristic parts of leaves of aloe plants. Provides an improved method of treating aloe vera plant leaves to prevent mixing It is something to do.

In summary, the present invention minimizes the concentration of undesirable components in the finished extract. Provides an improved method for producing various extracts of leaves of Aloe vera plants It is something.

The present invention also provides a composition for characterizing a particular part or segment of a leaf of an Aloe vera plant. the constituents that maximize the concentration of constituents that characterize other parts or segments of the leaf; Manufactures an extract of the leaves of the aloe vera plant that minimizes or eliminates harmful substances. The present invention provides a new and improved method for

The present invention also provides an aloe vera plant with a low concentration of yellow sap on the leaves of the aloe vera plant. provides a new and improved method for producing extracts from the leaves of Ru.

My preferred process has the following advantages over the prior art: 1. Since catalytic amounts of mild oxidizing agents are not used, oxidation of desired components is prevented. .

2. This process does not require heat, so it can be done at room temperature.

36 Young immature or damaged leaves can be used.

4. There is no need to use antioxidants to suppress catalytic oxidation.

5. β-globulin and α-globulin are removed by dialysis.

6. Both manual and mechanical extruders can be used.

7. This process allows the yellow sap to be collected, stored and preserved separately. (Yellow sap is prohibited as a laxative, but is recommended for use on undamaged skin.) It can be used as a sunscreen. Additionally, it imparts a brown color to the skin. ).

8. If yellow sap gets into the aloe vera gel, remove it. Can be done.

9. Content of the yellow sap, mineral content, color, taste, gluconcystin Providing aloe veraglut standardized in terms of strength and permeability (strength) can.

10. By isolating, dialyzing, and concentrating each component of aloe vera gel. can be concentrated to many times higher concentrations than found in natural juices. .

11, and by separating this fraction, it is possible to identify A new combination of aloe vera gel ingredients can be obtained.

12. Adding too many preservatives, fragrances or other GRAS substances If the batch is The case can be saved.

13. After removing selected aloe components by dialysis and chemically changing them, can be added.

14. Add the enzyme to aloe gel, let it react, and then separate it by ultrafiltration. Can be done.

Finally, the present invention provides a method for extracting active chemicals in aloe vera gel. It is. This active chemical is referred to below as Carrisyn (Trademark, Trademark). ). As mentioned above, kallidin is suitable for analytical chemistry and bioassay. The actual amount of acetylated mannose monomer measured and standardized using the method of It was found to be a non-degradable freeze-dried ordered linear polymer.

Drawing description Figures 1 and 2 show cut sections of Aloe vera leaves.

FIG. 3 is a schematic diagram of a preferred leaf cleaning device used in my method.

FIG. 4 shows a schematic diagram of a preferred apparatus for cutting and steeping Aloe vera leaves.

Figure 5 shows a preferred apparatus for shredding aloe cuts into fillets and crushing them. A schematic diagram is shown.

Figure 6 is a schematic of a preferred apparatus for fine homogenization and (optionally) filtration. Show the diagram.

Figure 7 shows the preferred use of a dialysis device to further separate the treated aloe material. 1 shows a schematic diagram illustrating.

Figures 8-13 show infrared spectra of various samples of kallidin.

Figure 14 shows the infrared radiation of raw aloe gel cast into a film. Showing the spectrum.

Figure 15 shows the infrared spectrum of one sample of Kallijin.

Figures 16-17 show ``When aloe gel is removed from the leaves and when aloe juice is added to the leaves.'' Calibration when the interval between extraction by call is 1 hour and 10 hours Infrared spectrum of a sample of gin is shown.

Figure 18 shows the reciprocal of the time the gel is present in alcohol (precipitation process). The yield of kallidin is plotted.

Figure 19 plots the yield of kallidin against the reciprocal of the total process time before precipitation. This is what was done.

Figure 20 plots the yield of kallidin against the reciprocal of the homogenization and filtration times. It is something that

Figure 21 shows the extraction of juice from Aloe ferox. Figure 2 shows an infrared spectrum of alcohol precipitation products.

Figure 22 shows the juice from Aloe africana. shows the infrared spectrum of the alcohol-precipitated product of the base.

Figure 23 is from A4ricana ferox. shows the infrared spectrum of the alcohol precipitation products of the juice.

Figure 24 is Aloedicot? Juice from (Aloe dichotoma> shows the infrared spectrum of the alcohol-precipitated product of the base.

Figure 25 shows the infrared spectrum of cellulose.

Figure 26 shows the infrared spectrum of polygalacturonic acid.

Figure 27 shows infrared spectra of glucomannan from Konjac plant. It shows the vector.

Figure 28 shows the infrared spectrum of dextran.

Figure 29 shows the infrared spectrum of guar gum.

Figure 30 shows the time course of release of 510r by topical reagents. human fibroblast Cell culture tissue was treated with 0.05% Granulex, Hibikku. ens (quadruple CI ens), Betadine or kallidin ( CDWG) and incubated for various times and the percentage of total radioactivity released. was measured. Data are averages of 3 to 5 separate measurements at each time point. be. Control cells (treated with medium only) lost a total of 51 c cells in 30 minutes. , released 3-5% of .

Figure 31 shows the effect of concentration on cell damage.

Cultured fibroblasts were treated with Hibiclex, Granulex, Betadine or Kali. gin (CDWG) at various concentrations for 15 minutes at 37°C]. Ta. The percentage of 51cr released was determined for each concentration. contrast The release of (from untreated cells) ranged from 1 to 5%. Data is separated from 3 to 5 times. is the average of each measurement.

Figure 32 shows the effect of CDWG and serum on chromium release. The cells are Medium only (shaded bar on the right) or medium containing kallidin (0.5%) (hatch) (bars with black dots) or a medium containing 10% tallow serum (bars with black dots) for 15 minutes. Incubated. Betadine, Granulex or Hibiclex (0,15 %) and the incubation was continued for an additional 15 minutes. The data is from 3 to 4 times. Average of independent experiments.

Detailed Description of Another Preferred Embodiment In accordance with these and other objects, the process of the present invention utilizes the characteristics of Aloe vera leaves. distinct parts of the body, specifically the yellow sap, the internal gluma matrix and the outer skin. , each exhibiting the characteristics peculiar to these parts, and the particularity of each of them. Features of the separable parts may aid in their use for specific applications or end-use products. and that the characteristics of another distinguishable part are unfavorable for such use. arising from the discovery and recognition that something is or is harmful. For example, inside Extracts from gel matrices are highly desirable for the production of cosmetics or skin care products. However, the yellow sap has some particularly harmful properties for such applications. I found out that they have a sexuality.

Furthermore, I noticed that the yellow sap and the sub-porti of the internal gel matrix ons) have characteristics characteristic of these subdivisions, and therefore that they discovered that the extracts of these two species have potentially distinguishable uses, and I recognized it. I have discovered that these distinguishable parts, some of these characteristics are also A summary of the and potential applications is as follows.

Part Detail Use Yellow sap (1) Sediment Laxative, fungicide, antibiotic o- gical), insecticides and sunscreen (2) Supernatant liquid: mucosal protective effect, sunscreen Internal gel (1) Viscous liquid penetrant, hypoallergenic agent Mad 1 Nox (hypoa lleraenic), wetting agent (2) Gel Ulcer protectant, cells 7-rLzyto Stimulant, wetting agent, wound healing (3) Interstitial fibers Natural preservative, hemostasis agent (4) Residual cell growth stimulant matrix Outer skin Insecticidal insect repellent, paper pulp fiber Based on the above knowledge and recognition, and depending on its intended use, the final extraction In order to optimize the quality and concentration of desired components in the output, the process of the present invention The leaves of the Aloe vera plant are first fractionated into specific distinguishable parts and subdivisions as described above. and for separating and isolating specific components of said subdivisions. There is. Specific details and characteristics of such processes can be found in the detailed description below. It will be easier to understand and recognize. The present invention also provides the above-mentioned specific components isolated by the process.

Note: Big 1 Danesu The extract produced by the process of the invention is preferably a mature, open-air grown Obtained from the lower outer leaves of grown Aloe vera plants. 2 year old plants are Always mature. However, the wider leaves of 4- to 5-year-old plants provide the desired extract They generally contain larger quantities of and are easier to handle. Norio, Texas The 4- to 5-year-old leaves of Aloe barbadensis Miller, which grows in Grande Valley, are Most preferred. These leaves generally weigh about 1.5 to 2.5 pounds each. be. Depending on the specific use or product desired, these leaves are removed from the plant. can be processed immediately after cutting, or they can be processed in various ways before being processed. Can be stored under suitable conditions for a period of time. Zarani, the concentration of various components in the leaves is influenced by seasonal fluctuations and environmental conditions to which the leaves are subjected. These are all should be considered according to the specific intended use to which the plant extract is to be directed. Must be.

The leaves are grown on the plant, preferably without damaging any part of the leaves prior to treatment. should be cut or removed near the base. preferably a pocket knife Cut the leaves at the base just above the stem using a small knife, less than 6 inches long, such as , to prevent clear cell gel leakage or gel contamination by yellow sap. Strip the leaves from the stems. Any damage to the leaves may result in an undesirable mixture of discernible parts. resulting in an undesirable mixture of leaf characteristic components.

After removal from the plant, the leaves are usually washed with a solution of a suitable detergent (for example we use York Olympic Pool Crawl 65 sold by Mikel (Dallas, Texas) ) (OLYHPICPOOL CHLOR65) is preferred. ) gently Cleaned by scrubbing or spraying.

Sometimes this cleaning is done using a soft brush.

After cleaning, the leaves are thoroughly rinsed in clean water to remove traces of the detergent solution.

The bottom of each leaf is white or light colored and the tip has a small sharp knife. Removed by careful cutting with a knife. substantially forming both ends of the leaf These parts can be processed separately to obtain a yellow sap from them, and this Yellow sap is produced by producing a product with ingredients that have the characteristics of the above yellow sap. is directed to the desired use.

The remaining portion of each Aloe vera leaf is then cut transversely into short pieces, preferably long pieces. Cut into 0.5 inch pieces and each piece can be hypertonic, isotonic or hypotonic. Place the pieces upright in an aqueous solution (preferably deionized water) to remove the yellow color from the pieces. Drain the colored juice. Alternatively, for use in other formulations with the above characteristics. In applications where it is desirable to collect yellow sap for Or stainless steel with a stainless steel wire mesh bottom. Place the container upright in a dry collection container, leave to drain, and contact with water. The leaves can be dialyzed with water.

The pieces are thus allowed to drain for about 20-30 minutes. cut off The pieces will eventually form a seal and stop drainage. The collected yellow sap is then suitable When left for a certain period of time, it separates into two subdivisions: a precipitate and a supernatant. . The yellow sap prevents good sunburn on intact skin (undamaged skin). useful for creating skin, giving skin an olive tanned tone, and It is also useful in the production of laxatives.

After completing the operation of removing the yellow sap from the cut leaf fragment, this fragment is then wire (i.e. household cheese) slicer or peeling blade (e.g. peeling Using a knife, remove the skin to form fillets; The integument, the skin of the leaf fragment and the layer immediately below this integument, is removed. leaf break The pieces can be frozen to facilitate this peeling operation. What remains after skinning This is the internal gel matrix part (fillet), and this part is inspected and attached. Exclude the skin or discolored parts and remove the remaining yellow sap from there. Clean it with your fingers to remove it. To facilitate the removal of this yellow sap residue A gentle spray of water, preferably deionized water (and alcohol-free) spray or submerge the gel matrix section in clean running water.

The resulting fillet (internal gel matrix) can then be drained for approximately 1 hour. Ru. During this drainage operation, a slimy film usually forms on the surface of this gel matrix. do. This coating is applied by gravity or by suitable means such as centrifugation. Collected with assistance. This collected film is the mucus subdivision.

The remainder of the gel matrix in the form of a gel matrix strip is then ground up and shredded or blended to destroy the interstitial fibers present within it, or Gel matrix strips are attached to wire mesh or filter screen for liquefaction. Can be forced through lean. This (q) material is then successively equalized. It can be refined. Alternatively, freeze this gel matrix strip and thaw it. and then mixed with fibers to produce a liquid substance (this substance is the gel fillet above). forming a subdivision). This material is then filtered to obtain the interstitial fiber fraction and , the above residual matrix subdivisions are left. In summary, for topical use during this time The quality fiber is removed from the internal gluma matrix area (fillet), while for internal use This fiber is left with the remainder of the fillet contents. for topical or internal use For this purpose, the appropriate gel fraction is homogenized, e.g. using a dairy milk homogenizer. (from the addition of stabilizers, preservatives * form 1 dyes or other GRAS additives) ), this gel fraction can be used for subsequent manipulations e.g. to reduce bacterial growth. Make it even more convenient. At this point in the process, the resulting extract is mass spectroscopic. chromatography, gas chromatography, high pressure liquid chromatography or other can be tested using known quality control techniques such as analytical methods.

The homogenized extract thus obtained typically has a pH of about 4-5. Preferably, it has a pH of about 4. homogenized to increase its stability Extracts typically contain citric acid, boric acid, ascorbic acid or or by adding other suitable substances (e.g. quench for internal use). Ascorbic acid and/or ascorbic acid are used; for ophthalmological applications, boric acid is used. ). Finally, this homogenized extract is subjected to irradiation, sonication, and controlled heating. using methods well known in the art or with antibacterial chemicals or bacteriostatic agents. chemical substances such as GRAS-cetyl alcohol, sodium benzoate or chemicals such as alcohol in amounts and concentrations well known in the art. It can be sterilized by adding using various combinations of the above It will be appreciated that extracts made according to the invention may also be sterilized. Dew. Various commercially acceptable stabilizers and additives may also be included, if necessary. It will also be appreciated that it can be added to homogenized gel extracts.

At this point, final quality control operations can be utilized and then the extract is bottled or blended with other ingredients. Extraction from internal gel matrix If the extract is produced as described here, it will contain a very low concentration of yellow sap. It is most suitable for skin care and cosmetic applications.

All steps in the process described herein are performed at about room temperature.

Various modifications and alternative improvements and variations of the disclosed processes and compositions of the invention Forms and equivalents will be apparent to those skilled in the art after reading the above general description. It will be. The following examples are merely illustrative, and such modifications, variations, etc. intended to limit the scope of the appended claims to cover equivalents or variations. It's not something I did.

Example 1 Method of preparing stabilized aloe gel fillets and homogenization 1. Place the pre-cleaned tank, mixer and attached equipment in 50% isopropyl alcohol. Disinfect with Cole (IPA) solution and rinse with hot deionized water to remove IPA.

2. Rinse the pump and attached hoses with 5% “HTH” chlorine swim pool solution. and then washed.

3. Disinfect this pump and the attached hose with a 50% isopropyl alcohol solution. . Flush the pump and attached hose with hot deionized water until all IPA is gone. I rinsed it off and washed it.

4. Clean the homogenizer, attached hoses and pump with 50% isopropyl alcohol. Disinfected with solution. Use this homogenizer and the attached hose with hot deionized water. I washed it under hot water until the IPA was gone.

Aloe barbadensi lambda mirror leaves collected from the Rio Grande Valley. Transfer to a refrigerated truck at 40-45°F within 8 hours and store at 40-45°F until processing. Refrigerate at 45°F to reduce decomposition.

Next, 20 to 60 bonds of the stored leaves are prepared in a preliminary aqueous solution of calcium hypochlorite at room temperature. It can be placed in a washing bath to virtually remove surface dirt from the leaves and remove surface bacteria on the leaves. I killed Leah.

An aqueous solution of calcium hypochlorite is approximately 98% calcium hypochlorite in 1ρ of water. Prepared by making a solution containing 0.125 g and 50 ppm of free chlorine. Ta. The leaves remained in the prewash bath for approximately 5 minutes.

Next, the virtually dirt- and bacteria-free leaves are harvested in Harlingen, Texas. Thompson Manufacturing Co., Ltd. Thompson made by Ring Company, Harlingen, EXAS) It was placed on the horizontal conveyor belt of the aloe washer. This Thompson Aloe The washer washes the leaves with room temperature water to remove surface dirt and hypochlorite calcium from the leaves. The aqueous solution of sium was removed. Visually inspect the leaf again and rub by hand if necessary. Removed surface dirt remaining on the leaf surface. The leaves were then rinsed with room temperature water. .

Next, remove the tip and base from each leaf and place the leaf into a funnel-shaped stem. Multiple pieces of stainless steel placed together on top of a stainless steel collector placed in a basket-shaped container. Each container also has a mesh bottom. ing. The yellow sap was allowed to drain from the leaves for approximately 30 minutes. yellow liquid The juice passes through the mesh bottom of the stainless steel basket and into a funnel shaped collector. was gathered in.

Collector's stainless steel basket-shaped container containing aloe leaves and then submerged into a second stainless steel container. This second step The stainless steel container has a continuous horizontal flow moving countercurrently to the above container. the container contains a room temperature water bath of rinsing water for about 30 minutes to 1 hour. The container is moved slowly by hand from one end to the other. This means that the yellow sap is Allow it to be excreted from the leaves. The leaves must be soaked in this solution for 30 minutes. Must be.

Then remove the leaves from this solution and use a sharp knife or wire cheese slicer to Remove the outer skin from each leaf using a Make a fillet. This aloe guru fillet is visually inspected and has a characteristic yellow color. Any contaminated aloe gel fillets or fillets detected by discoloration The let part was discarded.

The total amount of uncontaminated aloe gel fillets depends on leaf size and conditions. The weight of the leaves was 20-60% of the original leaf weight.

The uncontaminated aloe gel fillets were then placed in a 750-seat restaurant space. It was placed in a stainless steel waste disposal unit. This unit thickens the fillet but with the consistency of a free-flowing (coarsely homogenized) liquid Crushed to particle size. This stainless steel waste disposal unit is Emerging Electric Company (N-3i) nk- Next, this coarsely crushed aloe gel fillet is placed in a stainless steel holding vat. Moved. This holding bat is manufactured by Process Equipment Company (Process EQ). equipment corp, of Belding.

HiCiiQan) Model N (1100 gallon OVC, Syrian α408 It was 65-3. The coarsely ground aloe gel fillets are stored in this holding pad according to the intended use. and the final product oil to aloe fraction ratio, sodium benzoate, sol. GRAS preservation like potassium birate, methylparaben or propylparaben mixed with the agent. A mixture of methylparaben and propylparaben is recommended for topical use. A mixture of sodium benzoate and potassium benzoate is recommended for internal use. Ru.

For example, when making a topical formulation using 10% oil and 90% aloe fraction, The ratio of propylparaben to methylparaben is 1:9. Crushed aloe gelphy The weight concentration ratio of lets to preservatives is approximately 1000:1, which is suitable for food and cosmetic products. approximately the upper limit of FDA guidelines for preservatives.

The preservative solution in the holding vat is pre-filled with 10 gallons of the approved amount of GRAS preservative. It was prepared by adding water to deionized water.

From this holding vat, the crushed aloe gel fillet-preservative solution is transferred to the homogenizer. Transported by pump. The homogenizer is from Crepaco Food Equipment A. Crepaco Foo (Chicago, Illinois) dEquipment and Refrigeration Inc., o f　Chicag.

The serial number was 04-03 manufactured by 111inOiS. This homogenizer is a commonly used tank in dairy processes for milk homogenization. It belonged to Ip. Crushed aloe gel fillet-preservative solution is 200-10 ．． Finely homogenized at a pressure of 0OOpsi.

Finely homogenized aloe gel fillets from the homogenizer, tow preservative solution Pumped into stainless steel storage tank. This storage tank is S Equipment Co., Belding, Michigan (PrOCeSSEq uipment corp, of Be1din(],)lichioan ) Model N01000 Calo'10VC, C') 7/l/N (140086 It was 6-2. The total weight of the homogenized aloe glufillet-preservative solution is: It was 20-60% of the starting leaf weight. Next, if necessary, the homogenized product is Dialysis was performed using ultrafiltration.

3-7 disclose in further detail preferred embodiments of the process of the present invention. ing. Specifically, FIG. 3 discloses a leaf cleaning device. aloe vera leaves cleaning equipment (Thompson Manufacturing Co., Burlington, Texas) Hompson Manufacturing Company.

Hart 1 nqton, Texas) A was used, which first caused the leaves to become bulky. Pre-soaked in cut 4. Next, the leaf body α is placed on the conveyor belt 8 by hand. This pulls the leaves under the two brushes 9a and 9b. conveyor bell The housing 5 is connected to a second end by a motor and a pulley 6 extending from the housing 5. It is driven by a chain 7 which is rotating at . This is also Thompson Manufacture Provided by Alling. After passing the second brush 9b, the leaves become plush. The leaves are inspected at the end 10 of the conveyor belt 8 as they have been cleaned and washed. Thereby, it is visually inspected and determined whether the leaves are sufficiently clean. If the leaves If not clean enough, the leaves are placed in a vat 12 for rough cleaning. Ru. If the leaves are clean enough, they will be placed on the upward moving conveyor B. It will be done. On the conveyor B, each individual leaf is further washed with tap water by a sprayer 11. A step 13 for cleaning is provided. Conveyor B is Seagobe, Texas. Dallas Mill Wright Co., Ltd. , of Seagoville, TeXaS). rinsing jet The fogger 11 was manufactured by Key Plumbi of Seagoville, Texas. ng, Seagoville, Dingexas).

The stainless steel butt 12 is made of 316 stainless steel and is made of stainless steel. National Sheetmetal 11 Company (National 5sheet) in Dallas, KS Special by Metal Company of Dallas, EXAS) This is a made-to-order item.

After washing, the leaves are cut and soaked as shown in FIG. Conveyor B step After rising through the pipe 13, the clean raw material leaves α are provided with holes 15 for removing debris. It falls onto the raised tray 14. Tray 14 is from the National Championships in Dallas, Texas. National 5heet Ietal Company 316 stainless steel provided by This is a part of the cutting and dipping device C manufactured by Co., Ltd. This device is available in special order quantities. Tray 1 4, the leaf is cut by hand at both its ends, and the tip and base are inserted into the waste through the hole 15. The cut leaves β are then disposed of in a stainless steel container (not shown). Multiple stainless steel baskets, each with a wire mesh bottom. loaded into one of the blocks 16. These baskets 16 are then made of stainless steel. It is placed in a truck of rest steel type. This truck has trapezoidal funnel 17. This funnel 17 allows the yellow sap to pass through the basket to the bottom of the leaf. The liquid is drained from the bottom of the funnel 17 and falls to the bottom of the funnel 17. Remove the yellow sap regularly. and frozen for storage. The process of draining the yellow sap takes approximately 30 minutes.

After this step, the cut leaves β still remaining in the basket 16 are shaped like trapezoids. It is manually transferred to the water bath 18 at the position closest to the funnel 17. water in a countercurrent flow from the inlet water pipe 19 into the bath 18 at the point farthest from the trapezoidal funnel 17. and then exits through the outlet water pipe 20 at the point closest to the trapezoidal funnel 17. be done. The tray is manually moved gradually in the water bath in the direction away from the trapezoidal funnel 17. The cleaning process, in which the basket is placed in the water bath 18, takes about 1 hour.

After cleaning, the basket is placed on tray 21 for drying, which takes only a few minutes. continue. Including wire mesh, yellow sap discharge and automatic cleaning device, The entire assembly of FIG. 4 for the basket is again from Namjoon, Dallas, Texas. Made of custom made 316 stainless steel by National Sheet Metal be.

- After washing, the stack of cut leaves β in the basket 16 on the tray 21 is As shown in FIG. 5, it is moved to an area for rough cutting into fillets.

The skin 22 is removed from the fillet so that only substantially transparent material remains. . The remainder of the skin 22 is discarded. Next, the fillet γ is fed to a coarse grinder. placed in trough 23. This Trough 23 is located at the National Stadium in Dallas, Texas. It is made of 316 stainless steel made by Totometal. This crushing The machine is a model Nα5150-N sinker (Watso, Dallas, Texas). (Watson Food Service Industries, Inc.) (Watson Food 5) service Industries, Inc., Dallas.

lexas). After being roughly crushed by a crusher, it comes out of this crusher. The treated material γ' has a vertical half-shell tank made of 316 stainless steel. Portable Tank E (Pantone, Dallas, Texas) with a capacity of approximately 100 gallons. Perma-S, manufactured by Perma-S. an, Manufacture, distributed throughah Vanrone, Inc., Dallas S., Exas). Crushed powder The fillet γ' is a Belmarsan stirrer (model NQAAPH2) (also a Texas (Sold by Pantone, Dallas, State) in tank E. Ru.

After coarse homogenization, the material from tank E is subjected to fine homogenization and (optional) filtration. sent to a separate area. In Figure 6, the material from tank E is Lee Brand Glue Reliance Electric Reliance Motor Model NQB76Q2139M-VF (Reliance Motor Mode lNαB76Q2139t (-VF of the Re1ance Ele ctric Company.

Pump 26 (C1eveland, 0hio) driven by Crepaco, Inc., Dallas, Inc. Stainless steel centrifugal pump model Nα4 sold by AS) Fine Homogenizer F (Clebaco, Dallas, Texas model) with V-81 No. 30D13).

After micro-homogenization, this material was poured into a large 1ooo galvanized tube made of 316 stainless steel. vertical single-shell mixing tank G, distributed by Pantone, Dallas, Texas. (manufactured by Lumasan). 1j! Fillet △ crushed to 1iill is Pell Marsan Stirrer 126a (distributed by Pantone, Dallas, Texas) The mixture is stirred by a Permarsan model NCLAAPt12). Tank G middle Ingredients △ are for direct human consumption after flavoring and suitable preservatives have been added. can be sent directly to processing for beverage products. Or again in Tank G The material is removed and passed through a filter for pulp removal through the exhaust line 28. pump 27a forming one unit with a motor 27b. . Pump 27a and filter 27b are manufactured by Lomato diatomaceous earth filter (Da. Rasno Allen Recreation Company (Alfen Recreation Com) It forms part of the model Nα99-2138) sold by ing. The filtered material is then pumped into tank H. This tank H is A lid 25 can be provided in the same way as the link E.

In FIG. 7, the pulverized fillet Δ is partially filtered and the mixer 29 and pumped to the dialyzer by pump 30. mixer 2 9 is a Permarsan stirrer (Model No. sold by Pantone, Dallas, Texas). , AAPH2). Pump 30 is a superior stainless steel model. 5C345 Process Pump (Sveria Stainless, Delavan, Wis.) Made in Japan). The process pump 30 has a 4.4501’l)m band motor. 3 horsepower motor 30a (Cat N (lcM3559) of the Ba1dor Electric Company, Ft, Sm1th A rkansas) is attached. pumped by pump 30 The material is used in a dialysis unit J (not shown) with four filters 31 (not shown). Lomicon Model 4333 Limited, manufactured by Lomicon Co., Auburn, Massachusetts. Filtration system) (Romicon Model HF4SSSultrafi ltation system made by Romicon Inc. Woburn.

Massachusetts). Each filter has a filter housing 32. A portion of the material passes through the separation line 34 to the separation discharge line. It passes vertically to a point where it can be taken out by a tank 35. Other unseparated The collected material is either returned to the dialysis machine via recirculation return line 33 or sent to a separate return. It is returned to vat I through feed line 36 and recirculated.

The desired material depends on the desired aloe fraction and the desired end product. is obtained after processing through a separate discharge line 35 or in the vat I.

For example, if excess water and minerals need to be removed, water and minerals are separated using an ultrafilter and discharged through line 35; The desired aloe fraction can be returned to Vat I. This process completes the desired The product is dialyzed until the amount of salt and water is removed from the product in Vat I. Can be repeated by simply cycling through the unit. this The process can include two or more dialysis steps. For example, as explained earlier , salts, low-molecular acids, and undesired anthraquinones are removed in the first dialysis step. be able to. The undesired material is discharged through a separate discharge line 35 and the desired fraction Returned to Bat I. This process is the io, ooo Dalton obtained from Romicon. This is done by using an ultrafilter with a bore. Next 10,000 $50,000 Dalton ultrafilter also available from Romicon Replace the ultrafilter with an ultrafilter and repeat the dialysis process. This dialysis process The gel matrix polymer is now separated into two halves. The first fraction is io, ooo Consisting of a gel matrix polymer with a size of ~50,000 daltons, separated and discharged Discharged from line 35, the second fraction contains gelato larger than 50,000 Daltons. is made of polymeric liquid and is returned to vat I. This process A few minutes to several minutes depending on the amount of salt and water that needs to be removed from the product. It can continue over time. This means that topical aloe products with a prepared salt and water content, such as when used on broken skin. It is important because it is necessary. Salt content can cause burning pain and irritation.

In FIG. 7, the separation return line 36 is a Rubber Supply Company, Dall Tybon (T110On) tube (supplied by aS, REXAS) ( Made of food grade). Separate discharge line 35 is located at Pantoux, Dallas, Texas. The pipe is made of 316 stainless steel sold by Co., Ltd.

Daishu-2 Preparation of aloe vera for drinking. Aloe vera according to Example 1 using the specific preservatives listed below. Using 100 gallons (379 liters) of lagel: number! Kuai: 1. Two 100 gallon stainless steel tanks with mixers.

2. 1 i, ooo gallon stainless steel tank with mixer nine.

3. Homogenizer with pump.

4. Stainless steel screen.

5. One transfer pump with appropriate capacity.

Complete steps 1 and 2 below before adding raw aloe vera gel to the collection tank. Ru: 1. Add 40 gallons (151.4 liters) of deionized water.

2. Dissolve the following chemicals in deionized water with stirring in the collection tank (see below). See Section 13 below): Sodium Benzoate potassium sorbate Vitamin E 3. While continuing to stir, collect the raw aloe vera gel from the crushed material into a collection tank. for a total volume of 100 gallons (379 liters).

4. Move the tank to the blending area.

5. Place the next collection tank under the crusher outlet.

6. Set a pre-sterilized homogenizer to a pressure of 1.500 psi, Connect to collection tank.

7. Start the homogenizer and fill the i,ooo gallon stainless steel tank. Drain the product into an open stainless steel basket attached to the tank.

8. Start stirring when the product covers the mixer blades. stir Record the start and end times until the 1,000 gallon tank is empty (approx. 8 hours).

9. Continue stirring at low speed.

10. Dialyze the product to adjust the aloin content to 50 ppm or less, and if necessary to reduce excess acid.

11. Add vanilla flavor and cinnamon oil - mix for 20 minutes. Ru.

12. The product is ready for compounding. Mix immediately.

Required amount Deionized water (40,0 gallons) 151.4. Il Sodium Benzoate USP 37B (j glycine 3.0Kg Quen IUSP 416g Potassium sorbate υSP 1899 Vitamin E (1000 units) 1 capsule Raw material Aloe vera gel 379L (1 00 gallons) Cinnamon oil - natural W, S, 8.07X thought - Yu Aloe vera gel for producing aloe vera cosmetics prepared according to Example 1 100 gallons (379 liters) of finely homogenized aloe gel Made by Lomart, 960 Alabama Ave., Plutzklin, Que. Pumped into a stainless steel diatomaceous earth filter. This filter is It was a typical diatomaceous earth filter of the type used in swimming pools. Really Aloe gulf has been homogenized by a diatomaceous earth filter to remove all the fibrous material. Fillet - aloe gel fillet removed from preservative solution and free of pulp - A preservative solution was obtained.

A The following summer lI: 1. One 100 gallon stainless steel tank with attached mixer.

2. 1 i,ooo gallon stainless steel tank with mixer attached nine.

3. Homogenizer with pump attached.

4. Stainless steel screen.

5. One transfer pump of suitable capacity.

6. Pool filters and accessories.

7. Connecting hose and stainless steel tear fittings are included.

Preliminary work: 1. Pre-cleaned tank, mixer and mounting equipment with 50% IPA solution. Disinfect and rinse off the IPA using hot deionized water.

2. Drain the 5% HTH solution from the pump and attached hose.

Wash under running water.

3. Disinfect the pump and attached hose with 50% IPA solution. Pump and attached hose Rinse with hot deionized water until all inpropyl alcohol (IPA) is removed. cormorant.

4. Fill the homogenizer and attached hoses and pump with 50% isopropyl alcohol. Disinfect with kohl solution. Heat the homogenizer and attached hose until all IPA is gone. Rinse with deionized water.

5. Disinfecting the pool filter: (A) Make a 5% aqueous solution of chlorine HTH powder for swimming pools and soak the entire system for 20 minutes. circulate between.

Blending operation: Steps 1 and 2 before adding the stabilized raw aloe vera gel to the collection tank. conduct.

Add 1.50 gallons (189 liters) of deionized water.

2. Dissolve the following chemicals in deionized water with stirring in the collection tank: Methylparaben potassium sorbate 3. While continuing to stir, collect the stabilized raw material aloe vera gel from the grinder. Place in collection tank to bring total volume to 100 gallons (379 liters).

4. Move the tank to the blending area.

5. Set the pre-sterilized homogenizer to a pressure of 1,500 psig and Connect to collection tank.

6. Start the homogenizer and fill the 1,000 gallon stainless steel tank. Drain the product into an open stainless steel basket installed in the tank.

7. Start stirring when the product covers the mixer blades. started stirring Record the time.

8. Continue stirring at low speed for 20 minutes.

9. Add all 100 gallons produced to the 1000 gallon tank.

10. Leave the product overnight.

11. Preparation of pool filter: A. Rinse the disinfectant solution from the pool filter using hot deionized water.

8. Add IKg of diatomaceous earth to 10 gallons of deionized water and mix until suspended. .

Circulate the mixture through the pool filter until the C1 water is clear.

12. Use the product to flush excess water from the pool filter.

13. Circulate the product through the filter until the product is free of pulp. .

14. Mix for 30 minutes.

15. If necessary, perform dialysis to reduce the yellow sap to less than 50 ppm, and Keep the transparency below 150 milliosmoles.

Ingredients Required amount Deionized water (50 gallons) 189 L Methylparaben 6819 Potassium sorbate 379g Raw material Aloe Veragle 379[ (i.e. 100 gallons up to) Bujeongjang-A Manufacture of aloe cream Aloe vera gel 71.6 for producing aloe vera cosmetics prepared according to Example 3 Using a gallon (271 liters), use Star 1, counter motion stirrer and A suitably sized round-bottomed jacketed steel plate with a nylon scraper attached to it. 1 x Resteel kettle.

2. Comes with one suitably sized cylindrical jacket fitted with a variable speed mixer. Stainless steel tank.

3. 1 transfer pump.

4. Tyaon and/or polyethylene connection hose, Stainless steel fittings are included, as well as a suitably sized container and stainless steel. Clean kettles, tanks, mixers, pumps, hoses and and disinfect containers.

Preparation of aqueous phase: 1. Add the following substances to a cylindrical tank to make stabilized aloe vera gel for production. Start stirring when covers the mixing vane: Stabilized Aloe Vera Glu for Manufacturing Propylene glycol Allantoin Methylparaben GERMALL 115 (registered trademark) imidazolidinyl Urea (Sutton Laboratories, Chatsham, NJ 07928) ) (Sutton Laboratories, Chatham, NJ 07 928) Sorbitol 70 phosphoric acid 2. Heat to 75°C ± 5°C while stirring and maintain at this temperature.

3. Mix until all substances are dissolved.

Painting ratio: 1. Add the following substances to another kettle: glycerin stearate stearyl stearate Witskenol 163 (Wickham Product Co., Fukuenot, New York 12746) (Wickeno1163 (Wichham Prod) ucts, Inc., Huguenot, NY Lexamine p-13 (I Lurex Chemical Company, Philadelphia, Pennsylvania 1914B) ( Lexamine P-13 (Inolex Chemical Co.

Ph1ladelphia, PA 1914B>) Cetyl alcohol lanolin Mineral oil (light) (Keydor) (Delta Trubenz & K. Michals, Dallas, Texas Chemica! s, Dallas, TX 75 285)] dimethicone provilparaben vitamin e natural Lexane QX-3000 (Illex Chemical Company).

Philadelphia, PA 19148) (LEXEIN QX-3000 ( Inolex Chemical Co, Ph1ladelphia, P A19148)) 2.7 while stirring slowly until the substance is melted and homogenized. Heat to 5°C ± 5°C.

Preparation of emulsion: For successful emulsification, the oil and aqueous phases must be added at the same temperature during compounding. Must be. This process must occur without interruption.

1. Slowly add the aqueous phase to the oily phase while stirring slowly.

2. While continuing to stir slowly, immediately turn off the steam and drain the jacket. Start forced cooling using cold domestic water.

3. If necessary, fill to batch size using deionized water at 75°C ± 5°C.

4. Add flavor when the product has cooled to 40'C ± 5°C.

5. Stop stirring when cooled to 30°C and submit sample for approval.

Sei - A and the power of elutriation” stabilized for manufacturing Aloe vera gel (71.6 gallons > 271 Ky propylene glycol 17. 4Ng Allantoin 908g Methylparaben 690g Imidazolidinyl urea (Germol 115) 2.0y Sorbitol 70 9 08 g Phosphoric acid 1.05 ton oily phase Glyceryl stearate (Lexemal 515) (Irrex Chemical Co., Ltd., Fiery Ladelphia.

PA 19148) 35.6Nff Stearyl stearate (Rexol 5S) (Irex Chemical Co., Ltd., Fila) Delphia.

PA19148) 8.9Ng Dioctyl adipate (and) octyl stearate (and) octylpal Mitate (Winkenall 163. Witskam Products Co., Fuguenot, NY12746) Palmitamidopropyl dimethylamine on 8.9 (Lexamine P-13, Irtuks Chemical Company Philadelphia, PA19 148) 3.5Kg cetyl alcohol 3.5Kg Lanolin 3.5Ky Light Mineral Oil (Light) (Keydor) (Derri, Trubenz & Chemicals, Dallas, TX 75285) 3.5KI Dimethicone 908 8 Propylparaben 399g Vitamin E natural 363g Cocamidopropyldimonium 3637 hydroxypropylamino collagen (Lexane QX-3000. Ilux Chemical Co., Philadelphia, P. A19148) 109g Elias Fragrance #5394 (Elias Flagran Switzerland, Plutzklin.

NY 11203) 109g Example 5 Stabilized Aloe Vera Gel 115 Gel for Manufacturing Prepared by Example 3 anti-irritant agent (counter-1rrita) starting material (counter-1rrita Preparation of nt).

Equipment required: 1. 1 with counter motion stirrer and nylon scraper installed Stainless steel kettle with suitably sized round bottom jacket.

2. One suitably sized cylindrical tank equipped with a variable speed mixer.

3. 1 transfer pump.

4. Tyggon and/or polyethylene connection hose, A stainless steel tear-off piece is included, a suitably sized container and a stainless steel Steel fill pre-cleaned kettles, tanks, mixers, pumps, hoses and disinfect containers.

Carbo Ball Slurry II 1. Add deionized water to the mixing tank. Start the mixer.

2. Add all Carbomer 940 (Carbopol 940) (B, F, Gutdrich Chemical Group IL 60694) (Carbomer 940 (Carbopol 940) (B.F.

Goodrich Chemical Group, Chicago, IL 60694)) slowly. Continue mixing until solvated.

3. - Set up at night. Turn the mixer on and mix for 1 hour.

Preparation of aqueous phase: 1. Add the following substances to a cylindrical tank and stir when the deionized water covers the mixing vane. Start stirring: Deionized water (leaving 10 gallons to rinse the carbo ball slurry from the tank) put) stabilized aloe vera gel Propylene glycol Triethanolamine 99% Allantoin Methylparaben Potassium sonbate Urinary element 2. Heat to 70°C ± 5°C while stirring and maintain at this temperature.

3. Mix until all substances are dissolved.

4. Add carbopol slurry to form “fisheyes” Mix until all the ingredients are gone.

5. Rinse the tank with deionized water.

Preparation of oily layer: 1. Add the following substances to the kettle: Petroleum (White Protopet(R)) Digital Trubenz and Chemicals, Dallas.

TX 75285) (Petrolatum (White Protope) t) (De1℃a Sollovents and Chemicals, D allas, TX75285)) Mineral oil (carnation white ( (Registered Trademark)) (Delta Trubenz & Chemicals, Dallas, TX Mine) Ralnail (and) Lanolin Alcohol (Amercol L-101) ( Amerchol, Edison, NJ O8817) ((Amerchol L- 101) (Amerchal, Edison, NJ 08817)) Cetylated lanolin alcohol (Francol A [^ (registered trademark)) (Delta) Sonbenz & Chemicals.

Dallas, -TX 75285) Stearic Acid Triple Press essed) Glyceryl stearate (Lexemal 515) (Irtsukus Chemical Co., Ltd., FI) Radelphia, PA 19148) Cetyl Alcohol Dioctyl adipate (and) octyl stearate (and) Octyl palmitate (Uitskenol 163) (Uitsken Products Co., Ltd., Huegenot, NY 12746) White Oleic Acid USP Stearyl stearate (Lexol SS) (Illex Chemical Co., Ltd., Philadelphia, PA 19148) Propylparaben Methyl salicylate histamine dihydrochloride cinnamon oil 2. Heat to 70℃±5℃ while stirring slowly until the material is melted and homogenized. .

Preparation of emulsion: Note: For successful emulsification, the oily and aqueous phases must be added at the same temperature during blending. Must be. This process must occur without interruption.

2. While continuing to stir slowly, immediately turn off the steam, drain the jacket, and cool. Start forced cooling using household water.

3. Fill to batch size using deionization at 70°C ± 5°C if necessary.

4. Stop stirring when cooled to 35°C and submit sample for approval.

Deionized water (30 gallons) 113Ny Carbomer 940 (Carbopol (B.F. Gutdrich Chemical Group Chicago. IL 60694) 4 ．． 36 K’j aqueous phase Stabilized Aloe Vera Gel for Manufacturing (115 Gallons > 436Kg Deionized water (41.5 gallons) 157 NSF Propylene Glycol 54.5 kg Triethanolamine 99% 9.8Kg Allitein 653g Methylparaben 1.1Kg Potassium turudate 545g Urine element 21.8Kg oily phase Bethorium USP (White) (Delta Trubenz & Chemicals, Dallas.

TX 76285) 8.2KI Mineral Oil (Carnation White) (Delta Trubenz & K. Michals.

Dallas, TX 75285) 8.2Ng Mineral Oil (and) Lanolin Alcohol (Alcohol L-101> (Americol, Edison, NJ.0 8817) 7.6Kg Acetylated Lanolin Alcohol (Fancor ALA> (Delta Trubenz & Chemicals. Dallas, TX 75285) 7.6Kf stearic acid triple presto 23.5'j glyceryl steare (Lexemal 515) (Illex Chemical Co., Philadelphia PA) 1914B> - 10.3Ng Cetyl Alcohol 8.2Kg Dioctyl adipate (and) octyl stearate (and) octylpal Mitate (Witskenor 163) (Witsken Products, Huegenot , NY 12746) 10.3 'j White Olein @tJsP 1. INy stearyl stearate (Rexol SS) (Irex Chemical Co., Ltd., Philadelphia.

PA 19148) 1.1Kg Propylparaben 1. IKff Butylparaben 1.1 to 3 Eucalyptus oil 21.8Nff Methyl salicylate 136N$ Kampfre 21.8N9 Menthol at 21.8! F Histamine dihydrochloride 5459 Cinnamon oil 2189 support - support Preparation of freeze-dried aloe. Using 1 liter of aloe veragle prepared in Example 2, and start. Put the juice into a freeze-drying unit and freeze-dry aloe with approximately 459 g. I! ! miss

X Blood■-Yu Preparation of topical wound gel. Starting with aloe veraglut prepared according to Example 3 , concentrated to 190% by dialysis.

A Nezuku Seiyu 1. Two suitably sized stainless steel tanks with mixers.

2. 1 Romicon compression system.

3. 1 transfer pump 4. Tygon and/or polyethylene connecting hoses, stems stainless steel mounting hardware, suitably sized container and stainless steel manufactured filter.

i-Deaf Higashijo 1. Pre-cleaned tanks, mixers, pumps used during the manufacturing process, Disinfect hoses and containers.

2. Prepare Carbopol slurry one day before production.

3. Disinfect the pre-cleaned Romicon concentration system.

The entire system is washed with deionized water, filled with 37% hydrogen peroxide, and stored overnight.

Concentration of Aloe Veragle for Skin Wound Formulations for Treatment of Bedsores and Ulcers: 1. Rinse the concentrator with deionized water until the hydrogen peroxide is gone.

2. Pour the calculated amount of effluent necessary to concentrate the product into a 190% gel into aloe vera gel. separate from the file.

Carbopol slurry m: ], Add Aloe Vera Glue 190% to the mixing tank.

Start the mixer.

2. Add Carbomer 940 (Carbopol 940) to the water in the mixing tank (B, F , Gutdrich Chemical Group, Chicago IL 60694) slowly. . Continue mixing until solvated.

3. - Set up overnight. Start mixer and mix for 1 hour.

Mixing operation: 1. Add the following substances to a mixing tank of appropriate size, and when the blades are covered, start the

Aloe Veragle 190% [-glutamic acid Imidazolidinyl urea (Germol 1is) (Sutton Industrose, CH) Jatum, NJ 07928) Potassium sonbate Sodium benzoate Triethanolamine 99% 2. Calcium filtered through a 100 mesh stainless steel screen. Continue mixing while adding Bopol slurry.

3. Mix until there are no corn-like objects left.

4, p) 18 reading records. pH range 5.0+0.55, click if necessary Adjust pH using enoic acid or triethanolamine.

6. Record the pH.

7. When the product is completed, submit the sample for approval.

8. 100 mesh stainless steel strip when ready for filling. Pump through the clean into the filling hopper.

Concentrated Stabilized Aloe Vera Glu (5 cuts 90% (75 gallons)) 284 9 Carboma 940 (Carbopol 94o) (B, F, Goodrich Chemical Group Chicago, It 60694) 13.3Ng Concentrated Stabilized Aloe Belagle (5x 190%) (403 gallons > 1523 Ny panthenol 37.9 Ky Ingredients Required amount Allantoin 11.4 'I L-glutamic acid 5.7Kg Imidazolidinyl urea (Gelmol 115) (Saraton Laboratories, Cha Tam NJ 07928> 1.9pm Triethanolamine 99% 13.3Kg Potassium sorbate 945g Sodium benzoate 1.9? (g X dust ■-kun Start with 100 Ny of unwashed leaves taken directly from the field. 51.2K g aloe <7 youth from Thompson Manif 7 Kucharing Co. (Harlingen) Collected using a “Thompson Aloe Juice Extractor” manufactured by Co., Ltd., Texas, 78550). Melt.

be transferred. The resulting 51.2Nff aloe juice extract contained 0.1% methyl 1o with added parabens and a mixer.

Mixed for 20 minutes in a gallon stainless steel tank. This solution is then Romicon ultrafiltration equipment until the anthraquinone content is less than 50 ppm by weight. Dialyzed using

This gel is used to make the aloe cream of Example 4.

Separation and isolation process As mentioned above, in the fractionation process, aloe in the form of gel matrix strips The internal gel matrix portion of the filleted Vera leaf was homogenized and filtered to remove the interstitium. Remove the fiber subdivision (Sub-port 1on) and remove the remaining matrix subdivision. You can leave some minutes. This residual matrix subdivision is then processed to Separating, isolating and purifying the active chemical in Laglu, Kallidin® Can be done. To separate kallidin from the remaining matrix subdivision, a water-soluble lower Add an excess amount of aliphatic polar solvent to the remaining matrix subdivision. Then Karigi begins to precipitate from this mixture.

This solution should be maintained at a temperature sufficient to allow as much of the active ingredient as possible to precipitate out of solution. Yes, but let it sit for a long time, not long enough for the kallidin to start decomposing. after this time Decant the supernatant without shaking the precipitate, or place it in a sieve. Turn on and remove. The precipitate and remaining solution are then placed in a suitable centrifugal separator and collected. Let. After centrifugation, top? Decant and discard the liquid. If desired, this The pellet was washed with a small volume of fresh water-soluble lower aliphatic polar solvent, recollected, and the supernatant Discard the liquid again. The pellet is then lyophilized and dried overnight. obtained The product is a substantially non-degradable lyophilized form of kallidin. raw obtained The composition can be ground into a powder.

Another preferred process for separating and isolating kallidin includes the following steps.

Leaves from mature aloe vera plants grown outdoors, preferably before processing. You can remove or remove the plant from near the base without damaging any part of the leaf. is disconnected.

Preferably, the leaves are cut at the base of the plant, just above the stem (petiole), leaving clear cell buds. peeled from the stem to prevent gel leakage or contamination of the gel with yellow sap. .

After separation from the plant, the leaf base and tip are removed, and the cut leaves are fractionated. Peeled to form fillets as mentioned in the process.

The resulting fillet (internal gel matrix) is then liquefied to liquefy the internal gel matrix. grind, shred or blend the interstitial fibers present inside. Disassemble or force into the wire mesh or filter screen. pass through. The resulting liquefied internal gel matrix is then homogenized. in this way The resulting homogenized extract typically has a p.p. of about 4 to about 5, preferably about 4. I have H. The homogenized extract is then filtered to remove interstitial fibers. this The homogenized and filtered extract is then subjected to the same residual matrix subdivision as described immediately above. kallidin can be separated and isolated.

Another preferred process for separating and isolating kallidin involves the following steps: include.

Leaves of mature aloe vera plants grown outdoors are preferably decanted before processing. Remove or cut the plant from near the base without damaging any parts. do. Preferably, the leaves are cut at the base of the plant just above the stem (petiole) and cut into clear Peel the cells from the stem to prevent gel leakage or contamination of the gel with yellow sap. vinegar.

The leaves are then transferred by suitable means, such as the Aloe juice is crushed using the “Thompson Aloe Extruder” manufactured by Factoring Co., Ltd. push out the base.

The extruded aloe juice is then processed in the same manner as the residual matrix subdivision above. kallidin can be separated and isolated.

All steps of the process described herein involve freezing, which is preferably carried out at about -50'C. The disclosed processes and compositions of the present invention are carried out at about room temperature, except for the drying step. Various modifications, as well as alternative modifications, variations, and equivalents, are available upon reading the general description above. will be apparent to those skilled in the tobacco arts. The following examples are illustrative only. to extend the scope of the appended claims to cover such modifications, equivalents, or variations. It is not intended to be limiting.

Example 9 Process A, preliminary work for separating and isolating kallidin: 1. Pre-cleaned tank, mixer and mounting equipment with 50% isopropyl alcohol. Disinfection was performed with alcohol (IPA) solution and the IPA was rinsed off with hot deionized water.

2. Clean the pump and attached hoses with 5% “HTH” chlorine swimming pool solution. It was rinsed off and then rinsed with water.

3. Disinfect the pump and attached hose with a 50% isopropyl alcohol solution.

The pump and attached hoses were flushed with hot deionized water until free of IPA.

4. Insert the homogenizer and attached hoses and pump into 50% isopropyl alcohol. Disinfected with kohl solution. Heat the homogenizer and attached hose until all IPA runs out. Rinse with hot deionized water.

Harvesting Aloe barbadensi 2 mirror leaves collected from the Rio Grande Valley. Transfer to a refrigerated truck at 40-50°F within 8 hours to reduce decomposition. Stored under refrigeration at 40-50''F until further processing.

Next, store 20 to 60 bonds of leaves in a preliminary calcium hypochlorite aqueous solution at room temperature. in a cleaning bath to substantially remove surface dirt from the leaves and remove surface bacteria on the leaves. I killed Leah.

Calcium hypochlorite aqueous solution is 98% calcium hypochlorite in 1 liter of water. Approximately 0.1259 μm of chlorine was added to give a solution containing 500 μg of free chlorine. Leaves for about 5 minutes It remained in this prewash bath for a while.

The leaves, which are essentially free of dirt and bacteria, are then harvested by Tompso in Harlingen, Texas. Horizontal conveyor for “Thompson Aloe Washer” manufactured by N Manufacturing Co., Ltd. I placed it on top of Abert. This Tombson aloe tree has dirt on the surface from the leaves. The leaves were washed with room temperature water to remove the calcium hypochlorite aqueous solution. again Visually inspect this leaf and, if necessary, rub the leaf with your hands to remove any residue remaining on the leaf surface. Removed surface dirt. The leaves were then rinsed with room temperature water.

Next, remove the tip and base from the confectionery and place the leaf in a stainless steel funnel. A stainless steel collector, each with a mesh bottom, is placed on top of the stainless steel collector. It was placed in a steel basket type container. Yellow liquid juice for about 30 minutes I allowed it to drain from this confection. yellow sap in stainless steel basket It passed through the bottom of the mesh and was collected in a funnel-shaped collector.

A collection of stainless steel basket-type containers for storing aloe leaves. continuous horizontal flow of rinsing water that is separated from the container and moves countercurrently to the container was submerged in a second stainless steel vessel containing a room temperature water bath.

This container can be moved slowly by hand from one end of the container to the other for approximately 30 minutes to 1 hour. It will be done. This is done by removing the yellow sap from the leaves using a mackerel wire cheese slicer. Remove the outer skin from each leaf and extract aloe guru fillet from each leaf part. I made it. This aloe gel fillet is visually inspected and has a characteristic yellow discoloration. Contaminated aloe gel fillets or fillet parts detected by was thrown away. The total weight of an uncontaminated Aloe Gur fillet is determined by the size of the leaf. and 20-60% of the starting leaf weight depending on conditions.

The uncontaminated aloe gel fillets are then placed in a 750-seat restaurant space. Placed in a stainless steel waste disposal unit. This processing unit is a fillet of the consistency of a thick but free-flowing (coarsely homogenized) liquid. Crushed to average particle size. This stainless steel waste disposal unit is Emerging Electric Co., Inc. (Rasin, Wis.) Vision model NαSS-150-13, serial number Nα-115132. It was.

Next, this coarsely crushed aloe gel fillet is placed in a 100 gallon stainless steel container. Transferred to holding vat. This holding bat is made by Process & Co. in Belding, Michigan. Equipment manufactured by Modenoi (1100 gallon OVC, Syrianoi a 40 It was 865-3.

Pump the crushed aloe gel fillet solution from the holding vat into the homogenizer Transported. This homogenizer is manufactured by Crepaco Food Ike in Chicago, Illinois. Serial Na 04-03 manufactured by Equipment and Refrigeration Co., Ltd. there were. This homogenizer is an integral part of the dairy process for milk homogenization. It was a commonly used type. Approximately 1 cup of coarsely crushed aloe gel fillet solution , finely homogenized under a pressure of 500 ps i.

From the homogenizer, finely homogenized aloe gr fillet solution is poured into stainless steel. Pumped into steel storage tank. This storage tank is located in Verdy, Michigan. Model No. 0.1000 gallon OVC manufactured by Process Equipment Co., Ltd. .

The serial number was Nα40866-2. of homogenized aloe gel fillet solution The total weight was 20-60% of the weight of the raw leaves. Then if you need this homogeneous The converted product was dialyzed using ultrafiltration.

Next, the finely homogenized Aloe Gurfillet solution was added to the Leslie Interstitial fibers were removed by filtration using a diatomaceous earth filter model DE-48. . The interstitial fibers themselves are used as a filtration medium instead of diatomaceous earth; Long mesh, supported by a fabric filter support. Sufficient amount of fiber accumulated a few times before opening the outlet so that it can act as a filtration medium. Pumped through the filter for minutes.

20 gallons of filtered aloe gel fillet solution was then added to a 100 gallon tank. Pump and add 190 proof undenatured ethanol (ethyl alcohol, 190 proof.

u, s, p, , net, 54 gallon batch 1. D, #cT 185JO4, Tuscola, Illinois 61953, p, o, box 218 υ, S, Indus 80 gallons of aloe gel fillet (available from Trial Chemicals) added to the liquid.

This alcohol-gel solution was then immediately applied to an 18-8 stainless steel diameter tube. Several 11-quart pans measuring 10.5 inches and 8 inches tall (Deer, Illinois) Manufactured by Bloomfield Industries, Inc., Dallas, Texas 3712 Garway's Watson Food Service Equipment and Sabra (available from Izu).

This alcohol-glue solution was then allowed to stand for about 4 hours.

Next, pour the clear supernatant liquid, being careful not to shake up the precipitate that has settled to the bottom of the pan. While doing so, I extracted it with a decant or a siphon. Remove precipitate and remaining solution Each packet is then placed into four 1-bind stainless steel centrifuge packets. The precipitate and residual solution were transferred to the container.

This packet was then subjected to an IEC Centra-7 centrifugation m (021947゛saput). International Inn, 300 Second Ave., Secondham Heights, SE. Manufactured by Quipment Co., Ltd., 75050.

P.O., Box 1048, Guerand Braley, Texas.

available from American Scientific Products) (rEc Centra-γCentrifae (Internationala! Equ ipment Co,, 3002nd Avenue, Needham H eights.

)1assachUsetts 02194. AmeriCan 5cien tific　Pr0duCtS.

PO, Box 1048. Grand Prairie, Texas 75 050)) at 2000 x g for approximately 10 minutes.

After centrifugation, the supernatant was decanted and discarded. Next, the beret, fresh 190 Wash with proof undenatured ethanol and centrifuge again at 2000X 9 for about 10 minutes to collect. I met. After centrifugation, the supernatant was discarded again.

This pellet was then mixed with several pieces of 6007 Vertis (VIf?Tl5) lyophilized gel. Transferred to a jar and swirled around in liquid nitrogen until frozen. Next, this freeze-dried Welch Duo Seal Vacuum Pump (Model Nα1402.7523) (Available from 5 Telchi) 9. Birch Swimmer Coil Cooler (Model N) (16205-4350, in cooler acetone bath) and Bernas 18 port true Freeze drying equipment consisting of an empty drum manifold (model Nα6211-0350) installed on. All Vertis equipment is located at 75050 Guerand Braille, Texas. -.

P, 0. Box 1048. American Scientific Products? Available from The freeze-drying drum was filled with acetone maintained at -50°C.

Samples were lyophilized overnight to dryness and then weighed on a Mettler AE163 balance.

The remaining sample consisted of substantially non-degradable lyophilized kallidin.

Yield from 50 gallons of aloe vera gel was approximately 370-3809 kallidin .

x Calicin separation and isolation process Aloe collected from the Rio Grande Valley l<)I,t/<”>E,).

Transfer the Miller leaves to a refrigerated truck at 40-45°F within 8 hours of harvest to allow them to decompose. Stored frozen at 40-45°F until processing to reduce heat.

Next, the tip and base were removed from each leaf.

Then use a sharp knife or wire cheese slicer to slice each leaf. The outer skin was removed and aloe gel fillets were made from the famous confectionery parts.

Next, the aloe gel fillet is made of stainless steel for a 750-seat restaurant. I put it in the waste disposal machine. This waste processor handles fillets in a dense but free-flowing manner. coarsely ground to an average particle size of the consistency of the moving (roughly homogenized) liquid. Ta. This stainless steel waste disposal machine is manufactured by Emma from Racine, Wisconsin. - Model N manufactured by Sinn Electric Co., Ltd. - Shinku Erator Division It happened with o5S-150-13, serial number Nα115132. Next, this coarse crushing The Loegel fillets were transferred to a 100 gallon stainless steel holding vat.

This holding bat is manufactured by Process Equipment, Belding, Michigan. Model Nα100gao70VC, Syria/L/No, 40865-3 Tea Ivy.

Pump the crushed aloe gel fillet solution from the holding vat to the homogenizer did. This homogenizer is manufactured by Crebaco Food Equip in Chicago, Illinois. The serial number was Nα04-03 manufactured by Pement and Refrigeration Co., Ltd. . Homogenizers are commonly used in dairy processes for homogenization of milk. It was of the same type.

The coarsely crushed aloe glufillet solution was finely homogenized under a pressure of 1500 pS+. Ta.

Stain this finely homogenized aloe gel fillet solution from the homogenizer. Pumped into Ressteel storage tank. This storage tank is located in Bay, Michigan. Luding Process Equipment Model Nαi ooo gallon OV C, Syrian o, 40866-2. homogenized aloe gel fillet The total weight of the solution was 20-60% of the weight of the raw leaves. Next, if you need The homogenized product was dialyzed using ultrafiltration.

Next, the homogenized gel was filtered using a Reslow diatomaceous earth filter model DE-48. The filtered interstitial fibers were removed. These interstitial fibers themselves filter instead of diatomaceous earth. Used as a medium, the fibers are supported by a nylon mesh cloth filter support It was done. so that a sufficient amount of fiber can accumulate and act as a filtration medium , pump the gel through the filter for several minutes before opening the outlet.

Next, pump 20 gallons of filtered gel into the 100 gallon tank and Ron's 190 proof undenatured ethanol (ethyl alcohol 190 proof u, s, p, net, 54 gallon batch 1. D, #CT185 JO4, 61 953 Tuscola, Illinois.

P, O, Box 218υ, S, available from Industrial Chemicals. ) was added to this aloe gel fillet solution.

Apply this alcohol-gel solution to a 10.5 inch diameter 18-8 stainless steel several 8-inch-tall 1-1-1 pans (Bloomfu, Chicago, Illinois). Watson, manufactured by Fieldleys, 3712 Hanger Way, Dallas, Texas. ・Available from Food Service Equipment & Subrise) was immediately transferred to

This alcohol-gel solution was allowed to stand for about 4 hours.

Pour the clear supernatant liquid, being careful not to disturb the precipitate that has settled to the bottom of the pan. However, it was handled with a decant or a siphon. Next, this precipitate and the remaining solution are Transfer the liquid into four 1-bind stainless steel centrifuge packets, each packet About 5009 of the precipitate and the remaining solution were transferred to a container. Next, this packet is EC Centra-7 Centrifuge (2ndham Heights, Massachusetts 02194) 300 Second Avenue, manufactured by International Equipment Company.

75050 Guerand Braley, Texas, p. o. Box 1048.

(available from American Sun Scientific Products). Centrifugation was performed at 00×g for approximately 10 minutes.

After centrifugation, the supernatant was decanted and discarded. Next, the beret is freshly 190 pull The tube was washed with native ethanol and centrifuged again at 2000X for about 10 minutes.

After centrifugation, the supernatant was discarded again.

The pellets were then placed in several 600d Vertis freeze drying jars and liquid nitrogen I moved it around until it froze inside. Next, add this freeze-dried jar to Welch Duo. - Seal Shin P. 0. Box 35445, available from Sergeant Welch. ), Vertissmarshon coil cooler (Model Nα6205-435 0 cooler, in acetone bath) and Bernas 18 boat vacuum drum manifold (Model Nα6211-0350). All All Birtis equipment is located at P.G., Guerand-Braley, Texas 75050. 0. Bo x1048.

Obtained from American Scientific Products. freeze-dried dora The chamber was filled with acetone maintained at -50°C. Sample 1@Allow to dry, then It was weighed using a Mettler AE163 balance. The remaining sample is essentially non-degradable. It consisted of lyophilized kallidin. The yield from 50 gallons of aloe vera gel is , about 370-380 g of kallidin.

X straightness once Standard laboratory seedling-scale process to isolate and isolate calicin Approximately 50 pounds of Aloe Barbadensis Miller leaves washed and rubbed. , removed dirty, dried latex and other contaminants. Then each leaf The skin was removed and the entire fillet was placed in a large beaker (on ice).

Blend the whole aloe fillet in 1 to 5 liter batches in a Waring blender. ing blender). Blend the fillets twice at high speed for 2 minutes at room temperature. I ran it. This blended fillet was then cooled to 4°C during the blending operation. The generated bubbles disappeared.

Next, apply this blended aloe juice to 4 layers of Kotton (Cleveland Kotto). The fibrous cellulosic bulb was removed by filtration through a filtrate (Ton). Next, add the filtrate Approximately 4 liters of aloe juice was collected by passing it through 6 layers of cotton.

Next, pour this aloe juice into a large 5 gallon stainless steel container. It was. Add 16 liters of chilled ethanol to this filtered juice. ethanol reagent grade Cat. NαA995) was added. This ethanol , the aloe juice was slowly added while stirring. A flocculent precipitate formed.

The mixture was stirred for 15-30 minutes and left at room temperature for about 2 hours.

Next, decant the supernatant liquid and place the pellets in a small blender. Added 1 liter of deionized water to the render. Blend this mixture on low speed for a few minutes. to clean the pellet, then add 8 liters of nalgene condenser. I put it in tena. Roughly 4 liters of ethanol was added to this mixture, and the mixture was Stirred for 0 minutes. The formed precipitate was allowed to settle for about 2 hours.

Most of the supernatant liquid was decanted off and the resulting pellet was incubated at room temperature for 20 min. Centrifuge at 900X to remove the precipitate to facilitate decanting remaining solvent. It became a let.

The pellets were then placed in a freeze-drying flask and frozen overnight in a Vertis freeze dryer. It was dry.

The weight of the lyophilized powder was 10.99. The yield is 0.273% or It was 2.73xlO'g/d.

Characterization of Kalijin From a historical perspective, aloe vera has been used as a wound healing agent for centuries. It's been getting worse. It is also thought to be good for gastric disorders such as gastric ulcers, and for the treatment of arthritis. It has been considered to be effective in therapy. For example, Duke, James A., CRC Handbook of Medicinal Herbs.

CRC Press Boca Radon, Florida, 31 pages 1985: Henry, R. 0, Cosmetics and Toiletries, An Updated Review Nu of Aloe Vera: Al Art Publishing, Volume 94, pages 42-50 1979: and Morton, Schrier, F., Folk Use and ・Commercial Exploitation of Aloe Leaf Valp: Econo Mitsuku Botany 15 volumes 311-316 pages, 1961 (Duke, Jam es　A,,　CRCHandbook　ofMedical　Herbs,C RCPress, Inc., Boca Ratoo, Florida.

pp, 31. 1985; Henry, R., Cosmetics and Ding oiltries.

An Updated Review of Aloe Vera: A11 Ured Publishing Corp., Vol. 94. pp, 42- 50.1979; Horton, Julia F.

Folk Uses and C0mllerCial Exploitat ion of Al0e tearPulp: Economic Bota ny, Vol. 15. pp. 311-316.1961).

However, as mentioned above, the chemicals in aloe vera that are responsible for these special properties of aloe vera has not been identified and characterized so far.

Polysacca extracted from aloe vera using pharmaceutical screening methods It has now been discovered that Ride is the active chemical in Aloe Vera. This police Kallide is hereinafter referred to as Kallidin (registered trademark). Kallidin is essentially acetylated It is an ordered linear polymer of mannose monomers. protein, organic Other components such as acids, anthraquinones, vitamins and amino acids are It makes up less than 1% of gin. The concentration of kallidin in aloe vera is It was found to be about 0.05-0.3% by weight of the base. Collection of kallidin in leaves The amount or concentration depends on the degree of leaf maturity.

Using the whole Aloe vera plant as a starting material and using certain pharmacological models, When screened for activity, the plant was systematically divided into parts. So Each part was tested for pharmacological activity and inactive parts were discarded. next Divide the active part into smaller parts, discarding the inactive fraction each time and preserving the active fraction. This was carried out until the active substance kallidin was isolated. Activities of various components used rat ulcer protection and tissue-cultured human fibroblasts as a pharmacological model. Measured using stimulation.

Pharmacological data proving kallidin is the active chemical in aloe vera can be summarized as follows.

1. The dose-response of kallidin is It was the same as aloe vera juice containing gin.

2.Kallidin can be administered by various routes of administration: intravenous, intraperitoneal and oral. was effective in the ulcer protection model.

3. Kallidin is 100% effective in both pharmacological models.

4. Similar to kallidin from a completely different source, the konjac plant The substance, the chemical glucomannan, has shown several pharmacological effects.

Kallidin has been shown to be associated with burns, ulcers, and other wounds to the skin and gastrointestinal wall. It is known to increase the replication of fibroblasts in tissue culture by up to 300% in 48 hours. Laboratory studies have shown that there is an increase in

Kallidin was also shown to increase DNA synthesis in the nucleus of Fi fibroblasts.

And increased DNA synthesis increases metabolism, a fundamental step in the healing process. Increases activity and rate of cell replication.

Kallidin has been shown in controlled studies to increase the rate of healing in animals.

Kallidin has also been shown to be an effective treatment for gastric ulcers in animal studies. Ta. Laboratory rats, whose stomachs react similarly to human stomachs, were tested over a three-year period. It was. Kallidin is similar to or similar to current medications used to treat stomach ulcers. was found to be superior to that of Many of these products cause salt in the stomach. Acts as an acid suppressant. Kallidin works on a different principle, It does not alter the natural flow of acids.

As mentioned above, kallidin has a water-soluble lower aliphatic polarity in liquefied aloe <-y gel. It can be precipitated by adding a solvent, preferably ethyl alcohol. Ru. Powder of kallidin can then be made by freeze-drying and This freeze-dried product is then processed into a Mourinex coffee grinder (Da. (available from Dillard's, Las Vegas). It can be ground into powder. Kallidin powder is highly electrostatic and Depending on the oxidation state of some anthraquinone contaminants, the color ranges from yellowish white to pink. It is a purple amorphous powder.

Kallidin is stabilized and produced by freeze-drying, which removes water that causes hydrolysis. Qualitatively non-degradable. Freeze-dried aloe veraglut with prescribed oral strength lysine maintained its validity for two years. In lyophilized form, kallidin is stable for up to 10 years. It is thought that.

It turns out that heat and time are important factors in the production of powdered kallijin. won. Heat promotes the hydrolysis or decomposition of kallidin, resulting in The longer the lysine processing time, the more degradation occurs. Therefore, high molecular weight If lysine powder is desired, kallidin can be extracted from whole Aloe vera leaves the fastest. It is preferred to use a process that allows Low molecular weight kallijin powder If a powder is desired, kallidin can be extracted from the whole Aloe vera leaf in the slowest way. It is preferable to use a process that can

Rehydration of kallidin powder at a weight/volume concentration of 0.2-1% produces fresh aloe vera. A viscous ``gel'' resembling a la was formed again. Rejuvenation by rehydration of kallidin powder This gel-like consistency is attributed to kallidin's high molecular weight polysaccharide. It shows the nature of id. Generally, polysaccharides are degraded or hydrolyzed Then, its viscosity decreases.

Therefore, the viscosity of rehydrated kallidin powder gives an excellent indication of quality and ensures quality assurance. Can be used as a parameter.

Kallidin produced according to the process of the present invention is acetylated mannose monomer. -, preferably linked to each other by β(1→4) bonds, substantially non-degradable can be characterized as a lyophilized, ordered, linear polymer.

Various modifications and alternative modifications, variations and equivalents of the disclosed compositions of the present invention. etc. will be apparent to those skilled in the art after reading the above general description. next Examples (Examples (12-44) was carried out to roughly characterize and identify similar polysaccharides. below The examples are illustrative and may include supplements covering modifications or equivalents or variations of the above. It is not intended to limit the scope of the appended claims.

Wen Jeongfeng - Just Comparative High Performance Liquid Chromatography (HPLC) Commercially Available Glucoman Nan (polysaccharide isolated from the Japanese konjac plant, General Nu) (purchased from Region Center) in a pharmacological screening model. It showed some movements similar to Kalijin. This konnyaku mannan (hereinafter referred to as konnyaku mannan) lucomannan), which is used to characterize kallidin, is quite It turned out to be different.

Initial experiments were performed on glucomannan and kallidin in parallel. these Both polysaccharides were treated with 1 molar sulfuric acid (o, 01 SF polysaccharide). /2me sulfuric acid) at 110°C under reduced pressure for 24 hours. Hydrolysis High performance liquid chromatography (tlPLc) of the product Minex HPX-87C column; R, 1, detector: flow rate 0.6 d/min: elution elution medium (HPLC grade water) along with the presence of mannose and the glucose preparation. The eluted peaks are shown.

This “glucose” peak is a broad non-contrast peak, indicating that it is a modified It was shown that it contained a lot of glucose. performed at slightly different concentrations Acid hydrolysis gave similar results, with glucomannan showing distinct glucose and manno kallidin gives a distinct mannose peak and a glucose standard product. At least one peak migrating at the same retention time was given.

Next, these two types of polysaccharides were enzymatically hydrolyzed using a crude cellulase preparation. It was attached to. Commercial glucomannan was hydrolyzed and HPLC (same conditions) analysis shows well-separated glucose and mannose peaks and Three other peaks were observed that moved faster than lucose. Gee, Tory and These peaks showing polysaccharide and tetrasaccharide moieties are probably polysaccharides. This was thought to be due to branching or incomplete hydrolysis. Enzyme preparations are semi-purified preparations. Therefore, other glycosyl hydrolases are undoubtedly present, and this enzyme preparation Precise determination of the binding specificity of the protein could not be performed. Same enzyme treatment as kallidin A very different profile was observed on HPLC. this enzyme Hydrolysis leads to mannose, glucose and larger unhydrolyzed oligomers. At least two corresponding larger peaks were given. In fact, many polysacca Rides could be found in these larger molecular weight fragments. child This means that its composition (i.e. type of bond and degree of branching) is different from that of commercially available glucomas. This seems to indicate that it is significantly different from Nannan.

Acid hydrolysis is a common hydrolysis method, while enzymatic hydrolysis is a method of It is much more specific in terms of type.

Tomo Umaya - Tsuki Chemical hydrolysis of large polymeric saccharides to aid in characterizing caricin It was decomposed by the solution. Hydrolysis was performed by Alberginiim, P et al. , Nnevins, D.J.: English, P.D.: and Karl, ^ (1967) Carbohydrate Research, 5.340-345) (A lbersheim, P.; Nevins, D. J.: English , P, D; Karr, A (1967) Carbohydr, Res , 5.340-345) 2N trifle modified from the method developed by It was carried out in an oracetic acid solution (2N TEA). Dry the hydrolyzate in a vacuum oven The mixture was reconstituted with 0.0 IN H2304 and used for HPLC separation.

HPLC separation of hydrolyzate; Separation of hydrolysates was carried out using a Hewlett-Barackard liquid chromatograph model 108. 4-8 was used. Add bio-chemistry to the chromatograph to monitor eluates. A Rad Model 1170 refractive index detector was installed. Separation column is hydrogen type 0.65 x 30 cm bed filled with interaction cation exchange resin It was an interaction 0RH-801 organic acid column containing.

The mobile phase was 0.01 NH2SO4 at a temperature of 35° C. and a flow rate of 0.571 min. single The retention times of the chromatograms were determined using sugar and acid standards. Such a provision Galacturonic acid, glucose, mannose, galactose, The retention times of munose and arabinose are approximately 7.45.8.04.8. It was determined to be 69.8.86.9.20 and 9.78 minutes.

A hydrolyzate of the kallidin sample was separated under the same conditions and some of the sugars were shown below. were identified in various amounts.

Table 1 Batch #1 Trace 476 14279 - - 5818 Batch 113 584 Trace 16926 - -4538 Hatch Wing 3B 2329 Trace 15068 - - 5553 Hatch 15B 592 209 13158 - - 7888 batch 1i8B 1236 Trace 15049 - - 53 92 batch ti6B trace 329 12686 - - 7465C-70 1008 traces traces '13734 - - 6240 batch #4 traces 1 260 17800 - 535 7699 batch #8B Trace 286 1 3715--6677 (dialysis) C-613006 trace trace 13571 - - 5782 sucrose Octaacetate-2580---6407 Hoglucomannan 441 103 80 15050 - - 5553 (konnyaku) (Golf mannan and sucrose octaacetate from the umbrella konjac plant are It was included as a reference standard. ) Liquid chromatographic separation of these hydrolysates is This shows that kallidin is essentially mannose.

and varying amounts of galacturonic acid, glucose, rhamnose, and arabinose. and other sugars. Some of these sugars are present due to their low amounts in solution. It is difficult to measure accurately by this method. Presence of acetyl group as acetic acid Although the amount is large, it could not be completely attributed to kallidin. That's called The reason is that the membrane filter used to clarify this hydrolyzate solution is clogged. This is because they appear to have been made by adding various amounts of acetate to the matograph. Group Lucose was present in some kallidin samples analyzed by this hydrolysis method. It is noteworthy that the amount is extremely small. However, kallidin is similar to H2SO4. When hydrolyzed with mineral acids, the glucose content is reduced by the cellulose compounds present in the extract. It will probably be much higher due to quality hydrolysis.

The observation that kallidin is mostly mannose also suggests that this hydrolytic Limethylsilane (TH3) i-conductor gas chromatography/mass spectrometry It was also confirmed by GC/83 separation and identification. These means , showed that mannose accounted for more than 80% of the hydrolyzate.

Once upon a time Optical rotation: Kallidin is levorotatory with respect to the sodium D line at 589 nanometers. I found out. 25% methylmorpholine oxide in dimethyl sulfoxide solvent A 1% (W/V) kallidin solution dissolved in a solvent containing It gave a specific rotation of . This value was calculated using a 25% HMNO solution in DH3O as a blank. It was measured using The polarimeter used was PerkinElmer 2411 (C). Ta.

The optical rotation of the aqueous solution of the kallidin sample was measured using a PerkinElmer polarimeter model 2418. It was measured using C. This measurement was performed at a sodium line wavelength of 589 nm. I was disappointed. The sample cell was one with an optical path length of 1 decimeter (dm). direction Law: A total of 20 (ly) kallidin samples were accurately weighed, and a 200 d polypropylene sample was prepared. I put it in a bottle. Deionized water (ia, e megohm (HΩ), 100 d) In addition, the concentration was 0.2% (w/v).

This suspension was transferred to a laboratory line orbit shoer (jab-Line Orbit Shuker). 5haker) for 12 hours to completely dissolve (from the konjac plant) Commercially available glucomannan was partially dissolved).

A typical solution of kallidin, which is usually cloudy and viscous, was added to a 1.2 μm membrane filter. Luther (Uniflow# 46-02360゜Schuroscher & Schnee Le Company, Keene, NH) (υniflow $46-02360.5chliec her and 5channel Inc.

The mixture was filtered using Eene, NH). This filtrate was filtered through a 0.45 μm filter. (Uniflow #46-02340) to obtain a clear solution. . Optical rotation was measured using deionized water (18,6) 1Ω) as a blank.

The measured angle of optical rotation was converted to a comparative optical rotation as follows.

In the above formula [α]′5<= specific optical rotation α = measured rotation angle ■ = Volume of solution (ml) Q=weight of active substance in solution νd! (g) ↑=humidity temperature 25℃) λ=wavelength (589nm) The results are shown in Table 2.

1613006 -14.7±0.8 #　C701008-12,7±0.413B　-14,2±0.7 Oath 5B -9,5±0.2 Glucomannan -19.0±0.4 The optical rotation indicates that kallidin is levorotatory.

The rotation angle changes depending on the batch.

All of the above data indicate that kallidin is a polymer with β(1→4) bonds. It shows.

Example 15 Elemental analysis: Elemental analysis of organic compounds is a powerful tool in elucidating structure. Karijin is real Qualitatively organic and therefore able to follow this procedure. Karijin sample The batch was sent to Yatsuman Laboratory in Wheatridge, Colorado for elemental analysis. Huffman Laboratories Incorporated 1nWheat Ridge, Colorado). oxygen, carbon , hydrogen and nitrogen accounted for 80-90% of the structure of kallidin. Phosphorus and sulfur combine Together, they accounted for 0.5-2%. Table 3 shows the reported results.

Table 3 "j-'y7"k# Pi **mM MW fiLit Nidan> Hatsuchi 81 37.625.71 45.851.270.31 0.61 batch! ! 2 3 0.334.67 45.161.15 0.34 0.74 batch 113 2 9.584.23 44.300.46 0.28 0.33 hatch! 4 39 ．． 865.84 46.430.52 0.16 0.60 Batch #5 37. 455.76 46.290.81 0.18 0.68 Baychi 16 41.7 26.11 46.140.84 <, 30 0.48 Haiti $7 39.20 5.66 45.401.36 <, 30 0.54 Hiltop 36.455 ．． 35 44.071.51 0.44 1.02 Mitsushiel 41.575. 96 43.251.35 0.37 0.71TCX 33.475.034 4.531.570.461.25 For batch numbers 3, i4, #5 and #6 It is important that the nitrogen content is relatively low. These batches are made with denatured alcohol was created from. These physical appearances also differ from the majority of Kallidin samples. It was different. Roughly cell culture response of these batches (human fibroblast stimulation) was significantly lower than average.

Example 16 Emission spectrum analysis: Emission spectrum analysis was performed on the calicin sample. This analysis It was performed by Huffman Laboratories, Wheatridge, DE. Silver (Aq) All 68 elements from Zn to Zn were monitored. slightly Several types of elements were detected. These include calcium, sodium, silicon, and magnetism. They were lithium, manganese, copper, chromium, barium, iron and aluminum. ring Toxic metals listed by the Environmental Protection Agency (EPA), especially lead, molybdenum, and cobal. Cadmium, arsenic, selenium, and mercury have been detected by this semi-quantitative method. There wasn't.

X number -■ X-ray diffraction analysis: kallidin using a Philips Electronic Instrument X-ray diffractometer. A powder X-ray diffraction pattern was obtained. 2θ angle in the range of 5° to 60” per minute. The image was scanned at a speed of 2θ angle. The radiation is 15 milliamps and 3 2 (CuK2) to the copper from the tube operated at 5 kilovolts.

Many of the kallidin samples tested by X-ray diffraction analysis were appreciably crystalline. did not show gender. Kallidin was essentially amorphous. This kallidin amorphous properties can be destroyed by deacetylation, and the rehydrated form can also be destroyed by deacetylation. Thermogravimetric analysis: Decomposes slightly by air drying: This technique measures the change in weight loss with temperature. Produced using experimental seedlings Both the sample of Kallidin and the sample of Kallidin produced on a large scale were The analysis was performed using a Metosee thermogravimetric analyzer TA 3000 system. this system It consisted of a TC10 ATA processor and a 1G50 thermobalance. temperature range The temperature range was from room temperature (25°C) to 750°C.

Kallidin exhibited a single degradation pattern that led to the assumption of a possible simple structure. minutes The solution profile is that of glucomannan obtained from konjac plants. It was different from Le. Kallidin releases surface water and bound water between 50℃ and 210℃. The main loss peak was at 95°C. Most of the decomposition occurs between 210°C and 405°C. , the peak was around 325°C. Decomposition occurs roughly between 406°C and 735°C. It was. Oxidizable carbon disappeared between 630<0>C and 737<0>C. consists mostly of ash The residue accounted for more than 10-20% of the total weight. Table 4 shows some samples of Kalijin. The thermogravimetric decomposition profile of the sample is shown.

Table 4 San l-120 decomposition product oxidizable residue Sample # Pull Total Weight (Irtg>(y>(%)(%)( %) 606005 6.53 9.35 49.6611.596.0316. 4 7.70711009 5.60 6.55 40.0811.495.5 019.09 18.74430003 8.24 9.33 36.4310 ．． 764.6024.69 1B, 65620007 10.17 10.25 38.2110.273.6225.68 17.83701008 5.7 9 9.65 45.1010.555.0913.43 17.255310 04 4.96 8.92 4B, 9411.146.3811.98 13. 98613006 4.86 8.16 4B, 310.755.4015.4 1 16.38 (MEX 4 layer) 8.03 8.46 26.0411.553.46 31°61 25.84 (Man Batch 1) 9.22 58.2 12.70 4.70 3.52 10 ．． 74 (Manbatch 2 Acetyl) ? , 96 12.81 37.6310.166.84 15.28 18.9 7531004 7.74 5.45 66.94 6.893.69 15. 0 2.51 Glucomannan from konnyaku 30.6 9.82 11.3260.830 17.4 0.668 decomposition program Rofeel is unsure of the origin of the aloe leaf from which the Kallidin sample was made. It was observed that the results were almost the same. This profile is a konjac plant It was quite different from glucomannan. Manufactured batch date is some It really shows the difference. This is manufactured using a significantly different process. This was not surprising, since The precursor gel consists of kallidin It is clarified by filtration through a diatomaceous earth filter before being precipitated from the gel with coal. Ta.

X Healing Complex-U density: The density of calycin varies depending on the amount of inorganic salts precipitated with calydin.

The density of kallidin is also highly dependent on hydration. the product prior to freeze-drying. A fluffier, less dense product is obtained by adding additional water . In order to preserve the phlegm, an enzyme is added during its manufacture prior to freeze-drying. The following samples were prepared using standard methods using a tanol to water ratio of 4:1 (v/v). A sample was produced.

operation: 10m of pre-weighed dry kallijin powder! ! into a graduated cylinder, then The cylinder was weighed again to determine the total weight of kallidin added. all Weights were determined on a Metsy AE 163 analytical balance. Next, pop this cylinder. Stir at high speed for 10 seconds with a Vortex-Gen1e stirrer. Stir and then observe the volume of kallidin. The density is determined by the volume of dry kallidin powder (d ) per mass (g). The results are shown in Table 5.

Table--1 Batch: Ex-Ichie 11 0.074 #3 0.640 114 0.382 #613006 0.158 $701008 0.291 Valley 3B 0.561 15B 0.759 # 5B 0.626 7B each 0.683 8B each 0.499 Glucomannan 0.596 Dry kallidin powder 13 from batch 115B was dissolved in 50 d of deionized water.

This solution was then lyophilized. This dry kallidin has a density of 0.110 It was a fluffy white powder. Thus kallijin from batch 115B A dry powder is prepared by redissolving the 50 precepts in water and then freeze-drying this solution. The density of changed from 0.759 to 0.110.

Example 20 Solubility of kallidin: Caricin is sourced from aloe leaves, processed by filtration, ethanol precipitation and drying. exhibits varying solubility properties that are highly dependent on the extent of Carijin different batches of water The degree of solubility was observed. Different Kallidin batches with the same concentration (weight/volume) can create aqueous solutions (sols) that vary widely in viscosity and inorganic salt content. . Viscosity and inorganic salt content generally affect the solubility of a substance.

This study was conducted using Kallijin production batch 11 and batch $2. This 2 One batch has several characteristics, both in origin, processing and some chemical composition. have the difference. Solubility in water and other solvents may vary.

The solvents used were water, acetone, dimethyl sulfoxide (D) Lahydrofuran (THF), 0.9% sodium chloride and 0.1% benzoic acid It was sodium.

Method: A total of 18 polypropylene test tubes were set up. Each of the 6 bottles is a Karijin. The remaining 6 tubes were for the batch and the remaining 6 tubes were for the solvent blank.

0.049 of kallidin was weighed and added to each sample tube. Add 10ml of solvent A 0.4% (W/V) mixture was prepared. This mixture was stirred at room temperature for 5 hours. . This suspension was transferred to a centrifuge tube and centrifuged at 500 rpm for 50 minutes. . Solvent blanks were treated similarly.

The solution was decanted from the solids. Completely dries both solution and solid (precipitate) It was dried in an oven at a temperature below 50° C. until it was dry. The dried solids were weighed.

The solvent flask that produced the precipitate was also weighed. Subtract this weight from the sample weight. there was.

Table 6 result: Caricin was found to be more effective in water and in aqueous solution than in other solvents tested. It also had high solubility.

A compromise between the high viscosity in aqueous media and the extremely low solubility of kallidin in other solvents. A concentration of 0.4% (W/V) was selected for the purpose of cooperation. About 0.5% (W/V) of kallidin It is possible to dissolve it in water.

Inu 挌 and immediately koji once Canon Fenske clay mold (model 150. New Jersey, Union , Industrial Research Glassware Co. > (Cannon-FenS ke tVDe Viscometer (Node1150, Industry real Re5search Glassware Ltd,, Union, The viscosity was measured at 40° C. Kallijin 0.2% water in the viscometer Pour the solution (0.05% NaN5) into a water bath (Model SBG-1090, Blue -M, Electric Co., Blue Island, IL) for 10 minutes. next Solution flow was observed down to 0.1 seconds according to the manufacturer's instructions. caliber The ration factor is 0.04197 centistics/sec at 40°C. Ta.

The results are shown in Table 7.

Table 7 14 5.09 11613006 2.12 I C7010082,08 13B 2.02 15B 4.53 66B 1.75 ft 7B 1.55 188 1.97 Example 22 2 degrees ± Although the function of calcium in Aloe vera activity is not completely understood, The quality of Evera gel is sometimes based on its calcium and magnesium content . Since kallidin is the active substance in aloe vera gel, the calcium content is measured It was done.

The total calcium in Kallidin is determined by the Calcium Rapid Stat Kit (Rapid Stat Kit). Sir Medical St. Louis, No. (Calcium Rapid 5 tat kit (Lancer Medical.

It was measured spectroscopically using St. Louis, ) 10)). 0.2% potash A 50ul sample of aqueous solution (o, os% NaN5) was added to 3.0d of reagent. In addition to the blue calcium-methylthymol complex IB) l model 94 Measured at 612 nm using a 20 UV-visible spectrophotometer.

Bound calcium was determined by the following empirical method = 0.2% kallidin aqueous solution ( A 25 d sample of 0.05% NaN5) was placed in a dialysis bag and 18.6 ) Dialysis was performed overnight at room temperature against 10 liters of 1Ω deionized water. then the bag The contents were transferred to a lyophilization jar and lyophilized. Weigh the dry powder, then weigh 20 % nitric acid, 10% hydrochloric acid solution at 80° C. overnight. sample, atomic absorption spectroscopy Truck Laboratories in Denton, Texas to measure calcium by analysis I sent it to Leeds (choraCcaborator+es).

Note: All containers in the above operations are Nal (lene). Made of polypropylene, 1.0MHCU and deionized water (18,6) 1Ω ) and thoroughly washed.

The results are shown in Table 8.

Table 8 Freeze-dried Calcium (χ) Calcium (%) Solid overlay batch (dry kallidin) (dry Dried kallidin) (my>1 3.7 0.042 31.7 3 10.15 0.066 12.94 2.15 0.048 32.0 613006 6.85 0.060 20.6C7010086,650,0 6823,93B 6.50 0.074 20.75B 3.55 0.07 4 29.6 glucoma can be ignored Nnan Husband Healing Spot - Qian Nuclear Magnetic Resonance Spectrum (N)IR): N)IR is essential for elucidating molecular structure. This is a very powerful tool. NHR spectra are based on reference standards for chemical shifts. Caliplated using tramethylsilane, etc.).

N) Some solvents commonly used in IR spectroscopy (D20°DMSO, acetic acid This The technique was not successful in kallidin solution.

However, the excellent spectrum of kallidin as a solid is due to cross-polarization. It was obtained using the magic angle spinning (CP-)IAS) technique. In-phase power Lysine C-138) iR spectrum was obtained. This spectrum was taken using an IBM spectrometer. as well as some special accessories included with this IBM NHR spectrometer. I got it. According to this spectrum, C-138HH of the analyzed kallidin The peak of chemical reaction for tetramethylsilane (n+s> Regarding shifts, 20.54, 60.39, 71.33, 100.75. It was located at 171.78 and 180.75 ppm.

Tentatively these peaks are assigned as follows: 20.54 and C-4 and C -5 carbon). This assignment is probably correct given the size of this particular peak. 10 The peak at 0.75 ppm is probably due to (C-1 glycoside) (mag b e).

The peak at 171°78 is slightly lower than expected for ΩOOH, but It probably depends on the orientation of The last peak of 180.751)l)m is carbonyl carbon (C=O). Approximately 95pp corresponding to C-1 (reducing end) Some small peaks of m were also observed. May indicate C-1 of furanosyl unit I don't know why some peaks between 106 and 1109 pp are missing. is important.

These assignments are consistent with the structure of β(1→4)mannan.

X 旌桝-ha Infrared spectroscopy: Infrared spectroscopy is effective in identifying and characterizing kallidin. It was. IB)l IR/32 Fourier transform infrared spectrophotometer is suitable for this technique. used. This device uses software and a He-Ne laser to self-calibrate. This is proven by the standard spectrum of polystyrene film can do. Optical adjustment is sometimes used when the signal-to-noise ratio is low. is required.

Kallidin is ground into powder and mixed with infrared grade potassium bromide (KBr) powder. match. This mixture can be pressed into transparent discs and scanned with infrared light. I can do it.

0.2% (W/V) carigi exhibiting better spectral resolution than KBr disks. A new method has been developed to create transparent films of aqueous solutions. 40 karijin When scanning from 00 to 400 cm, the following functional groups are observed in the spectrum. The stretching vibration and corresponding bending vibration 1470-1460 cm-' of C- It was an H group. Other major functional groups observed were: (1) COO unit: its asymmetric and symmetric vibrations are 1600-155 respectively (2) Ester group ( GOOR): Its stretching vibration is 1746-1735 Cm-1 and 1250-12 It was observed between 40 cm-1.

(3) Amide group (CONH): The stretching vibration of the amide ■ and the bending vibration of the amide ■ The motions were observed around 1650-1645 and 1550-1540 cm-1, respectively. It was done. Table 9 lists the approximate vibrational frequencies (wavenumbers), corresponding functional groups and vibrational modes. is exemplified.

Table 9 -1 1100-10350-1-(.

C-0-CV (0-H) 2 3430-33900-HV (0-H) 3 2930-2800 C-HV (C-)(>4 1470-13800-Hδ(C-H)5 1600-155 0 Coo-.

6 1450-1400 V (C=O) symmetrical 7 1420-1410 0-C 131650-1645C0NHV7mit'I　C=0 9 1550-1540 C0 NHV amide ■10 1746-1735 C0 ORV (C=O) 11 1250-1240 C0ORV (C-0-C) 12 980-950　C-0゜ -CH2 Table 10 shows the absorption maxima of the peaks of various kallidin samples and the attribution of the peaks in Table 9. are shown below to show how they match.

Table 10 Karijin 531004 Karijin Hill Top Karijin Patch Cup 2 (cm-1) (cm-1) (cm-1>1591.5 1068.7 1585.734 14, 4 1037.8 3400.91072゜6 3420.2 1086 ．． 11246, 2 1244.2 1425.61740.0 1734.2 1037.81431.4 1635.8 1244.2603, 8 1653 ．． 2 1740.02930, 2 1375.4 605.780B, 3 4 72.6 679.01419, 8 2928.3 1558, 7 545.9 605.7 956.8 2924, 4 806.3 879, to According to infrared analysis of sil] appears to be necessary for polydisperse heteropolysaccharides with functional side chains. mosquito The lysine sample was fully O-acetylated and also deacetylated. Ace The carbonyl functionality and C-0-C stretch of the chilled kallidin sample are observed between 1748-1735 cm-' and 1246-1235 cm-', respectively. It was. The capoxylate carbonyl stretch of the deacetylated kallidin sample is , respectively 1600~1550o++ and 1450~1400cm- as follows. Located between 1: Table 11 Kallidin 531004 Kallidin 531004 Kallidin 531004 (deacetated) tylation　　〔acetylation〕 (cm””) Ccm-1) Ccm-1>1591.5 1431.4 124 4.23414.4 3400.9 1741.91072, 6 1032.0 1064.81246, 2 1062.9 3460.71740.0 10 91.8 1375.41431.4 875.8 164γ, 4603.8 1147.8 1433.32930, 2 1645.5 2932. , 280 6.3 2924.4 1541.3814, 1 956.8 615.4 603.8 When kallidin is deacetylated, the ester between 1248 and 1235 cm It was observed that there was no stretching vibration of Le C-0-C. The largest peak in the spectrum is It was also observed that there is a center at 1431 cm-1. Acetylation/deaeration of kallidin The degree of cetylation can be observed by these two peaks. actual , in this example, amide ■ is observed at about 16460, so 1431 The cm-1 peak (1560-1339 cm”) is mixed with the amide ■ peak. there was.

X dust cloud-pa Biological activity: Both in vivo and in vitro experiments have shown that kallidin promotes fibroblast proliferation. was observed. Facilitation by a factor of 2-3 compared to control is sometimes recorded It was done. Table 12 shows that kallidin at a concentration of 0.1% (W/V) for 12 hours Figure 1 shows affected fibroblast proliferation at different times.

These experiments were carried out under various conditions. Sample concentration 24 hours 48 hours 72 hours (w/v) % % 3 Ding CX Lab 7/25/85 0.1 90.7 162.2 163.8 H) 1 (TCX) o, i 1 04.3 139.7 129.5 batch 2 0.1 72.1 123.2 144.9 batch 3 0.1 75.5 130.9 128.4 batch 4 0 ．． 1 102.3 131.2 135.1 batch 5 0.1 79.3 11 5.0 129.2 x 6 0.1 57.7 130.9 113.3 x Chi7 0.1 65.1 110.6 120.2 Batch (Lab) 5/83 0.1 81.1 138.3 169.7 Manpatch IB O,1 125,6174,8114,8 Manpatch 2B 0.1 103.5 175 ．． 3 118.6 man patch 3B O, 1138, 9156, 4147, 0 man patch 3B (hydrate) 0.1 103.3 141.3 158.9 glucomannan 0. 1 103.3 69.9 108.6 (Koniya (plant) Control SCHO, 1100, Oioo, oioo.

was performed to evaluate the response of cells to various samples.

X group - average Acetyl group analysis The amount of O-acetyl groups in a series of different batches of kallidin was determined. prior to analysis Samples were then dried overnight with P2O5 at 40°C in a vacuum oven. Next, deionization Approximately 1ffi using water! A solution of F/d was prepared. The analytical operations used are Hestrin (column) containing colorimetric determination of iron-acetohydroxamic acid complex Mohestrin, Journal of Biological Chemistry (1949) , 180, 249-261) (Shlomo He5trin, J, Biol, Chem, (1949) 180.249-261). The operation was the same as the one given.

The results are as follows; Acetyl group per gram of kallidin 48 0, 23 5B 3.33 6B 2.28 88 2.32 108 1.68 531004 2, 97 613006 2, 77 701008 2.61 Complete acetylation 5.69 8B>100K 3.93 8B l0K-100K 2.98 333K -10K O, 10 3B<3K Not detected Methylated 8B not detected Relative activity values for 72 hours of eight 0.1% kallidin solutions measured on the same day (Fibroblast stimulation) and acetylation tables are as follows.

Table 14 Batch Millimoles of active Nα acetyl groups per gram of kallidin (%) 613006 2, 7700 136.0000701008 2.6100 95.50001 3.7200 113.2000 3 1, 3000 93.9000 4 4, 1500 104.7000 3B 2.1100 111.700048 0, 2300 74.4000 5B 3.3300 100.7000 This data indicates that the activity of kallidin is It is suggested that the amount of 0-acetyl groups per gram of carbon is directly proportional to the amount of 0-acetyl groups per gram of carbon. this is a surprising and unexpected result.

The table of viscosity values (centistokes) and acetylation for nine kallidin samples is as follows: That's right: Table 15 Batch 〇-viscosity per 1 g of kallidin (centiNα mmoles of acetyl groups) Stokes) 1 3, 7200 6.1000 3 1.3000 1.1800 4 4, 1500 5.0900 61300B 2.7700 2.1200701008 2, 6100 2. 08003B 2.1100 2.0200 5B 3.3300 4.5300 6B 2.2800 1.7500 8B 2.3200 1.9700 This data shows that viscosity is exponentially proportional to the degree of acetylation of kallidin. It shows. The following straight line of best fit was determined from the above data: Viscosity = g, 4g6xo (0,604x mmoles of acetyl group/g) The next experiment involved aloe leaves obtained from Hilltop Gardens in Harlingen, Texas. It was done using. The leaves were washed, cut, filleted, crushed, and homogenized (1 500 psi) and filtration (pulp filtration media). All processes in 1 It was completed in less than 20 minutes.

Part D 9:10am 3 liters of gel and 12 liters of 190 proof undenatured eta The nols were mixed vigorously in a 5 gallon polypropylene bottle. This process is , twice using a total of 6 liters of gel and 24 liters of ethanol.

5 liters of this mixture was added for 1 hour, 2 hours, 4 hours, 8 hours and 10 hours, respectively. Quickly place each of the five stainless steel pans labeled with time. I moved it. Another pan was used for the 24 hour test.

For example, if 1 hour has passed since the time when alcohol was added, attach a 1 hour label. The sample was concentrated by removing the alcohol with a siphon, and then Centrifuged. Wash the solid with fresh alcohol and decant this after centrifugation. did. The solid precipitate was placed in a lyophilization bottle, quickly frozen in liquid nitrogen, and lyophilized. I did drying. The process of removing alcohol and sending it to the freeze-drying process averages 50 to 60 It took a minute. Both labeled portions of each were treated in the same manner.

Table 16 Partoe research Sample time Nα (freeze-dried Time Dry weight Yield Yield Yield Rate Time Time put into the dryer＞(y＞(y/ fJ>(w/v)1 10:10a,m, 11:15a,m,,7176. 7176. 07182 11:10a, m, 12:10p, m,, 7643 ．． 7 & 43. 07644 1:10p, m, 2:15p, m,, 7748. 774B, 07758 5:10p, m, 6:10p, m,, 7903. 7 903. 079010 7:10p, m, 7:40p, m,, 8147. 8 147. 081524 9:10a, m, 10:05a, m,, 9373. 93732.0937 Bart■ The research in Part ■ is different from Part II in that purified aloe gel was subjected to time research. It's different. At X hours (X is 0, 1, 2, 4.8 and 10 hours respectively) between).

Mix 2 liters of purified gel with 8 liters of 190 proof undenatured alcohol. Ta. Stir the mixture and remove the alcohol by siphoning and centrifugation. It was allowed to settle for 4 hours beforehand. The solid precipitate was treated as described in part ①.

Table 17 - Examination of Kalijin's time course study on Bart's study Size exclusion chromatography (SEC): The average molecular weight distribution of calicin is 5ize Exclusion Chromatography oqraphy) (SEC). Liquid chromatograph, HP 79875A tlV-VIS detector automatic sampler 79841A and Hewlett Bakker featuring Bioland Model 1770 Refractive Index Detector It was a model 1084B. This column is Spheroguru. Beckman packed column 7, 5X 300M (Ding SK 2000 5w; Part N (1244282, 526 in serial Nα5). mobile phase is a 0.05% (W/V) sodium azide solution at a flow rate of 0.5rd! /min, warm The temperature was 30°C. Dextran from Sigma Chemical Co., St. Louis, Missouri Using the standard sample, the average molecular weight distribution was determined as follows. - Quitran Nα Lot Na Average molecular weight (min) D-537624F-02 982,000,00011,21D-139013F-042771,200 11,49D-4133114F-033540,60011,840-926 053F-02409,00017,71 The same conditions apply to the separation of kallidin The following three main peaks were observed.

Hilltop (lyophilized) fillet: Fraction Nα Retention time (minutes) % Total % - (dan" 1 minute - 110,8710,2 012,2 216,3817,7321,1 322,4856,0266,7 Hiltop instant process (short time): Good retention time (minutes) % total %-(3" 1 minute-111,8543,1446,8 216,5719,7421,4 322,0529,2431,7 Hiltsupbad I (1 hour): Otori Koho ■ Ishu Sara” Ben L % Total % (for 3 cars) 111.52 50.30 5 4.6 2 16.28 14.04 15.23 21.99 27.71 30.1 Hiltop Part II (4 hours): ■Savings: Hao JI Saki 1L package - % Total % (for 3 cars) 1 11.79 56. 42 58.32 16.33 15.30 15.83 21.98 25. 07 25.9 Hiltop Part ■ (10 hours): Fraction Nα Retention time (minutes) % Total % J - 111,1457,7360,4 216,3714,6615,5 322,0322,4423,7 Hiltsup part■ (24 hours): Hokoi Holding time (minutes) % Total % (3 cars)'l 11.41 45.75 50.52 16.55 19.35 21.43 21.28 25.43 28.1 When the initially prepared kallidin sample is separated under the same conditions as above, Three large bifurcations were also observed.

TCX batch #1 Fraction Nα Retention time (min) % Total % - (βff11 11.18 35.1 8 36.982 16.12 7.04 7.40 3 22.29 52.91 55.60 Mexican batch #2 Both minutes No. Holding time (minutes) % Total % (3 cars) 111.04 19.02 19.0 2 16.46 24.27 24.33 22.38 56.70 56.7 Based on the dextran standard, the average molecular weight distribution of the three halves of kallidin is as follows: It seemed like that.

Both #: 1 > 80,000 #2≧io, oo.

#3<1,000 Because dextran and kallidin polysaccharides are physically and chemically different, , only the average molecular weight distribution can be determined. For example, size separation is may be influenced by other factors such as ionic charge adsorption and/or partitioning of Ru.

Calcium content of kallijin: Although the function of calcium in aloe activity is not completely understood, This parameter was also observed in kallidin. Calcium content of kallijin Lancer is a commonly used constant colorimetric measurement of calcium in serum and urine. ・F11 was determined by Calcium Rapid Stat Kit. formed The deep blue calcium-methylthymol pull complex was prepared at 612 nanometers. Read with tar. In this example, for the measurement of absorbance, IB) lUV/VIS A spectrophotometer model 9420 was used. Calcium based on three calcium standards The calcium content of lysine was determined as follows.

Bartoe Sample # Calcium% 24 5.40 Part■ Sample # Calcium% infrared analysis Infrared (IR) spectroscopy as an analytical technique is based on the viscosity of an aqueous medium, which requires a This is particularly important in the analysis of kallidin, which presents problems in all analyzes that require It is.

Infrared analysis of kallidin was performed 9 to determine its functional groups. Equipment used was an IBI (Fourier transform infrared (FT-IR) spectrometer model 32. The condition of the device was: Resolution = 4 Number of scans = 32; Detector = Ding GS .

Since Kallidin is a powder, it can be easily converted into infrared grade potassium bromide (KBr>powder). This mixture can be pressed into discs. Next, this de scan the disk. However, the transparent fibril of 0.2% (W/V) kallidin aqueous solution A new technique for creating lums has been developed. This film uses the KBr disc method. It also gives an excellent infrared spectrum of kallidin. Caliton at 4000cm-'? When scanning up to 400 cm-', the following characteristic absorption vibrations were observed: Table 18 Peak # Wave number (cm-1> Functional group Vibration mode i 1100-1035 Q -H C-O-Cδ(0-H) 2 3430-3390 0-HV (0-H>3 2930-2800 0-H V(C-H)4 1470-1380 0-Hδ(C-H)5 1600-15 50 Coo-.

5 1450-1400 V (C=O) symmetrical 7 1420-1410 C-0 81650-1645C0N) (amide engineering C=0 9 1550-1540 C0NH amide ■δ(N-1-() 10 1746-1735 C0ORV (C=O) 11 12501240 C 0ORV(C-0-C)12 980-950 G-0゜ 0-CI-12 Research oscillations in Partoe and Bart ■ using the characteristic infrared frequencies mentioned above The number absorption maximum was as follows: 艮--U Arranged in order of IR peak maximum wave number #5 (ca-1) intensity of parts ■ and ■ 1 hour 4 hours 8 hours 10 hours 24 hours 1 1074.5 1068.7 1068.7 1587.6 1068.72 1039.8 1037.8 1037.8 3370.1 1037.83 3393.2 3412.5 3412.5 1076.4 3408.64 1589.5 1242.3 1244.2 1037.8 1242.35 1425.6 1740.0 1738.1 1421.7 1591.56 1244.2 1373.5 1375.4 1246.2 1375.47 1738.1 1B37.8 1635.8 2042.9 1421.7F3 603.8 1423.6 1425.6 1738.1 1738.19 2926゜4 603.8 601.9 603.8 669.410 2039.0 2924.4 29 24.4 2928.3 2924.411 960.7 960.7 958 ．． 7 960.7 603.8 part ■ 0 hours zero 1 1074.5 3397.1 1072.6 1068.7 1068.7 2 3393.2 1072.6 1037.8 3408.6 1037.8 3 1593.4 1593.4 3404.8 1244.2 3420.2 4 1244.2 1560.6 1244.2 1734.2 1244.2 5 1419.8 1244.2 1597.3 1635.8 1734.2 5 1734.2 1419.8 1558. γ 1653.2 1635.8 7 2039.0 1736.2 1419.8 1375.4 1653.2 F3 638.5 2924.4 1375.4 2922.5 1375.4 9 474.5 472.6 1740.0 472.6 472.610 2 927.4 607.7 605.7 1558.7 1419.811 96 0.7 960.7 669.4 1419.8 1558.712 873. 9 875.8 2924.4 1541.3 605.7603.8 292 4.4 541.3 Infrared scan of kallidin, deacetylated kallidin and acetylated kallidin The vibrations of the carboxylate carbonyl bond and ester carbonyl were identified from It was done. The carbonyl functional group and C-0-C stretch of acetylated kallidin are each 1 It was found between 748-1735 cm-' and 1246-1235 cm-'.

Deacetylated kallidin carboxylate carbo- The expansion and contraction are respectively 1600-1550cm and 1450-1400cm-' The absorption of two peaks is shown.

Table 21 Sample Nα 1450-1400 12480-1240 Absorption 4. 58 5. 584. 998 2. 607. 600. 988 4. 289. 487 1.685 8. 2B9. 450 1.557 10. 387. 326 0.84224. 248. 308 1.242 Part ■ 0. 278. 475 1.709 1. 496. 513 1.034 2 N/A N/A N/A 4’, 452. 497 1.108. 437. 524 1.20 10. 291. 436　1.498Approximately 35 gallons of aloe veragle processed for signature studies. The process from washing to filtration takes about 1 hour and 20 minutes. Continued. This study shows that no appreciable degradation of the gel occurs within this time. Indicated.

The alcohol precipitation process is an important factor in the quality of kallidin produced. was shown. The yield of kallidin increases with increasing precipitation time in alcohol. However, the quality of kallidin produced does not seem to be ideal for cell proliferation. It is. A time period of 4 to 5 hours in alcohol is not accompanied by undesirable decomposition. This method is considered to be optimal for increasing high yields.

In the study of part ■, if alcohol precipitation occurs immediately, kallidin The highest yield was observed. If alcohol is added within 4 hours If the Ta.

Infrared spectroscopy provides the best method for determining the quality of kallidin produced. can be provided. This infrared spectrum has a characteristic strong ester carbonyl absorption peak. However, deacetylated kallidin does not give such a peak.

It is believed that the degree of deacylation can be determined by this technique. This hand The only problem with the method is that the presence of the amide affects the value of the carboxylate absorption peak. This means that there are times when it is given.

Fujou (sha) - Fallen Figures 8 to 13 show Aloe barbadensis mirror produced by the process of the present invention. Infrared spectra of six different samples of kallidin obtained from plants are shown. ing. As is easily understood, the spectrum of each sample has a wavenumber of approximately 1γ50. It shows 5 to 7 well-separated peaks from 1 to 1250 wavenumbers. fruit As discussed in Example 24, these peaks indicate the following functional groups: RCOO-, R-NHCOCH3 and R-00CCI-13゜kallidin. It is these functional groups that are thought to confer activity. Also, Figures 8 to 1 Figure 3 shows leaves of Aloe barbadensis Miller plant produced by the process of the present invention. This indicates that certain Karrijin products are manufactured from.

Example 29 Figure 14 shows the infrared spectrum of raw material aloe vera gel cast into a film. are doing. This spectrum also has 5 waves from about 1150 wave numbers to about 1250 wave numbers. Shows ~7 well resolved peaks. Therefore, Figure 14 shows that kallidin is Provides evidence that the presence of a raw material gel from the Loevera plant .

X dust property - average Figure 15 shows the infrared spectrum of one sample of kallidin. noticed As shown, there is a characteristic well-divided i! between the wave numbers of 1750 and 1250! 6 Pi There are almost no tracks. This Kallijin sample was made using recycled alcohol. It is precipitated from gel or diacetate, and this causes the precipitation of kallidin. It is possible that it was lowered for some reason.

Example 31 Figures 16 and 17 are prepared as quickly as practicable after producing the aloe juice. It is important to extract kallidin from aloe juice using alcohol to It is shown that. Figure 16 shows that the aloe juice was removed 1 hour after making the aloe juice. This is a spectrum of a kallidin sample extracted from yeast using alcohol. . Figure 17 shows the amount of alkaline from aloe juice 10 hours after producing aloe juice. This is a spectrum of a kallidin sample extracted using Cole. 175G wave The characteristic well-separated peaks between wavenumbers 1 and 1250 are shown in the spectra of It shows a significantly higher absorption level compared to the spectrum in Figure 17. Ru. Therefore, kallidin can be extracted faster from aloe juice using alcohol. It is thought that the quality of the kallijin produced will improve as the amount of water increases.

Example 32 Figures 18 to 20 were created from the data shown in Table 22 below.

Table 22 Various processes during the production of some produced Kallidin batches time required Leaf treatment homogenization filtration alcohol Batch Manufacturing date I18 Hours Hours Addition 1 hour (minutes) (minutes) (minutes ) (minutes) 5B 1/10/86 146 30 63 106B 1/24/86 11 3 25 1058B 2/18/86 143 26 44 139B 2/ 26/86 103 32 29 15 Table 22 Continued 1 1210 8.26 255 3.922 1095 9.13 3B4 2.603 790 12.7 700 1.434 790 12.7 53 5 1.875 870 11.5 526 1.906 660 15.2 695 1.447 1035 9.66 345 2.901B 283 3 5.3 117 8.542B 3γ5 26.7 76 13.163B 2 70 37.0 252 3.974B 960 10.4 542 1.85 5B 250 40.0 255 3.926B 250 40.0 253 3.957B 285 35.0 144 6.948B 227 44.1 230 4.359B 240 41.7 167 5.9910B 255 39.2 185 5.41 Table 22 Continued Time 2 (minutes) (9> (w/v) 1 160 6.3 36.7 0.0542 105 9.5 Bo, 0 0 ．． 1063 246 4.1 71.1 0.0754 2B9 3.7 22 ．． 0 0.0235 272 3.7 17.8 0.0186 2B5 3, 5 15.3 0.0167 81 12.3 88.7 0.0981B 2 9 34.5 34.3 0.0652B 32 31.3 13.8 0.0 413B 167 6.0 177.3 0.1174B 392 2.6 8 1.8 0.0545B 93 10.8 145.0 0.0966B 13 0 7.7 120.3 0.0747B 50 20.0 184.9 0. 1228B 70 14.3 109.8 0.0739B 61 16.4 188.7 0.12510B 179 5.6 151.5 0.100 Note 1 : Gel from which the first drop of alcohol was obtained after cutting the first aloe vera confection. The total time until added to.

Note 2 Total time required for homogenization and filtration of Nigel only.

As shown in Figure 18, the yield of kallidin depends on the time the gel remains in the alcohol. I don't think there is a connection. As shown in Figure 19, the yield of kallidin before precipitation is It is inversely proportional to the total processing time. As shown in Figure 20, the yield of kallidin is homogenized. and is inversely proportional to the filtration time. Therefore, to maximize the yield of kallidin, The total processing time before precipitation, more specifically the homogenization and filtration times, should be kept as short as possible. It is possible.

absenteeism Figure 21 shows the red color of the alcoholic precipitation product of juice from Aloe ferrotus. The outer spectrum is shown. This spectrum ranges from a wavenumber of about 1750 to about 1250 It shows several well-separated peaks over the wavenumber of . Therefore, aloe The alcohol precipitation product of the juice from Ferotx has similar activity to kallidin. will have.

X banner and Figure 22 shows the red color of the alcohol precipitation product of juice from Aloe africana. The outer spectrum is shown. This spectrum ranges from a wavenumber of about 1750 to about 1250 It shows several well-separated peaks over the wavenumber of . Therefore, aloe The alcohol precipitation product of juice from Africana has similar activity to kallidin. There will be more.

Example 35 Figure 23 shows Africana ferrotucus, or Aloe ferrotucus and Aloe vera Infrared spectrum of alcohol precipitation products of juice from Africana hybrids It shows.

This spectrum has several types of waves ranging from about 1750 wavenumbers to about 1250 wavenumbers. It shows well-separated peaks. Therefore, Z furikana The alcoholic product of Euse would have similar activity to kallidin.

Example 36 Figure 24 shows the red color of the alcohol precipitation product of juice from Aloe dichotoma. The outer spectrum is shown. This spectrum ranges from a wavenumber of about 1750 to about 1250 It shows several peaks towards the wave number of . Aloe from Dichotoma The alcohol precipitation product of Euse may have some activities similar to kallidin. cormorant.

Example 37 Figures 25-29 show cellulose, polygalacturonic acid, from konjac plant Infrared spectra of glucomannan, dextran and guar gum, respectively. It shows. These spectra range from about 1750 wavenumbers to about 1250 wavenumbers. However, as shown in Figure 8, the sharp peaks of kallidin are shown. Not isolated peaks. Therefore, cellulose, polygalacturonic acid, koni Glucomannan, dextran or guar gum from the nutmeg plant are combined with kallidin. It is unlikely that they have similar activity.

Example 38 Modification of the rheological properties of kallidin: The modifications of kallidin discussed below may improve one or more of the following rheological properties of kallidin: Can affect two or more: absorption, permeability, specificity, solubility, viscosity, stability, active or target tissue. These modifications are important for pharmaceutical targeting.

The infrared spectrum of the modified form of kallidin was then measured. Modified form of mosquito Lysine is inferred to have similar biological activity to unmodified kallidin. .

A. Kalijin hydrogen peroxide treatment i. oo g of kallidin (light pinkish brown, lot #8B) was deionized for 500 d. Placed in a 1 gram round bottom flask with water and a 1 inch stir bar. This mixture is Stir at room temperature for 30 minutes, then at low temperature (4°C) for 30 minutes to form a slightly viscous, A heterogeneous mixture containing some undissolved kallidin was created. This intense stirring Add 5d of 30% Town 02 solution (Mallinkrodt, ΔR Gray) to the mixture at 4°C. #5240) (Mallinckrodt, AR grade, 852 40) S: Added directly. There were no observed changes or gas evolution. this The mixture was gently stirred at 4°C for 16 hours. No physical changes were observed.

Solid Sodium Bisulfite (granular, Barker, #1-3556) (4.588 9, 1.0 equivalent with respect to H2O2) was dissolved in 820 of 307. The resulting transparency The solution was added dropwise to the stirred reaction mixture over a period of 20 minutes. During the addition, the reaction pH Measure with pH paper and add sodium bicarbonate solution (1 molar, Fisher, #S -233) was used to maintain the pH at 7±0.5. After the addition is complete, bring the mixture to ambient temperature. Stir for 1 hour at 35°C and concentrate under reduced pressure on a rotary evaporator at 35-40°C. The volume was approximately 80 d. This concentrate was poured into a pre-cleaned dialysis tube (approximately 30 d long, dried). Cylinder diameter 28.6. Spectlabor Medical Industries, Inc. Los Angeles, CA# 132633, 2000) IW cutoff) (Spec trapor Medical Industry. , Inc.

Los Anaeles, CA 1132B33, 2000 MW cut of 4) and added to 6 g of deionized water at 4°C in an Ersinmeyer flask. Dialysis was carried out for 24 hours with water exchanged every 6 hours. Open this tube and pour out the contents. Freeze-dry under vacuum for 12 hours to obtain a yellowish white (off-white) powder of 42 oz. I got 3.

B. 1,00 g of kallidin (bright peach) during sodium periodate cleavage of kallidin brown, lot #8B) in 1.5 g containing 500 d of deionized water and a 1 inch stir bar. 1! into a round bottom flask. This mixture was heated at ambient temperature for 30 minutes, then at low temperature ( Stir for 30 minutes at 4°C to form a slightly viscous mixture containing some undissolved kallidin. A heterogeneous mixture was obtained. This was followed by 10 d of sodium periodate in deionized water. Solution (99% 7) Ltt:I)y Chi Chemical Co., #21, 044- 8: 5 9 m3, 180/5 mol% based on the average molecular weight of the monomer) was vigorously heated for 15 minutes. Add with stirring. This mixture was stirred at 4°C for 15 hours. the solution is uniform It turned a bright pink color. Measure the pH with pH test paper and keep it at pH ± 0.5. Ta.

I understand. Solid Na BN2 (630 mg, Aldrich Chemical Co., #19, 807-2, 980%) was added little by little over 30 minutes while stirring vigorously. Ta. After the addition was complete, the mixture was stirred at ambient temperature for 3 hours. Excess Na BN2 2 0d acetone (distilled, 99.9+% HPLC grade, Aldrich 9 Chemikaf companies.

#27, 072-5> was inactivated at air temperature for 2 hours. Is the pH greater than 8? It was. Therefore, a 50% HOAc solution (Fisher Chemical Co., HOAc reagent grade) was used. The pH was adjusted to 7 with a

The acetone was evaporated and the solution was heated to 35°C to 40°C on a rotary evaporator. The solution was concentrated under reduced pressure to about 80 d. Pour the residue into a dialysis tube (approximately 30 cm long, directly into the drying cylinder) Diameter 28.6#WN Spectraper Medical Industries, Los Angeles Luss, CA#132633, 2000) IW cutoff) (previously washed with water). Transfer this residue to 6 g of Ersinmeyer flask. Dialysis was performed for 24 hours against deionized water in a microcosm with water exchange every 6 hours.

The contents of the tube were removed and lyophilized under vacuum for 12 hours to yield 470 mg of a light brown color. A powder was obtained.

*Experimental operations were performed by J. M. Boitz “Periodic acid oxidation of carbohydrates” Atbar Inst Carpohydrate Chemistry 11: 1-41 (1956) (J.) 1. BOffitt “Periodate Oxidation o f　Carbohydrate　”Adv.

Corbohyrate Chemistry 11: 1-41 (1956 )).

C. Phosphorylation of kallidin Sodium dihydrogen phosphate monohydrate (Mathison Colema) in 30 d of deionized water N & Bell, CB 742; 577 mg) and hydrogen phosphate ditetate lium heptahydrate (Fisher sci.

S-373, Lot #855049837mg>, add i, oog to the solution. Lysine (light pink powder, lot #8B) was added. Heterogeneous mixture 35 Stirring at °C for 30 minutes gave a light pink-brown paste. Roast the water at about 40℃. Distillation was carried out under reduced pressure using a tally evaporator. Roughly remove the residue using an oil pump. 0. It was dried at lmHg pressure for 3 hours. Powder the hard solid with a glass rod, 1 It was placed in an fJ round bottom flask and then placed in a preheated oil bath at 155°C. . Argon ( (pre-purified) was conducted in a gentle flow. After 3 hours, bring the mixture to ambient temperature. slurried in 50 d of deionized water and prewashed with deionized water. Dialysis tube (length 30cmz drying cylinder diameter 28.6M, Spectraper Medical Industries, Inc., Los Angeles CA# 132633.200 I put it in the OHW cutoff). This mixture was placed in a 61 Ersinmeyer flask. water in a cold room (4℃) for 24 hours while changing the water every 6 hours. Dialyzed.

The contents of the tube at a pH of approximately 7 were removed and lyophilized under vacuum for 12 hours to produce 865#Ig. A brown solid was obtained.

*Experimental operations were performed by E, F, and Patsial Phosphation with In-Auger. Nick Phosphate Salts”.

Methods in Carpohydrate Chemistry VO1, IV, page 29 4-296.1964; Edited by R.L. Whissey, Academic Press, Ni. New York (E, F.

Pa5cha l l l “Phosphation with Inorgan icPhosphate 5 alts”, Methods in Car bohydrateChemistry Vol, IV, pp 294- 296.1964; R,L.

Old 5tler, Ed, Academic Press, New Yor Partially adopted from k).

For other operations, G, A, Tauru and R, L, Hoitus Sea Methods Carbo Hydrate Chemistry, 6°408 (1972) (G, A, Towl e and R, L, W old 5tler, Methods Carbohydra See Te Chem, 6, 408 (19722).

D. Partial methylation of kallidin* 1.009 kallidin (light pinkish brown, lot #8B) was powdered in a mortar and pestle. , then 75d acetone (Fisher Sci, A-18, Royt #856 136. vaporized). Add Na to deionized water while stirring vigorously. OH pellet (Mallinkrodt AR grade 7708. Lot KVH3) (H allinckrodt AR grade, 7708. Lot KVH3) Obtained by dissolving. 4.4d of 30% NaOH solution and 2.2m! ! sulfuric acid di Methyl (Fisher) was added at the same time. These reagents were divided into 4 equal parts at 15 minute intervals. and added. After the addition was complete, the mixture was stirred for 15 minutes and then rotary evaporated. The mixture was evaporated under reduced pressure using a rotor. Slurry the residue in 50ml of deionized water and deionize it. Dialysis tubing pre-washed with water (3 ocm long, dry cylinder diameter 28.6 sq. Pectraper Medical Industries, Inc. 1 Los Angeles, CA#1 32633. 200 OMW cutoff). 6g of residue 5% NaHCO3 solution (NaHCO3 powder,) in jar flask - 1”/sha , S-233, Ot# 722534), change the solution every 6 hours. The mixture was dialyzed at 4°C for 24 hours. Next, this residue was passed through a 61 Ersinmeyer flask. Dialyzed against deionized water for 36 hours at 4°C with water changes every 6 hours in a Ta.

The contents of the tube at approximately pH 7 were removed and lyophilized in a vacuum oven for 12 hours to form a light brown powder. 759m(] was obtained.

*Experimental procedures were performed by E, L, Hirst and E, Percivalpapolisacchucharide. Fractionation of chilled and methylated products - Method in Carbohydrate Chemistry Tory.

Vol, V, pages 287-296.1965; Edited by R, L, Whissey, Academic Press, New York (E,L.

Old rst and E, Percival,”) lethylation of Polysaccharides and Fractionation of 　1ethylated　Products”, 　Methods　i n Carbohydrate Whistler, Ed,, Acade+n ic Press, New York).

Also, Sho N, Howarth, Journal of Chemical Society 107: 8-160 (1915), (W, N. Haworth.

See also J. Chem. Soc. 107: 8-16 (1959) stomach.

E, Carboxymethylation of kallidin i, oo y kallidin (light pinkish brown solid, lot #8B) in a mortar and pestle. Powder, then 15d isopropyl alcohol (AC3 reagent grade, Aldo Rich Chemical Co., 919, 076-4). While stirring vigorously, 1&! 0.5(] of Na 0f-1 in deionized water (R grade to Mallinckrodt) Code, 7708. Loft 97708KVH3>t 5 minute chip 721 addition Engineering, immediately 1&! 140m of deionized water! ! Brome vinegar r1i<eastma Nkodak #964) was added. This mixture was heated at room temperature under an argon atmosphere for 800 min. Stir for a minute. The organic solvent was distilled under reduced pressure using a rotary evaporator at a bath temperature of 30'C. I left. The resulting brown paste was diluted with 30 ml of deionized water and the mixture was diluted with 20 ml of deionized water. % aqueous acetic acid (prepared from Fisher's wood vinegar [AC3 reagent grade A-38] neutralized to pH 7 using Dialysis of the product prewashed with deionized water Pipe (length 30 cm, drying cylinder diameter 28.6 mm, Spectraper Medical Industries, Inc., Los Angeles, CA# 132633.20 001 (W cutoff). This product was stored in a cold room (4°C) every 6 hours. 2 to 6 ρ of deionized water in an Ersinmeyer flask while changing the water. I underwent dialysis for 4 days.

The contents of the tube were removed and lyophilized under vacuum for 12 hours to give a brown color of 609/ff9. A brittle solid was obtained.

*Experimental operations were conducted by Hans Biink and Die Macro Molexie Hemi-122. : 271-274 (1969) (HansVink, Die Hakr Omolekulare Chemie 122:271-274 (196 Partly adopted from 9)).

F, kallidin @oxidation* Dimethylformamide (Baker Analyzed Reagent Grade, 3- 9221) in the sulfur trioxide-trimethylamine complex (Aldrichke Mikafu 1 company, 13,587-9.

1.003 of kallidin (white powder) was added to a suspension of O'C (white powder) with vigorous stirring. Powdered in a pestle and mortar) all at once added. After about 5 minutes, the mixture became very viscous and difficult to stir.

The mixture was kept at 0°C for 24 hours without stirring, then diluted with 50 d of deionized water. Dialysis tubing (length 30 cm, dry cylinder diameter), previously washed with deionized water 28.6M, Spectlabor Medical Industries, Inc. 2 Los Angeles vinegar.

CA# 132633.20001-IW cutoff). this mixture 10% NaHCO3 aqueous solution (solid pea) in a master Ersinmeyer flask. HCO3, Fisher 3Ci, AC3 certified, prepared from S-233 ) at 4°C for 24 hours, then dialyzed at 0°C with water changes every 6 hours. Dialyzed against deionized water at ~4°C for 48 hours.

The contents of the tube were removed and lyophilized under vacuum for 12 hours to give 575 m9 of a hard brown solid. I got it.

* Experimental operations were performed by R, L, Whistler and Spencer “Triethylamine III”. Sulfation by oxidized sulfur complex”Methods incarpohydrate chemis Tory.

Vol, IV, page 297-298.1964; R, I-, Whissy ed., Akadatetsumi Press, New York (R.L.

Whistler and W, W, 5pencer in “5ulfat ion by Triethylamine 5ulfur Trioxide Complex”.

)1methods in Carbohydrate Chemistry, Vol, IV.

pp, 297-298.1964; R.L. Whistler, Ed. , Academic Press, New York).

For other operations, please refer to R. Story, ■, pages 426-429, 1972 (R, L, Whistle r, Methods CarbohydrateChem, VI, pp, 426-429.1972).

G, cross-linking of kallidin with epichlorohydrin 1.00 g of kallidin (bright Powder the pink solid, lot #8B) in a mortar and pestle, then deionize the 20 (7!) The suspension was suspended in water. While stirring vigorously, add epichlorohydrin (0.40 g).

Aldrich Chemical Company 99%, gold label #24, 069-9) rare NaOH in 2.5 d of deionized gas water ( Mallinckrodt AR grade, 7708, Lot 7708KVHs O. 5 g) (7) Add and evaporate the solution. Heating the mixture at 60°C for 80 minutes, then Cooled to temperature. Adjust the pH to 20% and acetic acid aqueous solution (Fischer wood vinegar I ACS). 7 using reagent grade A-38). this reaction The mixture was added to a dialysis tube (30 cm long + dry cylinder) previously rinsed with deionized water. Diameter 28.6m, Spectlabor Medical Industries, Los Angeles Los Angeles, CA#132633, 2000 M-cutoff). anti Add 6 g of Ercin to the reaction mixture in a cold room at 4°C with water changes every 6 hours. Dialysis was performed for 24 hours against deionized water in a Mayer flask.

The contents of the tube were removed and freeze-dried under vacuum for 12 hours to produce 485 m3 of light brown A solid was obtained.

H. Kallidin cross-linked with formalin 1,00’J of kallidin (light pinkish brown solid, lot #8B) in a mortar and pestle. Powdered and then suspended in 200 d of deionized water. This suspension contains 37% of the three precepts. Limarin solution (Baker Analyzed Reagent Grade, Lot #33674, Manufacture #2106.

37% formaldehyde) and then 2.5d of 0.59% formaldehyde in deionized water. Na OH (Mallinkrodt AR grade, 7708, lot # 7708 KVHS> Wokaeta. The compound (7) a was vigorously stirred at 60°C for 80 minutes.

The mixture was cooled to room temperature and 20% aqueous acetic acid (Fischer glacial acetic acid, ACS) was added. The pH was adjusted to 7 using reagent grade, A-38).

This mixture was poured into a dialysis tube (length 30 cm dry cylinder diameter 28.6#).

Spectlabor Medical Industries, Los Angeles, CA #132B33, 2000MW cutoff).

This mixture was poured into a 6 g Ersinmeyer flask with water changes every 6 hours. Dialyzed against deionized water at 4°C.

The contents of the tube were removed and freeze-dried under vacuum for 12 hours to produce 518 m3 of white/brown material. A thick solid was obtained.

Example 39 Comparison of common topical agents for wound treatment: Cytotoxicity to cultured human fibroblasts Several methods commonly used to clean wounds using cultures of human fibroblasts The cytotoxicity of topical agents was determined.

The aim was to develop wound gels containing kallidin with different action mechanisms. It was compared with some standard cleaning agents. These standard cleansers are effective for cleaning the epidermis. designed to reduce bacterial damage and tissue breakdown secondary to barrier rupture. ing. Using release of radiolabeled chromium and uptake of trivan blue dye Cytotoxicity was measured. Cultured fibroblasts are exposed to high concentrations of potassium such as 0.5%. Undamaged by Jin. In contrast, Povidone-Yo Uronium (Betadine), Trypsin and Balsam of Peru (Granurex), Chlor Hexidine (Hiviclens), or hydrogen peroxide, is removed from labeled cells. c, was released. Betadine, Hibiclens and Granulex are also Lipan blue enhanced staining, but kallidin treatment did not. It was. According to these in vitro tests, kallidin is suitable for skin and 01 wound treatment. appears to be safe.

Cell culture. Human skin @ fibroblasts from newborn foreskin explants and during caesarean section Grown from adult skin samples taken from the lower abdomen. This tissue is fat and subcutaneous connective tissue were removed and chopped into 2 cubic particles, small (25c+ f) placed in a culture flask. 5% beef tallow serum (HaZa ltOn) 200m! (Minimum basal medium supplemented with glutamine and 1% antibiotics) ()IE) Ri (Inland Laboratories) was added and the culture was The temperature was maintained at 37°C under a 5% CO2 atmosphere.

A mixture of keratinocytes and fibroblasts emerges from the edges of this tissue within a few days. It was long. Keratinocytes could not divide, but fibroblasts could not divide after 16-21 days. They proliferated inward to form a monolayer of elongated cells with a characteristic whorled pattern.

All cultures were passaged at least 3 times and up to 10-15 passages before use. used in

Topical agents. in the treatment of wounds and pressure sores to measure cytotoxicity in in vitro systems. Some commonly used products include standardized cultures of human fibroblasts. added directly to. Povidone-iodine solution (Betadine), hydrogen peroxide, Granulec Chlorhexidine and Chlorhexidine (Hibiclens) are commonly used drugs with different mechanisms of action. It is a cleaning agent.

These compounds (0,001-0.5%) were added to cultured fibroblast cells. The cytotoxic effect was compared with kallidin.

Cell processing. Lightly exposed to Pax-EDTA (PUCkS-EDTA) solution (5 After ~10 min), separate the cells from the confluent cell monophase using 0.25% trypsin. I let go. The suspended cells were pelleted by centrifugation and washed once with fresh medium. Afterwards, they were resuspended in HEH supplemented with glutamine, antibiotics and 1% tallow serum.

Count the cells using an electronic cell counter (Coulter Electronics, Hiary, Floating). (Coulter Electronics, Hialeah, El ) and diluted and adjusted as required for individual experiments. This cell is 51Cr (1μCi/ae) and one well in a multi-well plate with 24 wells. It was placed at a density of 105 per bottle. Return the plate to the incubator for 18 hours. Ta. At the beginning of each experiment, remove the radioactive medium by aspirating and fill each well with fresh (IEl−) Washed 4 times with 1+1% tallow baby serum. HEI (alone or by species of test product) Add IEH (containing various dilutions) to the replicate wells and gently incubate the plate for 1-30 minutes. Incubated. Collect the medium at the end of the incubation period and collect the released It was saved for radioactivity measurement. Cells in each well were treated with 0.1M NaOH. Lyse by adding 0.5 ml of Triton X-100 (1%) and sample the lysate. The tube was taken out for radioactivity measurement.

Measurement of cytotoxicity. Cytotoxicity incubated with a variety of different chemicals quantified by the release of radioactive chromium from labeled fibroblasts. release% is the amount of radioactivity in the medium supernatant divided by the sum of the radioactivity of both the medium and the cells. It was calculated by

Another measurement of cytotoxicity was performed by staining with trypan blue.

Cells were incubated with each test reagent for 15 minutes. Tripumble - (1%) into each well and incubation continued for approximately 5 minutes. Ta. This sample was examined with an optical microscope, and the Nikon This photo was taken with the N35 Cocoon Camera.

Cytotoxicity was determined by the percentage of cells stained with trypan blue relative to control (untreated) cells. Evaluation was made by determining the fraction.

The results of this measurement are shown in Table 23 below.

Table 23 by Triban blue staining method Measured cytotoxicity of topical formulations Betadine 0.01% 15 minutes 100 Hibiclence 0.01 15 minutes 10 0 Granulex 0.01 // 15 minutes 100 Kalijin 0.01 1 5 minutes 5 Medium alone 15 minutes 1 Incubate cultured cells for 15 minutes with each reagent or medium alone. , trypan blue (1%) was applied, and after 5 minutes the number of stained nuclei was counted. Expressed as a percentage of total cell nuclei.

Time course of cell damage. Direct cell damage from topical agents occurs quickly, and was observed by an increase in the release of 51Cr. Medium alone or 10% tallow baby blood Cultured fibroblasts treated with the supernatant were incubated for 5 to 60 minutes to completely remove the cultured fibroblasts. Less than 5% of the ROM label was released. In contrast, 0.05% betadine, granules Cells treated with Glucose or Hibiclens shed 55-62% of their total label within 5 minutes. Released. A 10 or 15 minute incubation is required for all of these reagents. The amount released was slightly larger, but longer incubations (30 minutes) did not increase the release of radioactivity. Kalijin (CDW) Cells treated with G > (0,05%) can be incubated for as long as 30 minutes. Less than 5% of the total label was released during the reaction (Figure 30).

Effect of concentration on cell damage. Various reagents in the range of 0.005 to 0.05% The concentrations were tested for cytotoxicity. As shown in Figure 31, Granulex and Hibiclence (0,01%) is 25% of the total chromium label from fibroblasts and 15% was released. At the lowest concentrations of betadine (0,005% and 0.01%) The release by media was not greater than that by medium alone. However, 0.015% Cells exposed to tagine released more than 70% of their total radioactivity. 0.5 Concentrations of up to % kallidin resulted in more release than medium alone.

Similar results were obtained when examining cell damage using trypan blue staining.

0.01% Betadine, Hibiclens or Granulex as shown in the table. The 15 minute incubation used killed 100% of the cells. same concentration of force Incubation with lysine killed only 5%. 0.01% concentration Hydrogen peroxide severely damages these cells, judging by the changes they make in their morphology. Ta. However, trypan blue staining with this reagent is difficult to measure due to its decolorizing effect. I couldn't.

Effect of concomitant reagents. Whether kallidin has a protective effect in some experiments To determine the It was done. Its cytotoxic effects were demonstrated in medium alone and in medium containing 10% tallow serum. It was measured. Figure 32 shows that although kallidin alone does not damage cells, it 51 released by Hibiclens or Betadine during a minute incubation. c indicates that the amount of , was not changed. Similarly, fetal bovine serum in the culture medium did not alter the cytotoxic effects of these reagents.

Example 40 An 83-year-old female patient, TB, developed an ulcer 23 m in diameter on the lateral edge of her left foot. this The ulcer appeared for several months and did not respond to several therapeutic regimens.

The wound was treated with the product of Example 3 and Example 3 using a treatment schedule of 4 times a day. 7 products. Soak a clean wound with the product of Example 3 for 15 minutes. Ta. Excess product was absorbed from the wound using dry sterile 4X4 gauze.

The product of Example 7 was then applied to cover the wound and allow the wound to dehydrate between dressing changes. applied in an amount sufficient to prevent

The progress of wound healing can be monitored by taking photographs at intervals and measuring the area of the wound defect. I measured it. Table 24 shows the progress of wound closure.

Table 24 Progress of wound healing wound area Days (square inches) Percentage of healing 1 1.24 0.00 28 0.51 58.87 77 0.29 76.61 83 0, 12 90.32 97 0, 00 100.00 The epidermal wound was substantially closed in 12 weeks and completely closed in 14 weeks.

Example 41 The 32-year-old patient had a history of ulcerative colitis for many years. Currently, she is on 4omH of prednisone (predn i 5One), 3g of Asulfidine, 50m3 6-mercaptopurine and did not respond to daily regimen consisting of Flagyl.

She continued to have abdominal pain and had bloody stools 4 to 8 times a day. she has an intravenous drip I received it. Endoscopic diagnosis shows mild hepatic or severe progressive disease with transverse muscle ulceration. It showed membranous ulceration. This patient takes 50mH of potassium four times a day in addition to her other medications. He was given gin and sent home. Within a week her symptoms had virtually disappeared. abdomen It is slightly painful when removed, and an endoscopy reveals a slightly engorged mucous membrane that has healed. It was. The patient was gradually weaned off other medications and his disease picture continued to improve. this The patient is currently continuing Kallidin as the sole drug at this time. physical examination All examinations and symptoms were recorded as normal.

Similar responses to ulcerative colitis and Crohn's disease Five other cases were recognized. - One patient cut the kallidin capsule. ()Ta. At 4 weeks, mild symptoms began to return (defecation with mild abdominal discomfort). has increased. ) and she restarted the drug. In 3 days she was totally normal. The symptoms of defecation returned.

X directness-M Many AIDS patients receive high doses of kallidin for long periods without toxicity or side effects. received the dose. In these AIDS patients, there is a reduction and disappearance of clinical symptoms and infection. Increased ratio of T-4 and T-8 lymphocytes and extinction of T-4 with decreased chance of An increase in logarithm was observed. Kallidin may be antiviral or immunomodulatory in patients It is suggested that it has an effect.

Stimulation of lymphocytes in these patients was observed. Kallidin is immune to this This suggests that it may be involved in regulation.

Fu Jing-Chang - Zhang Tic douloureux, or neuralgia of the fifth cranial nerve, is 1 Characterized by attacks of severe and intolerable pain in one or more branches of the trigeminal nerve Can be attached. This pain is generally transient and attacks occur in certain areas of the face, so-called Contact with the rigger zone may result in fish and shellfish.

The cause and treatment of this painful disease are unknown. to treat this disease Some attempts have had little or no success. Various treatments include sedatives. Painkillers, phenytoin, peripheral removal and degassing of the involved nerve branches passing through the skull. Lianganglion involves injecting 98% alcohol.

A more radical procedure is to sever the sensory roots of the nerves adjacent to the ganglion. It permanently deprives the patient of sensation in the area served by the severed nerve. Ru. Another recent treatment attempt is carbamazepine and phenolifluoride. This method uses multiple injections. However, these injections can numb the pain and It can be troublesome due to serious side effects.

None of the treatments proposed so far are favorable.

A 43-year-old woman was diagnosed with tic dollarro.

The areas of decreased pain included the 1st and 3rd parts of the lateral trigeminal nerve.

This patient experienced pain when brushing or sun-combing her hair on the right side. she diazepam (Valium), antihistamines, painkillers, Planolol hydrochloride (Inderal) and phenobarbita I tried to treat it with a tube, but it didn't work. This patient has been suffering from this disease for some time. He said that there was never a dull day. The proposed treatment is the product of Example 2 The plan was to take 1 to 2 ounces of the drug daily for 3 months. After this period, this treatments were evaluated.

The patient's pain was virtually gone within two weeks of starting treatment. she is number He said he felt good all week. But then she went on a two-week trip, during which she did no real work. The product of Example 2 was not taken. The symptoms and pain returned during this trip. deer When she resumed this medication, the pain disappeared within a few days. the next few weeks, She felt good again.

After drinking this juice daily for more than 6 months without any problems, she experienced pain. They report being able to brush and comb their hair without causing any irritation.

Her condition has improved and she feels better than ever, Hilltop said. leaves (15,9 lbs.) washed, filleted and ground in a Waring blender and then filtered through 8 layers of cotton cloth. Next, apply this gel to four 11 quart stainless steel plates. Transfer to a steel pan and add cold USP grade ethyl alcohol to each The capacity ratio is 2:1, 3:1.4:1. and added at a ratio of 5:1. The amount is It can be summarized as below: Ratio (ethanol ethyl : Aloe gel) Amount of gel Amount of alcohol 2: 1 500 precepts 1000 power 3: 1 500d 1500d 4: 1 1670&! 6680d5: 1 500a2 2500m The precipitate was allowed to settle for 4 hours, then the remaining alcohol-gel solution was carefully decanted. sampled and stored in a separate container. This precipitate was collected using a TEC Centra-7 centrifuge. centrifuged at 280 rpm for 10 minutes, washed with alcohol, and then centrifuged under the same conditions. It was centrifuged again. Transfer the pellets to a 600 d jar and freeze with liquid nitrogen. , lyophilized overnight.

Additional alcohol was added to the supernatant from the 2:1 ratio and allowed to settle overnight at room temperature.

The remaining supernatant was also allowed to settle at room temperature overnight.

The next day, remove the pellet from the 2:1 ratio precipitated with additional alcohol and The precipitate was collected from the supernatant as described. In this case, the pellets are placed in a freeze-drying jar. - Approximately 5-10m when moving! ! of water was added.

result: The results of the first 4 hours of alcohol precipitation can be summarized as follows: Ratio (ethanol yield% (kallidin) : aloe gel) Yield (g) O/g gel) 2: 1. 051B, 010 3: 1. 3847. 077 4: 1 1.945. 116 5: 1. 6675. 134 After adding more ethyl alcohol, the 2:1 supernatant liquid had a concentration of roughly 178 my. produced lysine. 1@supernatant with ratios of 3:1 and 4:1 by just precipitation The sap produced approximately 89 m3 and 105 m5 respectively. The 5:1 ratio is , after initial isolation produced little precipitate. Therefore, we did not re-collect .

For the second precipitation from a 2:1 ratio (3:1), 5 to 10 The centrifuge packet was rinsed with water from d. This is similar to the density produced by other samples. A low-density white fluffy kallidin sample that differs markedly from the high gray kallidin sample. produced gin.

summary Kallidin is based on the fact that kallidin is composed of approximately 80% mannose. As evidenced by Example 13, it is primarily mannan. mannan man The north monomer is primarily β(1→4) as evidenced by Example 14. connected by bonds. Elemental analysis, IR, NMR and acetyl group analysis The results show that kallidin has O-acetyl, N-acetyl and carboxylate functionalities. It shows that a group exists.

The time to process kallidin prior to precipitation affects the overall quality and yield of kallidin. was shown to be important for quantity. The gel is allowed to stand before the kallidin is precipitated. The longer the time for the enzymes in the gel to act on kallidin, the more group as well as the glycosidic bond (β(1→4)), thus reducing its activity. It's small and disappears.

From the results of the above experiments and all the evidence, the following structure appears in the above formula: R1=-CH 20H, -COO'-, or -C ■ 200C ■ 3: R2 = -OH, -00C CH3, or -NHCOCH3; R3=-OH, -00CCH3, or - NHCOCH3; and n=2 to about 50,000° Mainly mannan chains contain other substituted simple pentose or hexose monomers. You can

The production, isolation and characterization of kallidin involved novel and non-obvious manipulations and techniques. I was reading. In this technical field that was incorrect or inaccurate for various reasons As evidenced by other previous work, kallidin contains the active substances in aloe nonobvious and novel from previous attempts by others to isolate and characterize .

FIG.2 Release analysis 1 P 凰 Send degree VL nine degrees vIt, roundness ν and range Matagago friend superiority barking feet , L degree 9L roundness FIG. 19 Ca±, Fishing/I:: Bungiku 1st IM Ishige y' Rose, Itre Nimono 1/T(,EL(MIN,-Li) FIG. 20 vL busy return -ayJI, busy skin MaL view rate Gomaru return Roumaru return Sakamarudo 91L L friend Likelihood Slope level Closed (46 tons) FIG. 30 ＊＊１≦−゛Note゛・)ㅅ11 FIG, 3f 51C & people, 24 international search report

Claims (61)

[Claims] 1. Producing aloe gel substantially free of anthraquinones from the leaves of aloe plants. In a method: a) washing aloe leaves with a disinfectant solution to remove substantially all surfaces; b) removing at least the first wash from said washed leaves; removing the end of 1; c) Draining, storing and collecting the anthraquinone-rich sap from the cut and washed leaves. and d) removing the integument from said leaves to substantially contain the anthraquinone. to produce a gel that does not method including. 2. Producing aloe gel substantially free of anthraquinones from the leaves of aloe plants. In a method: a) washing aloe leaves with a disinfectant solution to remove substantially all surfaces; b) Crushing the washed aloe leaves; and and c) dialyzing the crushed leaves to substantially remove anthraquinone from the remaining fraction of the crushed leaves. Chemically extracting and separating the free gel method including. 3. Producing aloe gel substantially free of anthraquinones from the leaves of aloe plants. In a method of: a) crushing the leaves of an aloe plant; and b) dialyzing the crushed leaves. to convert the residual fraction of crushed leaves into an aloe gel that is substantially free of anthraquinones. to remove and separate the method including. 4. Produce a substantially anthraquinone-free juice from the leaves of the aloe plant In the method: a) Clean the aloe leaves with a disinfectant solution to substantially remove surface dirt and bacteria. b) removing a first end and a second end from said washed leaf; c) Draining, storing and collecting the anthraquinone-rich sap from the washed and cut leaves. to gather; d) aloe gelaf substantially free of anthraquinones by removing the outer skin from said leaves; manufacturing a fillet; and e) grinding said substantially anthraquinone-free aloe gel fillets; and homogenized to produce aloe juice substantially free of anthraquinones. How to include. 5. Claims further comprising filtering the aloe juice to remove fibrous substances. For the production of aloe juice substantially free of anthraquinones according to scope 4. How to do it. 6. At least one selected from the group consisting of preservatives, fragrances, adjuvants, carriers and colorants. Adding one kind to the aloe gel fillet or the aloe juice. Aloe juice substantially free of anthraquinones according to claim 4 contained in method of manufacturing. 7. The preservative is selected from methylparaben, propylparaben and butylparaben. substantially anthraquinone-free according to claim 5 selected from the group consisting of How to make aloe juice. 8. The juice is irradiated, freeze-dried, sonicated, or pasteurized. substantially in accordance with claim 5, whereby said juice is preserved. A method of producing aloe juice free of intraquinone. 9. The aloe juice is filtered by ultrafiltration to permeate the aloe juice. The practice according to claim 5 further comprising the step of adjusting pressure and sorting by molecular weight. A method for producing aloe juice qualitatively free of anthraquinones. 10. A product made by the method of claim 4. 11. A product made by the method of claim 5. 12. A product made by the method of claim 6. 13. A product made by the method of claim 7. 14. A product made by the method of claim 8. 15. A product made by the method of claim 9. 16. The substantially anthraquinone-free aloe gel fillet is Claim 4 further comprising the step of washing in a tumble washer prior to crushing. A method of producing aloe juice substantially free of anthraquinones according to the invention. 17. a) Aloe juice 0.1 to 100 weight substantially free of anthraquinones amount%; b) allantoin 0-2% by weight; c) panthenol 0-6% by weight; d) Adjuvant or carrier A composition comprising. 18. a) Aloe juice 0.1 to 100 weight substantially free of anthraquinones amount%; b) food preservative; c) citric acid; and d) Vitamin E A beverage composition comprising. 19. In a method for extracting active chemicals in aloe plants from leaves of aloe plants : a) Clean aloe leaves with a disinfectant solution to remove virtually all surface dirt and germs. b) removing at least a first end from said washed leaf; thing; c) draining and storing the anthraquinone-rich sap from the cut and washed leaves; , collecting; d) removing the integument from said leaves to substantially contain the anthraquinone; e) producing a gel fillet substantially free of anthraquinone; The aloe gel fillet is ground and homogenized to contain the solubilized material and substantially anthracite. Producing aloe juice free of non-containing substances; f) Add a water-soluble lower aliphatic polar solvent to this aloe juice to analyze active chemicals. to form a heterogeneous solution; g) Remove water-soluble lower aliphatic polar solvent and solubilized substances from this heterogeneous solution and precipitate isolating the active chemical; and h) a method comprising drying the precipitated active chemical. 20. In a method for extracting active chemicals in aloe plants from leaves of aloe plants : a) Clean aloe leaves with a disinfectant solution to remove virtually all surface dirt and bacteria. b) Crushing the washed aloe leaves; c) Dialyzing the crushed leaves to substantially remove anthraquinone from the remaining fraction of the crushed leaves. chemically removing and separating the gel that does not contain; d) The substantially anthraquinone-free aloe gel is ground and homogenized to make it soluble. Producing an aloe juice containing compounds and substantially free of anthraquinones; e) Add a water-soluble lower aliphatic polar solvent to this aloe juice to analyze active chemicals. causing a heterogeneous solution to form; f) Remove and precipitate the water-soluble lower aliphatic polar solvent and solubilized substances from this heterogeneous solution. isolating the active chemical; and g) a method comprising drying the precipitated active chemical. 21. In a method for extracting active chemicals in aloe plants from leaves of aloe plants : a) Clean aloe leaves with a disinfectant solution to remove virtually all surface dirt and bacteria. b) Crushing the washed aloe leaves; c) Grinding and homogenizing crushed aloe leaves to produce aloe juice containing solubilized matter. thing; d) Add a water-soluble lower aliphatic polar solvent to this aloe juice to analyze active chemicals. causing a heterogeneous solution to form; e) Remove the water-soluble lower aliphatic polar solvent and solubilized substances from this heterogeneous solution and precipitate isolating the active chemical; and f) a method comprising drying the precipitated active chemical. 22. In a method for extracting active chemicals in aloe plants from leaves of aloe plants : a) Clean the aloe leaves with a disinfectant solution to substantially remove surface dirt and bacteria. b) grinding said washed aloe leaves; c) ground aloe leaves; filtering to remove fibrous substances; d) Homogenize ground aloe leaves to obtain anthraquinone-rich aloe containing solubilized matter. e) producing a water-soluble lower aliphatic polar solution in this aloe juice; adding a medium to precipitate the active chemical, thereby forming a heterogeneous solution; f) Remove and precipitate the water-soluble lower aliphatic polar solvent and solubilized substances from this heterogeneous solution. isolating the active chemical; and g) A method comprising drying the precipitated active chemical. 23. In a method for extracting active chemicals in aloe plants from aloe plant vegetables : a) crushing the leaves of the aloe plant to extrude the aloe juice containing the solubilized matter; b) Add a water-soluble lower aliphatic polar solvent to this aloe juice to analyze active chemicals. causing a heterogeneous solution to form; c) Remove and precipitate the water-soluble lower aliphatic polar solvent and solubilized material from this heterogeneous solution. isolating the active chemical; and d) A method comprising drying the precipitated active chemical. 24. In a method for extracting active chemicals in aloe plants from leaves of aloe plants : a) Clean aloe leaves with a disinfectant solution to remove virtually all surface dirt and bacteria. b) removing the outer skin from the leaves to produce aloe gel fillets; to do; c) Grind and homogenize the aloe gel fillet to obtain aloe juice containing solubilized matter. d) Adding a water-soluble lower aliphatic polar solvent to this aloe juice; precipitating the active chemical, thereby forming a heterogeneous solution; e) Remove the water-soluble lower aliphatic solvent and the solubilized material from this heterogeneous solution and collect the precipitated active material. isolating the sex chemicals; and f) a method comprising drying the precipitated active chemical. 25. irradiating the precipitate, thereby sterilizing and preserving the active chemical; A method for extracting active chemicals in aloe plants according to claim 19. 26. Add 4 times the volume of the water-soluble lower aliphatic polar solvent to 1 time the volume of aloe juice. in an aloe plant according to claim 19 which additionally precipitates said active chemical. How to extract active chemicals. 27. The water-soluble lower aliphatic polar solvent may be methanol, ethanol or propano. active chemistry in the aloe plant according to claim 19 selected from the group consisting of A method of extracting substances. 28. Claim 27, wherein the water-soluble lower aliphatic polar solvent is ethanol. Follow the method of extracting active chemicals in aloe plants. 29. The active chemical is removed before the water-soluble lower aliphatic polar solvent is removed. Claims that can be precipitated from a water-soluble lower aliphatic polar solvent and aloe juice for about 4 hours. A method for extracting active chemicals in aloe plants according to clause 19 of the scope of the invention. 30. Claim 1, wherein the precipitated active chemical is dried by freeze-drying. A method for extracting active chemicals in aloe plants according to paragraph 9. 31. A product manufactured by the method of claim 19. 32. A product manufactured by the method of claim 20. 33. A product manufactured by the method of claim 21. 34. A product manufactured by the method of claim 22. 35. A product manufactured by the method of claim 23. 36. A product manufactured by the method of claim 24. 37. A product manufactured by the method of claim 25. 38. A product manufactured by the method of claim 26. 39. A product manufactured by the method of claim 27. 40. A product manufactured by the method of claim 28. 41. A product manufactured by the method of claim 29. 42. A product manufactured by the method of claim 30. 43. Virtually non-degradable freezing of substantially acetylated mannose monomers A composition comprising a dried ordered linear polymer. 44. Claim No. 1, wherein the monomers are interconnected by β (1→4) bonds. Composition of item 43. 45. In a composition comprising the ethanol precipitation product of the juice of the aloe vera plant , wherein the precipitated product is substantially non-degradable and is lyophilized. 46. The ordered linear copolymer is irradiated with gamma rays or microwaves. A composition according to claim 43. 47. The ordered linear copolymer is irradiated with gamma rays or microwaves. A composition according to claim 44. 48. The ordered linear copolymer is irradiated with gamma rays or microwaves. A composition according to claim 45. 49. Composition comprising the ethanol precipitation product of the juice of the Aloe ferox plant wherein the precipitated product is substantially non-degradable and is lyophilized. Composition. 50. A composition comprising the ethanol precipitation product of the juice of the Aloe africana plant wherein the precipitated product is substantially non-degradable and is lyophilized. Composition. 51. Contains ethanol precipitation products of Africana ferox plant juice In the composition, the precipitated product is substantially non-degradable and is not lyophilized. composition. 52. Composition comprising ethanol precipitation product of juice of Aloedicotoma plant wherein the precipitated product is substantially non-degradable and is lyophilized. thing. 53. Ethanol precipitation products of aloe vera plant juice were oxidized with hydrogen peroxide. A composition according to claim 45. 54. The ethanol precipitation product of aloe vera plant juice is sodium periodate. 46. A composition according to claim 45, wherein the composition is cleaved with a polyamide. 55. The ethanol precipitation product of aloe vera plant juice is sodium dihydrogen phosphate. Phosphorylated with a mixture of um-hydrate and disodium hydrogen phosphate heptahydrate A composition according to claim 45. 56. Ethanol precipitation products of aloe vera plant juice were treated with dimethyl sulfate. 46. A composition according to claim 45, wherein the composition is methylated. 57. Ethanol precipitation products of aloe vera plant juice are bromoacetic acid and vinegar 46. A composition according to claim 45 which is carboxymethylated with an acid. 58. The ethanol precipitation products of aloe vera plant juice are sulfur trioxide-trimester. According to claim 45 sulfated by thylamine complex Composition. 59. The ethanol precipitation product of aloe vera plant juice is epichlorohydrin. 46. A composition according to claim 45, wherein the composition is crosslinked by: 60. Ethanol precipitation products of aloe vera plant juice are treated by formalin. A composition according to claim 45 which is crosslinked. 61. Substantially non-degradable freezing having repeating monomers represented by the formula: Compositions containing dried ordered linear polymers: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ In the above formula, R1 is -CH2OH, -COO- or -CH2OOCCH3. R2 is -OH, -OOCCH3 or -CH2OO Can be selected from the group consisting of CCH3; R3 is selected from the group consisting of -OH, -OOCCH3 or -CH2OOCH3 and n is from 2 to about 50,000.