SI9600104A - Procedure for the extraction and isolation of natural antioxidants of the labiates (labiatae) - Google Patents

Abstract

A new process for obtaining and isolation of antioxidant oral compounds (Labiatae) which consists of: 1. extraction of ground vegetable material with organic solvent, 2. partial solvent evaporation, 3. the isolation of the antioxidant components that is carried out a) steam distillation, drying and extraction with gases above the critical point; or b) extraction of aromatic of concentrate components with gases above the critical point. The products obtained are odorless and tasteless and are therefore high quality antioxidants, useful e.g. v pharmaceuticals, cosmetics and the food industry.

Description

PROCEDURE FOR THE OBTAINING AND INSULATION OF NATURAL ANTIOXIDIDES USTNATIC (Labiatae)

The present invention relates to a novel process for the production and isolation of natural oral antioxidants (Labiatae). The products obtained are odorless and tasteless and therefore are of high quality antioxidants, useful e.g. in the pharmaceutical, cosmetics and food industries.

BACKGROUND OF THE INVENTION

Oils and fats belong to the group of lipids. They are distinguished mainly by origin and, to a lesser extent, by melting points. At room temperature, oils are liquid and are mostly extracted from plants, while fats are solid or semi-solid and are usually of animal origin (RiegeVs Handbook of Industrial Chemistry, ll ^th Edition, Van Nostrand Reinhold Company Inc., 1983). Due to external factors (oxygen, light, heat, ...), fats deteriorate relatively quickly, greatly reducing their physiological and nutritional value. The most common cause of fat breakdown is auto-oxidation. The biological changes caused by enzymes and microorganisms are small compared to autoxidation.

Unsaturated fatty acids, such as oleic, linoleic and linoleic acid, have one or more double bonds in the molecule, which are oxidized to hydroperoxides under the influence of atmospheric oxygen. Lipid oxidation is an autocatalyzed chain radical reaction. The degree of auto-oxidation depends on:

* levels of unsaturation of fatty acids, * presence of free fatty acids, * surface exposed to oxygen, * partial pressure of oxygen, * temperature and * catalysts present.

The oxidation of fatty acids, in addition to the unpleasant odor and taste, also results in:

* formation of toxic polymers, * discoloration of various pigments (chlorophyll, hemoglobin), * loss of nutritional and physiological value of fatty acids (destruction of oxidatively sensitive vitamins and essential fatty acids), * structural changes and * other chemical changes.

The primary products of auto-oxidation are monohydroperoxides and diperoxides. Diperoxides are formed by the oxidation of unsaturated fatty acids having more than two methyl groups between two double bonds. Hydroperoxides are odorless and colorless, but their decomposition produces volatile products. In addition to hydroperoxide, an epodioxide is formed, where two oxygen molecules are embedded in the cyclic form of peroxide. Hydroperoxides form aldehydes, ketones, mono- and dicarboxylic acids, keto-, epoxy- and hydroxy acids. Subsequent reactions result in methyl ketones and at high temperatures dimers, polymers and condensates. These products adversely affect the odor and taste of fat, which is generally referred to as the notion of embers, but also destroys vitamins, tocopherols and carotenoids.

There are several ways of preventing or preventing. auto-oxidation retention:

* by blocking access and removing oxygen, * by storing fats in low temperatures and in a dark place, * by removing metal ions * and by adding antioxidants.

Antioxidants are organic substances that are added to fats to slow down their oxidation and thus prolong the shelf life of fat. Once all of the added amount of antioxidants is consumed, the process of auto-oxidation of unsaturated fatty acids continues. Care should be taken not to add too many antioxidants as they have a pro-oxidative effect.

Antioxidants must meet the following requirements:

* they must be effective at low concentrations, * they must not be volatile and toxic, * they must be odorless and tasteless, * they must not alter the organoleptic properties of the product, * they must have a lower redox potential than the compound to be protected and * must physiologically indifferent compounds.

Antioxidants prevent auto-oxidation through two reactions:

First reaction: Antioxidant (AH) gives hydrogen (H) that binds to free radical peroxide (ROO ') or fatty acid radical (R').

AH + ROO - "ROOH + A" or

AH + R RH + A

Second reaction: the antioxidant radical (A ') binds to ROO' or R '

A + ROO- »ROOA or

A + R -> RA

The effects of antioxidants can be prolonged by:

* store fats and fatty foods in dark places or in light-free packaging * store fats at low temperature * prevent as much oxygen as possible during production and especially during packaging (vacuum packaging, inert gases), * no use copper or rusted devices to release metal ions.

As antioxidants cannot eliminate the oxidation products produced before their addition, it is advisable to add them to the product as early as possible. Antioxidants also have no effect on the irradiation caused by fat hydrolysis. This type of irradiation occurs in the presence of moisture and enzymes. They are avoided by removing all the moisture from the fat and destroying the lipases with temperature.

Phenolic compounds account for the largest proportion of antioxidants, while the rest are aromatic amines, organo-sulfur and phosphorus compounds (Ullman's Encyclopedia of Industrial Chemistry, VCH Verlagsgesellschaft, Weinheim, 1985, Vol. A3, Antioxidants, p 91.)

Synthetic antioxidants:

The most commonly used synthetic antioxidants are:

1. 3- tertiary butyl - 4 - hydroxy anisole - BHA,

2. 2,6 - butyric acid - p - hydroxy toluene - BHT,

3. 2 - tertiary butyl hydroquinone - TBHQ and

4. Propyl gallate (PG).

Antioxidants-Natural:

Many herbs and spices contain antioxidant components, but it is only sensible to isolate them with higher amounts of these components. Rosemary, sage, pepper, nutmeg, cocoa, coffee, green tea, olive leaves, mustard, black pepper, cloves are used as a source of natural antioxidants (Bracco U., Lbliger J., Viret L., Production and Use of Natural Antioxidants, JAOCS, June 1981, p. 686).

Natural antioxidants are various chemical compounds, and the most widely used group of compounds that have proven antioxidant efficacy are phenolic type compounds. It is these compounds that contain the Labiatae extracts. Natural antioxidants include tocopherols and carotene.

Oral Antioxidants (Labiatae):

Common rosemary, sage, thyme belong to the lipstick family. Lips are one or perennial herbs, semi-shrubs or shrubs. The family of about 3500 plants (eg peppermint, sage, lemon balm) is widespread in all climates, especially in the Mediterranean and Central Asia. In the lips there is a high content of isoprenoid substances in the form of monoterpenes (essential oils), sesquiterpenes, diterpenes (bitterness) and triterpenes. Dried leaves contain 1.5 to 2.5% of essential oil, the main components of which are cineole, borneol and camphor.

Whole plants can be used for extraction, but the content of the individual components in the leaves is highest, so leaf extraction is the best option. In addition, the essential oil also contains borniacetate, α and 0 pinene. Some essential oil components have anti-oxidizing properties. Therefore, it is desirable to minimize the content of essential oils in antioxidant extracts. In addition, essential oils give products containing oral extracts an undesirable dominant odor and taste.

So e.g. important are the ingredients of common rosemary (rosmarinic acid), bitter (carnosol, carnosolic acid), triterpenes (ursolic acid), flavones (luteolin, apigenin, ...). The antioxidant effect of rosemary, however, is caused by phenolic diterpenes. The most effective is carnosol, from which, by oxidation, the slightly less effective carnosic acid is produced. The antioxidant effects of rosemary, rosemarydiphenol and rosemaryninone are also active. The chemical formulas of these substances are shown in Figure 1. The chemically related compounds are also the antioxidants of sage and thyme.

The literature (J.Lblinger, Lipid Technology April-June 1991 pp. 58) and a number of patent documents describe fat stabilization processes and preparations (e.g.

EP 658316 Al 950621 - Nestle, JP 55018436800208 - Tokyo Tanabe Co., JP 55018437800208 - Tokyo Tanabe Co., JP 57159874 A2 821002 - Takasago Perfumeiy, JP 590309279 A2 840303 - Hasegawa) or cosmetic products (CH patent N 6452929) . The use of Labiatae cell cultures for the production of higher antioxidant substrates is described in JP 62032889 A2 870212 Hasegawa.

Rosemarydiphenol isolation from rosemary is described in US patent 4638095 A 870120 Research Corp.

The use of alcohols for the isolation of antioxidant components is described in JP 59075978 A2 840428 - Hasegawa and in J.W. Wu et al. JAOCS vol 59, No. 8, pp. 339.

The processes described in U.S. Patent Files 3950266 - Rutgers Research, U.S. Pat. No. 4,876,735 Kalamazoo use a low boiling solvent in the first step, which is then removed from the evaporation of the extract solution, and then in the second stage, the aromatic components are separated by the addition of a highly volatile solvent. The obtained products have relatively low antioxidant activity.

US patent 4877635 describes the process of isolating antioxidants with organic solvents, and the process of separating non-antioxidant components is quite complex.

Antioxidant extraction is also possible with fatty acid glycerides (Cg-C ^); e.g. EP 639336 Al 95022 - Nestle ter US 3732111730508 - Campbell Soup. Co., and using oils with the addition of non-polar solvents (EP 507064 A2 921007). In this connection we also refer to the article by Tateo, Fellin M. Perfumer & Flavorist Vol 13, Dec. 1988, PP ⁴⁹ ·

All the extracts obtained by known organic solvents or glycerides known to date have a number of additives that give the product an undesirable dominant odor and taste. have low antioxidant activity.

The process described in patent DE 4207739 SKW Trostberg is performed by extracting aromatic components from rosemary with supercritical CO ₂ at pressures between 80 and 300 bar and temperatures between 10 and 80 ° C. The remaining plant material containing antioxidant compounds is extracted with alcohol (1-4C atoms), non-polar hydrocarbons having 5 to 7C atoms, then the extract is decolourised with activated carbon, the solvent is evaporated and the resulting product is treated with water. This process requires relatively large extractors for supercritical extraction of rosemary at relatively high pressures and temperatures. The process according to patent EP 93103772 - SKW is carried out in the same way as in patent DE 4207739 SKW Trostberg, and the separation of bitter components from the antioxidant concentrate is carried out with methanol.

The use of supercritical CO ₂ is described in an article by Z. Djarmati et al. JAOCS Vol 68, No 10.1991, pp 731. and in JP 06065573 A2 940308 Heisei and Norac EP 0 454 097 Al (Norac technologies), USP 5,120,558 (Norac technologies).

The isolation process using supercritical CO ₂ produces quality products with a relatively high content of antioxidants, but components that give the product an undesirable dominant odor and taste are still present, and relatively large and therefore expensive SC extraction devices are required.

DESCRIPTION OF THE IMPLEMENTATION CASE

Now we have found that we are solving the above problems with a new process for the production and isolation of natural oral antioxidants (Labicitaej, consisting of:

1. extraction of ground vegetable material with an organic solvent,

2. partial solvent evaporation,

3. isolation of the antioxidant components carried out by

a) steam distillation, drying and extraction with gases above a critical point; or

b) extraction of aromatic components from the concentrate with gases above the critical point.

The solvents used in the process may be alcohols having 1-5 C atoms, esters (such as ethyl acetate, butyl acetate, etc.), ketones (such as acetone), hydrocarbons and their halogenated derivatives, cyclic hydrocarbons.

The solvent / solid ratio is between 2 and 30 liters of dissolved 1 kg of plant material (lips).

The extraction times are between 1 and 20 hours and the extraction temperature is between 10 and 90 ° C. Extraction can be carried out in two steps.

The activated carbon / solution ratio is between 1 and 250 g / L. Activated charcoal has an active surface (between 500m ² / g and 2500m ² / g).

Mixing times are between 1 and 20 hours.

The evaporation pressures are between P _abs = 0.05 and 0.95 bar.

The following gases are useful as high pressure extraction media: C0 ₂ , N ₂ O, hydrocarbons having 1-10 C atoms, freons, or mixtures thereof.

Gas extraction pressures are between 10 and 450 bar.

Extraction temperatures with gases between 0 and 95 ° C.

The gas extraction flows are between 0.01 and 150 kg of gas / h / kg of extracted matter.

Several different solvents were used to obtain the antioxidant components of rosemary in accordance with EC84 / 344 / EEC, and the procedure was carried out as follows:

The dried rosemary (moisture content is 5-11%) was ground in an inert atmosphere and then dosed into the extractor. We added 2 - 30 liters of solvent per 1 kg of rosemary and extracted for 1-20 hours at a temperature between 10 and 90 ° C. After the first extraction stage was completed, the second extraction stage was started, under similar conditions as the first stage. The whole extract solution was filtered to remove any particulate matter. The filtrate was run in a vacuum evaporator (P _abs ~ 0.15-0.70 bar), where 50 to 70% of the solvent was evaporated, which can be returned to the process. Activated charcoal was added to the concentrate, ie 1-250 g of activated charcoal per liter of concentrate. It was stirred for 1-20 hours and then the concentrate was filtered.

The residue of the solvent and the aromatic component could be removed in two ways:

1. The filtrate was then extracted with CO ₂ at pressures between 10 and 450 bar and a temperature between 0 and 95 ° C and a CO ₂ flow between 0.01 and 150 kg / h / kg of the extract solution. The separator separated CO ₂ and an extract containing mostly ethanol and rosemary essential oils. The extractor (residue after extraction) gave the final product, which was light brown in color, odorless and tasteless.

The scheme of the proposed technological process for the production of antioxidants from rosemary is shown in FIG. 2.

2. After evaporation, the concentrate was run into a distillation column in which steam distillation was carried out. Distillate containing acetone, water and essential oil was discarded. The distillation residue was filtered off and the precipitate was then dried in an oven at temperatures between 20 and 80 ° C. The extract was further treated with supercritical CO ₂ (at pressures between 10 and 450 bar and temperatures between 0 and 95 ° C and a CO ₂ flow between 0.01 and 150 kg / h / kg of extract), removing the remaining essential oils and some dyes.

The scheme of the proposed technological process for the production of antioxidants from rosemary is shown in FIGURE 3.

The effect of the solvent on the content of the antioxidant components was determined by thin layer chromatography and the antioxidant activity by determining the peroxide number of oils to which the product obtained by the process of the invention was added.

Several different plant materials from the mouthpiece group were used to obtain the antioxidant components, but the antioxidant effectiveness of the extracts obtained from rosemary, sage and thyme was highest.

The results of comparative analyzes of some known products with products obtained by our process showed that the antioxidants obtained by our process are more effective than synthetic and of the two known antioxidants (RAPS & Co, DKulmbach and Nutrilo Gesellschaft fur Lebensmitteltechnologie m.b.H., D-Cuxhaven). From this, we can conclude that our products contain more phenolic diterpenes than these two known extracts. Based on our analyzes, we have determined that our extracts contain 27% by weight of polyphenols. Typically, the percentage of polyphenols in the extracts is between 2.8 and up to 29.5% by weight. From this we can conclude that our process gives high quality extracts. Their advantage is that they are completely odorless and tasteless. In addition, the process is simpler and more energy efficient.

The process of the invention is illustrated by the following Examples, but by no means limited thereto.

Example 1

The dried rosemary (residual moisture content 7.1%) is ground in an inert atmosphere and then dosed into the extractor. Add ethanol 201 per 1 kg of rosemary and extract for 15 hours at 60 ° C. After the first extraction stage is completed, start the second extraction step under the same conditions as the first stage.

The whole extract solution is filtered to remove any particulate matter. The filtrate was taken up in a vacuum evaporator (P _abs «0.45 bar) where 60% ethanol was evaporated. Activated carbon concentrate 50 g of activated carbon per liter of concentrate. The mixture was stirred for 20 hours and then the concentrate was filtered.

The filtrate is then extracted with CO ₂ at a pressure of between 250 bar and a temperature of 65 C and a flow of CO ₂ between 20 kg / h / kg of the extract. The separator separates CO ₂ and an extract containing mostly ethanol and rosemary essential oils. The extractor (residue after extraction) gave the final product (9.2% by weight by weight of dry rosemary), which is light brown in color, odorless and tasteless.

Example 2

The dried sage (residual moisture content 6.9%) is ground in an inert atmosphere and then dosed into the extractor. Add 20 l of ethanol per 1 kg of sage and extract for 15 hours at 60 ° C. After the first extraction stage is completed, start the second extraction step under the same conditions as the first stage.

The whole extract solution is filtered to remove any particulate matter. The filtrate was taken up in a vacuum evaporator (P _abs «0.45 bar) where 60% ethanol was evaporated. Activated carbon concentrate 50 g of activated carbon per liter of concentrate. The mixture was stirred for 20 hours and then the concentrate was filtered.

The filtrate is then extracted with CO ₂ at a pressure between 250 bar and a temperature of 65 C and a flow of CO ₂ between 20 kg / h / kg solution. The separator separates CO ₂ and an extract containing mostly ethanol and sage essential oils. The extractor (residue after extraction) gave the final product (8.3 wt% by weight of dry sage), which is light brown in color, odorless and tasteless.

Example 3

The dried thyme (residual moisture content of 7.2%) is ground in an inert atmosphere and then dosed into the extractor. 201 ethanol per 1 kg of thyme is added and extracted for 15 hours at 60 C. After the first extraction stage is completed, the second extraction stage is started, which takes place under the same conditions as the first stage.

The whole extract solution is filtered to remove any particulate matter. The filtrate was taken up in a vacuum evaporator (P _abs ~ 0.45 bar), where 60% ethanol was evaporated.

Activated carbon concentrate 50 g of activated carbon per liter of concentrate.

The mixture was stirred for 20 hours and then the concentrate was filtered.

The filtrate is then extracted with CO ₂ at a pressure between 250 bar and a temperature of 65 C and a flow of CO ₂ between 20 kg / h / kg solution. The separator separates CO ₂ and an extract containing mostly ethanol and thyme essential oils. The extractor (residue after extraction) gave the final product (7.9 wt% by weight of dry thyme), which is light brown in color, odorless and tasteless.

Example 4

The dried rosemary (residual moisture content 7.1%) is ground in an inert atmosphere and then dosed into the extractor. 221 acetone is added per 1 kg of rosemary and extracted for 13 hours at a temperature of 50 C. After the first extraction stage is completed, the second extraction stage is started, under the same conditions as the first stage.

The whole extract solution is filtered to remove any particulate matter. The filtrate was taken up in a vacuum evaporator (P _abs «0.55 bar), where 56% acetone was evaporated. Activated charcoal 55 g of activated charcoal per liter of concentrate is added to the concentrate. The mixture was stirred for 12 hours and then the concentrate was filtered.

The filtrate was then extracted with CO ₂ at a pressure of 250 bar and at a temperature of 55 C and a CO ₂ flow rate of 20 kg / h / kg of extract. The separator separates CO ₂ and an extract containing mostly acetone and rosemary essential oils. The extractor (residue after extraction) yields the final product (9.3% by weight by weight of dry rosemary), which is light brown in color, odorless and tasteless.

Example 5

The dried rosemary (residual moisture content 7.1%) is ground in an inert atmosphere and then dosed into the extractor. Add 17 liters of acetone per 1 kg of rosemary and extract for 20 hours at 50 C. After the first extraction stage is completed, begin the second extraction step under the same conditions as the first stage. The solution is filtered to remove any particulate matter. The filtrate was taken to a blender, where 120 g of activated charcoal per liter of extract was added. The mixture was stirred for 10 hours. The filtrate was filtered and the filtrate was taken to a vacuum evaporator (P _abs = 0.50 bar), where approximately 70% of the solvent was evaporated. The concentrate is led into a distillation column in which steam is distilled. Distillate containing acetone, water and essential oil is discarded. The distillation residue was filtered off and the precipitate was then dried in an oven at 80 C. The dried extract was further treated with supercritical CO ₂ (400 bar, 60 C) to remove any remaining essential oils and some colorants. The weight of the final product is 8.2% by weight based on the weight of the dry rosemary, which is light brown in color, odorless and tasteless.

Example 6

The dried thyme (residual moisture content of 7.2%) is ground in an inert atmosphere and then dosed into the extractor. Add 17 liters of acetone per 1 kg of thyme and extract for 20 hours at 50 C. After the first extraction stage is completed, start the second extraction step under the same conditions as the first stage. The solution is filtered to remove any particulate matter. The filtrate was taken to a blender, where 120 g of activated charcoal per liter of extract was added. The mixture was stirred for 10 hours. The filtrate was filtered and the filtrate was taken to a vacuum evaporator (P _abs = 0.55 bar), where approximately 70% of the solvent was evaporated. The concentrate is led into a distillation column in which steam is distilled. Distillate containing acetone, water and essential oil is discarded. The distillation residue was filtered off and the precipitate was then dried in an oven at 80 C. The dried extract was further treated with supercritical CO ₂ (400 bar, 60 C) to remove any remaining essential oils and some colorants. The weight of the final product is 8.1% by weight, based on the weight of dry thyme, which is light brown in color, odorless and tasteless.

Example 7

The dried sage (residual moisture content 6.9%) is ground in an inert atmosphere and then dosed into the extractor. Add 17 liters of acetone per 1 kg of sage and extract for 20 hours at 50 ° C. After the first extraction stage is completed, start the second extraction step under the same conditions as the first stage. The solution is filtered to remove any particulate matter. The filtrate was taken to a blender, where 120 g of activated charcoal per liter of extract was added. The mixture was stirred for 10 hours. The filtrate was filtered and the filtrate was taken to a vacuum evaporator (P _abs = 0.55 bar), where approximately 70% of the solvent was evaporated. The concentrate is led into a distillation column in which steam is distilled. Distillate containing acetone, water and essential oil is discarded. The distillation residue was filtered off and the precipitate was then dried in an oven at 80 C. The dried extract was further treated with supercritical CO ₂ (400 bar, 60 C) to remove any remaining essential oils and some colorants. The weight of the final product is 8.0% by weight based on the weight of the dry sage, which is light brown in color, odorless and tasteless.

Example 8

The dried rosemary is ground in an inert atmosphere and then dosed into the extractor. Add 17 liters of hexane per 1 kg of rosemary and extract for 20 hours at

C. After the first extraction stage is completed, we begin with the second extraction stage, which takes place under the same conditions as the first stage. The solution is filtered to remove any particulate matter. The filtrate was taken to a blender, where 120 g of activated charcoal per liter of extract was added. The mixture was stirred for 10 hours. The filtrate was filtered and the filtrate was led to a vacuum evaporator (P _abs = 0.55 bar), where approximately 67% of the solvent was evaporated. The concentrate is led into a distillation column in which steam is distilled. Distillate containing hexane, water and essential oil is discarded. The distillation residue was filtered off and the precipitate was then dried in an oven at 80 C. The dried extract was further treated with supercritical CO ₂ (430 bar, 60 C) to remove any remaining essential oils and some colorants. The weight of the final product is 7.8% by weight relative to the weight of the dry rosemary, which is light brown in color, odorless and tasteless.

Example 9

The dried rosemary (residual moisture content 7.1%) is ground in an inert atmosphere and then dosed into the extractor. Add 17 liters of ethyl acetate per 1 kg of rosemary and extract for 20 hours at 45 C. After the first extraction stage is completed, begin the second extraction step under the same conditions as the first stage. The solution is filtered to remove any particulate matter. The filtrate was taken to a mixer, where 120 g of activated charcoal per liter of extract was added. The mixture was stirred for 10 hours. The filtrate is filtered and the filtrate is taken to a vacuum evaporator (Pabs = 0 _> 55 bar) where approximately 70% of the solvent is evaporated. The concentrate is led to a distillation column in which steam is distilled. Distillate containing ethyl acetate, water and essential oil is discarded. The distillation residue is filtered off and the precipitate is then dried in an oven at 80 C. The dried extract is further treated with supercritical CO ₂ (440 bar, 60 C) to remove any remaining essential oils and some dyes. The weight of the final product is 8.5% by weight based on the weight of the dry rosemary, which is light brown in color, odorless and tasteless.

Example 10

The dried sage (residual moisture content 6.9%) is ground in an inert atmosphere and then dosed into the extractor. Add 17 liters of cyclohexane per 1 kg of sage and extract it for 20 hours at 50 C. After the first extraction stage is completed, begin the second extraction step, which takes place under the same conditions as the first stage. The solution is filtered to remove any particulate matter. The filtrate was taken to a blender, where 120 g of activated charcoal per liter of extract was added. The mixture was stirred for 10 hours. The filtrate was filtered and the filtrate was taken to a vacuum evaporator (P _abs = 0.55 bar), where approximately 55% of the solvent was evaporated. The concentrate is led into a distillation column in which steam is distilled. Distillate containing cyclohexane, water and essential oil is discarded.

The distillation residue is filtered off and the precipitate is then dried in an oven at 80 C. The dried extract is further treated with supercritical CO ₂ (450 bar, 60 C) to remove any remaining essential oils and some dyes. The weight of the final product is 8.4% by weight relative to the weight of the dry sage, which is light brown in color, odorless and tasteless.

Example 11:

Figure 4 shows the results of measurements of peroxide numbers for our (extract obtained by the procedure as in Example 5) and two competing extracts (RAPS & Co, DKulmbach sample-KOl and Nutrilo Gesellschaft fur Lebensmitteltechnologie m.b.h., D-Cuxhaven sample-KO2). Our extract was also added in the form of a mixture of acet.PIN (37.5% acetone extract, 37.5% ascorbic acid and 25% ascorbyl palmitate). The active substance content of all samples was 0.02%.

Of all the oil extracts, our acetone extract has the most effective antioxidant activity. A PIN formulation containing an emulsifier and synergist is even more effective by 20-30%. From these measurements we can also deduce the shelf life of this vegetable oil. The sample without the added antioxidant exceeded the peroxide count (10 mmolOg) after five hours (i.e., 5 months at room temperature). The oil that would add 0.02% of our extract would have a shelf life of 7-8 months, and competitive-RAPS and Nutrilo would have 5-5.5 months.

Example 12:

We have added our antioxidant ROS.PIN to our salami dressing mix. This antioxidant contained 37.5% ethanol extract of rosemary, 37.5% ascorbic acid and 25% ascorbyl palmitate. 80 g of this mixture was added per 100 kg of sausages. Since these sausages contain between 35-40% of fat, the additive of our antioxidant with regard to fat content was 0.075%.

Sausage samples with and without added antioxidant were first analyzed weekly (4 weeks) and then 3x at intervals of 1 month.

Analyzes were carried out by slicing 20 g of sausage (10 mm wide, 3 mm thick) and then extracted with a Soxhlet apparatus. Dichloromethane was used as the solvent, which, after extraction, was evaporated on a rotavapor. The quantitative content of fat and the peroxide number of extracted fat were determined. The results of measurements of peroxide numbers of extracted fats are shown in Figure 5. Sausage samples were also cut into pieces after one and two weeks of aging and left to stand in the air for 5 days. The fats were then extracted and peroxide numbers were determined. The difference between sausage with and without added antioxidant was large: 1.96 with added antioxidant and 19.8 mmolO ^ kg without antioxidant (1 week sample),

The difference between sausage with and without added antioxidant was large: 1.96 with added antioxidant and 19.8 mmolO ^ g without antioxidant (sample after week 1), and 7.02 with added antioxidant and 30.6 mmolO ^ g without antioxidant (sample after week 2) .

Example 13

Rosemary extract can be used in concentrated form. However, to expand the application range, it can be mixed with various substances to achieve optimum effect.

Excipients may be:

* emulsifiers (ascorbyl palmitate, lecithin, glycerides), * synergists (ascorbic acid, citric acid), * other antioxidants (tocopherols) and * alcohols (ethanol, propylene glycol).

The following are various mixtures based on rosemary antioxidant extract according to the invention.

ROS.CON

Composition

Description

Use

Dosage

Declaration:

Storage pure rosemary extract light brown-yellow powder meat and sausage

0.02-0.1% depending on the fat content is added to the seasoning mixture or directly into the product natural spice or. spice extract for 4 months in sealed packaging at room temperature, protected from light and moisture

ROS.OLEO

Composition

Description

Use

Dosage Rosemary extract (1-50% by weight) and vegetable oil dark yellow viscous liquid, water-insoluble, fat soluble meat, sausages, poultry, soup bases, chips, pasta, pastries, fresh and frozen seafood, mayonnaise, dressings , sauces, sweets, cake icing, pharmaceuticals and cosmetics

0.02-0.1% depending on the fat content can be added directly to the product while stirring, dispersed into spices or flavors, then mixed into the product, into frying oil, sprayed after dilution with vegetable oil or animal fat, which is especially suitable when oxidation is taking place on the surface or when the product is of irregular shape

Declaration: natural spice or. spice extract, vegetable oil

Storage for 4 months in a sealed container, protected from light, at a temperature between 20-30 C

ROS.EM.OLEO

Composition Rosemary extract, vegetable oil, lecithin, mono- and diglycerides

Description Dark yellow viscous liquid, water-dispersible and fat soluble

Use of meat, sausages, basics for soups, chips, flakes, mayonnaise, dressings, pasta, fried, fresh and frozen seafood, liquid smoke

A dosage of 0.01-0.1% based on fat content is added during the process to the aqueous phase under vigorous and continuous stirring, sprayed after dilution with water, added to phosphate solution or injected

Declaration Natural Spice spice extract, vegetable oil, mono- and diglycerides (E471, E472), lecithin (E322)

Storage for 4 months in sealed containers, at room temperature, protected from light

ROS.MALDEX

Ingredients Rosemary extract, maltodextrin

Description yellow powder

Use of meat, sausages, poultry

The dosage of 0.1-0.5% depending on the fat content is added directly to the meat or spice mixture

Declarationsnatural seasoning spice extract, saccharide

Storage for 3 months in a sealed container, in a cool, dry place

ROS.PROGLI

Composition: rosemary extract, propylene glycol

Description yellow-brown liquid miscible with water at pH-7-10, partially soluble in fats

Use in phosphate systems, cereals, potatoes, chips, flakes, basics Dosage for soups and other low fat foods 0.02-0.1% based on fat content Declaration: Storing added during the process to the aqueous phase under vigorous and continuous stirring, dispersed after dilution with water, added to phosphate solution or injected with natural spices or. spice extract, propylene glycol 6 months in a sealed container in a cool, dry place

ROS.E.SLAD

Composition Description Use Dosage tocopherol - vitamin E, rosemary extract, sugars yellow powder, unstable in alkaline sausages, meat, mayonnaise, dressings 0,2% based on fat content, mix directly into the product or into the spice mixture

Declarations of Tocopherol-Vitamin E, Natural Seasoning spice extract, sugars

Storage for 6 months in sealed original packaging, in a dry and cool place

ROS.E.OLEO

Composition tocopherol - vitamin E, rosemary extract, Description vegetable oil yellow viscous fluid, unstable in alkaline Use oils and fats, liquid and viscous foods Dosage 0,2% based on fat content, added together with other liquid components

Declaration of Tocopherol - Vitamin E, Natural Seasoning spice extract, vegetable oil

Storage for 4 months in sealed containers, in a dark and dry place, at a temperature between 20-30 C

Dosage 0.4% based on fat content Declaration: add together with other components during mixing natural spice or. spice extract, α-tocopherol-vitamin E, Storing ascorbic acid (E300), sodium ascorbate (E301), sugars for 6 months in sealed containers, dry, dark and cold

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ROS.E.EM Composition Description Use Dosage vitamin E, rosemary extract, mono- and diglycerides brown viscous paste, water-dispersible meat products, mayonnaise, dressings 0.2% by weight of fat is added along with other components Declaration: natural spice or. spice extract, α-tocopherol-vitamin E, emulsifiers (E471 and E472 a + b) ¹ Storing 4 months in sealed containers, in dry, cool and dark

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ROS.E.AP Composition vitamin E, rosemary extract, citric acid, Description Use Dosage ascorbyl palmitate, lecithin brown viscous paste, oil soluble meat products, mayonnaise, dressings 0.2% based on fat content Declaration: added along with other components natural spice or. spice extract, α-tocopherol-vitamin E, Storing citric acid (E330), emulsifiers (E304, E322) 4 months in sealed containers, in dry, cool and dark

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ROS.ALC Composition Description Use Dosage rosemary extract (20%), ethanol (80%) dark yellow liquid drinks, juices (for color stabilization) 0.1-0.15% added while stirring Declaration: natural spice or. spice extract, ethanol

Storage for 6 months in sealed containers, in a cool and dark place

ROS.AP.ALC

Ingredients rosemary extract, ascorbyl palmitate, ethanol

Description Dark yellow liquid

Use of meat, drinks, juices

Dosage 0.25%

Declaration: natural spice or. spice extract, emulsifier (E304), ethanol

Storage for 4 months in sealed containers, in a cool and dark place

ROS.AP.C (ROS.PIN)

Composition Rosemary extract, ascorbic acid, ascorbyl palmitate

Description yellow powder

Use of meat, sausages, seafood, poultry

0.0575% dosage is added together with other components

Declaration: natural spice or. spice extract, emulsifier (E304), ascorbic acid (E300)

Storage for 4 months in sealed containers in a dry, cool and dark place

DESCRIPTION PICTURE

Figure 1: a) carnosol, b) carnosolic acid, c) rozmanol,

d) rosemary diphenol; e) rosemary quinone.

Figure 2: Flowchart for the production of antioxidants from rosemary without distillation

Figure 3: Flowchart for the production of antioxidants from rosemary by distillation

Figure 4: Peroxide numbers of ours (Example 5) and competing extracts. Figure 5: Peroxide fat content extracted from sausages.

For

PINUS, d.d.

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Claims (14)

• PATENT APPLICATIONS A method for the production and isolation of antioxidant oral compounds (Labiatae), characterized in that it consists of: 1. extraction of ground vegetable material with an organic solvent, 2. partial solvent evaporation, 3. isolation of the antioxidant components carried out by a) steam distillation, drying and extraction with gases above a critical point; or b) extraction of aromatic components from the concentrate with gases above the critical point. Process according to claim 1, characterized in that the solvents are alcohols having 1-5 C atoms, esters such as e.g. ethyl acetate, butyl acetate, ketones such as e.g. acetone, hydrocarbons and their halogenated derivatives, cyclic hydrocarbons. Process according to claim 1, characterized in that the solvent / solid ratio is between 2 and 30 L solvents / kg of plant material (mouthpieces). Method according to claim 1, characterized in that the extraction times are between 1 and 20 hours. Process according to claim 1, characterized in that the extraction temperature is between 10 and 90 ° C. Process according to claim 1, characterized in that the extraction process is two-stage. Method according to claim 1, characterized in that the activated carbon / solution ratio is between 1 and 250 g / L and the activated carbon has an active surface between 500 m ² / g and 2500 m ² / g. Method according to claim 1, characterized in that the mixing times are between 1 and 20 hours. Method according to claim 1, characterized in that the evaporation pressures between P _abs = 0.05 and 0.95 bar. Process according to claim 1, characterized in that C0 ₂ , N ₂ O, hydrocarbons having 1-10C atoms, freons, or mixtures thereof are used as high pressure extraction gases. Process according to claim 1, characterized in that the extraction pressures with gases are between 10 and 450 bar. Process according to claim 1, characterized in that the extraction temperatures with the gases are between 0 and 95 ° C. Method according to claim 1, characterized in that the gas extraction flows are between 0.01 and 150 kg of gas / h / kg of the extracted substance. The antioxidants obtained according to claims 1 to 13. For PINUS, d.d. ABSTRACT PROCEDURE FOR THE OBTAINING AND INSULATION OF NATURAL ANTIOXIDIDES USTNATIC (Labiatae) A new process for the production and isolation of antioxidant oral compounds (Labiatae) has been developed, consisting of: 1. extraction of ground vegetable material with an organic solvent, 2. partial solvent evaporation, 3. isolation of the antioxidant components carried out by a) steam distillation, drying and extraction with gases above a critical point; or b) extraction of aromatic components from the concentrate with gases above the critical point. The products obtained are odorless and tasteless and therefore are of high quality antioxidants, useful e.g. in the pharmaceutical, cosmetics and food industries.