CN101849973A - Total triterpenoid extract of inonotus obliquus with hypoglycemic function and pharmaceutical preparation thereof - Google Patents
Total triterpenoid extract of inonotus obliquus with hypoglycemic function and pharmaceutical preparation thereof Download PDFInfo
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- CN101849973A CN101849973A CN 201010197991 CN201010197991A CN101849973A CN 101849973 A CN101849973 A CN 101849973A CN 201010197991 CN201010197991 CN 201010197991 CN 201010197991 A CN201010197991 A CN 201010197991A CN 101849973 A CN101849973 A CN 101849973A
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Abstract
The invention discloses a total triterpenoid extract of inonotus obliquus with a hypoglycemic function, which is prepared through the following steps: (1) crushing and sieving inonotus obliquus, adding ethanol aqueous solution for immersion, carrying out ultrasonic extraction, separating the supernatant, and concentrating and drying the supernatant, thereby obtaining ethanol extract; (2) dispersing the ethanol extract into distilled water, oscillating and extracting with petroleum ether, and separating the extract from water-phase residues generated in extraction; and (3) oscillating and extracting the water-phase residues with n-butanol, separating the extract, mixing, concentrating and drying, thereby obtaining the total triterpenoid extract of the inonotus obliquus with the hypoglycemic function, wherein the content of the triterpenoid is 70-300mg/g. the total triterpenoid extract of the inonotus obliquus with the hypoglycemic function provided by the invention can obviously reduce the fasting blood glucose level of mouse with hyperglycemia, can significantly ease the diabetes symptoms of the mouse with hyperglycemia, and has no significant toxicity to major organs.
Description
Technical field
The invention belongs to pharmaceutical field, particularly relate to a kind of total triterpenoid extract of inonotus obliquus and pharmaceutical preparation thereof with hypoglycemic activity.
Background technology
Diabetes are a kind of common incretion metabolism diseases that genetic predisposition is arranged, and its pathophysiological change is the absolute or relative deficiency of insulin secretion, causes metabolism disorders such as sugar, fat, protein.At present, diabetes have become the commonly encountered diseases and the frequently-occurring disease of the many countries in the whole world, and its mortality rate has occupied the 3rd after tumor, the cardiovascular, and what suffer from more than 90% in the global diabetics is type 2 diabetes mellitus, i.e. non-insulin-dependent diabetes mellitus (NID.DM).In addition, the present fashion trend of diabetes is rejuvenation, and WHO predicts had only 26% to surpass 65 years old in 2025 in the diabetic population of developing country, and is 62% in this numeral of developed country.China is owing to large population base, and at present absolute number of patients occupies first place in the world, and wherein the overwhelming majority is a type 2 diabetes mellitus.According to the WHO prediction, in the period of the 1995-2025, Chinese diabetes increasing degree will reach 68%, and it is the first that rate of increase occupies the whole world, and the absolute number of China's diabetics will break through 5,000 ten thousand when the time comes.Along with the continuous propelling of the modernization of Chinese medicine and internationalization process, extract its effective ingredient or monomer from Chinese medicine in recent years, it is developed to antidiabetic medicine becomes the research focus.
Inonqqus obliquus is a kind of medicinal fungi, mainly be distributed in the cold district of 45 °-50 ° of north latitude, contain multiple composition, have multiple pharmacologically actives such as antitumor, antioxidation, blood sugar lowering, antiinflammatory, pain relieving, anti-platelet aggregation, antiviral, enhancing human body immunity power based on polysaccharide, triterpenes, steroidal class and polyphenol compound.Since 16th century, the resident of states such as Russia, Poland, Finland just begins to eat, and is used to prevent and treat multiple diseases such as gastrointestinal tract, diabetes, cancer.Nineteen fifty-five, medical courses in general institute in Moscow announced that Inonqqus obliquus is a cancer-resisting substance, and the government permission Inonqqus obliquus can be used for the pharmaceuticals exploitation.The U.S. extremely pays close attention to the immunoenhancement result of Inonqqus obliquus, and is made into healthy accesary foods.The research worker of Japan is spoken highly of this medicine among the people, and as hepatocarcinoma, the therapeutic agent that acquired immune deficiency syndrome (AIDS) and 0-157 escherichia coli poison, and the patent of applying for multinomial relevant Inonqqus obliquus think that Inonqqus obliquus is a kind of " catholicon " inonotus obliquus sclerotium.Close all reports that adopt ultrasonic method from Inonqqus obliquus, to obtain the total triterpenes chemical compound and develop the hypoglycemic drug preparation but up to the present still find no.
Summary of the invention
The object of the present invention is to provide a kind of total triterpenoid extract of inonotus obliquus clear and definite, that toxic and side effects is little that acts on hypoglycemic activity.
Second purpose of the present invention provides a kind of pharmaceutical preparation with total triterpenoid extract of inonotus obliquus of hypoglycemic activity.
Technical scheme of the present invention is summarized as follows:
A kind of total triterpenoid extract of inonotus obliquus with hypoglycemic activity, make with following method:
(1) with the Inonqqus obliquus grinding and sieving, adding volumetric concentration in the ratio of 1g: 8-12mL is that the ethanol water of 70%-95% soaks and carries out ultrasonic extraction after 1-3 hour, the number of times that extracts is 2-4 time, each 30-120 minute, merge extractive liquid,, separation of supernatant, and will obtain ethanol extract after the supernatant concentration drying;
(2) described ethanol extract is scattered in the distilled water, uses petroleum ether oscillation extraction 3-5 time then, each 2-4 hour, extract is separated with the water raffinate;
(3) with n-butyl alcohol oscillation extraction 3-5 time of described water raffinate, each 2-4 hour, isolate extract, merge concentrated and drying, promptly obtaining triterpene content is the total triterpenoid extract of inonotus obliquus with hypoglycemic activity of 70-300mg/g.
Described hyperacoustic power is 100-150W.
The amount ratio of ethanol extract and distilled water is 1g: 20-50mL in the described step (2).
The consumption of described step (2) PetroChina Company Limited. ether is 10-30 a times of ethanol extract weight.
The consumption of the middle n-butyl alcohol of described step (3) is 10-30 a times of ethanol extract weight.
Described preparation has the total triterpenoid extract of inonotus obliquus of hypoglycemic activity by 25-70 weight portion described and the pharmaceutic adjuvant of 30-75 weight portion is made.
The dosage form of described pharmaceutical preparation is granule, capsule, tablet, powder, drop pill, suspending agent or suppository.
Total triterpenoid extract of inonotus obliquus with hypoglycemic activity of the present invention has tangible reduction fasting blood glucose level to the hyperglycemia mice, obviously alleviates the effect of hyperglycemia mice diabetic symptom, and main organs is not had overt toxicity.
Description of drawings
Fig. 1 is the thin-layer chromatogram with total triterpenoid extract of inonotus obliquus of hypoglycemic activity.
The specific embodiment
The present invention is further illustrated below in conjunction with specific embodiment.
Embodiment 1
(1) the 100g Inonqqus obliquus is pulverized the back and cross 30 mesh sieves, with the 800mL volumetric concentration is that the ethanol water of 70% concentration soaked 2 hours, ultrasonic extraction is 2 times then, hyperacoustic power is 150W, each 30min, merge extractive liquid,, the centrifugalize supernatant, and will obtain ethanol extract 6.462g after the supernatant concentration drying;
(2) ethanol extract is scattered in 20 times of weight in the distilled water of ethanol extract, uses 10 times of weight then, extract is separated with the water raffinate in the petroleum ether oscillation extraction of ethanol extract 3 times each 2 hours;
(3) with above-mentioned water raffinate with 10 times of weight in the n-butyl alcohol oscillation extraction of ethanol extract 3 times, each 2 hours, isolate extract, then extract is merged and concentrate and drying, promptly obtain the total triterpenoid extract of inonotus obliquus that 4.294g has hypoglycemic activity, it is carried out assay, and triterpene content is 100.270mg/g.
See Fig. 1 (unfolding condition: petroleum ether: acetone=wherein some a was the total triterpenoid extract of inonotus obliquus with hypoglycemic activity in 4: 1, and some b is Inonqqus obliquus alcohol standard substance)
Embodiment 2
(1) the 100g Inonqqus obliquus is pulverized the back and cross 30 mesh sieves, with the 1000mL volumetric concentration is that the ethanol water of 80% concentration soaked 3 hours, ultrasonic extraction is 3 times then, hyperacoustic power is 100W, each 2 hours, merge extractive liquid,, the sucking filtration separation of supernatant, and will obtain ethanol extract 10.977g after the supernatant concentration drying;
(2) ethanol extract is scattered in 30 times of weight in the distilled water of ethanol extract, uses 20 times of weight then, each 3 hours, extract is separated with the water raffinate in the petroleum ether oscillation extraction of ethanol extract 4 times;
(3) with above-mentioned water raffinate with 20 times of weight in the n-butyl alcohol oscillation extraction of ethanol extract 4 times, each 3 hours, merge concentrated extract and drying then, can obtain the total triterpenoid extract of inonotus obliquus that 6.304g has hypoglycemic activity, triterpene content is 115.736mg/g.
Embodiment 3
(1) the 100g Inonqqus obliquus is pulverized the back and cross 30 mesh sieves, with the 1200mL volumetric concentration is that the ethanol water of 95% concentration soaked 1 hour, ultrasonic extraction is 4 times then, hyperacoustic power is 100W, each 1 hour, merge extractive liquid,, the centrifugalize supernatant, and will obtain ethanol extract 4.391g after the supernatant concentration drying;
(2) above-mentioned ethanol extract is scattered in 50 times of weight in the distilled water of ethanol extract, uses 30 times of weight then, each 4 hours, extract is separated with the water raffinate in the petroleum ether oscillation extraction of ethanol extract 5 times;
(3) with above-mentioned water raffinate with 30 times of weight in the n-butyl alcohol oscillation extraction of ethanol extract 5 times, each 4 hours, merge concentrated extract and drying then, can obtain the total triterpenoid extract of inonotus obliquus that 3.840g has hypoglycemic activity, triterpene content is 75.000mg/g.
Annotate: wherein the cubage formula is
The total triterpenoid extract of inonotus obliquus with hypoglycemic activity of each embodiment is met acetic anhydride-strong sulfuric acid response, and chloroform-strong sulfuric acid response and trichloroacetic acid reaction are positive.Thin-layer chromatographic analysis contains triterpenoid compound Inonqqus obliquus alcohol in this extract, its content is analyzed through HPLC and is 2-50mg/mL.
Embodiment 4
Have the hypoglycemic activity of the total triterpenoid extract of inonotus obliquus of hypoglycemic activity to the hyperglycemia mice
Mode inducing mouse by the lumbar injection alloxan produces diabetic symptom.Behind the injection alloxan hydrate 72h, measure the blood glucose value (blood derives from mouse tail vein) of mice with test kit, the mice that blood glucose value is higher than 11.1mmol/L is divided into 4 groups at random, 8 every group.First group is the diabetic mice contrast; Second group is the positive contrast medicine of glibenclamide group; The 3rd group is the total triterpenoid extract of inonotus obliquus with hypoglycemic activity of the present invention, and be that the total triterpenoid extract of inonotus obliquus with reducing blood sugar and blood fat effect is raised in the mice filling every day.The 4th group is the normal mouse matched group, every day normal diet.Testing sample all with 0.4% sodium carboxymethyl cellulose (CMC-Na) aqueous solution suspendible after, by irritating the stomach pin sample solution is poured into the mice gastric by force, first group and irritate the 4th group of every day and raise the CMC-Na aqueous solution.Write down diet, drinking-water and body weight every day.Irritate after the stomach 20 days, get blood until dead mouse, immediately mice is dissected then, take out liver, thymus and spleen rapidly and weigh from eyeball of mouse.The centrifugal acquisition serum of mouse blood is measured blood sugar content, total cholesterol level, content of triglyceride, hdl concentration and Mouse Liver glycogen content in the mice serum.
The result shows, the inductive diabetic mice polydipsia of alloxan, polyphagia, poor growth, spleen index and thymus index are lower than normal group mice, and the total triterpenoid extract of inonotus obliquus with hypoglycemic activity can effectively improve the diabetic mice polydipsia, the symptom of polyphagia, body weight, spleen index and thymus index all increase to some extent than diabetic model group.
As shown in table 1, the lumbar injection alloxan causes the blood sugar level of diabetic mice to increase more than 2 times.The total triterpenoid extract of inonotus obliquus that the filling stomach has hypoglycemic activity is after 20 days, and fasting blood glucose level has had reduction (p＜0.01) significantly, and fasting blood sugar is reduced to 3.96mmol/L from initial 10.26mmol/L.The hepatic glycogen content of total triterpenoid extract of inonotus obliquus mice with hypoglycemic activity illustrates that than diabetic model group height the total triterpenoid extract of inonotus obliquus with hypoglycemic activity can promote the conversion of blood glucose to liver glycogen.
Table 1 has the influence of the total triterpenoid extract of inonotus obliquus of hypoglycemic activity to blood glucose in diabetic mice level and hepatic glycogen content
Compare with diabetic model group ##:p＜0.01; Compare with the normal control group *: p＜0.01
The result who measures blood lipid level shows that the total cholesterol level of diabetic model group mice, content of triglyceride are than normal control group height, and hdl concentration is lower than normal control group.After the filling stomach had the total triterpenoid extract of inonotus obliquus of hypoglycemic activity, total cholesterol level, the content of triglyceride of diabetic mice decreased, and hdl concentration obviously increases (p＜0.05), increases to 1.77mmol/L from 1.02mmol/L.These data show that the total triterpenoid extract of inonotus obliquus with hypoglycemic activity has good effect for reducing blood fat to diabetic mice.
Table 2 has the influence of the total triterpenoid extract of inonotus obliquus of hypoglycemic activity to the diabetic mice blood lipid level
#:p＜0.05, compare with diabetic model group ##:p＜0.01; *: p＜0.05, compare with the normal control group * *: p＜0.01
Embodiment 5
Granule: (by weight percentage)
Total triterpenoid extract of inonotus obliquus 25% with hypoglycemic activity of the present invention
Corn starch 65%
Sucrose 10%
It is that soft material is made in 70% ethanol water moistening that above-mentioned raw materials is added an amount of volumetric concentration, crosses 14 order nylon mesh and granulates, and wet granular is in 60 ℃ of dryings, dried granule is crossed 14 mesh sieve granulate, goes fine powder, packing then after No. 4 sieve sieves, sealing, packing gets product.
Embodiment 6
Capsule: (by weight percentage)
Total triterpenoid extract of inonotus obliquus 70% with hypoglycemic activity of the present invention
Corn starch 5%
Dextrin 10%
Microcrystalline Cellulose 10
Micropowder silica gel 5%
It is that soft material is made in 70% ethanol water moistening that above-mentioned raw materials is added an amount of volumetric concentration, crosses 20 order nylon mesh and granulates, and wet granular is in 60 ℃ of dryings, and dried granule is crossed 14 mesh sieve granulate, and the gained granule pours into capsule, makes finished product.
Embodiment 7
Tablet: (by weight percentage)
Total triterpenoid extract of inonotus obliquus 32% with hypoglycemic activity of the present invention
Corn starch 32%
Dextrin 25%
Sucrose 5%
Pulvis Talci 5%
Magnesium stearate 1%
Get total triterpenoid extract of inonotus obliquus, starch, dextrin, the sucrose with hypoglycemic activity of the present invention, mixing is made granule.Dry below 60 ℃, add 5% Pulvis Talci, 1% magnesium stearate, mixing, compacting is in blocks, and sugar coating or film-coat had both got finished product.
The preparation of powder, drop pill, suspending agent or suppository is to get the total triterpenoid extract of inonotus obliquus with hypoglycemic activity of the present invention to add that suitable pharmacy acceptable auxiliary method routinely makes.
Claims (7)
1. total triterpenoid extract of inonotus obliquus with hypoglycemic activity is characterized in that making with following method:
(1) with the Inonqqus obliquus grinding and sieving, adding volumetric concentration in the ratio of 1g: 8-12mL is that the ethanol water of 70%-95% soaks and carries out ultrasonic extraction after 1-3 hour, the number of times that extracts is 2-4 time, each 30-120 minute, merge extractive liquid,, separation of supernatant, and will obtain ethanol extract after the supernatant concentration drying;
(2) described ethanol extract is scattered in the distilled water, uses petroleum ether oscillation extraction 3-5 time then, each 2-4 hour, extract is separated with the water raffinate;
(3) with n-butyl alcohol oscillation extraction 3-5 time of described water raffinate, each 2-4 hour, isolate extract, merge concentrated and drying, promptly obtaining triterpene content is the total triterpenoid extract of inonotus obliquus with hypoglycemic activity of 70-300mg/g.
2. the total triterpenoid extract of inonotus obliquus with hypoglycemic activity according to claim 1 is characterized in that described hyperacoustic power is 100-150W.
3. the total triterpenoid extract of inonotus obliquus with hypoglycemic activity according to claim 1 is characterized in that the amount ratio of middle ethanol extract of described step (2) and distilled water is 1g: 20-50mL.
4. the total triterpenoid extract of inonotus obliquus with hypoglycemic activity according to claim 1, the consumption that it is characterized in that described step (2) PetroChina Company Limited. ether are 10-30 times of ethanol extract weight.
5. the total triterpenoid extract of inonotus obliquus with hypoglycemic activity according to claim 1, the consumption that it is characterized in that n-butyl alcohol in the described step (3) are 10-30 times of ethanol extract weight.
6. the pharmaceutical preparation of the total triterpenoid extract of inonotus obliquus with hypoglycemic activity of one of claim 1-5 is characterized in that described preparation has the total triterpenoid extract of inonotus obliquus of hypoglycemic activity by 25-70 weight portion described and the pharmaceutic adjuvant of 30-75 weight portion is made.
7. the pharmaceutical preparation with total triterpenoid extract of inonotus obliquus of reducing blood sugar and blood fat effect according to claim 6, the dosage form that it is characterized in that described pharmaceutical preparation is granule, capsule, tablet, powder, drop pill, suspending agent or suppository.
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| CN102579511A (en) * | 2012-03-13 | 2012-07-18 | 天津大学 | Method for integrally extracting steroidal compounds, polysaccharides and polyphenols from inonotus obliquus |
| CN103665080A (en) * | 2013-12-05 | 2014-03-26 | 杭州华东医药集团新药研究院有限公司 | Triterpenoid compounds and application thereof in diabetes treatment drugs |
| CN103690572A (en) * | 2013-12-17 | 2014-04-02 | 杨庆贺 | Preparation method of inonotus obliquus tea powder |
| CN104004106A (en) * | 2014-04-28 | 2014-08-27 | 杨庆贺 | Method for extracting effective substances in inonotus obliquus and preparation method for teabag |
| CN105663178A (en) * | 2016-03-25 | 2016-06-15 | 沈阳工学院 | Method for extracting inonotus obliquus total triterpenes |
| CN106309509A (en) * | 2016-08-19 | 2017-01-11 | 广州滋得洛夫生物科技有限责任公司 | Inonotus obliquus polyphenol complex and preparation method thereof |
| CN107125376A (en) * | 2016-02-26 | 2017-09-05 | 上海仙草堂保健食品有限公司 | A kind of Inonotus obliquus electuary（Tea） |
| CN109453209A (en) * | 2018-11-22 | 2019-03-12 | 杭州娃哈哈科技有限公司 | A kind of composition inhibiting tumor cell proliferation |
| CN110075132A (en) * | 2019-05-08 | 2019-08-02 | 山西大学 | A kind of method of extraction purification Inonotus obliquus triterpene |
| CN110367533A (en) * | 2019-08-16 | 2019-10-25 | 王远彤 | A kind of alimentation composition that treating diabetes and its application |
| CN112401239A (en) * | 2020-12-07 | 2021-02-26 | 广州正广生物科技有限公司 | Composition for reducing blood sugar and preparation method and application thereof |
| CN112790382A (en) * | 2020-12-31 | 2021-05-14 | 吉林省桦树茸国际健康产业集团有限公司 | Preparation method of blood sugar regulating extract and health food prepared by same |
| CN112899323A (en) * | 2019-12-04 | 2021-06-04 | 中国农业大学 | Method for promoting inonotus obliquus to produce hypoglycemic active polysaccharide by adding pretreated birch sawdust |
| CN113633668A (en) * | 2021-09-20 | 2021-11-12 | 黑龙江八一农垦大学 | High-efficiency extraction method of total triterpenoids of inonotus obliquus and preparation method of fine powder of total triterpenoids |
| CN115181680A (en) * | 2020-11-18 | 2022-10-14 | 山西运奕道生物科技有限公司 | Active substance extraction and blood sugar reducing application in inonotus obliquus culture |
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| CN101422491A (en) * | 2008-11-11 | 2009-05-06 | 天津大学 | Preparation method of inonotus obilquus as alpha-glucosidase inhibitors |
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| CN102579511B (en) * | 2012-03-13 | 2013-11-27 | 天津大学 | Method for integrally extracting steroidal compounds, polysaccharides and polyphenols from inonotus obliquus |
| CN102579511A (en) * | 2012-03-13 | 2012-07-18 | 天津大学 | Method for integrally extracting steroidal compounds, polysaccharides and polyphenols from inonotus obliquus |
| CN103665080A (en) * | 2013-12-05 | 2014-03-26 | 杭州华东医药集团新药研究院有限公司 | Triterpenoid compounds and application thereof in diabetes treatment drugs |
| CN103665080B (en) * | 2013-12-05 | 2015-06-10 | 杭州华东医药集团新药研究院有限公司 | Triterpenoid compounds and application thereof in diabetes treatment drugs |
| CN103690572A (en) * | 2013-12-17 | 2014-04-02 | 杨庆贺 | Preparation method of inonotus obliquus tea powder |
| CN103690572B (en) * | 2013-12-17 | 2017-01-04 | 杨庆贺 | Preparation method of inonotus obliquus tea powder |
| CN104004106A (en) * | 2014-04-28 | 2014-08-27 | 杨庆贺 | Method for extracting effective substances in inonotus obliquus and preparation method for teabag |
| CN107125376A (en) * | 2016-02-26 | 2017-09-05 | 上海仙草堂保健食品有限公司 | A kind of Inonotus obliquus electuary（Tea） |
| CN105663178A (en) * | 2016-03-25 | 2016-06-15 | 沈阳工学院 | Method for extracting inonotus obliquus total triterpenes |
| CN106309509B (en) * | 2016-08-19 | 2019-05-31 | 广州滋得洛夫生物科技有限责任公司 | Inonotus obliquus polyphenol complex compound and preparation method thereof |
| CN106309509A (en) * | 2016-08-19 | 2017-01-11 | 广州滋得洛夫生物科技有限责任公司 | Inonotus obliquus polyphenol complex and preparation method thereof |
| CN109453209A (en) * | 2018-11-22 | 2019-03-12 | 杭州娃哈哈科技有限公司 | A kind of composition inhibiting tumor cell proliferation |
| CN110075132A (en) * | 2019-05-08 | 2019-08-02 | 山西大学 | A kind of method of extraction purification Inonotus obliquus triterpene |
| CN110367533A (en) * | 2019-08-16 | 2019-10-25 | 王远彤 | A kind of alimentation composition that treating diabetes and its application |
| CN112899323A (en) * | 2019-12-04 | 2021-06-04 | 中国农业大学 | Method for promoting inonotus obliquus to produce hypoglycemic active polysaccharide by adding pretreated birch sawdust |
| CN115181680A (en) * | 2020-11-18 | 2022-10-14 | 山西运奕道生物科技有限公司 | Active substance extraction and blood sugar reducing application in inonotus obliquus culture |
| CN115181680B (en) * | 2020-11-18 | 2023-09-05 | 山西运奕道生物科技有限公司 | Active substance extraction and blood sugar reducing application in inonotus obliquus culture |
| CN112401239A (en) * | 2020-12-07 | 2021-02-26 | 广州正广生物科技有限公司 | Composition for reducing blood sugar and preparation method and application thereof |
| CN112790382A (en) * | 2020-12-31 | 2021-05-14 | 吉林省桦树茸国际健康产业集团有限公司 | Preparation method of blood sugar regulating extract and health food prepared by same |
| CN113633668A (en) * | 2021-09-20 | 2021-11-12 | 黑龙江八一农垦大学 | High-efficiency extraction method of total triterpenoids of inonotus obliquus and preparation method of fine powder of total triterpenoids |
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