Preparation method of timosaponin A III

A technology of timosaponin and timosaponin, which is applied in the field of enzymatic hydrolysis to prepare timosaponin AⅢ, can solve the problems of low timosaponin AⅢ content and inability to prepare timosaponin AⅢ in large quantities.

Active Publication Date: 2014-01-29
SHANGHAI UNIV OF T C M
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AI Technical Summary

Problems solved by technology

[0006] In view of the low content of timosaponin AⅢ in Anemarrhena medicinal material, the traditional acid hydrolysis extraction method cannot produce a large amount of timos

Method used

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  • Preparation method of timosaponin A III
  • Preparation method of timosaponin A III
  • Preparation method of timosaponin A III

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] Weigh 250g Anemarrhena medicinal material, heat and reflux extraction with 50% ethanol twice, each time for 2 hours, the extract is recovered under reduced pressure and the ethanol is concentrated under reduced pressure to 625mL, then added to the treated D101 macroporous resin, washed with water for 3 times the column volume Impurities were removed, followed by elution with 3 times column volume of 20% ethanol, 3 times column volume of 35% ethanol and 3 times column volume of 50% ethanol, collecting the 50% ethanol eluted part, concentrating and drying under reduced pressure to obtain The total timosaponins of timosaponin BⅡ with a purity of 58% were 11.7 grams. Add 2260 U of β-glucosidase to the total saponins of Anemarrhena, add it to 500 mL of acetic acid buffer solution with a pH of 4.50, react in a water bath at 50°C for 1.0 hour, and extract with n-butanol three times (the dosages are 500 mL, 500mL, 500mL), the combined n-butanol extract was washed three times wi...

Embodiment 2

[0104] Weigh 500g Anemarrhena medicinal material, heat and reflux extraction with 70% ethanol for 3 times, each time for 2 hours, extract the extract under reduced pressure, recover the ethanol and concentrate under reduced pressure to 1200mL, add to the processed HPD100 macroporous resin, wash with 3 times the column volume Remove impurities, then use 3 times column volume of 20% ethanol, 3 times column volume of 35% ethanol, 3 times column volume of 50% ethanol and 3 times column volume of 70% ethanol to elute, collect 50% ethanol and The fraction eluted with 70% ethanol was concentrated and dried under reduced pressure to obtain 19.8 g of total timosaponins with a purity of 51% of timosaponin BII. Add the total saponins of Anemarrhena to 6080U of β-glucosidase and mix, add 800mL of acetic acid buffer solution with pH 4.80, react in a water bath at 50°C for 2.0 hours, and extract with n-butanol 3 times (the dosages are 1L, 1L, 1L), the combined n-butanol extracts were washed...

Embodiment 3

[0106]Weigh 1 kg of Anemarrhena medicinal material, heat and reflux with 60% ethanol for 2 times, each time for 2 hours, extract the extract under reduced pressure to recover the ethanol and concentrate it to 3L under reduced pressure, add it to the processed AB8 macroporous resin, wash with water for 3 times the column volume Impurities were removed, followed by elution with 3 times column volume of 20% ethanol, 3 times column volume of 35% ethanol and 3 times column volume of 50% ethanol, collecting the 50% ethanol eluted part, concentrating and drying under reduced pressure to obtain The total timosaponins of timosaponin BⅡ with a purity of 60% are 61.2 grams. Add 22400 U of β-glucosidase to the total saponins of Anemarrhena, mix it with 1.5 L of acetic acid buffer solution with a pH of 4.80, react in a water bath at 50°C for 3 hours, and extract with n-butanol for 3 times (the dosage is 1.5 L, 1.5L, 1.5L), the combined n-butanol extracts were washed twice with 0.1mol / L NaO...

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Abstract

The invention discloses a preparation method of timosaponin A III. The method is used for preparing timosaponin A III by using commercially-available traditional Chinese medicine rhizoma anemarrhenae decoction slices or freshly collected rhizoma anemarrhenae roots and stems or fibrous roots as materials through a combined process of alcohol extraction, enzymolysis, extraction, resin and silicagel column chromatography, and crystallization. The purity of timosaponin A III prepared by the method disclosed by the invention is higher than 90%, therefore, the preparation method is suitable for largely preparing timosaponin A III.

Description

technical field [0001] The invention relates to a method for preparing timosaponin AⅢ by enzymolysis. Background technique [0002] The traditional Chinese medicine Anemarrhena is the dried rhizome of Anemarrhena asphodeloides Bunge, which is mainly produced in Hebei, Inner Mongolia and other places. It has the effects of clearing away heat and purging fire, nourishing yin and moistening dryness. The chemical components of Anemarrhena are mainly saponins and flavonoids, among which steroidal saponins are the main components, and their content in rhizomes is about 6%. At present, more than 40 steroidal saponins have been found. [0003] The content of timosaponin BⅡ in Anemarrhena is the highest, and its structure is as follows: figure 1 . The 2010 edition of "Chinese Pharmacopoeia" stipulates that Anemarrhena medicinal materials contain no less than 3.0% timosaponin BⅡ; while literature reports (Determination of mangiferin and timosaponin BⅡ in Anemarrhena medicinal materi...

Claims

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Application Information

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IPC IPC(8): C12P33/20
Inventor 张彤刘艳平杨轶舜丁越蔡贞贞王如峰
Owner SHANGHAI UNIV OF T C M
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