CN1872870A - Method for preparing oleanolic acid from wrinkled papaya - Google Patents

Abstract

This invention relates to a method extracting oleanolic acid from Chaenomeles seines. The method comprises: (1) soaking pressed Chaenomeles seines residue (40 mesh) in water at a weight ratio of 1:4 for twice (3 h for each time), and soaking in water at a weight ratio of 1:3 for a third time; (2) adding pectinase, cellulase and hemicellulase at a weight ratio of 1:2:3 (the total amount is 1000 u/100 g Chaenomeles seines residue), enzymolyzing at 40 deg.C for 3 h to decompose cellulose and pectin, and filtering or pressing to obtain a filter cake; (3) adding 95% ethanol solution to the filter cake at a weight ratio of 1:4, extracting at 40 deg.C and 0.065 atm for 3 times (2 h for each time), and incorporating the extracts; (4) separating and purifying with a filtration membrane and then with an adsorption resin to obtain the final product. The method has such advantages as simple process, low cost, high yield and high product purity.

Description

A kind of method that from cockle pawpaw, prepares Oleanolic Acid

Technical field:

The present invention relates to a kind of method that from cockle pawpaw, prepares Oleanolic Acid, belong to the fruit processing technique field.

Background technology:

Pawpaw is a conventional Chinese medicine, and its former plant is Chaenomeles (Chaenomeles Lindl.) plant cockle pawpaw [Chaenomelesspeciosa (Sweet) Nakai], claims chaenomeles lagenaria, Chaenomeles pawpaw etc. again, and all there is cultivation the southern china various places.Its flower can be for viewing and admiring; Fruit medicine, can wind dispelling strong, Shujin, analgesia detumescence among the peoplely are usually used in soaking medicinal liquor; Plant benevolence and be rich in oleic acid and linolic acid.

One one of the Pharmacopoeia of the People's Republic of China (calendar year 2001 version) is the dry maturescent fruit of rosaceous plant chaenomeles lagenaria Chaenomeles speciosa (Sweet) Nakai 45 pages of pawpaws of recording of text.Have another name called cockle pawpaw.Its composition contains saponin, oxysuccinic acid, tartrate, citric acid, vitamins C, flavonoid, tannin.Its function with cure mainly flat liver Shujin and stomach dampness elimination.Be used for the arthritis with fixed pain caused by dampness contraction, the heavy pain of waist knee joint acid, the convulsion of vomiting and diarrhoea, beriberi oedema.

China is the origin distribution center of Chaenomeles (Chaenomeles Lindl) plant.The Chaenomeles plant is complete to belong to 5 kinds, and except that the fruit of Japanese Floweringquince kind, all the other 4 kinds all originate in China.Cockle pawpaw fallen leaves, half evergreen shrubs, branch tool thorn, flowers give birth to, and 3～5 every bunch, open or and the Ye Tongfang of first leaf.The wealthy ovum circle of blade is to long elliptic-lanceolate.Fruit is a operatic circle, tool fragrance, shrinkage after the pericarp drying.Early the numerous look of spring flower is gorgeous, autumn in summer fruit fragrance, can for view and admire, medicinal and edible.The cockle pawpaw resource is widely distributed in China, main product in Shandong, province such as Jiangsu, Gansu, Sichuan, Yunnan; Suitable temperate zone, the subtropical climate district be born in, happiness sun, anti-slightly the moon, cold-resistant, heat, adaptability is strong, and little acid, slight alkalinity soil all can adapt to.

Main medicinal ingredients is an Oleanolic Acid in the pawpaw, and Oleanolic Acid is the pentacyclic triterpene compound, and molecular formula is C ₃₀H ₄₈O ₃Pure product Oleanolic Acid is a white, needle-shaped crystals, and fusing point is 308～310 ℃, specific optical rotation [a] ²⁰ _D=+73.3 ° of (C=0.15, C ₃₀H ₄₈O ₃) water insoluble, dissolve in methyl alcohol, ethanol, acetone and chloroform.Modern science experimental study report, Oleanolic Acid have and protect liver effect of reducing enzyme levels, reducing blood lipid, antiplatelet aggregative activity and to the provide protection of chromosome damage, have application promise in clinical practice.

Extract the method for medicinal ingredients Oleanolic Acid from pawpaw, existing percolation, soxhlet extraction, cold soaking extraction method and alkali extraction and acid precipitation, alkali dissolved salt are analysed methods such as purifying.But still there are shortcomings such as the Oleanolic Acid yield of complex process, cost height, extraction is low, purity is low in these methods.

Method of the present invention is not seen the open report identical with present method through Yunnan Province's Office for Scientific and Technical Information literature search.

Summary of the invention:

The object of the present invention is to provide that a kind of technology is simple, cost is low, the yield height of extract Oleanolic Acid, the method that from cockle pawpaw, prepares Oleanolic Acid that purity is high.

Method of the present invention comprises extraction, isolation and purification operation, is specially:

A, to get the back granularity of squeezing the juice be 40 purpose pawpaw slags, puts into water immersion 2 times by 1: 4 ratio of mass percent, each 3 hours; Carrying out the 3rd time in mass percent in 1: 3 ratio in water again soaks, the prozyme that adds polygalacturonase, cellulase and hemicellulase simultaneously respectively, ratio is 1: 2: 3, add-on is a 1000u/100 gram pawpaw slag, in temperature is to carry out enzymolysis under 40 ℃ 3 hours, filter or squeeze after decomposing fiber and pectin, filter cake or filter residue are standby;

B, be that 40 ℃, pressure be 0.065 normal atmosphere under decompression extraction 2 hour in 1: 4 ratio of mass percent in temperature with 95% ethanol with filter cake, re-extract is 3 times respectively, and merges 3 alcohol extraction liquids standby;

C, alcohol extraction liquid is earlier carried out ultrafiltration with ultra-filtration membrane, abandon out the macromolecular alcohol solubility impurity, will abandon assorted alcohol extraction liquid again and carry out membrane concentration, reclaim ethanol, obtain paste;

D, adopt resin absorption to separate the paste of gained to separate with 200 order silica gel column chromatographies, recrystallization, promptly get the pure product of Oleanolic Acid.

The yield that the present invention and existing Soxhlet are extracted, the diacolation extraction method obtains Oleanolic Acid sees Table relatively down.

Material quantity (kilogram)	Dry weight (kilogram)	Method	Yield (%)
				100.0 100.0 100.0	16.5 16.2 16.6	This law Soxhlet is extracted diacolation and is extracted	88.7 59.5 72.8

The present invention compared with prior art has that technology is simple, cost is low, the yield of extract Oleanolic Acid and a purity advantages of higher.

Description of drawings:

Accompanying drawing is a process flow diagram of the present invention.

Embodiment:

Pawpaw slag with 100 kilograms of bright pawpaws (abandoning seed and flesh) is a raw material, carries out extraction, the isolation and purification of Oleanolic Acid in the pawpaw.

Detailed process is as follows:

1, getting the back granularity of squeezing the juice is 40 purpose pawpaw slags, puts into water in 1: 4 ratio of mass percent and soaks each 3 hours 2 times; Carrying out the 3rd time in mass percent in 1: 3 ratio in water again soaks, (ratio is 1: 2: 3 to add the prozyme of polygalacturonase, cellulase and hemicellulase simultaneously respectively, add-on is a 1000u/100 gram pawpaw slag), in temperature is to carry out enzymolysis under 40 ℃ 3 hours, filter or squeeze after decomposing fiber and pectin, filter cake or filter residue are standby;

2, be 0.065 normal atmosphere under decompression extraction 2 hour in 1: 4 ratio of mass percent at 40 ℃ of temperature, pressure with 95% ethanol with filter cake, re-extract is 3 times respectively, and merges 3 alcohol extraction liquids standby;

3, alcohol extraction liquid is carried out ultrafiltration with the conventional ultrafiltration film earlier, abandon out the macromolecular alcohol solubility impurity, will abandon assorted alcohol extraction liquid again and carry out conventional membrane concentration, reclaim ethanol, obtain paste;

4, the paste of gained is adopted conventional resin (as D101) fractionation by adsorption and silica gel (200 order) chromatographic separation, recrystallization, promptly get the pure product of 50.6g Oleanolic Acid.

Claims (1)

1, a kind of method for preparing Oleanolic Acid from cockle pawpaw comprises extraction, isolation and purification operation, it is characterized in that:

A, to get the back granularity of squeezing the juice be 40 purpose pawpaw slags, puts into water immersion 2 times by 1: 4 ratio of mass percent, each 3 hours; Carrying out the 3rd time in mass percent in 1: 3 ratio in water again soaks, the prozyme that adds polygalacturonase, cellulase and hemicellulase simultaneously respectively, ratio is 1: 2: 3, add-on is a 1000u/100 gram pawpaw slag, in temperature is to carry out enzymolysis under 40 ℃ 3 hours, filter or squeeze after decomposing fiber and pectin, filter cake or filter residue are standby;

B, be that 40 ℃, pressure be 0.065 normal atmosphere under decompression extraction 2 hour in 1: 4 ratio of mass percent in temperature with 95% ethanol with filter cake, re-extract is 3 times respectively, and merges 3 alcohol extraction liquids standby;

C, alcohol extraction liquid is earlier carried out ultrafiltration with ultra-filtration membrane, abandon out the macromolecular alcohol solubility impurity, will abandon assorted alcohol extraction liquid again and carry out membrane concentration, reclaim ethanol, obtain paste;

D, adopt resin absorption to separate the paste of gained to separate with 200 order silica gel column chromatographies, recrystallization, promptly get the pure product of Oleanolic Acid.