CN102091107B - Method for extracting total triterpene of centella asiatica by enzymic process - Google Patents
Method for extracting total triterpene of centella asiatica by enzymic process Download PDFInfo
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- CN102091107B CN102091107B CN201110058021A CN201110058021A CN102091107B CN 102091107 B CN102091107 B CN 102091107B CN 201110058021 A CN201110058021 A CN 201110058021A CN 201110058021 A CN201110058021 A CN 201110058021A CN 102091107 B CN102091107 B CN 102091107B
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Abstract
The invention provides a method for extracting total triterpene of centella asiatica by an enzymic process, which comprises: deactivating endogenous enzymes of centella asiatica; and performing enzymic extraction. The extraction steep is high, the process conditions are mild, the extraction rate is high, the original properties of active ingredients are retained, and the low energy consumption is lowered. By using membrane separation, purification and concentration, organic solvent residue can be avoided effectively, less waste water is produced in the process, the purity of the separated total triterpene of centella asiatica is high, and the extraction rate is high.
Description
Technical field
The present invention relates to medical technical field, particularly a kind of method of Enzymatic Extraction Herba Centellae total glycosides.
Background technology
Herba Centellae [Centella asiatica (L.) Urban] is the dry herb or the whole herb with root of dicotyledon Umbelliferae [Umbelliferae].Being distributed widely in various places on the south the Yangtze river basin, is area among the people, Guangdong Chinese herbal medicine commonly used, and China is medically with Herba Centellae external and existing more than the 2000 year history of curing the disease for oral administration.Herba Centellae is cold in nature, bitter in the mouth, suffering, has the effect of clearing away heat-damp and promoting diuresis, promoting blood circulation and hemostasis, removing toxic substances and promoting subsidence of swelling.The herb master contains triterpenes, polyacetylene olefines, compositions such as volatile oil.The triterpenes main component comprises asiaticoside (Asiaticoside, molecular formula C
48H
78O
19, molecular weight 959.15CAS No:16830-15-2), asiaticoside (Madecassoside, molecular formula C
48H
78O
20, molecular weight 975.12CAS No:34540-22-2), domino asiaticoside (Terminoloside, molecular formula C
48H
78O
20, molecular weight 975.12CAS No:125265-68-1).They belong to the pentacyclic triterpene saponin of ursane type, are the main components of Herba Centellae pharmacologically active, and its structural formula is distinguished as follows:
Research shows, Herba Centellae total glycosides has anti-hypertrophy, fibrosis, antidepressant, anticancer, antioxidation and improves effect such as intelligence, in addition, asiaticoside to skin trauma, eliminate cicatrix etc. tangible curative effect arranged.Therefore, extraction separation Herba Centellae total glycosides and asiaticoside product have great importance to Herba Centellae industrialization and Herba Centellae resource efficiently.
At present, the technology of extraction separation Herba Centellae total glycosides and asiaticoside has some documents and patent report from Herba Centellae, and patent 200810303151.9 discloses a kind of Asiaticoside and preparation method thereof, adopts water or the hot diafiltration of low-carbon alcohols, hot reflux and supersound extraction; Patent 200710157180.4 discloses a kind of method of utilizing macroporous adsorbent resin to prepare Herba Centellae total glycosides, adopts soak with ethanol to extract; Patent 200710061217.3 discloses a kind of Herba Centellae extract and preparation method thereof, adopts soak with ethanol to extract; Patent 200910152717.7 discloses the method for distilling of asiatic centella total saponins and the application in field of medicaments thereof, adopts ethanol ultrasonic extraction.The method for distilling of immersion and diafiltration needs the doubly amount of solvent big, and extraction time is long, and production efficiency is low.Hot reflux is extracted, and a large amount of heat energy need be provided, and has increased the energy consumption of producing, and has also increased production cost simultaneously greatly.The supersound extraction extraction efficiency is higher, needs the input of increase equipment.
Bibliographical information is also arranged with the asiaticoside in the enzyme extraction Herba Centellae, method directly adopts cellulase at 40 ℃, and PH6.0 carries out enzymolysis and extraction.But because the Herba Centellae raw material itself contains endogenous enzyme, this enzyme has destruction or Degradation to Herba Centellae total glycosides in cryogenic aqueous solution, and the content of Herba Centellae total glycosides is reduced.
Summary of the invention
To the deficiency of prior art, the purpose of this invention is to provide a kind of method of Enzymatic Extraction Herba Centellae total glycosides.This method extraction rate is fast, and process conditions are gentle, and extraction ratio is high, not only helps to keep original character of active ingredient, and cuts down the consumption of energy.
A kind of method of Enzymatic Extraction Herba Centellae total glycosides comprises that adding cellulase degradation extracts, and filters, concentrate, and the microwave drying step, different with prior art is: before enzymolysis and extraction, set up deactivation Herba Centellae endogenous enzyme, and continued the step that insulation is extracted.
A kind of method of Enzymatic Extraction Herba Centellae total glycosides comprises following concrete steps:
(1) water of 5 times of amounts of adding Herba Centellae Radix Glycyrrhizae in extractor; Earlier water is warmed up to 100 ℃, the Herba Centellae raw material is dropped in the extractor again the endogenous enzyme in the Herba Centellae is carried out deactivation, temperature is controlled at 95-100 ℃; The insulation of deactivation continued is extracted 60min for 100 ℃, collects extracting solution;
(2) in extractor, add the water of 15 times of amounts of Herba Centellae Radix Glycyrrhizae again, coolant-temperature gage is controlled at 40-50 ℃, PH is transferred to 6.0, adds cellulase, makes that enzyme activity reaches 20-40u/ml in the water liquid, and enzymolysis 1.5 hours is collected extracting solution;
(3) merge extractive liquid, filters, and filtrating is carried out multistage membrance separation remove impurity, concentrate;
(4) the Herba Centellae total glycosides concentrated solution is used vacuum microwave drying, microwave pressure :-0.08MPa, temperature: below 60 ℃, pulverize and obtain the Herba Centellae total glycosides powder.
The endogenous enzyme inactivation time is 30min in the Herba Centellae raw material of step (1), is used effectively for making heat energy, extracts 60min for 100 ℃ so continue insulation,
The Herba Centellae total glycosides of this method extraction separation, the content of its Herba Centellae total glycosides are 30-60%, more than the Herba Centellae total glycosides extraction rate reached to 99%.
This method adopts the method for Enzymatic Extraction Herba Centellae total glycosides, and cellulase can be degraded and destroys the cell wall RF the cellulose in the Herba Centellae cell, makes that effective ingredient discharges to greatest extent in the cell, thereby improves the Herba Centellae total glycosides extraction ratio.
The invention has the advantages that:
1) the present invention adopts high temperature that the Herba Centellae endogenous enzyme is carried out deactivation, has effectively protected effective ingredient not to be degraded or destroys, and has improved the extraction ratio of asiaticoside greatly, has simplified operation sequence;
2) utilize cellulase that the cellulose in the Herba Centellae cell is degraded, destroy cell wall, the Herba Centellae total glycosides in the cell is discharged to greatest extent, Enzymatic Extraction speed is fast, and process conditions are gentle, and extraction ratio is high;
3) enforcement of the present invention has changed the high energy consumption of traditional extraction, the method for expensive, inefficient Herba Centellae total glycosides, and the insulation of deactivation continued is extracted, and had both played the purpose of deactivation, and heat energy is fully used.
The specific embodiment
Through embodiment the present invention is described further below.
Embodiment 1
Take by weighing Herba Centellae Radix Glycyrrhizae 50kg; 250L pure water in extractor is heated to 100 ℃, after the heating Herba Centellae Radix Glycyrrhizae is dropped in the extraction pot and carries out the deactivation of Herba Centellae endogenous enzyme; In the inactivation process; Temperature is controlled at 95-100 ℃ in the extraction pot, and the insulation of deactivation 30min continued is extracted 60min for 100 ℃, collects extracting solution; In extraction pot, add the 750L pure water again, coolant-temperature gage is controlled at 40-50 ℃, PH is transferred to 6.0, adds cellulase, makes that enzyme activity reaches 20-40u/ml in the water liquid, and enzymolysis 1.5 hours is discharged extracting solution, collects extracting solution; Merge extractive liquid, with the membrance separation that molecular weight 1500dalton crosses in extracting solution elder generation, is held back away soluble protein in the solution, larger molecular organics, the collection filter liquor; Filter liquor adopts the membrance separation of molecular weight 800dalton again, removes micromolecule inorganic matter and small organic molecules such as ferrum in the solution, calcium, magnesium, but the moisture of filtering simultaneously plays spissated effect, operating pressure: 0.05-0.20MPa; Reuse vacuum microwave drying (pressure :-0.08MPa, temperature: below 60 ℃) drying, pulverizing promptly gets.
Through detecting, Herba Centellae total glycosides content is 58.5%, and raw material Herba Centellae total glycosides extraction ratio is 99.2%.Utilize membrane separation purification to concentrate and can effectively avoid the organic solvent residual thing, the waste water that produces in the process is few, and isolates the high Herba Centellae total glycosides of purity, has practiced thrift energy consumption simultaneously.
Embodiment 2
Take by weighing Herba Centellae Radix Glycyrrhizae 500kg; 2500L pure water in extractor is heated to 100 ℃, after the heating Herba Centellae Radix Glycyrrhizae is dropped in the extraction pot and carries out the deactivation of Herba Centellae endogenous enzyme; In the inactivation process; Temperature is controlled at 95-100 ℃ in the extraction pot, and the insulation of deactivation 30min continued is extracted 60min for 100 ℃, collects extracting solution; In extraction pot, add the 7500L pure water, coolant-temperature gage is controlled at 40-50 ℃, PH is transferred to 6.0, adds cellulase, is that enzyme activity reaches 20-40u/ml in the water liquid, and enzymolysis 1.5 hours is discharged extracting solution, emits extracting solution; Merge extractive liquid, filters, and with the membrance separation that molecular weight 1500dalton crosses in filtrating elder generation, holds back away soluble protein in the solution, larger molecular organics, the collection filter liquor; Filter liquor adopts the membrance separation of molecular weight 800dalton again, removes micromolecule inorganic matter and small organic molecules such as ferrum in the solution, calcium, magnesium, but the moisture of filtering simultaneously plays spissated effect, operating pressure: 0.05-0.20MPa; Reuse vacuum microwave drying (pressure :-0.08MPa, temperature: below 60 ℃) drying, pulverizing promptly gets.
Through detecting, Herba Centellae total glycosides content is 59.4%, and raw material Herba Centellae total glycosides extraction ratio is 99.6%.
Claims (1)
1. the method for an Enzymatic Extraction Herba Centellae total glycosides comprises that adding cellulase degradation extracts, and filters, and concentrates and the microwave drying step, it is characterized in that: before cellulase degradation extracts, set up deactivation Herba Centellae endogenous enzyme, and continued the step that insulation is extracted; Said deactivation Herba Centellae endogenous enzyme; And the step that continues the insulation extraction is: the water that in extractor, adds 5 times of amounts of Herba Centellae Radix Glycyrrhizae earlier; And water is warmed up to 100 ℃, and the Herba Centellae raw material being dropped in the extractor carry out the endogenous enzyme deactivation again, temperature is controlled at 95-100 ℃; The insulation of deactivation continued is extracted 60min for 100 ℃, collects extracting solution.
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| CN108392503A (en) * | 2018-05-29 | 2018-08-14 | 重庆医药高等专科学校 | A kind of method of total saposins in extraction Sabia parviflora Wall.ex Roxb |
| CN114886815B (en) * | 2022-05-30 | 2023-06-20 | 广东美宝化妆品有限公司 | Method for extracting purely natural allergy-relieving anti-aging component from centella asiatica |
| CN114984071A (en) * | 2022-07-05 | 2022-09-02 | 广西昌洲天然药业有限公司 | Method, device, equipment and medium for extracting asiaticoside from centella asiatica |
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Non-Patent Citations (1)
| Title |
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| 樊亮等.用酶解预处理-微波法提取积雪草总苷.《华东理工大学学报（自然科学版）》.2009,第35卷(第4期),第592页右栏1.3项下. * |
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Denomination of invention: A kind of method for enzymatic extraction of centella asiatica total glycosides Effective date of registration: 20220830 Granted publication date: 20120905 Pledgee: Bank of China Limited by Share Ltd. Guilin branch Pledgor: GUILIN PLANT INGREDIENT Co.,Ltd. Registration number: Y2022450000144 |
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