CN1057300C - Glucoside compound of catechine and preparation method and use thereof - Google Patents
Glucoside compound of catechine and preparation method and use thereof Download PDFInfo
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Abstract
The present invention relates to a glucoside compound of catechin, a preparation method thereof and an application thereof. The compound provided by the present invention is dextrogyrate catechin-3-O-glucoside and has a structural formula (I). The compound is obtained by separating from PotentillaviscosaJ. Den and discloses a preparation method. By the experimental verification of medicine effect, the compound has obvious anti-liver damage function and can be used for preparing medicine for curing hepatitis. The present invention has good prospect for the comprehensive development and the use of the potentillaviscosaJ. Den as a wild resource and provides a possibility for preparing new medicine for curing the hepatitis.
Description
The invention belongs to field of medicaments, relate to a kind of glycoside compound of catechin and the method and the purposes of preparation thereof.
Potentilla viscosa Donn (Potentilla viscosa J.Den.) belongs to the Rosaceae, it is a kind of wild herb, raised growth is in China the Northeast, main vitellarium is Baicheng, Jilin Province and Yan Biandiqu, all be to be used for treating hepatitis by among the people for many years, and clinical proof have certain curative effect as a kind of herbal medicine.But the research of the pharmacology of relevant potentilla viscosa Donn and effective constituent does not appear in the newspapers so far.Potentilla viscosa Donn is as a kind of wild resource that has pharmaceutical use, and its comprehensive utilization especially research and development aspect deep processing is blank always, as Chinese materia medica big country not talkative be not one sorry.
The inventor starts with from the anti-liver injury studied on Pharmacology of Inhibiting to potentilla viscosa Donn, effective active composition is wherein studied exploration, isolated a kind of glycoside compound of catechin, and found that by pharmacodynamics test this compound has obvious anti-liver injury effect.
One of purpose of the present invention provides a kind of compound with anti-liver injury.It is the method for this compound of feedstock production that another object of the present invention provides with the potentilla viscosa Donn.A further object of the present invention is to propose the purposes of this compound aspect the treatment hepatopathy.
Compound provided by the invention is a kind of glycoside compound of catechin, has following structure:
The chemical name of this compound is: CGS.
Compound CGS provided by the invention is to separate to obtain from rosaceous plant potentilla viscosa Donn (Potentilla viscosaJ.Den.).
The separation preparation of the compound CGS among the present invention realizes by following process:
(1), the raw material potentilla viscosa Donn pulverize is added 60～80% acetone soln room temperature lixiviate 2～3 times, each extraction time 12～36 hours, the consumption of acetone soln is 3～8 times of material quantity, and the vat liquor of each time filters the back and merges, and reclaims acetone and is controlled in and carry out concentrating under reduced pressure below 50 ℃;
(2), filtering and concentrating liquid, filtrate enters Sephadex LH-20 molecular sieve chromatography post, first water wash-out discards elutriant, uses the methanol aqueous solution gradient elution again, collects the elutriant of 10 → 60% methanol solution, the concentrating under reduced pressure drying;
(3), desciccate enters Sephadex LH-20 molecular sieve chromatography post after with a small amount of 70% dissolve with ethanol solution once more, with 70% ethanolic soln wash-out, collects the elutriant that contains the tannin composition through the thin-layer chromatography inspection, carries out the concentrating under reduced pressure drying;
(4), desciccate is dissolved in the less water, enters the MCI-gel chromatographic column, water → 50% methanol solution gradient elution is collected and is merged 5～50% methanol solution elutriant, concentrating under reduced pressure drying;
(5), desciccate enters Sephadex LH-20 molecular sieve chromatography post after with a small amount of 95% dissolve with ethanol solution, uses 95% ethanol elution, collects the elutriant that contains the tannin composition through the thin-layer chromatography inspection, merge and carry out the concentrating under reduced pressure drying;
(6), enriched product is dissolved in less water, enters the MCI-gel chromatographic column, water → 50% methanol solution gradient elution is crossed post, wash-out 2～3 times repeatedly, collects the elutriant that contains the tannin composition through the thin-layer chromatography inspection, merges and carries out the concentrating under reduced pressure drying;
(7), desciccate enters Sephadex LH-20 molecular sieve chromatography post after with a small amount of 95% dissolve with ethanol solution, use 95% ethanol elution, segmentation receives elutriant, and carries out thin-layer chromatography and detect, the corresponding elutriant that collection has same spot carries out crystallization and recrystallization.
The product that obtains through above process is a colourless acicular crystal, identifies that by thin-layer chromatography structure is shown in (I) formula.
Repeatedly use chromatographic column (Sephadex LH-20 molecular sieve chromatography post and MCI-gel chromatographic column) among the present invention, the specification of post is decided according to the application of sample amount, under the general condition, the height and the diameter ratio of chromatographic column are about 10～20: 1, and the application of sample amount is about 1/50 of molecular sieve amount.
Product of the present invention is the tannin compound, need detect intermediate product in the sepn process at any time and whether contain the tannin composition, discards the non-tanning substance composition, and detection method can be conventional or any feasible method.Method is preferably: adopt the silica gel G chromatographic sheet, benzene-ethyl formate-formic acid is made developping agent, and ratio is 1～2: 7: 1, and launch, take out, dry, make developer with 1% iron trichloride methanol solution, show sap green spot person and be the tannin composition.
The tannin compound is met high temperature and easy polymerization, so in the sepn process, the elutriant behind each post is excessively reclaimed the temperature of solvent and concentrate drying all to be no more than 50 ℃ for well, so all adopt decompression operation, reclaim in the dissolving agent process, can drip propyl carbinol and prevent bumping.Dissolving can be adopted ultrasonic processing method when meeting difficulty.The purpose of recrystallization is to be further purified product, and is purer as product, can save this step.
Figure 1 shows that the schematic flow sheet of the inventive method.
The invention still further relates to the medical usage of this compound, namely prove that by effect experiment this compound has obvious anti-liver injury effect, can be used for preparing the medicine for the treatment of hepatitis.
By to the effect experiment in the various hepatic injury situations of animal mouse, find: CGS is to mouse carbon tetrachloride (CCl4) and the glutamic-pyruvic transaminase (SGPT) that causes of thioacetamide (TAA) raises and the reduction of liver glycogen has inhibitory action; To carbon tetrachloride (CCl4) and the reduction of thioacetamide (TAA) poisoning mice hepatic tissue SOD activity and MDA (MDA) content raise obvious inhibitory action arranged; Can obviously increase carbon tetrachloride (CCl4) and thioacetamide (TAA) poisoning mice liver cell pigment P-450 content; Can obviously reduce alcoholism murine liver tissue MDA (MDA) content; Can obviously shorten the mouse yellow Jackets length of one's sleep and increase hepatomicrosome cytochrome P-450 content; Rising has obvious reducing effect to ethionine induced mice liver tg; This compound also can promote normal or carbon tetrachloride (CCl4) poisoning mice liver drainage bromine sulphur peptide sodium (BSP) function; Promote the mouse choleresis.
The effective dose of this compound is 100～200mg/kg body weight. About the method for effect experiment and result detail display in an embodiment.
Comprehensive conclusions can find out that the compound CGS truly has obvious anti-liver injury effect, and its mechanism of action is by following several respects; (1) lipoid peroxidization resistant; (2) keep nucleic acid, protein and sugar unit metabolism normalization; (3) strengthen liver detoxification and excretory function; (4) promote the hepatic secretion function.
In addition, the inventor finds also that in research process the compound CGS has the effect of obvious anti-common cold virus simultaneously.
Compound CGS provided by the invention is to separate preparation from natural plants, and remarkable in the effect that protects the liver, falls aspect enzyme, cholagogic, the removing jaundice, so this compound has good medical value, particularly be used for preparing the newtype drug for the treatment of hepatitis, good prospect will be arranged.
In a word, the present invention finds and separates a kind of unique bioactive ingredients---CGS that protects the liver, falls enzyme, cholagogic, removing jaundice effect that has first from potentilla viscosa Donn, filled up the blank of relevant potentilla viscosa Donn study of active components, can drive and promote that the comprehensive development and utilization of this wild resource will produce larger Social benefit and economic benefit to the further research of the medical value of potentilla viscosa Donn. And CGS is used for the research of the medical usage of anti-hepatitis, also provides possibility for preparing a kind of new drug for the treatment of hepatitis.
Preparation embodiment 1
Take from right drying, and potentilla viscosa Donn root 7.46kg through pulverizing, add the lixiviate 3 times under room temperature of 60% acetone soln, 3 times used acetone soln is respectively 25800ml, 22500ml, 22500ml, extraction time was respectively 36 hours, 18 hours, 12 hours, 3 times vat liquor filters the back respectively and merges, reclaim acetone down and concentrate vat liquor in 40 ℃, filter out impurity, (post is high about 2 meters in SephadexLH-20 chromatographic column top to get filtrate, about 20 centimetres of diameter), with about 4000ml washing decon, use 10 → 100% successively more earlier, gradient is that 10% methanol aqueous solution carries out gradient elution, and the methanol usage of each gradient is 2000ml, collect 10～60% meoh eluate, 40 ℃ of following concentrating under reduced pressure dryings; Desciccate adds a small amount of 70% alcoholic solution after vibration dissolving on the ultrasonic apparatus, add Sephadex LH-20 chromatographic column top, with 70% ethanol stepwise elution, elutriant is with the silica gel g thin-layer plate inspection, is that benzene-ethyl formate-formic acid of 1: 7: 1 is developping agent with ratio, and every section elutriant is launched, takes out, dries, spray is with 1% iron trichloride methanol solution, show some sap green spots on the thin layer plate, merge the Rf value greater than 0.50 elutriant part, 40 ℃ of following concentrating under reduced pressure dryings; Desciccate is dissolved in and is added on MCI-gel chromatographic column top (high about 1 meter of post in the less water, 7 centimetres of diameters), add 0 → 40% successively, gradient is each 300ml wash-out of methanol aqueous solution of 5%, the elutriant that merges 5～40% methanol aqueous solutions, 40 ℃ of following concentrating under reduced pressure dryings; Product is dissolved in a small amount of 95% ethanol, is added on Sephadex LH-20 chromatographic column top, uses 95% ethanol elution, collects the elutriant contain the tannin composition with method, 40 ℃ of following decompression recycling ethanols and concentrate drying; Desciccate is dissolved in the less water, enters MCI-ge l chromatographic column, and water → methanol solution gradient elution three times so repeatedly, is collected the elutriant that contains the tannin composition, 40 ℃ of following concentrating under reduced pressure dryings; Product is dissolved in a small amount of 95% ethanol, is added on Sephadex LH-20 chromatographic column top, uses 95% ethanol elution, detects, merges the elutriant that tool is educated same spot with method with silica gel g thin-layer plate, and 40 ℃ of following concentrating under reduced pressure are placed a few days, separate out colourless crystallization.Weigh 1.77 the gram.Measure 227-228 ℃ of mp, [α]
18 D-13.3 ℃ of [Me
2CO-H
2O (1: 1); C10].
Fig. 2 provides the hydrogen spectrum nucleus magnetic resonance figure of this product:
1HNMR(Me
2CO-D
6+D
2O)：δ2.73(2H，d-like，J＝5Hz，H＝4)，4.32(1H，d，J＝7Hz，glc，H-1)，4.92(1H，d，J＝6Hz，H-2)，5.89(1H，d，J＝2Hz，H-6)，6.00(1H，d，J＝2Hz，H-8)，6.64(1H，dd，J＝2.8Hz，H-6｀)，6.76(1H，d，J＝8Hz，H-5｀)，6.84(1H，d，J＝2Hz，H-2｀)。
The CGS of this crystallized product for having formula (I) structure.
Preparation embodiment 2
Potentilla viscosa Donn root 3kg, add the lixiviate 3 times under room temperature of 60% acetone soln, 3 times used acetone soln is respectively 9000ml, 9000ml, 9000ml, extraction time was respectively 24 hours, 24 hours, 40 hours, 3 times vat liquor filters the back respectively and merges, reclaim acetone down and concentrate vat liquor in 40 ℃, filter out impurity, get filtrate in Sephadex LH-20 chromatographic column top, with about 2000ml washing decon, use 20 → 70% successively more earlier, gradient is that 10% methanol aqueous solution carries out gradient elution, and the methanol usage of each gradient is 800ml, collect 30～60% meoh eluate, 40 ℃ of following concentrating under reduced pressure dryings; As follows embodiment 1, the crystallization that obtains is faint yellow, crystallization is dissolved in hot water carries out recrystallization, promptly gets about 2.5 grams of pure product.
Preparation embodiment 3
Get bright potentilla viscosa Donn root 2.5kg, take advantage of the fresh-cut flakiness, add the lixiviate 3 times under room temperature of 80% acetone soln, 3 times used acetone soln is respectively 15000ml, 12000ml, 12000ml, extraction time was respectively 36 hours, 24 hours, 24 hours, and other are with embodiment 1.Obtain product 2.46 grams.
Experimental example
Carry out the test of pesticide effectiveness of animal mouse with CGS (CGS), verify the effect of this compound aspect anti-liver injury.This compound is made into desired concn with physiological saline before test.
Test one,
Mouse is divided into two batches, and every batch is divided into 4 groups, and 10 every group, be administered once to 3,4 groups of mouse subcutaneous injections by the dosage of 200mg/kg and 100mg/kg every day respectively in the male and female half and half, 3 day, and 1,2 group of injection is with volume physiological saline.After the last administration 1 hour, 1 group of mouse is pressed 10ml/kg abdominal injection soya-bean oil, and to first mouse, 2～4 groups all by 10ml/kg abdominal injection 0.1% tetracol phenixin (CCl
4) soybean oil solution; To second batch of mouse, 2～4 groups all by 50ml/kg abdominal injection thioacetamide (TAA).After the fasting 16 hours, disconnected neck is put to death animal, gets blood and hepatic tissue, carries out following index determining:
1, measures thymus nucleic acid (DNA), Yeast Nucleic Acid (RNA), protein, the fat and sugar unit content of gpt (SGPT) and hepatic tissue, the results are shown in Table 1.
2, measure the superoxide-dismutase (SOD) and mda (MDA) content of hepatic tissue, the results are shown in Table 2.
3, measure hepatomicrosome part cytochrome P-450 content, the results are shown in Table 3.
Test two,
Mouse is divided into 4 groups, and 10 every group, male and female half and half, be administered once to 3,4 groups of mouse subcutaneous injections by the dosage of 200mg/kg and 100mg/kg every day respectively in 3 days, and 1,2 group of injection is with volume physiological saline.Fasting is 8 hours after the last administration, presses 15ml/kg for 1 group and irritates stomach physiological saline, all irritates stomach 50% ethanol by 15ml/kg for 2～4 groups, and fasting is after 12 hours, and disconnected neck is put to death animal, gets hepatic tissue, measures mda (MDA) and content of triglyceride, the results are shown in Table 4.
Test three,
Mouse is divided into 3 groups, and 10 every group, male and female are pressed the dosage of 200mg/kg and 100mg/kg respectively to 2,3 groups of mouse subcutaneous injection administration secondaries (every injection in 12 hours once) in half and half, 24 hour, and 1 group of injection is with volume physiological saline.After the last administration 1 hour, all mouse peritoneal is injected vetanarcol (50mg/kg), normal temperature writes down every mouse righting reflex loss down to the timed interval (being the length of one's sleep) of recovering, and in mouse disconnected neck execution in awake back 2 hours, get hepatic tissue, measure hepatomicrosome part cytochrome P-450 content, the results are shown in Table 5.
Test four,
Mouse is divided into 4 groups, and 10 every group, male and female half and half, be administered once 1,2 group of injecting normal saline 10ml/kg every day respectively to 3,4 groups of mouse subcutaneous injections by the dosage of 200mg/kg and 100mg/kg in 3 days.After the last administration 6 hours, press 10ml/kg for 1 group and irritate stomach physiological saline, all irritate the stomach ethionine by 250ml/kg for 2～4 groups, after 24 hours, disconnected neck is put to death animal, gets hepatic tissue, measures content of triglyceride, the results are shown in Table 6.
Test five,
Mouse is divided into two batches, and first is divided into 2 groups, 10 every group, male and female half and half, 1 group of injecting normal saline 10ml/kg presses 200mg/kg subcutaneous injection administration for 2 groups, and injection every day once in 3 days, fasting is 16 hours after the last administration, give whole mouse tail vein injection bromine sulphur peptide sodium (BSP) 100mg/kg, disconnected neck is put to death animal after 15 minutes, gets blood system from serum, survey the BSP hold-up, the result is shown in first hurdle of table 7.
Second batch of mouse is divided into 3 groups, and 10 every group, half and half, the 3 group of male and female are administered once by the 200mg/kg subcutaneous injection every day in 3 days, and other 2 groups of injections are with volume physiological saline.After the last administration 1 hour, press 10ml/kg abdominal injection 0.1% CCl for 2,3 groups
4Soybean oil solution, 1 group of injection are with the volume soybean oil solution, and fasting was given whole mouse tail vein injection bromine sulphur peptide sodium (BSP) 100mg/kg after 16 hours, and disconnected neck is put to death animal after 15 minutes, gets blood system from serum, surveys the BSP hold-up, and the result is shown in second hurdle of table 7.
Test six,
Mouse is divided into 3 groups, and 10 every group, male and female half and half, be administered once to 2,3 groups of mouse subcutaneous injections by the dosage of 200mg/kg and 100mg/kg every day respectively in 3 days, and 1 group of injection is with volume physiological saline.After the last administration 24 hours, mouse is used etherization, under xiphoid-process, open the abdominal cavity, the ligation biliary pore is sewed up stomach wall, immediately 2,3 groups are administered once again, after 2 hours, break cervical vertebra and put to death animal, win gall-bladder, weigh after blotting, break gall-bladder and make bile flow to end, weigh after blotting gall-bladder with filter paper, it is poor as choleresis to take second place with two, and the result is as shown in table 8.
Table 1 X ± SD
| Group (mg/kg) | SGPT (u/100ml) | Glycogen (mg/kg) | DNA (mg/g) | RNA (mg/g) | Protein (mg/g) | Fat (mg/g) |
| 1 contrast | 182±33.1 | 51±10.1 | 2.6±0.61 | 29±3.2 | 220±36.5 | 68±7.2 |
| 2 CCl 4 | 711±67.9 ΔΔΔ | 12±3.5 ΔΔΔ | 2.1±0.32 | 26±2.5 Δ | 182±29.7 Δ | 74±8.8 |
| 3 CGS200+CCl 4 | 621±58.7 | 28±4.2 ... | 2.3±0.29 | 30±3.9 . | 232±28.5 . | 62±6.6 .. |
| 4 CGS100+CCl 4 | 650±50.2 . | 21±5.1 ... | 2.2±0.31 | 28±3.7 | 229±31.6 . | 65±6.9 . |
| 1 contrast | 186±35.2 | 54±10.9 | ||||
| 2 TAA | 543±91.0 ΔΔΔ | 29±9.4 ΔΔΔ | ||||
| 3 CGS200+TAA | 394±61.2 … | 46±12.8 .. | ||||
| 4 CGS100+TAA | 415±78.4 .. | 38±9.7. |
Table 2 X ± SD
| Group (mg/kg) | SOD (u/g wet tissue) | MDA (mg/g) |
| 1 contrast | 28±6.75 | 69±11.2 |
| 2 CCl 4 | 17±3.39 ΔΔΔ | 231±47.6 ΔΔΔ |
| 3 CGS200+CCl 4 | 24±4.58 .. | 147±21.4 ... |
| 4 CGS100+CCl 4 | 22±4.0 . | 179±25.9 .. |
| 1 contrast | 29±6.78 | 71±11.8 |
| 2 TAA | 21±3.77 Δ | 142±27.2 ΔΔΔ |
| 3 CGS200+TAA | 25±3.12 . | 80±19.6 … |
| 4 CGS100+TAA | 24±3.23 | 96±21.4 .. |
Table 3 X ± SD
| Group (mg/g) | P-450 (nmol/mg albumen) |
| 1 contrast | 083±0.193 |
| 2 CCl 4 | 0.63±0.102 ΔΔ |
| 3 CGS200+CCl 4 | 0.77±0.163 . |
| 4 CGS100+CCl 4 | 0.67±0.142 |
| 1 contrast | 0.84±0.199 |
| 2 TAA | 0.70±0.147 |
| 3 CGS200+TAA | 0.82±0.035 . |
| 4 CGS100+TAA | 0.73±0.072 |
Table 4 X ± SD
| Group (mg/kg) | MDA (nmol/kg wet tissue) | Triglyceride level (mg/g wet tissue) |
| 1 contrast | 69±23.9 | 31±49.2 |
| 2 ethanol | 150±48.1 ΔΔ | 62±7.86 ΔΔΔ |
| 3 CGS200+ ethanol | 81±35.7 .. | 48±4.99 .. |
| 4 CGS100+ ethanol | 88±36.2 .. | 55±7.21 |
Table 5 X ± SD
| Group (mg/kg) | The length of one's sleep (minute) | P-450 (nmol/mg albumen) |
| 1 contrast | 62.2±10.81 | 0.79±0.080 |
| 2 CGS200 | 48.4±6.97 .. | 0.97±0.097 ... |
| 3 CGS100 | 52.5±7.82 . | 0.90±0.092 . |
Table 6 X ± SD
| Group (mg/kg) | Triglyceride level (mg/g hepatic tissue) |
| 1 contrast | 25±6.72 |
| 2 ethionines | 55±9.87 ΔΔΔ |
| 3 CGS200+ ethionines | 36±6.68 ... |
| 4 CGS100+ ethionines | 40±7.86 .. |
Table 7 X ± SD
| Group (mg/kg) | Serum BSP hold-up (μ g/0.1ml) |
| 1 contrast | 1.36±0.311 |
| 2 CGS200 | 0.72±0.282 ΔΔ |
| 1 contrast | 1.27±0.318 |
| 2 CCl 4 | 3.01±0.452 |
| 3 CGS200+CCl 4 | 2.54±0.427 . |
Table 8 X ± SD
| Choleresis (mg/10g body weight) | P | |
| 10.1±4.51 | ||
| 15.4±5.04 . | ＜0.05 | |
| 13.2±4.85 | ＞0.05 |
Annotate:
1, in above each table, X ± SD: mean value ± coefficient of variation
ΔP＜0.05;
The Δ ΔP＜0.01:
Δ Δ ΔCompare with control group P＜0.001;
.P＜0.05:
..: P＜0.01;
...P＜0.001 and the 2nd group of comparison separately;
2, the measuring method of each index is in the above test: SGPT is by improvement KingShi method; Component separating such as hepatic tissue DNA, RNA, protein and fat are pressed the Rowland method; DNA, RNA and protein content press Burton, Schjeide respectively and the Lowry method is measured; Hepatic tissue MDA content is pressed the TAB method and is measured; The separation of SOD and mensuration are pressed the Geller method and are measured; Hepatic mitochondria cytochrome P-450 content is pressed the Mitoma method and is measured.
The above test of pesticide effectiveness can prove that CGS (CGS) is to CCl
4Cause that with TAA mouse SGPT rising has obvious restraining effect, and SOD activity in the energy enhance liver, to CCl
4, TAA and alcoholism mouse liver lipid peroxidation product---mda (MDA) content has obvious reduction effect; CGS (CGS) not only can increase CCl
4With TAA poisoning mice liver glycogen content, and can obviously suppress CCl
4The disorder of poisoning mice liver rna and protein synthesis, raise also inhibited to ethionine induced mice liver tg, show that CGS has inhibition and normalizing effect to liver injury mouse liver tissue metabolism: in addition, CGS can obviously shorten the mouse vetanarcol length of one's sleep and increase hepatomicrosome cytochrome P-450 content, show that this compound has inducing action to hepatomicrosome " drug metabolizing enzyme ", thereby strengthen the function of detoxification of liver; CGS also can obviously promote mouse liver excretory function and choleresis function.
Claims (5)
1, a kind of glycoside compound of catechin, feature are that this compound is the CGS with following structure.
2, a kind of preparation method with compound CGS of the described structure of claim 1, feature is that this compound is separated from potentilla viscosa Donn (Potentilla viscosa J.Den.), and this separation method is realized by following process:
(1), the raw material potentilla viscosa Donn pulverize is added 60～80% acetone soln room temperature lixiviate 2～3 times, each extraction time 12～36 hours, the consumption of acetone soln is 3～8 times of material quantity, and the vat liquor of each time filters the back and merges, and reclaims acetone and is controlled in and carry out concentrating under reduced pressure below 50 ℃;
(2), filtering and concentrating liquid, filtrate enters Sephadex LH-20 molecular sieve chromatography post, first water wash-out discards elutriant, uses the methanol aqueous solution gradient elution again, collects the elutriant of the methanol solution of 10 60%, the concentrating under reduced pressure drying;
(3), desciccate enters Sephadex LH-20 molecular sieve chromatography post after with a small amount of 70% dissolve with ethanol solution once more, with 70% ethanolic soln wash-out, collects the elutriant that contains the tannin composition through the thin-layer chromatography inspection, carries out the concentrating under reduced pressure drying;
(4), desciccate is dissolved in the less water, enters the MCI-gel chromatographic column, water → 50% methanol solution gradient elution is collected and is merged 5～50% methanol solution elutriant, concentrating under reduced pressure drying;
(5), desciccate enters Sephadex LH-20 molecular sieve chromatography post after with a small amount of 95% dissolve with ethanol solution, uses 95% ethanol elution, collects the elutriant that contains the tannin composition through the thin-layer chromatography inspection, merge and carry out the concentrating under reduced pressure drying;
(6), enriched product is dissolved in less water, enters the MCI-gel chromatographic column, water → 50% methanol solution gradient elution is crossed post, wash-out 2～3 times repeatedly, collects the elutriant that contains the tannin composition through the thin-layer chromatography inspection, merges and carries out the concentrating under reduced pressure drying;
(7), desciccate enters Sephadex LH-20 molecular sieve chromatography post after with a small amount of 95% dissolve with ethanol solution, use 95% ethanol elution, segmentation receives elutriant, and carries out thin-layer chromatography and detect, the corresponding elutriant that collection has same spot carries out crystallization and recrystallization.
3, the preparation method of compound according to claim 2, be characterised in that, detecting the method that contains the tannin composition in the elutriant is: adopt the silica gel G chromatographic sheet, benzene-ethyl formate-formic acid is made developping agent, ratio is 1～2: 7: 1, launch, take out, dry, make developer, show sap green spot person and be the tannin composition with 1% iron trichloride methanol solution.
4, the preparation method of compound according to claim 2 is characterised in that, the temperature that the elutriant behind each post is excessively reclaimed solvent and concentrate drying all is no more than 50 ℃.
5, a kind of purposes with compound CGS of the described structure of claim 1, feature are that this compound can be used for preparing the medicine for the treatment of hepatitis.
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| CN97125804A CN1057300C (en) | 1997-12-23 | 1997-12-23 | Glucoside compound of catechine and preparation method and use thereof |
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| CN1057300C true CN1057300C (en) | 2000-10-11 |
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| CN107823286A (en) * | 2017-12-15 | 2018-03-23 | 延边大学 | Potentilla viscosa Donn extract and its application |
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| CN111166731A (en) * | 2020-01-14 | 2020-05-19 | 延边大学 | Application of kaurine-type diterpenoids derived from Nymphalus chinensis in inhibiting fat accumulation |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5176786A (en) * | 1974-12-26 | 1976-07-02 | Sekisui Plastics | SUKURYUUKON BEA |
| JPS6040883A (en) * | 1984-03-28 | 1985-03-04 | 株式会社日立製作所 | Sealed container for hermetic compressor and its manufacturing method |
| JPH07179489A (en) * | 1993-12-21 | 1995-07-18 | Mitsui Norin Kk | New polyphenol glycoside |
-
1997
- 1997-12-23 CN CN97125804A patent/CN1057300C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5176786A (en) * | 1974-12-26 | 1976-07-02 | Sekisui Plastics | SUKURYUUKON BEA |
| JPS6040883A (en) * | 1984-03-28 | 1985-03-04 | 株式会社日立製作所 | Sealed container for hermetic compressor and its manufacturing method |
| JPH07179489A (en) * | 1993-12-21 | 1995-07-18 | Mitsui Norin Kk | New polyphenol glycoside |
Non-Patent Citations (1)
| Title |
|---|
| YUKUGAKU ZASSHI 105 7 1985.1.1 MUKAKAMI CHEMICAL AND CHEMO-TAXONOWICAL STUELIES OF FILICES * |
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