CN101574381A - Extractive of effective part of chloranthus glaber and preparation method thereof - Google Patents
Extractive of effective part of chloranthus glaber and preparation method thereof Download PDFInfo
- Publication number
- CN101574381A CN101574381A CNA2009101149065A CN200910114906A CN101574381A CN 101574381 A CN101574381 A CN 101574381A CN A2009101149065 A CNA2009101149065 A CN A2009101149065A CN 200910114906 A CN200910114906 A CN 200910114906A CN 101574381 A CN101574381 A CN 101574381A
- Authority
- CN
- China
- Prior art keywords
- preparation
- extractive
- effective
- acid
- effective part
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention relates to a phenolic acid effective part extracted from the traditional Chinese medicine-chloranthus glaber and preparation method thereof. Phenolic acid compound contained in the effective part includes rosmarinic acid, protocatechuic acid, isofraxidin and caffeic acid, and the total content of the phenolic acid compound in the extractive is no less than 50 percent. The chloranthus glaber is extracted by water or ethanol, extracting solution is processed by high-speed centrifugation, centrifugation solution is processed by ultrafiltration by an organic composite film, the pH value of ultrafiltrate is regulated to range from 1 to 7, and the ultrafiltrate is applied to a resin column to be absorbed; according to column volume, 2 to 3 times water is used for elution, and then 10 to 70 percent of ethanol with the pH value ranging from 8 to 14 is used for elution; and ethanol elution solution is collected, concentrated and dried, thereby obtaining the extractive of the effective part.
Description
Technical field
The present invention relates to a kind of effective ingredient in Chinese extract, phenolic acid effective kind part that extracts in particularly a kind of therefrom medicinal herbs Corallium Japonicum Kishinouye and preparation method thereof and its application in the adjuvant therapy medicaments of preparation antibacterial-anti-inflammatory drug and cancer.
Background technology
Herba Pileae Scriptae (Sarcandra glabra (Thunb.) Nakai) is the Chloranthaceae plant, has another name called Herba Sarcandrae, Herba Pileae Scriptae, synthetism lotus, Ramulus Sambuci Williamsii etc.Whole plant for medical use, have effects such as anti-inflammation, heat-clearing and toxic substances removing, expelling wind and removing dampness, promoting blood circulation and stopping pain, the synthetism of stimulating the menstrual flow, be used for the treatment of various diseases associated with inflammation, rheumatic arthritis, sore swollen toxin, traumatic injury, fracture etc., in recent years also be used for the auxiliary treatment of cancer, cancer of pancreas, gastric cancer, rectal cancer, hepatocarcinoma, esophageal carcinoma etc. are had obvious curative effects.
Though the chemical compound that part is got from Herba Pileae Scriptae has been proved to be certain pharmacologically active, but the medical substance basis that does not up to the present also have complete clear and definite Herba Pileae Scriptae, the formula sarcandra preparation of selling on the market all belongs to the crude extract preparation, as " " ZHONGJIEFENG PIAN ", " XUEKANG KOUFUYE " recorded of Chinese pharmacopoeia 2005 editions, " ZHONGJIEFENG ZHUSHEYE " that " FUFANG CAOSHANHU HANPIAN " and " the Sanitation Ministry medicine standard " 14 recorded; Therefore, the exploitation effective part of chloranthus glaber preparation that consumption is little, curative effect is high, effective ingredient is clear and definite is significant to the medical value that promotes the Chinese medicine Herba Pileae Scriptae.
Research to effective part of chloranthus glaber at present concentrates on flavone mostly, coumarin and organic acid chemical compound, research to phenolic acid compound is very few, publication number be CN 1879672A patent disclosure a kind of with caffeic acid 3, it is a kind of that 4-dihydroxy benzenes ethyl ester is the Herba Sarcandrae effective site total polyphenols of effective ingredient and patent disclosure that publication number is CN 101020637A is the method that feedstock production contains the rosmarinic acid total polyphenols with the Herba Sarcandrae, but the former described effective ingredient caffeic acid 3,4-dihydroxy benzenes ethyl ester does not have it to separate the bibliographical information that obtains so far from Herba Pileae Scriptae, the latter is except that rosmarinic acid, there is not other phenols component in the clear and definite total polyphenols, and the crucial effective site preparation method that is that both adopt is comparatively loaded down with trivial details, the enforcement difficulty is higher, is not easy to industrialized great production.
According to lot of documents, the higher rosmarinic acid of content has the multiple pharmacological action identical or close with the Herba Pileae Scriptae herb in the Herba Pileae Scriptae phenolic acid compound.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, a kind of new Herba Pileae Scriptae phenolic acid effective kind part and preparation method thereof is provided, be beneficial to industrialized great production; Another object of the present invention is exactly the exploitation effective part of chloranthus glaber preparation that consumption is little, curative effect is high, effective ingredient is clear and definite.
Extractive of effective part of chloranthus glaber disclosed by the invention is the phenolic acid effective kind part that therefrom extracts in the medicinal herbs Corallium Japonicum Kishinouye, and contained phenolic acid compound comprises protocatechuic acid, caffeic acid, isofraxidin and rosmarinic acid in the described effective site.
Described Chinese medicine Herba Pileae Scriptae is the dry Herb of Chloranthaceae plant plait Corallium Japonicum Kishinouye Sarcandra glabra (Thunb.) Nakai.
The total content of phenolic acid compound 〉=50% in the described extractive of effective part of chloranthus glaber.
The present invention finishes by the following technical programs.Comprise phenolic acid compound separation and purification in the Herba Pileae Scriptae, and the chemical constituent of gained effective part extract has been carried out deep research, its structure is identified; The phenolic acid compound Study on extraction; The content assaying method research of total phenolic acid; Phenolic acid compound antibacterial and anti-inflammation functions research in the Herba Pileae Scriptae, detailed process and result are as follows:
One, the separation of phenolic acids chemical constituent in the extractive of effective part of chloranthus glaber
Extractive of effective part of chloranthus glaber after being dissolved in water, is used ethyl acetate, n-butanol extraction successively, and extract concentrating under reduced pressure respectively gets acetic acid ethyl dissolution part and n-butyl alcohol dissolving part.
Acetic acid ethyl dissolution partly uses silica gel column chromatography (100-200 order) to carry out crude separation, again by methods such as silica gel column chromatography (200-300 order), RP-18 reversed phase column chromatography, gel filtration chromatography and recrystallization further separate, purification, wherein 4 phenolic acid compounds have finally been identified, they are respectively 3,4-resorcylic acid, 3,4-dihydroxycinnamic acid, isofraxidin and rosmarinic acid, all the other phenolic acid compounds also need further separation and purification.
Two, the evaluation of phenolic acids chemical constituent in the extractive of effective part of chloranthus glaber
3,4-resorcylic acid [being protocatechuic acid (3,4-Dihydroxybenzoic acid) again]: molecular formula C
7H
6O
4, white particulate crystal (methanol) is soluble in methanol; UV (MeOH) λ
Max(log ε): 222 (3.92), 343 (3.94) nm; ESIMS m/z:307[2M H]
, 189[M+Cl]
, 153[M H]
, 109[M COOH]
1H NMR (400MHz, CDCl
3): δ 7.32 (1H, d, J=2.0Hz, H-2), 7.27 (1H, dd, J=2.0,8.0Hz, H-6), 6.76 (1H, d, J=8.0Hz, H-5).Structural formula is as follows:
3,4-dihydroxycinnamic acid [having another name called caffeic acid (Caffeic acid)]: molecular formula C
9H
8O
4, faint yellow cluster crystal (MeOH); UV (MeOH) λ
Max: 326,300,217nm; ESI-MS m/z:181[M+Na]
+, molecular mass 180;
1HNMR (Acetoned
6, 400mHz) δ: 8.25 (1H, brs, H-4), 8.15 (1H, brs, H-3), 7.55 (1H, d, J=15.6Hz, H-8), 7.16 (1H, d, J=2.0Hz, H-2), 7.04 (1H, d, J=8.0,2.0Hz, H-6), 6.87 (1H, d, J=8.0Hz, H-5), 6.27 (1H, d, J=16.0Hz, H-7);
13CNMR (Acetone-d
6, 100mHz) δ: 127.3 (C-1), 116.0 (C-2), 146.0 (C-3), 148.4 (C-4), 122.1 (C-5), 115.4 (C-6), 145.6 (C-7), 114.8 (C-8), 167.9 (C-9).Structural formula is as follows:
Isofraxidin (Isofraxidin): molecular formula C
11H
10O
5, yellow needle (acetone) is soluble in methanol, acetone; UV (MeOH) λ
Max(log ε): 222 (4.25), 266 (4.04), 326 (2.35) nm;
1H NMR (400MHz, CDCl
3): δ 7.90 (1H, d, J=9.6Hz, H-3), 7.02 (1H, s, H-5), 6.24 (1H, d, J=9.6Hz, H-4), 3.82 (6H, s, 2 OCH
3).Structural formula is as follows:
Rosmarinic acid (Rosmarinic acid): molecular formula C
18H
16O
8,
D 20+ 142 (c1.5, MeOH), pale yellow powder is soluble in methanol; UV (MeOH) λ
Max(log ε): 221 (4.07), 291 (4.04), 327 (4.17) nm; ESIMS m/z:719[2M H]
, 359[M H]
197,161;
1H NMR (400MHz, C
5D
5N): δ 8.01 (1H, d, J=16.0Hz, H-7), 7.51 (1H, d, J=2.0Hz, H-2), 7.49 (1H, d, J=2Hz, H-2 '), 7.20 (1H, d, J=8.0Hz, H-5), 7.15 (1H, d, J=8.0Hz, H-5 '), 7.05 (1H, dd, J=8.0,1.6Hz, H-6), (7.03 1H, dd J=8.0,1.6Hz, H-6 '), 6.67 (1H, d, J=16.0Hz, H-8), 5.92 (1H, dd, J=4.0,8.4Hz, H-8 '), 3.57 (1H, dd, J=4.0,14.4Hz, H-7 ' a), 3.48 (1H, dd, J=8.4,14.4, H-7 ' is b);
13C NMR (100MHz, C
5D
5N): δ 126.8 (C-1), 116.0 (C-2), 147.7 (C-3), 150.6 (C-4), 118.0 (C-5), 122.2 (C-6), 146.2 (C-7), 114.8 (C-8), 167.4 (C-9), (129.3 C-1 '), 116.7 (C-2 '), 147.3 (C-3 '), (146.6 C-4 '), 116.6 (C-5 '), 121.1 (C-6 '), (37.8 C-7 '), 74.9 (C-8 '), 173.4 (C-9 ').Structural formula is as follows:
The present invention also utilizes high performance liquid chromatography that the gained effective part extract is further analyzed, and has successfully belonged to above-mentioned 4 phenolic acid compounds having identified on the gained chromatogram.
Another technical problem to be solved by this invention is the preparation method that discloses above-mentioned extractive of effective part of chloranthus glaber, and described preparation method comprises the following steps:
Herba Pileae Scriptae water or ethanol extraction, the extracting solution high speed centrifugation, wherein the medicinal liquid of ethanol extraction reclaims ethanol earlier, after fully dissolving with the water of equivalent then, leave standstill, filter, the filtrate high speed centrifugation, centrifugal liquid is carried out ultrafiltration with organic hybrid films, and the ultrafiltrate adjust pH is 1～7, last macroporous adsorptive resins absorption, first water elution with 2～3 times of column volumes, reuse pH is 8～14 10%～70% ethanol elution, collects ethanol elution, concentrates, is drying to obtain effective part extract.
In the extractive of effective part of chloranthus glaber preparation process of the present invention, the used organic hybrid films of ultrafiltration can be polysulfones, acetate fiber class, politef class or polypropylene type organic hybrid films, and its molecular cut off is 5000～200000; Described resin column is selected from macroporous resin or polyamide, and wherein the used macroporous adsorbent resin of macroporous adsorptive resins can be HPD-100, HPD-400, HPD-500 or AB-8.
Extractive of effective part of chloranthus glaber of the present invention can be used in the preparation antibacterial-anti-inflammatory drug.
Extractive of effective part of chloranthus glaber of the present invention can be used as the pharmaceutical composition or the preparation of active constituents of medicine and pharmaceutically acceptable pharmaceutic adjuvant composition anti-inflammation, and the dosage form of described pharmaceutical composition or preparation comprises injection, tablet, capsule, granule, drop pill, oral liquid, ointment or gel.
Extractive of effective part of chloranthus glaber of the present invention (extract A) and the pharmacodynamics comparative study test of the total water extract of Herba Pileae Scriptae (extract B) aspect anti-inflammation:
1. two kinds of extract on Carrageenan cause the influence of mice foot swelling
Get body weight and be 60 of the mices of 18～22g, male, the conventional raising after 3 days, by the body weight random packet, irritate stomach Herba Pileae Scriptae extract A 30g/kg respectively, Herba Pileae Scriptae extract B 30g/kg, prednisone 10mg/kg and equivalent distilled water, twice of every day, continuous 7 days, only caused inflammation in the right back sufficient subcutaneous injection 1% carrageenin 0.05ml/ of mice in 1 hour after the last administration, causing scorching back 4 hours, take off left and right sides metapedes with length hair intersection and weigh, with left and right sides metapedes weight difference as the swelling degree.The results are shown in Table 1.
Two kinds of extract on Carrageenan of table 1 Herba Pileae Scriptae cause the influence (X ± SD) of mice foot swelling
| Group | Dosage (crude drug in whole g/kg) | Number of animals | Swelling degree (mg) |
| Contrast | - | 15 | 93.82±18.49 |
| Prednisone | 10mg/kg | 15 | 37.12±14.10 * |
| Extract A | 30 | 15 | 64.77±26.01 * |
| Extract B | 30 | 15 | 67.58±24.27 * |
Compare with matched group:
*P＜0.01.
As known from Table 1, prednisone and Herba Pileae Scriptae A, B extract all can significantly suppress carrageenin and cause the mice foot swelling, compared significant difference with matched group, and the effect of extract A slightly are better than extract B.
2. two kinds of extract Dichlorodiphenyl Acetates cause the influence of mouse peritoneal permeability
Get body weight and be 40 of the mices of 20～22g, male and female half and half, be divided into 4 groups at random, irritate stomach extract A 30g/kg respectively, extract B 30g/kg, prednisone 10mg/kg and equivalent distilled water, every day twice, continuous 5 times, after the last administration 1 hour, the blue normal saline solution 10ml/kg of mouse mainline 0.5% ivens, lumbar injection 0.5% acetic acid 10ml/kg put to death animal after 30 minutes simultaneously, divided the flushing mouse peritoneal three times with the 6ml normal saline, flushing liquor is through centrifugal (2000rpm * 10min), get supernatant and survey its optical density at the 590nm place, the blue solution of ivens of using 10ug/ml is simultaneously measured its OD value as standard, and with this concentration of calculating ivens orchid in each sample, the result checks with t.
Two kinds of extract Dichlorodiphenyl Acetates of table 2 Herba Pileae Scriptae cause the influence (X ± S) of mouse peritoneal permeability
| Group | Dosage (crude drug in whole g/kg) | Number of animals | Ivens orchid (μ g/ml) | The P value |
| Contrast | - | 10 | 4.6±1.9 | - |
| Prednisone | 10mg/kg | 10 | 2.3±1.4 | ＜0.01 |
| The A extract | 30 | 10 | 2.7±1.1 | ＜0.05 |
| The B extract | 30 | 10 | 2.8±1.0 | ＜0.05 |
As seen from Table 2, Herba Pileae Scriptae A, B extract all have inhibition acetic acid to cause the effect that the mouse peritoneal permeability increases, and with matched group significant difference are arranged relatively, and the action intensity of two kinds of extracts is suitable.
3. two kinds of external influences of extract to the parts of fine bacteria growing
Test organisms is inoculated in plain agar culture medium flat plate or blood plate by nutritional need, cultivates 18 hours for 37 ℃, scrapes the lawn that takes a morsel and is emulsifiable in meat soup, than its concentration of turbid correction, does 100,000 times of dilutions again, makes its final concentration suitable 10
4Cfu/ml meat soup (beta hemolytic streptococcus is used serum broth).
Determination of tube method minimal inhibitory concentration (MIC): during experiment medicine is used doubling dilution after sterilizing, with aseptic meat soup (beta hemolytic streptococcus the is used serum broth) dilution of opposing doubly, the concentration series that makes the A sample is 10,5.0,2.5,1.25,0.625,0.313g crude drug in whole/ml, and the concentration series of B sample is 10,5.0,2.5,1.25,0.625,0.313 crude drug in whole/ml.Establish the contrast of blank and positive drug simultaneously, with the Cmin of asepsis growth as MIC.
Plating is measured minimal bactericidal concentration (MBC): the test tube of getting asepsis growth is inoculated in the plate to be cultivated, and is minimal bactericidal concentration with the concentration of the antibacterial that do not grow, the results are shown in Table 3.
Annotate: positive drug is selected different positive drug for use because of strain is different, what wherein staphylococcus aureus and beta hemolytic streptococcus were selected for use is penicillin, and what escherichia coli were selected for use is gentamycin, and Candida albicans is not selected positive drug for use in this experiment.
The external influence of two kinds of Herba Pileae Scriptae extracts of table 3 to the part antibacterial
This experiment record extract A to the MIC of staphylococcus aureus, escherichia coli, beta hemolytic streptococcus and Candida albicans all less than a minute extract B, and MBC except escherichia coli also all less than extract B.
No matter above-mentioned pharmacodynamics comparative study result of the test proof extractive of effective part of chloranthus glaber of the present invention all is better than the total water extract of Herba Pileae Scriptae in the effect aspect the antibiotic still antiinflammatory.
Extractive of effective part of chloranthus glaber of the present invention and its production and application compared with prior art has following advantage:
(1) the present invention is clear and definite first Herba Pileae Scriptae phenolic acid effective kind part, the phenolic acid compound total content reaches more than 50% in the gained effective part extract, and 4 chemical compounds have wherein been identified, be respectively protocatechuic acid, caffeic acid, isofraxidin and rosmarinic acid, wherein the rosmarinic acid that content is the highest has multiple and the same or analogous pharmacological action of Herba Pileae Scriptae herb according to the lot of documents report, as antitumor, anti-inflammation, inhibition blood platelet reduction, adjusting immunity of organism etc.; No matter pharmacodynamics comparative study result of the test proof extractive of effective part of chloranthus glaber of the present invention all is better than the total water extract of Herba Pileae Scriptae in the effect aspect the antibiotic still antiinflammatory.
(2) preparation method of extractive of effective part of chloranthus glaber provided by the invention has also changed the extracting mode that effective site extraction in the past adopts the multiple separation means of multiple organic solvent to combine, the Herba Pileae Scriptae water extraction, filter the back and cross macroporous adsorptive resins absorption, use the finite concentration ethanol elution, eluent concentrates, be drying to obtain, method is simple, safety, and cost is low, pollute for a short time, be suitable for industrialized great production.
Description of drawings
The high-efficient liquid phase chromatogram of Fig. 1 extractive of effective part of chloranthus glaber
A---protocatechuic acid
B---caffeic acid
C---isofraxidin
D---rosmarinic acid
The specific embodiment
The invention will be further described below in conjunction with specific embodiment, but the present invention is not limited in cited embodiment.
Embodiment 1
Herba Pileae Scriptae medicinal material coarse powder 5kg adds 10 times of water gagings, decocts 2 times, each 2 hours, merge decoction liquor, decoction liquor high speed centrifugation, centrifugal liquid molecular cut off are that 200000 polysulfone membrane is carried out ultrafiltration, the ultrafiltrate adjust pH is 1.0, last HPD-100 macroporous adsorptive resins absorption, with the water elution of 2～3 times of column volumes, reuse pH is 14.0 70% ethanol elution earlier, collect ethanol elution, concentrating under reduced pressure, vacuum drying promptly get effective part extract 20.56g.In rosmarinic acid, wherein contain total phenolic acid 62.52%.
Embodiment 2
Herba Pileae Scriptae medicinal material coarse powder 5kg adds 10 times of water gagings, decocts 2 times, each 2 hours, merge decoction liquor, decoction liquor high speed centrifugation, centrifugal liquid molecular cut off are that 150000 cellulose acetate film carries out ultrafiltration, the ultrafiltrate adjust pH is 2.0, last HPD-400 macroporous adsorptive resins absorption, with the water elution of 2～3 times of column volumes, reuse pH is 10.0 50% ethanol elution earlier, collect ethanol elution, concentrating under reduced pressure, vacuum drying promptly get effective part extract.In rosmarinic acid 18.25g, wherein contain total phenolic acid 59.11%.
Embodiment 3
Herba Pileae Scriptae medicinal material coarse powder 5kg adds 10 times of water gagings, decocts 2 times, each 2 hours, merge decoction liquor, decoction liquor high speed centrifugation, centrifugal liquid molecular cut off are that 20000 poly tetrafluoroethylene carries out ultrafiltration, the ultrafiltrate adjust pH is 4.0, last HPD-500 macroporous adsorptive resins absorption, with the water elution of 2～3 times of column volumes, reuse pH is 11.0 30% ethanol elution earlier, collect ethanol elution, concentrating under reduced pressure, vacuum drying promptly get effective part extract 16.85g.In rosmarinic acid, wherein contain total phenolic acid 58.12%.
Embodiment 4
Herba Pileae Scriptae medicinal material coarse powder 5kg adds 10 times of water gagings, decocts 2 times, each 2 hours, merge decoction liquor, decoction liquor high speed centrifugation, centrifugal liquid molecular cut off are that 10000 polypropylene screen carries out ultrafiltration, the ultrafiltrate adjust pH is 6.0, last AB-8 macroporous adsorptive resins absorption, with the water elution of 2～3 times of column volumes, reuse pH is 8.0 10% ethanol elution earlier, collect ethanol elution, concentrating under reduced pressure, vacuum drying promptly get effective part extract 15.50g.In rosmarinic acid, wherein contain total phenolic acid 56.73%.
Embodiment 5
Herba Pileae Scriptae medicinal material coarse powder 5kg, the ethanol that adds 10 times of amounts 60%, extract 2 times, each 1 hour, merge extractive liquid, reclaims ethanol, after fully dissolving with the water of equal volume then, leave standstill, coarse filtration, filtrate high speed centrifugation, centrifugal liquid molecular cut off are that 150000 polysulfone membrane is carried out ultrafiltration, the ultrafiltrate adjust pH is 2.0, last HPD-100 macroporous adsorptive resins absorption, with the water elution of 2～3 times of column volumes, reuse pH is 10.0 60% ethanol elution earlier, collect ethanol elution, concentrating under reduced pressure, vacuum drying promptly get effective part extract 17.64g.In rosmarinic acid, wherein contain total phenolic acid 61.87%.
Embodiment 6
Herba Pileae Scriptae medicinal material coarse powder 5kg, the ethanol that adds 10 times of amounts 70%, extract 2 times, each 1 hour, merge extractive liquid, reclaims ethanol, after fully dissolving with the water of equal volume then, leave standstill, coarse filtration, filtrate high speed centrifugation, centrifugal liquid molecular cut off are that 100000 cellulose acetate film carries out ultrafiltration, the ultrafiltrate adjust pH is 3.0, last HPD-400 macroporous adsorptive resins absorption, with the water elution of 2～3 times of column volumes, reuse pH is 9.0 50% ethanol elution earlier, collect ethanol elution, concentrating under reduced pressure, vacuum drying promptly get effective part extract.In rosmarinic acid 17.37g, wherein contain total phenolic acid 61.52%.
Embodiment 7
Herba Pileae Scriptae medicinal material coarse powder 5kg, the ethanol that adds 10 times of amounts 80%, extract 2 times, each 1 hour, merge extractive liquid, reclaims ethanol, after fully dissolving with the water of equal volume then, leave standstill, coarse filtration, filtrate high speed centrifugation, centrifugal liquid molecular cut off are that 20000 poly tetrafluoroethylene carries out ultrafiltration, the ultrafiltrate adjust pH is 4.0, last HPD-500 macroporous adsorptive resins absorption, with the water elution of 2～3 times of column volumes, reuse pH is 11.0 30% ethanol elution earlier, collect ethanol elution, concentrating under reduced pressure, vacuum drying promptly get effective part extract 18.75g.In rosmarinic acid, wherein contain total phenolic acid 60.77%.
Embodiment 8
Herba Pileae Scriptae medicinal material coarse powder 5kg, the ethanol that adds 10 times of amounts 90%, extract 2 times, each 1 hour, merge extractive liquid, reclaims ethanol, after fully dissolving with the water of equal volume then, leave standstill, coarse filtration, filtrate high speed centrifugation, centrifugal liquid molecular cut off are that 5000 polypropylene screen carries out ultrafiltration, the ultrafiltrate adjust pH is 5.0, last polyamide absorption, with the water elution of 2～3 times of column volumes, reuse pH is 8.0 40% ethanol elution earlier, collect ethanol elution, concentrating under reduced pressure, vacuum drying promptly get effective part extract 18.68g.In rosmarinic acid, wherein contain total phenolic acid 59.39%.
Embodiment 9 ultraviolet visible spectrophotometry are measured the content of total phenolic acid in the extractive of effective part of chloranthus glaber
(1) selection of maximum absorption wavelength λ max:
Rosmarinic acid (reference substance) is dissolved in methanol, and to be prepared into concentration be that the solution of 55 μ g/ml is as standard solution, getting effective part extract again is dissolved in methanol to be prepared into concentration is that the solution of 250 μ g/ml is as need testing solution, get each 0.5ml of standard solution and need testing solution respectively, be diluted to 5ml with dehydrated alcohol, add 2ml 0.3% sodium lauryl sulphate, add 1ml developer (0.6% ferric chloride: 0.9% potassium ferricyanide=1: 0.9) shake up again, 5min is placed in the dark place, add 0.1mol/L hydrochloric acid and be settled to 25ml, shake up, 20min is placed in the dark place.Then with blank solvent as blank, in the 420-900nm wave-length coverage, scan, obtain abosrption spectrogram separately.The result shows that reference substance and sample all have strong absorption at 750～760nm place, so select λ max=756nm.
(2) content assaying method of total phenolic acid
Rosmarinic acid 5.2mg is got in the preparation of standard substance, uses dissolve with methanol, is settled to 25ml, shakes up.Get wherein that 0.5ml is diluted to 5ml with dehydrated alcohol, add 2ml 0.3% sodium lauryl sulphate, add 1ml developer (0.6% ferric chloride: 0.9% potassium ferricyanide=1: 0.9) shake up again, 5min is left standstill in the dark place, add 0.1mol/L hydrochloric acid and be settled to 25ml, shake up, 20min is left standstill in the dark place.
Extractive of effective part of chloranthus glaber 0.04g is got in the preparation of test sample, uses water dissolution, is settled to 100ml; Get wherein that 0.5ml is diluted to 5ml with dehydrated alcohol, add 2ml 0.3% sodium lauryl sulphate, add 1ml developer (0.6% ferric chloride: 0.9% potassium ferricyanide=1: 0.9) shake up again, 5min is left standstill in the dark place, add 0.1mol/L hydrochloric acid and be settled to 25ml, shake up, 20min is left standstill in the dark place.
Get above-mentioned standard substance and test sample respectively and measure its absorbance at the 756nm place, parallel assay 3 times is averaged, and calculates the content of total phenolic acid.
The preparation of embodiment 10 tablets
Get effective part extract 100g of the present invention (embodiment 1 sample, down together), starch 80g, dextrin 5g mix homogeneously, add 10% starch slurry system soft material, granulate 60～70 ℃ of aeration-dryings, 16 mesh sieve granulate with 14 order nylon screens, add magnesium stearate 1.5g, carboxymethyl starch sodium 5g mixing is pressed into 1000, and coating promptly.
Embodiment 11 capsular preparations
Get effective part extract 100g of the present invention, starch 78g, magnesium stearate 2g mixing directly is filled to 1000 with Autocapsulefillingmachine, and polishing is promptly.
The preparation of embodiment 12 drop pill
Get effective part extract 10g of the present invention, drop in the polyethylene glycol 6000 of 32g heating and melting, be stirred to dissolving, be transferred in the reservoir, airtight and insulation is regulated drop pill machine drop quantitative valve at 80～90 ℃, splash into from top to bottom in 10～15 ℃ the liquid paraffin, make 1000 altogether, the drop pill that forms is drained and the erasing liquor paraffin body, be drying to obtain.
The preparation of embodiment 13 oral liquids
Get effective part extract 20g of the present invention, mix with Mel 300g, sucrose 50g, sodium benzoate 2g and distilled water 300ml, be heated to 85～90 ℃, stir and make dissolving, insulation 30min filters, and the filtrate thin up stirs evenly to 1000ml, embedding, and sterilization is promptly.
The preparation of embodiment 14 granules
Get effective part extract 10g of the present invention, dextrin 20g, sucrose 100g and ethanol are an amount of, and mixing is crossed 10 mesh sieves and made granule, in 60～70 ℃ of dryings, and granulate, packing is promptly.
The preparation of embodiment 15 injections
Get effective part extract 100g of the present invention, add the injection water and make dissolving in right amount, 0.02% the active carbon that adds amount of preparation stirs 5～10min, filters, filtrate is diluted to about 10 liters, adds sodium chloride adjusting osmotic pressure and oozes to waiting, and regulates pH7.5～8.0, ultrafiltration, embedding, sterilization is promptly.
Claims (9)
1. extractive of effective part of chloranthus glaber is characterized in that: this extract is the phenolic acid effective kind part that therefrom extracts in the medicinal herbs Corallium Japonicum Kishinouye, and contained phenolic acid compound comprises rosmarinic acid, protocatechuic acid, isofraxidin, caffeic acid in the described effective site.
2. extractive of effective part of chloranthus glaber according to claim 1 is characterized in that: phenolic acid compound total content 〉=50% wherein.
3. the preparation method of extractive of effective part of chloranthus glaber according to claim 1 and 2, it is characterized in that: this method comprises the following steps:
(1), Herba Pileae Scriptae water or ethanol extraction, the extracting solution high speed centrifugation, centrifugal liquid is carried out ultrafiltration with organic hybrid films;
(2), the ultrafiltrate adjust pH is 1～7, last resin column absorption, first water eluting, reuse pH are 8～14 10%～70% ethanol elution;
(3), collect ethanol elution, concentrate, be drying to obtain effective part extract.
4. the preparation method of extractive of effective part of chloranthus glaber according to claim 3, it is characterized in that: the used organic hybrid films of ultrafiltration is selected from polysulfones, acetate fiber, politef or polypropylene type, and its molecular cut off is 5000～200000.
5. the preparation method of extractive of effective part of chloranthus glaber according to claim 3 is characterized in that described resin is selected from macroporous resin or polyamide, and wherein macroporous adsorbent resin is selected from HPD-100, HPD-400, HPD-500 or AB-8.
6. pharmaceutical composition or preparation that Herba Pileae Scriptae phenolic acid effective kind part according to claim 1 and pharmaceutically acceptable pharmaceutic adjuvant are formed.
7. preparation according to claim 6 comprises injection, tablet, capsule, granule, drop pill, oral liquid, ointment or gel.
8. the application of Herba Pileae Scriptae phenolic acid effective kind part extract according to claim 1 in the preparation antibacterial-anti-inflammatory drug.
9. the application of Herba Pileae Scriptae phenolic acid effective kind part extract according to claim 1 in the adjuvant therapy medicaments of preparation cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2009101149065A CN101574381A (en) | 2009-02-02 | 2009-02-02 | Extractive of effective part of chloranthus glaber and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2009101149065A CN101574381A (en) | 2009-02-02 | 2009-02-02 | Extractive of effective part of chloranthus glaber and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101574381A true CN101574381A (en) | 2009-11-11 |
Family
ID=41269542
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2009101149065A Pending CN101574381A (en) | 2009-02-02 | 2009-02-02 | Extractive of effective part of chloranthus glaber and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101574381A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102727542A (en) * | 2012-07-17 | 2012-10-17 | 福建中医药大学 | Ointment with function of relieving swelling and pain |
| CN102772456A (en) * | 2012-08-13 | 2012-11-14 | 桂林百里香生物科技有限公司 | Manufacturing method of sarcandra glabra extraction extractum |
| CN106967029A (en) * | 2016-01-14 | 2017-07-21 | 广东省中医院 | One kind prepares isofraxidin method from Chinese medicinal material of sarcandra glaber |
-
2009
- 2009-02-02 CN CNA2009101149065A patent/CN101574381A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102727542A (en) * | 2012-07-17 | 2012-10-17 | 福建中医药大学 | Ointment with function of relieving swelling and pain |
| CN102772456A (en) * | 2012-08-13 | 2012-11-14 | 桂林百里香生物科技有限公司 | Manufacturing method of sarcandra glabra extraction extractum |
| CN102772456B (en) * | 2012-08-13 | 2014-06-11 | 南宁智阳科技咨询管理有限公司 | Manufacturing method of sarcandra glabra extraction extractum |
| CN106967029A (en) * | 2016-01-14 | 2017-07-21 | 广东省中医院 | One kind prepares isofraxidin method from Chinese medicinal material of sarcandra glaber |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101732417B (en) | Preparation method and application of ion pair mixture of macleaya cordata total alkaloid | |
| CN100545164C (en) | The preparation technology of disodium cantharidinate | |
| CN1872106A (en) | Application of wild basil circle leaves in treating disease of virulence cold | |
| CN102805762A (en) | Chinese magnoliavine fruit polysaccharide extract and preparation method and application thereof | |
| CN1723981A (en) | Novel use of extractive of Momordica grosvenori as adjuvant drug for preparing medicine | |
| CN114832022B (en) | Preparation of Phellinus linteus fruiting body phenol active substances and application thereof in regulating intestinal flora and uric acid metabolism | |
| CN101664446A (en) | Relinging extractum and preparation method and application | |
| JP2010528063A (en) | Method and use for obtaining an extract containing sequoyitol from a plant belonging to the genus Rhododendron, soybean, genus Ginkgo | |
| CN101574381A (en) | Extractive of effective part of chloranthus glaber and preparation method thereof | |
| CN101849950A (en) | Application of rotundic acid in preparing blood lipid regulating medicines | |
| CN102988529B (en) | Preparation method and novel application of total phenolic acid in cherry stones | |
| CN1947747B (en) | Traditional Chinese medicine composition containing luteolin and capsule of sweeping forsythia and its preparation method and use | |
| CN1233378C (en) | Chinese medicine composition for treating oral cavity and throat diseases and its preparing method | |
| CN104910291B (en) | A kind of jackfruit leaf polyose and its preparation method and application | |
| CN1772089B (en) | Freeze dried oldenlandia powder for injection and its preparation method | |
| CN101606977B (en) | Preparation method and application of clove leaf total iridoid glycoside | |
| CN101125163A (en) | Plant extract and preparation with antitumor efficacy | |
| CN1165545C (en) | Method for separating and extracting total flavone from goldenrain tree plant and its application | |
| CN107260810A (en) | The Preparation method and use of high-purity clove leaf total iridoid glycoside extract and its preparation | |
| CN102240314B (en) | Medicine for treating urinary tract infection | |
| CN102008534B (en) | Antitubercular pharmaceutical composition containing balloonflower root extract | |
| CN102093380B (en) | Cyclic icaritin aglycon as well as preparation method and application of cyclic icaritin aglycon | |
| CN103880678A (en) | Benzoic acid derivative as well as preparation and hypoglycemic application thereof | |
| CN101633661B (en) | Process for preparing sodium cantharidinate | |
| CN101099756B (en) | Anti-tumor traditional Chinese medicinal composition and preparation method and medicinal preparation thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20091111 |