CN104257768A - Application of total anthraquinone with various ingredients in stable and uniform proportion and composition of total anthraquinone to treatment of pancreatitis - Google Patents
Application of total anthraquinone with various ingredients in stable and uniform proportion and composition of total anthraquinone to treatment of pancreatitis Download PDFInfo
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Abstract
The invention relates to an application of total anthraquinone with various ingredients in a stable and uniform proportion and a composition of the total anthraquinone to treatment of pancreatitis. The total anthraquinone comprises free anthraquinone, combined anthraquinone, anthrone, sugar, tannin or a mixture of the materials.
Description
Technical field:
The present invention relates to a kind of therapeutic use of Chinese medicine extract preparation, particularly a kind of total Radix Et Rhizoma Rhei anthraquinone of various component ratio stable uniform and compositions thereof are used for the treatment of pancreatitis.
Background technology:
Radix Et Rhizoma Rhei is one of conventional Chinese medicine, is the dry root welding technology of Polygonaceae herbaceos perennial sorrel, Rheum tanguticum or Rheum officinale.
Radix Et Rhizoma Rhei bitter in the mouth, cold in nature, have and rush down lower attack, clearing away heat-fire, removing pathogenic heat from blood and toxic substance from the body, effect of eliminating blood stasis and inducing menstruation.Modern chemistry research proves that the main chemical compositions of Radix Et Rhizoma Rhei is anthraquinone analog compound; The extraction separation method of effective rhubarb component mainly contains: decocting method (water extraction): decocting method is the traditional extraction process of effective rhubarb component, because the polarity of dissociated anthraquinone class is little, thus not good to the extraction effect of dissociated anthraquinone constituents with decocting method; And its combined anthraquinone glycoside polarity is comparatively large compared with its aglycon, therefore available water is extracted.Alcohol extracting method: because the polar contribution of active component in Radix Et Rhizoma Rhei is very wide, therefore with regard to the extraction of general anthraquinone class, the efficiency of alcohol extracting method will apparently higher than decocting method.Alcohol extracting method comparatively conventional at present has ethanol refluxing process and percolation.
Radix Et Rhizoma Rhei complex chemical composition, chemical constitution is by least existing 136 kinds that illustrate, but its main component is anthraquinone analog compound, total content is 2% ~ 5%, wherein free hydroxyanthraquinone compounds only accounts for 1/10 ~ 1/5, being mainly chrysophanol, emodin, aloe-emodin, physcione and chrysophanic acid etc., is the main antimicrobial component of Radix Et Rhizoma Rhei.Associativity anthraquinone derivative is glucoside (i.e. chrysophanol monoglucoside, emodin monoglucoside, aloe-emodin monoglucoside, Rhein Glucoside etc.) and the dianthracene ketoside class of dissociated anthraquinone, be Radix Et Rhizoma Rhei mainly rush down lower composition, wherein dianthracene ketoside is: Sennoside A, B, C, D, E etc.The isomer each other of Sennoside A and sennoside B; Sennoside C and sennoside D isomer each other.In addition the two glucosides still containing emodin, aloe-emodin and chrysophanol.Radix Et Rhizoma Rhei is also containing tannins about 5%, and wherein having galloyl glucose, gallic acid, d-catechin and tetrarin etc., is astringing to arrest bleeding effective ingredient.
The drug effect of Radix Et Rhizoma Rhei and total Radix Et Rhizoma Rhei anthraquinone has report in many documents, and main effect has:
(1) furuncle and phyma and stomatitis, (2) leg ulcer (ecthyma) under, (3) intestinal tympanites, (4) infectious eczematoid dermatitis, (5) (6) acute gastroduodenal of coughing is hemorrhage, (7) chronic renal insufficiency, (8) postoperative hemorrhage (9) of having tooth pulled out is burnt, (10) acute necrotizing enteritis causes enteroparalysis, (11) biliary tract ascarid, (12) acute icterohepatitis, (13) the stomach cancer hemorrhage, (14) acute pancreatitis, (15) tonsillitis (acute suppurative tonsillitis), (lS) mumps, (17) neonate is not sucked the breast, (18) chronic prostatitis, (19) epistaxis, (20) chronic constipation, (21) infantile anorexia, (22) hepatic encephalopathy, (23) Acute Cerebral Hemorrhage, (24) hemobilia, (25) brain traumatic intracranial hemorrhage, (26) hyperlipemia, (27) chronic renal failure, (28) acute cholecystitis, (29) acute ileus, (30) abscess of appendix, (31) tension vesicle, (32) retroperitoneal hematoma, (33) herpes zoster, (34) ovulation function imbalance, (35) acute gonorrhea, (36) psoriasis, (37) acute soft tissue injury, (38) paronychia, (39) lymphoid tuberculosis, (40) old simple obesity, (41) Apoplectic Constipation, (42) difficulty in urination, (43) Complicated by Extensive Burn disease is controlled in advance, (44) hepatitis gravis, (45) epidemic hemorrhagic fever digestive tract hemorrhage.
In a word, Radix Et Rhizoma Rhei and anthraquinones extract thereof have diversified drug effect, are one of Chinese medicine that Recent study is maximum.
The medicine prepared with Radix Et Rhizoma Rhei has gone on the market for many years, and existing rhubarb preparation has xinqingning tablet and new clear Yiganning capsule, and its main component is Radix Et Rhizoma Rhei.There is heat-clearing and toxic substances removing.Blood circulation promoting and blood stasis dispelling, laxation effect.For interior solid heat, larynx swells, toothache, conjunctival congestion, constipation, dysentery, infective inflammation, the cards such as fever.
Main active because of Radix Et Rhizoma Rhei is total Radix Et Rhizoma Rhei anthraquinone, therefore there is report using total Radix Et Rhizoma Rhei anthraquinone as ingredient and corresponding extraction separation method more, total Radix Et Rhizoma Rhei anthraquinone extracts the rhizome from Radix Et Rhizoma Rhei, its color shape is brownish black extractum or brown powder crystallization, is the mixture such as dissociated anthraquinone, combined anthraquinone, anthrone, sugar, tannin.
The total Radix Et Rhizoma Rhei anthraquinone preparation gone on the market has total Radix Et Rhizoma Rhei anthraquinone capsule, and its main component is Radix Et Rhizoma Rhei extract.[function cures mainly] is used for the treatment of damp-heat jaundice hepatitis.
Chinese Patent Application No.: 200810107381.8 applyings date: 2008-11-13 discloses a kind of preparation method for the treatment of damp-heat jaundice hepatitis total Radix Et Rhizoma Rhei anthraquinone capsule, wherein the Main Ingredients and Appearance of total Radix Et Rhizoma Rhei anthraquinone capsule is Radix Et Rhizoma Rhei active ingredient compositions total Radix Et Rhizoma Rhei anthraquinone, and press total Radix Et Rhizoma Rhei anthraquinone 18%-22%, the proportioning of starch 78%-82% is prepared
The indication of the present inventor to total Radix Et Rhizoma Rhei anthraquinone preparation is studied, and finds unexpectedly, and total Radix Et Rhizoma Rhei anthraquinone preparation of the present invention may be used for pancreatitic treatment, and achieves gratifying effect.
Summary of the invention:
The invention provides a kind of new treatment use of total Radix Et Rhizoma Rhei anthraquinone preparation, total Radix Et Rhizoma Rhei anthraquinone of the present invention comprises Radix Et Rhizoma Rhei anthraquinone material prepared by any one method, comprises dissociated anthraquinone, combined anthraquinone, anthrone, sugar, tannin etc. or their mixture.The total Radix Et Rhizoma Rhei anthraquinone mentioned in following Chinese patent, Radix Et Rhizoma Rhei anthraquinone or Radix Et Rhizoma Rhei anthraquinone analog derivative:
Total Radix Et Rhizoma Rhei anthraquinone of the present invention, most preferably adopts total Radix Et Rhizoma Rhei anthraquinone prepared by following methods: its preparation method is as follows:
Step 1,
Get Radix Et Rhizoma Rhei 1000g, according to the percolation under Chinese Pharmacopoeia version in 2010 annex I0 fluid extract and extractum item, with ethanol as solvent, flood after 48 hours, with the speed slowly percolation of per kilogram medicated powder 1 ~ 1.5ml per minute, collect the liquid 5000ml that just filters for subsequent use; Continue percolation, collect continuous liquid 10000ml decompression recycling ethanol to 60 DEG C relative density 0.90 ~ 0.9 of filtering, let cool, leave standstill, filter, obtain precipitate I, filtrate l is for subsequent use;
Step 2,
Get liquid of just filtering and add 50% ethanol dilution to alcohol content 75%, alkalize to liquid of filtering without gallic acid speckle with 65% alcoholic solution of 10% sodium hydroxide, leave standstill 12 hours, get supernatant salt acid for adjusting pH value to 6.0 ~ 7.0, decompression recycling ethanol to 60 DEG C relative density 1.00 ~ 1.02, lets cool, leave standstill, filter, obtain precipitate l and filtrate 2
Step 3,
Filtrate 2 and filtrate l merge, be evaporated to 80 DEG C of relative densities 1.40 ~ 1.45, add the alcohol reflux 2 times of 6 times of thick paste weight, each 0.5 hour, filter, filtrate is alkalized to solution without gallic acid speckle with 65% alcoholic solution of 10% sodium hydroxide, leave standstill 12 hours, filter, filtrate adjust ph to 6.0 ~ 7.0, decompression recycling ethanol, being concentrated into 80 DEG C of relative densities is 1.45 ~ 1.48, and merge with above-mentioned precipitate 1, weighed weight, add 10 times amount 0.5% sodium hydrate aqueous solutions to dissolve, concentrated hydrochloric acid adjust ph to 6.0 ~ 7.0, add the gelatin of 0.2 times of consolidated material weight, heating, being concentrated into 80 DEG C of relative densities is 1.40 ~ 1.45, slowly add 4 times amount ethanol while hot, heated and stirred is separated out to gelatin, filter, collecting precipitation and filtrate, precipitation adds proper amount of boiling water, heating makes dissolving, 80 DEG C of relative densities are made to be 1.40 ~ 1.45, slowly add 4 times amount ethanol while hot, heated and stirred is separated out to gelatin, filter, collecting precipitation and filtrate, precipitate repetitive operation again 1 time, merge 3 filtrates, decompression recycling ethanol to 60 DEG C relative density 0.95 ~ 0.97, let cool, filter, collect precipitate II and filtrate 3,
Step 4,
Filtrate 3 checks that tannin is to negative, adds 4 times amount alcohol settling and eliminates residual gelatin, filters, reclaims ethanol, evaporate to dryness, 85 DEG C, 0.07MPa drying under reduced pressure, pulverizes, adds 6 times of dry substance w ethanol and reflux 2 times, each 0.5 hour, merge ethanol, decompression recycling ethanol, obtains residue
Step 5,
Residue and precipitate I, precipitate II merge, and mixing, less than 65 DEG C drying under reduced pressure, obtain total Radix Et Rhizoma Rhei anthraquinone of the present invention.
Total Radix Et Rhizoma Rhei anthraquinone of the present invention, its detection method is as follows:
Its [character] is brownish yellow powder, feeble QI, mildly bitter flavor.
Its [discriminating] method is as follows: get total Radix Et Rhizoma Rhei anthraquinone 0.1g of the present invention, adds chloroform 4Oml supersound process lO minute, and filter, filtrate is as need testing solution.Separately get aloe-emodin, chrysophanol, physcione, emodin, chrysophanic acid reference substance, add dissolve with methanol, make every 1ml respectively containing 0.2mg, 0.1mg, 0.1mg, 0.1mg, the solution solution in contrast of 0.2mg, test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VIB), draw above-mentioned two kinds of test samples and each 5 ~ 10 μ l of reference substance solution put respectively in same with Sodium Tvlose be adhesive silica gel H lamellae on (newly activating), the upper solution of placing with less than 10 DEG C, petroleum ether (30 ~ 60 DEG C) Ethyl formate one formic acid (15:5:1) is for developing solvent, launch below 25 DEG C, take out, dry, inspect under putting ultra-violet lamp (365nm).In test sample chromatograph, on the position corresponding to reference substance chromatograph, the fluorescence speckle of aobvious same color.
Its [inspection] method is as follows:
The inspection of gallic acid: get total Radix Et Rhizoma Rhei anthraquinone fine powder 25mg of the present invention, put in 5ml measuring bottle, adds methanol 4 ~ 5ml supersound process and makes dissolving in 30 minutes, and cooling, adds methanol dilution to scale, filters, gets subsequent filtrate, as need testing solution.Separately get gallic acid reference substance, add methanol and make the solution of every 1ml containing 0.1mg, product solution in contrast, according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VIB) test, draw each 5 μ l of above-mentioned two kinds of solution and put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-formic acid (5:4:1) for developing solvent, launch, take out, dry, spray with 1% ferric chloride alcoholic solution.In test sample chromatograph, on the position corresponding to reference substance chromatograph, the spot colors of appearance should be deeper than the color of reference substance speckle.
The inspection of ponticin: get total Radix Et Rhizoma Rhei anthraquinone 0.2g of the present invention, add methanol 2ml, warm macerating 10 minutes, lets cool, and gets supernatant 10ul point on filter paper, launch with 45% ethanol, take out, dry, place 10 minutes, inspect under putting ultra-violet lamp (365nm), lasting bright purple fluorescence must not be shown.
Its [assay] method is as follows:
(1), general anthraquinone
The preparation of reference substance solution
Get emodin reference substance and be about 10mg, accurately weighed, put in 100ml measuring bottle, add methanol dilution to scale, shake up, to obtain final product.
The preparation of standard curve: precision measures reference substance solution 0ml, 0.1ml, 0.2ml, 0.4ml, 0.8ml, 1.2ml, 1.6ml put in l0ml tool plug test tube, be evaporated to dry, residue respectively precision adds 0.8% magnesium acetate methanol solution l0ml and makes dissolving, shake up, with first part for blank, according to ultraviolet one visible spectrophotometry (Chinese Pharmacopoeia version in 2010 annex VA), absorbance is measured at the wavelength place of 510nm, take absorbance as vertical coordinate, concentration is abscissa, drawing standard curve.
Algoscopy: get this product and be about 40mg, accurately weighed, put in flask at the bottom of 100ml garden, add 8% hydrochloric acid solution 20ml, then add chloroform 20mL and to put in boiling water bath reflux 1 hour, cooling, put in separatory funnel, with a small amount of chloroform washing container to colourless, be incorporated in separatory funnel, divide and get chloroform layer, acid solution uses chloroform extraction 3 times (20ml, 15ml, 10ml) again, merge chloroform extraction liquid to put in 100ml measuring bottle, add chloroform and be diluted to scale, shake up.Precision measures 0.4ml and puts in 10ml tool plug test tube, be evaporated to dry, residue respectively precision adds 0.8% magnesium acetate methanol solution l0ml and makes dissolving, shakes up, with first part for blank, according to ultraviolet one visible spectrophotometry (Chinese Pharmacopoeia version in 2010 annex VA), measure absorbance at the wavelength place of 510nm, measure absorbance in accordance with the law, read the amount of general anthraquinone in test sample from standard curve, calculate, to obtain final product.
This product is pressed dry product and is calculated, containing general anthraquinone with emodin (C
l5h
loo
5meter, should be 50%-60%).
(2), dissociated anthraquinone
The preparation of reference substance solution
Get emodin reference substance and be about 10mg, accurately weighed, put in 100ml measuring bottle, add methanol dilution to scale, shake up, to obtain final product.
The preparation of standard curve: precision measures reference substance solution 0ml, 0.1ml, 0.2ml, 0.4ml, 0.8ml, 1.2ml, 1.6ml put in lOml tool plug test tube, be evaporated to dry, residue respectively precision adds 0.8% magnesium acetate methanol solution lOml and makes dissolving, shake up, with first part for blank, according to ultraviolet one visible spectrophotometry (Chinese Pharmacopoeia version in 2010 annex VA), absorbance is measured at the wavelength place of 510nm, take absorbance as vertical coordinate, concentration is abscissa, drawing standard curve.
Algoscopy: get this product and be about 40ml, accurately weighed, put in 100ml conical flask, add chloroform and be about 60ml supersound process (power 250W, frequency 25KHz) 40 minutes, let cool, filter, rinse filter and conical flask with chloroform, be incorporated in filtrate, filtrate is transferred in 100ml measuring bottle, adds chloroform and is diluted to scale, shake up.Precision measures 0.4ml and puts in lOml tool plug test tube, be evaporated to dry, residue respectively precision adds 0.8% magnesium acetate methanol solution lOml and makes dissolving, shakes up, with first part for blank, according to ultraviolet one visible spectrophotometry (Chinese Pharmacopoeia version in 2010 annex VA), measure absorbance at the wavelength place of 510nm, measure absorbance in accordance with the law, read the amount of dissociated anthraquinone in test sample from standard curve, calculate, to obtain final product.
This product is pressed dry product and is calculated, containing dissociated anthraquinone with emodin (C
l5h
loo
5) meter, should be 48%-58%.
(3), aloe-emodin, chrysophanic acid, emodin, chrysophanol, physcione total amount: measure according to high performance liquid chromatography (China is to allusion quotation version in 2010 annex VID).
Chromatographic condition and system suitability with 18 a heatable brick bed base silicon a heatable brick bed bonded silica gels for filler; With methanol-0.1% phosphoric acid solution (85:15) for mobile phase: determined wavelength is for 440nm.Number of theoretical plate calculates should be not less than 3000 by emodin peak.
Aloe-emodin is got in the preparation of reference substance solution, chrysophanic acid, emodin, chrysophanol, physcione reference substance are appropriate, accurately weighed, add methanol and make the mixed solution of every 1ml containing aloe-emodin, chrysophanic acid, each 10 μ g of emodin, physcione 20 μ g, chrysophanol 50 μ g, to obtain final product.
The preparation of need testing solution: get this product and be about 15mg, accurately weighed, put in 100ml measuring bottle, add methanol appropriate, supersound process (power 250W, frequency 25KHz) 30 minutes, make it dissolve, take out, let cool, add methanol dilution to scale, shake up, filter, precision measures subsequent filtrate 10ml (residual filtrate is for subsequent use), put in flask, fling to solvent, add 8% hydrochloric acid solution 20ml and add chloroform 20ml reflux 2 hours again, let cool, put in separatory funnel, with a small amount of chloroform washing container, be incorporated in separatory funnel, divide and get chloroform layer, acid solution extracts 3 times with chloroform jolting again, when each 10, merge chloroform liquid, decompression and solvent recovery is to dry, residue adds methanol to be made dissolving and is transferred in 10ml measuring bottle, add methanol to scale, shake up, filter, get subsequent filtrate, obtain.
Algoscopy is accurate respectively draws reference substance solution and each 20 μ l injection liquid chromatographies of need testing solution, measures, to obtain final product.
This product is pressed dry product and is calculated, containing aloe-emodin (C
15h
30o
5), chrysophanic acid (C
15h
8o
6), emodin (C
15h
lOo
5), chrysophanol (C
15h
lOo
4), physcione (C
16h
12o
5) total amount must not be less than 50%.
(4) free aloe-emodin, free chrysophanic acid, Free Emodin, free chrysophanol, Free Emodin methyl ether total amount measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VID).
Measure according to high performance liquid chromatography (China is to allusion quotation version in 2010 annex VID).
Chromatographic condition and system suitability with 18 a heatable brick bed base silicon a heatable brick bed bonded silica gels for filler; With methanol-0.1% phosphoric acid solution (85:15) for mobile phase: determined wavelength is for 440nm.Number of theoretical plate calculates should be not less than 3000 by emodin peak.
Aloe-emodin is got in the preparation of reference substance solution, chrysophanic acid, emodin, chrysophanol, physcione reference substance are appropriate, accurately weighed, add methanol and make the mixed solution of every 1ml containing aloe-emodin, chrysophanic acid, each 10 μ g of emodin, physcione 20 μ g, chrysophanol 50 μ g, to obtain final product.
The preparation of need testing solution: get this product and be about 15mg, accurately weighed, put in 100ml measuring bottle, add methanol appropriate, supersound process (power 250W, frequency 25 KHz) 30 minutes, makes it dissolve, take out, let cool, add methanol dilution to scale, shake up, filter, a part for filtrate is as need testing solution.
Algoscopy: accurate absorption reference substance solution and each 20 μ l injection liquid chromatographies of need testing solution respectively, measure, to obtain final product.
This product is pressed dry product and is calculated, containing free aloe-emodin (C
15h
30o
5), free chrysophanic acid (C
15h
8o
6), Free Emodin (C
15h
lOo
5), free chrysophanol (C
15h
lOo
4), Free Emodin methyl ether (C
16h
12o
5) total amount must not be less than 40%.
Total Radix Et Rhizoma Rhei anthraquinone of the present invention, wherein,
Containing general anthraquinone in emodin, be 50%-60%.
Containing dissociated anthraquinone in emodin, be 48%-58%.
Calculate, containing aloe-emodin (C by dry product
15h
30o
5), chrysophanic acid (C
15h
8o
6), emodin (C
15h
lOo
5), chrysophanol (C
15h
lOo
4), physcione (C
16h
12o
5) total amount be no less than 50%.
Calculate by dry product, containing free aloe-emodin (C
15h
30o
5), free chrysophanic acid (C
15h
8o
6), Free Emodin (C
15h
lOo
5), free chrysophanol (C
15h
lOo
4), Free Emodin methyl ether (C
16h
12o
5) total amount be no less than 40%.
Total Radix Et Rhizoma Rhei anthraquinone of the present invention can be prepared into tablet, the oral pharmaceutical forms such as capsule, also can be prepared into external and injectable drug form, the tablet prepared as the component containing following part by weight or capsule:
Preparation method is as follows:
1) total Radix Et Rhizoma Rhei anthraquinone, microcrystalline Cellulose, mix homogeneously;
2) by the dissolve with ethanol of polyvidone with 95% (v/v), the povidone solution (polyvidone of 10 parts of weight is dissolved into the polyvidone alcoholic solution obtaining 100 parts of volumes in the ethanol of 95%) containing 10% is prepared into;
3) povidone solution of 10% is added in said mixture stir, soft material processed, cross 18 mesh sieve granules, dry, add magnesium stearate after particle drying and mix, fill 1000 capsules.Or upper tablet machine presses 1000, then coating.
The present invention is unexpected in the process of research total Radix Et Rhizoma Rhei anthraquinone finds that it has therapeutical effect to pancreatitis, and core of the present invention is for this reason, provides a kind of use of approved drugs for nonapproved uses, namely prepares one with total Radix Et Rhizoma Rhei anthraquinone and is used for the treatment of pancreatitic medicine.
Below by way of experimental data, novelty teabag of the present invention is described.
Acute pancreatitis (AP) is one of clinical common emergency case.Its onset is anxious, and state of an illness weight, complication is many, and mortality rate is high.In recent years, Chinese medicine attempted the treatment participating in primary disease from too many levels, obtained remarkable progress.
Experiment 1, single oral gavage total Radix Et Rhizoma Rhei anthraquinone is to the protective effect of Severe Acute Pancreatitis SAP damage of intestines.
Method: adopt the ductus pancreaticus that drives in the wrong direction to inject 5% sodium taurocholate and prepare dog Model of Severe Acute Pancreatitis; animal is divided at random sham operated rats, model group, administration group; 6h administration treated animal gavage total Radix Et Rhizoma Rhei anthraquinone 20mgkg-1 after modeling; before operation consent, administration, after administration, different time points gets the change of serum amyloid enzymatic activity in hematometry peripheral blood, tumor necrosis factor-alpha (TNF-α) content and endotoxin content, and before administration, after administration, different time points gets diamine oxidase activity change in hematometry portal vein.
Result: administration group is compared with model group, after administration, 0 arrives 6h serum amyloid enzymatic activity without obvious reduction, in peripheral blood cachectin (TNF-α) level upon administration 5h significantly reduce (P < 0.05), in portal vein, diamine oxidase is active significantly reduces (P < 0.05), but in peripheral blood, endotoxin content significantly raises (P < 0.05) on the contrary.
Conclusion: total Radix Et Rhizoma Rhei anthraquinone, by alleviating body inflammatory reaction and enterocyte damage, improving intestinal microcirculation, plays the protective effect to Severe Acute Pancreatitis SAP damage of intestines.
Experiment 2, rhubarb anthraquinone material is at the distribution in vivo of acute pancreatitis in rats model
Method: employing normal SD rats (normal group, n=5), Magainin lumbar injection set up acute pancreatitic model rat (model group, n=5) as experimental subject.Total Radix Et Rhizoma Rhei anthraquinone (20gkg-1) gavage 2 groups of rats 1 time weekly, after 2 h
Result: emodin, aloe-emodin, chrysophanic acid and the chrysophanol concentration in acute pancreatitic model rat pancreas is higher than other histoorgans, and in pancreas, kidney, the in-house concentration of spleen apparently higher than normal group, therefore, in treatment pancreatitis, the oral total Radix Et Rhizoma Rhei anthraquinone that gives is effective method.
Detailed description of the invention:
Further illustrate the present invention by the following examples, but not as limitation of the present invention.
Embodiment 1
The preparation method of the total Radix Et Rhizoma Rhei anthraquinone capsule of invention is as follows:
1) total Radix Et Rhizoma Rhei anthraquinone, microcrystalline Cellulose, mix homogeneously;
2) by the dissolve with ethanol of polyvidone with 95% (v/v), the povidone solution (polyvidone of 10 parts of weight is dissolved into the polyvidone alcoholic solution obtaining 100 parts of volumes in the ethanol of 95%) containing 10% is prepared into;
3) povidone solution of 10% is added in said mixture stir, soft material processed, cross 18 mesh sieve granules, dry, add magnesium stearate after particle drying and mix, fill 1000 capsules.
Embodiment 2
Formula is
Preparation method is as follows:
1) total Radix Et Rhizoma Rhei anthraquinone, microcrystalline Cellulose, mix homogeneously;
2) by the dissolve with ethanol of polyvidone with 95% (v/v), the povidone solution (polyvidone of 10 parts of weight is dissolved into the polyvidone alcoholic solution obtaining 100 parts of volumes in the ethanol of 95%) containing 10% is prepared into;
3) povidone solution of 10% is added in said mixture stir, soft material processed, cross 18 mesh sieve granules, dry, add magnesium stearate after particle drying and mix, press 1000.
Embodiment 3
Formula is
Preparation method is as follows:
1) total Radix Et Rhizoma Rhei anthraquinone, microcrystalline Cellulose, mix homogeneously;
2) by the dissolve with ethanol of polyvidone with 95% (v/v), the povidone solution (polyvidone of 10 parts of weight is dissolved into the polyvidone alcoholic solution obtaining 100 parts of volumes in the ethanol of 95%) containing 10% is prepared into;
3) povidone solution of 10% is added in said mixture stir, soft material processed, cross 18 mesh sieve granules, dry, add magnesium stearate after particle drying and mix, fill 1000 capsules.
Claims (6)
1. total Radix Et Rhizoma Rhei anthraquinone and the application of compositions in the pancreatitic medicine of preparation treatment thereof.
2. application according to claim 1, wherein said total Radix Et Rhizoma Rhei anthraquinone comprises Radix Et Rhizoma Rhei anthraquinone material prepared by any one method, comprises dissociated anthraquinone, combined anthraquinone, anthrone, sugar, tannin etc. or their mixture.
3. application according to claim 1, wherein said total Radix Et Rhizoma Rhei anthraquinone is the total Radix Et Rhizoma Rhei anthraquinone adopting following methods to prepare:
Step 1,
Get Radix Et Rhizoma Rhei 1000g, according to the percolation under Chinese Pharmacopoeia version in 2010 annex I0 fluid extract and extractum item, with ethanol as solvent, flood after 48 hours, with the speed slowly percolation of per kilogram medicated powder 1 ~ 1.5ml per minute, collect the liquid 5000ml that just filters for subsequent use; Continue percolation, collect continuous liquid 10000ml decompression recycling ethanol to 60 DEG C relative density 0.90 ~ 0.9 of filtering, let cool, leave standstill, filter, obtain precipitate I, filtrate l is for subsequent use;
Step 2,
Get liquid of just filtering and add 50% ethanol dilution to alcohol content 75%, alkalize to liquid of filtering without gallic acid speckle with 65% alcoholic solution of 10% sodium hydroxide, leave standstill 12 hours, get supernatant salt acid for adjusting pH value to 6.0 ~ 7.0, decompression recycling ethanol to 60 DEG C relative density 1.00 ~ 1.02, lets cool, leave standstill, filter, obtain precipitate l and filtrate 2
Step 3,
Filtrate 2 and filtrate l merge, be evaporated to 80 DEG C of relative densities 1.40 ~ 1.45, add the alcohol reflux 2 times of 6 times of thick paste weight, each 0.5 hour, filter, filtrate is alkalized to solution without gallic acid speckle with 65% alcoholic solution of 10% sodium hydroxide, leave standstill 12 hours, filter, filtrate adjust ph to 6.0 ~ 7.0, decompression recycling ethanol, being concentrated into 80 DEG C of relative densities is 1.45 ~ 1.48, and merge with above-mentioned precipitate 1, weighed weight, add 10 times amount 0.5% sodium hydrate aqueous solutions to dissolve, concentrated hydrochloric acid adjust ph to 6.0 ~ 7.0, add the gelatin of 0.2 times of consolidated material weight, heating, being concentrated into 80 DEG C of relative densities is 1.40 ~ 1.45, slowly add 4 times amount ethanol while hot, heated and stirred is separated out to gelatin, filter, collecting precipitation and filtrate, precipitation adds proper amount of boiling water, heating makes dissolving, 80 DEG C of relative densities are made to be 1.40 ~ 1.45, slowly add 4 times amount ethanol while hot, heated and stirred is separated out to gelatin, filter, collecting precipitation and filtrate, precipitate repetitive operation again 1 time, merge 3 filtrates, decompression recycling ethanol to 60 DEG C relative density 0.95 ~ 0.97, let cool, filter, collect precipitate II and filtrate 3,
Step 4,
Filtrate 3 checks that tannin is to negative, adds 4 times amount alcohol settling and eliminates residual gelatin, filters, reclaims ethanol, evaporate to dryness, 85 DEG C, 0.07MPa drying under reduced pressure, pulverizes, adds 6 times of dry substance w ethanol and reflux 2 times, each 0.5 hour, merge ethanol, decompression recycling ethanol, obtains residue
Step 5,
Residue and precipitate I, precipitate II merge, and mixing, less than 65 DEG C drying under reduced pressure, to obtain final product.
4. application according to claim 3, wherein said total Radix Et Rhizoma Rhei anthraquinone contains general anthraquinone in emodin, for 50%-60%, containing dissociated anthraquinone in emodin, be 48%-58%, calculate by dry product, total amount containing aloe-emodin, chrysophanic acid, emodin, chrysophanol, physcione is no less than 50%, calculate by dry product, the total amount containing free aloe-emodin, free chrysophanic acid, Free Emodin, free chrysophanol, Free Emodin methyl ether is no less than 40%
Its [character] is brownish yellow powder, feeble QI, mildly bitter flavor,
Its [discriminating] method is as follows: get total Radix Et Rhizoma Rhei anthraquinone 0.1g of the present invention, add chloroform 4Oml supersound process lO minute, filter, filtrate is as need testing solution, separately get aloe-emodin, chrysophanol, physcione, emodin, chrysophanic acid reference substance, add dissolve with methanol, make every 1ml respectively containing 0.2mg, 0.1mg, 0.1mg, 0.1mg, the solution solution in contrast of 0.2mg, test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VIB), draw above-mentioned two kinds of test samples and each 5 ~ 10 μ l of reference substance solution put respectively in same with Sodium Tvlose be adhesive silica gel H lamellae on (newly activating), the upper solution of placing with less than 10 DEG C, petroleum ether (30 ~ 60 DEG C) Ethyl formate one formic acid (15:5:1) is for developing solvent, launch below 25 DEG C, take out, dry, inspect under putting ultra-violet lamp (365nm), in test sample chromatograph, on the position corresponding to reference substance chromatograph, the fluorescence speckle of aobvious same color,
Its [inspection] method is as follows:
The inspection of gallic acid: get total Radix Et Rhizoma Rhei anthraquinone fine powder 25mg of the present invention, put in 5ml measuring bottle, add methanol 4 ~ 5ml supersound process and make dissolving in 30 minutes, cooling, add methanol dilution to scale, filter, get subsequent filtrate, as need testing solution, separately get gallic acid reference substance, add methanol and make the solution of every 1ml containing 0.1mg, product solution in contrast, test according to thin layer chromatography (Chinese Pharmacopoeia version in 2010 annex VIB), draw each 5 μ l of above-mentioned two kinds of solution to put respectively on same silica gel g thin-layer plate, with toluene-ethyl acetate-formic acid (5:4:1) for developing solvent, launch, take out, dry, spray is with 1% ferric chloride alcoholic solution, in test sample chromatograph, on the position corresponding to reference substance chromatograph, the spot colors occurred should be deeper than the color of reference substance speckle, the inspection of ponticin: get total Radix Et Rhizoma Rhei anthraquinone 0.2g of the present invention, add methanol 2ml, warm macerating 10 minutes, let cool, get supernatant 10ul point on filter paper, launch with 45% ethanol, take out, dry, place 10 minutes, inspect under putting ultra-violet lamp (365nm), lasting bright purple fluorescence must not be shown,
Its [assay] method is as follows:
(1), general anthraquinone
The preparation of reference substance solution
Get emodin reference substance and be about 10mg, accurately weighed, put in 100ml measuring bottle, add methanol dilution to scale, shake up, to obtain final product,
The preparation of standard curve: precision measures reference substance solution 0ml, 0.1ml, 0.2ml, 0.4ml, 0.8ml, 1.2ml, 1.6ml put in l0ml tool plug test tube, be evaporated to dry, residue respectively precision adds 0.8% magnesium acetate methanol solution l0ml and makes dissolving, shake up, with first part for blank, according to ultraviolet one visible spectrophotometry (Chinese Pharmacopoeia version in 2010 annex VA), absorbance is measured at the wavelength place of 510nm, take absorbance as vertical coordinate, concentration is abscissa, drawing standard curve
Algoscopy: get this product and be about 40mg, accurately weighed, put in flask at the bottom of 100ml garden, add 8% hydrochloric acid solution 20ml, add chloroform 20mL again and to put in boiling water bath reflux 1 hour, cooling, put in separatory funnel, with a small amount of chloroform washing container to colourless, be incorporated in separatory funnel, divide and get chloroform layer, acid solution uses chloroform extraction 3 (20ml again, 15ml, 10ml), merging chloroform extraction liquid puts in 100ml measuring bottle, add chloroform and be diluted to scale, shake up, precision measures 0.4ml and puts in 10ml tool plug test tube, be evaporated to dry, residue respectively precision adds 0.8% magnesium acetate methanol solution l0ml and makes dissolving, shake up, with first part for blank, according to ultraviolet one visible spectrophotometry (Chinese Pharmacopoeia version in 2010 annex VA), absorbance is measured at the wavelength place of 510nm, measure absorbance in accordance with the law, the amount of general anthraquinone in test sample is read from standard curve, calculate, obtain,
(2), dissociated anthraquinone
The preparation of reference substance solution
Get emodin reference substance and be about 10mg, accurately weighed, put in 100ml measuring bottle, add methanol dilution to scale, shake up, to obtain final product,
The preparation of standard curve: precision measures reference substance solution 0ml, 0.1ml, 0.2ml, 0.4ml, 0.8ml, 1.2ml, 1.6ml put in lOml tool plug test tube, be evaporated to dry, residue respectively precision adds 0.8% magnesium acetate methanol solution lOml and makes dissolving, shake up, with first part for blank, according to ultraviolet one visible spectrophotometry (Chinese Pharmacopoeia version in 2010 annex VA), absorbance is measured at the wavelength place of 510nm, take absorbance as vertical coordinate, concentration is abscissa, drawing standard curve
Algoscopy: get this product and be about 40ml, accurately weighed, put in 100ml conical flask, add chloroform and be about 60ml supersound process (power 250W, frequency 25KHz) 40 minutes, let cool, filter, filter and conical flask is rinsed with chloroform, be incorporated in filtrate, filtrate is transferred in 100ml measuring bottle, add chloroform and be diluted to scale, shake up, precision measures 0.4ml and puts in lOml tool plug test tube, be evaporated to dry, residue respectively precision adds 0.8% magnesium acetate methanol solution lOml and makes dissolving, shake up, with first part for blank, according to ultraviolet one visible spectrophotometry (Chinese Pharmacopoeia version in 2010 annex VA), absorbance is measured at the wavelength place of 510nm, measure absorbance in accordance with the law, the amount of dissociated anthraquinone in test sample is read from standard curve, calculate, obtain,
(3), aloe-emodin, chrysophanic acid, emodin, chrysophanol, physcione total amount: measure according to high performance liquid chromatography (China to allusion quotation version in 2010 annex VID),
Chromatographic condition and system suitability with 18 a heatable brick bed base silicon a heatable brick bed bonded silica gels for filler; With methanol-0.1% phosphoric acid solution (85:15) for mobile phase: determined wavelength is for 440nm, and number of theoretical plate calculates should be not less than 3000 by emodin peak,
Aloe-emodin is got in the preparation of reference substance solution, chrysophanic acid, emodin, chrysophanol, physcione reference substance are appropriate, accurately weighed, add methanol and make the mixed solution of every 1ml containing aloe-emodin, chrysophanic acid, each 10 μ g of emodin, physcione 20 μ g, chrysophanol 50 μ g, obtain
The preparation of need testing solution: get this product and be about 15mg, accurately weighed, put in 100ml measuring bottle, add methanol appropriate, supersound process (power 250W, frequency 25KHz) 30 minutes, make it dissolve, take out, let cool, add methanol dilution to scale, shake up, filter, precision measures subsequent filtrate 10ml (residual filtrate is for subsequent use), put in flask, fling to solvent, add 8% hydrochloric acid solution 20ml and add chloroform 20ml reflux 2 hours again, let cool, put in separatory funnel, with a small amount of chloroform washing container, be incorporated in separatory funnel, divide and get chloroform layer, acid solution extracts 3 times with chloroform jolting again, when each 10, merge chloroform liquid, decompression and solvent recovery is to dry, residue adds methanol to be made dissolving and is transferred in 10ml measuring bottle, add methanol to scale, shake up, filter, get subsequent filtrate, obtain,
Algoscopy is accurate respectively draws reference substance solution and each 20 μ l injection liquid chromatographies of need testing solution, measures, to obtain final product,
(4) free aloe-emodin, free chrysophanic acid, Free Emodin, free chrysophanol, Free Emodin methyl ether total amount measure according to high performance liquid chromatography (Chinese Pharmacopoeia version in 2010 annex VID),
Measure according to high performance liquid chromatography (China is to allusion quotation version in 2010 annex VID),
Chromatographic condition and system suitability with 18 a heatable brick bed base silicon a heatable brick bed bonded silica gels for filler; With methanol-0.1% phosphoric acid solution (85:15) for mobile phase: determined wavelength is for 440nm, and number of theoretical plate calculates should be not less than 3000 by emodin peak,
Aloe-emodin is got in the preparation of reference substance solution, chrysophanic acid, emodin, chrysophanol, physcione reference substance are appropriate, accurately weighed, add methanol and make the mixed solution of every 1ml containing aloe-emodin, chrysophanic acid, each 10 μ g of emodin, physcione 20 μ g, chrysophanol 50 μ g, obtain
The preparation of need testing solution: get this product and be about 15mg, accurately weighed, put in 100ml measuring bottle, add methanol appropriate, supersound process (power 250W, frequency 25 KHz) 30 minutes, make it dissolve, take out, let cool, add methanol dilution to scale, shake up, filter, a part for filtrate is as need testing solution
Algoscopy: accurate absorption reference substance solution and each 20 μ l injection liquid chromatographies of need testing solution respectively, measure, to obtain final product.
5. application according to claim 1, wherein said total Radix Et Rhizoma Rhei anthraquinone compositions is the tablet of total Radix Et Rhizoma Rhei anthraquinone, capsule.
6. application according to claim 5, wherein said total Radix Et Rhizoma Rhei anthraquinone capsule, its formula is as follows:
Preparation method is as follows:
1) total Radix Et Rhizoma Rhei anthraquinone, microcrystalline Cellulose, mix homogeneously;
2) by the dissolve with ethanol of polyvidone with 95% (v/v), the povidone solution (polyvidone of 10 parts of weight is dissolved into the polyvidone alcoholic solution obtaining 100 parts of volumes in the ethanol of 95%) containing 10% is prepared into;
3) povidone solution of 10% is added in said mixture stir, soft material processed, cross 18 mesh sieve granules, dry, add magnesium stearate after particle drying and mix, fill 1000 capsules, or upper tablet machine presses 1000, then coating.
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| CN113424795A (en) * | 2021-05-24 | 2021-09-24 | 四川大学华西医院 | Construction method and application of acute pancreatitis animal model |
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