CN103163272A - Quality control method of infant spleen tonifying medicament - Google Patents
Quality control method of infant spleen tonifying medicament Download PDFInfo
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Abstract
The invention discloses a quality control method of an infant spleen tonifying medicament. According to the method, hawthorn and licorice in the medicament are identified by using layer chromatography respectively, and the hesperidin content of the medicament is measured by using liquid chromatography. In the established quality control method, the medicinal material identification method is mature, feasible, high in specificity, negative and non-interfering; the content measurement method is easy to operate, high in precision and high in reproducibility; and by adopting the quality control method, the quality of the medicament can be accurately and stably controlled to adapt to industrialized stable production of the medicament.
Description
Technical field
The present invention relates to a kind of Chinese medicine for infant spleen-tonifying, particularly relate to the method for quality control of this medicine.
Background technology
It is the medication of paediatrics class that children's joins the art Jianpi Wan, is used for the spleen and stomach in children weak, and indigestion is lean and haggard, lassitude.
Children's joins the existing operative norm of art Jianpi Wan and includes in the 5th of the Sanitation Ministry medicine standard Traditional Chinese medicine historical preparation.Formulated following content in this quality standard.
Get this product, put microscopically and observe: irregular branched agglomerate is colourless, meets chloral hydrate liquid and dissolves; Hyphae colorless or light brown, diameter 4-6 μ m.Connect latex dust diameter 12-15 μ m, contain fine particle shape thing.The pericarp lithocyte is single be dispersed in or minority in groups, lilac red, redness or yellowish-brown, similar round or polygon.Plant the long 80-150 μ of skin palisade cells m.Around fibrous bundle, parenchyma cell contains the calcium oxalate prismatic crystal, forms crystal fiber.
Check, should meet relevant every regulation under the pill item.
Above-mentioned quality standard exists more serious defective: at first, this standard is not carried out the quantitative measurement of active constituent content for main medicine; Secondly, the composition medicine more than 50% does not all carry out the chemistry discriminating.
Chinese medicine preparation steady quality, controlled and have that to produce repeatability be the premise that medicine has pharmacology and clinical effectiveness repeatability, can traditional Chinese medicine quality controlled and whether the traditional Chinese medicine quality standard is scientific, is the important restriction factor that affects industrialization of Chinese medicine.Quality controllability is the important indicator that medicine is estimated, and quality standard is the imbody of drug quality controllability.
Summary of the invention
The method of quality control that the purpose of this invention is to provide a kind of above-mentioned infant spleen-tonifying medicine is stablized controlled with the drug quality of guaranteeing to produce.
The method of quality control of infant spleen-tonifying medicine provided by the invention is suitable for the medicine that is prepared from by following bulk drug: Radix Codonopsis, stir-baked RHIZOMA ATRACTYLODIS MACROCEPHALAE with soil, Radix Glycyrrhizae (processed with honey), bran stir-baked SEMEN EURYALES, soil are fried that Semen Lablab Album, Rhizoma Dioscoreae (parched with bran), soil are fried Semen Nelumbinis, dried orange peel, fried hawthorn, bran is fried Medicated Leaven, are fried Fructus Hordei Germinatus, Poria cocos, soil are fried coix seed.Described method of quality control comprises discrimination method and content assaying method.
Wherein, described discrimination method comprises following discriminating:
1) get described medicine 10g, add zeyssatite 5g, grind well, the 40ml that adds diethyl ether, ultrasonic processing 15 minutes, filter, discard ether solution, the dregs of a decoction add ethyl acetate 40ml, and ultrasonic processing 15 minutes filters, the dregs of a decoction are standby, and filtrate evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution; Separately get hawthorn control medicinal material 1g, with the standby control medicinal material solution of legal system; Separately get the ursolic acid reference substance, add methyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution; According to the thin-layered chromatography test, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, reference substance solution 2 μ l put respectively on same silica gel g thin-layer plate, and take toluene: ethyl acetate: formic acid=20:4:0.5 is as developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, be heated to the spot colour developing at 105 ℃ clear, in the test sample chromatogram, with reference substance chromatogram corresponding position on, aobvious identical aubergine spot;
2) get 1) in residue after ethyl acetate extraction, fling to ethyl acetate, add methyl alcohol 30ml, add hot reflux 1 hour, and filtered the filtrate evaporate to dryness, residue adds water 40ml makes dissolving, extracts 3 times with water-saturated n-butanol, each 20ml, merge normal butyl alcohol liquid, with the saturated water washing of normal butyl alcohol 3 times, each 15ml, normal butyl alcohol liquid is recycled to dried, residue adds methyl alcohol 5ml makes dissolving, as need testing solution; Another extracting Radix Glycyrrhizae control medicinal material 1g adds methyl alcohol 20ml, is made in the same way of control medicinal material solution; According to the thin-layered chromatography test, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate, take methenyl choloride: acetone: formic acid=8:1:0.1 launches as developping agent, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with aluminum trichloride solution, put under the 365nm ultraviolet lamp and inspect, in the test sample chromatogram, with control medicinal material chromatogram corresponding position on, the fluorescence spot of aobvious same color.
Described content assaying method comprises the mensuration of content of hesperidin.
The present invention is specifically with the content of the described aurantiamarin of following high effective liquid chromatography for measuring:
1) chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent, take methyl alcohol: water=40:60 as mobile phase; Detect wavelength 283nm; Number of theoretical plate calculates by aurantiamarin and is not less than 3000;
2) preparation of reference substance solution: get the aurantiamarin reference substance appropriate, accurately weighed, add methyl alcohol and make the reference substance solution that every 1ml contains 50 μ g;
3) preparation of need testing solution: get described medicine, porphyrize, mixing is got 0.8g, accurately weighed, add zeyssatite 0.4g, grind well, put in tool plug conical flask, precision adds methyl alcohol 25ml, close plug, weighed weight, the ultrasonic processing of power 200W, frequency 40kHz 30 minutes lets cool, weighed weight is again supplied the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, and get final product;
4) draw respectively each 10 μ l of reference substance solution and need testing solution, the injection liquid chromatography is measured and get final product.
The present invention has set up the method for quality control of infant spleen-tonifying medicine, in the method for setting up, the discrimination method mature and feasible of medicinal material, specificity is strong, and is negative noiseless, the content assaying method processing ease, precision is high, favorable reproducibility uses method of quality control of the present invention can accurately, stably control the drug quality that children's joins the art Jianpi Wan, to adapt to the industrialization steady production of medicine.
Embodiment
Embodiment 1: the thin layer of hawthorn is differentiated.
Get children's and join art Jianpi Wan 10g, add zeyssatite 5g, grind well, the 40ml that adds diethyl ether, ultrasonic processing 15 minutes filters, and the dregs of a decoction add ethyl acetate 40ml, and ultrasonic processing 15 minutes filters, and filtrate evaporate to dryness, residue add methyl alcohol 2ml dissolving, as need testing solution; With the standby negative sample solution of legal system; Separately get hawthorn control medicinal material 1.0g, with the standby control medicinal material solution of legal system; Separately get the ursolic acid reference substance, add methyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution.According to thin-layered chromatography (" appendix VI B of Chinese pharmacopoeia version in 2010) test, draw negative sample solution 10 μ l, need testing solution 10 μ l, control medicinal material solution 5 μ l, reference substance solution 2 μ l put respectively on same silica gel g thin-layer plate, take toluene-ethyl acetate-formic acid (20:4:0.5) as developping agent, launch, take out, dry, spray the ethanol solution of sulfuric acid with 10%, be heated to the spot colour developing at 105 ℃ clear, put under daylight and inspect.
As a result, in the test sample chromatogram, with reference substance and control medicinal material chromatogram corresponding position on, the aubergine spot of aobvious same color on negative sample chromatogram corresponding position, is speckless, and illustrate that feminine gender is noiseless, specificity is strong.
Embodiment 2: the thin layer of Radix Glycyrrhizae is differentiated.
Get children's and join art Jianpi Wan 10g, add zeyssatite 5g, grind well, 40ml adds diethyl ether, ultrasonic processing 15 minutes filters, and the dregs of a decoction add ethyl acetate 40ml, ultrasonic processing 15 minutes, filter, the dregs of a decoction add methyl alcohol 30ml, add hot reflux 1 hour, filter, filtrate evaporate to dryness, residue add water 40ml makes dissolving, extracts 3 times with normal butyl alcohol, each 20ml merges normal butyl alcohol liquid, washes with water 3 times, evaporate to dryness, residue add methyl alcohol 5ml makes dissolving, as need testing solution; With the standby negative sample solution of legal system; Another extracting Radix Glycyrrhizae control medicinal material 1g is made in the same way of control medicinal material solution.According to thin-layered chromatography (" appendix VI B of Chinese Pharmacopoeia 2005 version) test, draw need testing solution 10 μ l, control medicinal material solution 6 μ l, negative sample solution 10 μ l, put respectively on same silica gel g thin-layer plate, take methenyl choloride-acetone-formic acid (8:1:0.1) as developping agent, launch, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with aluminum trichloride solution, puts under ultraviolet lamp (365nm) and inspects.
In the test sample chromatogram, with control medicinal material chromatogram corresponding position on, have three places corresponding, aobvious identical blueness, yellow green, green fluorescence spot respectively from top to bottom, negative noiseless, specificity is strong.
Embodiment 3: the mensuration of content of hesperidin.
Children's joins the art Jianpi Wan and is comprised of the 13 flavor Chinese medicines such as Radix Codonopsis, the bighead atractylodes rhizome, Radix Glycyrrhizae, Gorgon fruit, Semen Lablab Album, Chinese yam, Semen Nelumbinis, dried orange peel, and primary standard is without the assay item.For the better monitoring of controlling the quality of the pharmaceutical preparations and realizing technological process, the aurantiamarin in selected dried orange peel is as the assay index.
1, instrument and reagent.
Waters2695-2487 high efficiency liquid phase spectrometer, chromatographic work station (U.S. Waters company); Aurantiamarin reference substance (lot number 0721-200010, Nat'l Pharmaceutical ﹠ Biological Products Control Institute); Methyl alcohol: chromatographically pure (α Cygni friend fine chemicals company limited); Water: pure water; It is pure that zeyssatite and other reagent are analysis.
Children's joins the art Jianpi Wan.
2, chromatographic condition.
Chromatographic column: YMC-Pack ODS-A(4.6mm * 250mm); Mobile phase: methanol-water (40:60); Flow velocity: 1.0ml/min; Detect wavelength: 283nm; Column temperature: 35 ℃.Number of theoretical plate calculates by aurantiamarin and is not less than 3000.
3, the preparation of reference substance solution.
Get the aurantiamarin reference substance appropriate, accurately weighed, add methyl alcohol and make the reference substance solution that every 1ml contains 50 μ g.
4, the preparation of need testing solution.
Get children's and join art Jianpi Wan porphyrize, mixing is got 0.8g, accurately weighed, add zeyssatite 0.4g, grind well, put in tool plug conical flask, precision adds methyl alcohol 25ml, close plug, weighed weight, ultrasonic processing (power 200W, frequency 40kHz) 30 minutes lets cool, weighed weight is again supplied the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, and get final product.
5, the mensuration of content of hesperidin.
Precision is drawn reference substance solution and each 10 μ l of need testing solution respectively, and the injection liquid chromatography is measured, and be get final product.
6, negative sample test.
Join art Jianpi Wan preparation prescription ratio in children's, the negative control sample of dried orange peel is removed in the preparation of simulation process method for making.Precision takes negative control sample 3g, makes negative need testing solution by the need testing solution preparation method.Under chromatographic condition, record respectively the chromatogram of aurantiamarin reference substance, test sample and negative control product, the collection of illustrative plates result shows: with the corresponding section of aurantiamarin retention time, occur without chromatographic peak in negative control product chromatogram, namely under the condition determination of determining, in preparation prescription, other flavour of a drug are noiseless to the mensuration of aurantiamarin, and the method specificity is strong.
7, the investigation of linear relationship.
Get aurantiamarin reference substance storing solution (measuring bottle sealing, freezing preservation, concentration 0.1002mg/ml), put to room temperature, check whether less loss of solution, shake up, filter.Accurate reference substance solution 2,4,6,10,15, the 20 μ l of drawing, twice of each parallel sample introduction, measure peak area, take the average peak area integrated value as ordinate, aurantiamarin reference substance sample size is that horizontal ordinate calculates regression equation, and getting regression equation is y=1815.7x-14240, r=0.99995, show that aurantiamarin sample size and peak area in the 200.4-2004ng scope are good linear relationship, see Table 1.
8, precision test.
The accurate need testing solution of drawing, continuous sample introduction 6 times, each 10 μ l, measurement result such as table 2, RSD show that less than 3% precision is higher, and reliability is strong, and method is feasible.
9, stability test.
Get children's and join art Jianpi Wan 3g, accurately weighed, add zeyssatite 1.5g, grind well, to put in 50ml tool plug conical flask, precision adds methyl alcohol 25ml, close plug, weighed weight, ultrasonic processing 1 hour after letting cool, is supplied less loss weight with methyl alcohol, shakes up, and filters, and gets subsequent filtrate.Draw need testing solution 10 μ l 0,1,2,4,6,8 hour precision respectively, the injection liquid chromatography is measured, and the results are shown in Table 3, and need testing solution is stable in 8 hours, and the RSD value is less than 3%.
10, replica test.
Get same lot number sample (lot number: 081016) 0.8g, accurately weighed, extract by the need testing solution preparation method respectively, assay calculates.Average content is 1.561mg/ml as a result, and RSD is 1.63%, the results are shown in Table 4, shows that the method reappearance is good.
11, average recovery test.
Precision takes aurantiamarin reference substance 9.22mg, adds methyl alcohol and dissolves and be diluted to 100ml, shakes up, and filters, and makes reference substance solution, and every ml contains aurantiamarin 0.0922mg.
The accurate reference substance solution 6.5ml that draws puts in the 50ml conical flask, and evaporate to dryness is with standby 6 parts of legal system.Take 6 parts of the same batch samples of known content (1.561mg/g), every part of about 0.4g, accurately weighed, add zeyssatite 0.2g, grind well, be transferred in conical flask, precision adds methyl alcohol 25ml respectively, close plug, weighed weight, ultrasonic processing 30 minutes lets cool, and supplies the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, make need testing solution.
Accurate above-mentioned reference substance solution 5 μ l, need testing solution 10 μ l, injection liquid chromatography, the mensuration drawn.Calculate recovery rate, result such as table 5, the average recovery rate of aurantiamarin are that 97.34%, RSD is 1.27%.
Claims (2)
1. the method for quality control of an infant spleen-tonifying medicine, described infant spleen-tonifying medicine is the medicine that is prepared from by following bulk drug: Radix Codonopsis, stir-baked RHIZOMA ATRACTYLODIS MACROCEPHALAE with soil, Radix Glycyrrhizae (processed with honey), bran stir-baked SEMEN EURYALES, soil are fried that Semen Lablab Album, Rhizoma Dioscoreae (parched with bran), soil are fried Semen Nelumbinis, dried orange peel, fried hawthorn, bran is fried Medicated Leaven, are fried Fructus Hordei Germinatus, Poria cocos, soil are fried coix seed, it is characterized in that the discrimination method in described method of quality control comprises following discriminating:
1) get described medicine 10g, add zeyssatite 5g, grind well, the 40ml that adds diethyl ether, ultrasonic processing 15 minutes, filter, discard ether solution, the dregs of a decoction add ethyl acetate 40ml, and ultrasonic processing 15 minutes filters, the dregs of a decoction are standby, and filtrate evaporate to dryness, residue add methyl alcohol 2ml makes dissolving, as need testing solution; Separately get hawthorn control medicinal material 1g, with the standby control medicinal material solution of legal system; Separately get the ursolic acid reference substance, add methyl alcohol and make the solution that every 1ml contains 1mg, in contrast product solution; According to the thin-layered chromatography test, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, reference substance solution 2 μ l put respectively on same silica gel g thin-layer plate, and take toluene: ethyl acetate: formic acid=20:4:0.5 is as developping agent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, be heated to the spot colour developing at 105 ℃ clear, in the test sample chromatogram, with reference substance chromatogram corresponding position on, aobvious identical aubergine spot;
2) get 1) in residue after ethyl acetate extraction, fling to ethyl acetate, add methyl alcohol 30ml, add hot reflux 1 hour, and filtered the filtrate evaporate to dryness, residue adds water 40ml makes dissolving, extracts 3 times with water-saturated n-butanol, each 20ml, merge normal butyl alcohol liquid, with the saturated water washing of normal butyl alcohol 3 times, each 15ml, normal butyl alcohol liquid is recycled to dried, residue adds methyl alcohol 5ml makes dissolving, as need testing solution; Another extracting Radix Glycyrrhizae control medicinal material 1g adds methyl alcohol 20ml, is made in the same way of control medicinal material solution; According to the thin-layered chromatography test, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate, take methenyl choloride: acetone: formic acid=8:1:0.1 launches as developping agent, take out, dry, spray is heated to the spot colour developing at 105 ℃ clear with aluminum trichloride solution, put under the 365nm ultraviolet lamp and inspect, in the test sample chromatogram, with control medicinal material chromatogram corresponding position on, the fluorescence spot of aobvious same color.
2. the method for quality control of infant spleen-tonifying medicine according to claim 1 is characterized in that content assaying method in described method of quality control is the content with following high effective liquid chromatography for measuring aurantiamarin:
1) chromatographic condition and system suitability: take octadecylsilane chemically bonded silica as filling agent, take methyl alcohol: water=40:60 as mobile phase; Detect wavelength 283nm; Number of theoretical plate calculates by aurantiamarin and is not less than 3000;
2) preparation of reference substance solution: get the aurantiamarin reference substance appropriate, accurately weighed, add methyl alcohol and make the reference substance solution that every 1ml contains 50 μ g;
3) preparation of need testing solution: get described medicine, porphyrize, mixing is got 0.8g, accurately weighed, add zeyssatite 0.4g, grind well, put in tool plug conical flask, precision adds methyl alcohol 25ml, close plug, weighed weight, the ultrasonic processing of power 200W, frequency 40kHz 30 minutes lets cool, weighed weight is again supplied the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, and get final product;
4) draw respectively each 10 μ l of reference substance solution and need testing solution, the injection liquid chromatography is measured and get final product.
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| CN109342609B (en) * | 2018-12-21 | 2021-04-20 | 国药控股星鲨制药（厦门）有限公司 | Method for identifying chemical components of traditional Chinese medicine fluid extract of compound three-dimensional calcium D-pantothenate syrup |
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