CN101422588A - Quality control method of medicine for treating gynecologic disease - Google Patents

Abstract

The invention relates to a quality control method of a plant Chinese herbal medicine compound preparation and the invention aims at providing a specific quality control method of medicaments treating gynecologic diseases. The quality control method consists of the following steps of: 1. identification: first, tanshinone IIA is identified by using thin-layer chromatography (TLC); and second, tetrahydropalmatine is identified by using TLC. 2. inspection: the relative density is not lower than 1.30(an appendix seven A of subdivision one of Chinese pharmacopoeia); and others accord with the various related regulations of soft extract terms (an appendix I F of subdivision one of Chinese pharmacopoeia). 3. content etermination; first, red paeonia contents are determined according to HPLC (an appendix VID A of subdivision one of Chinese pharmacopoeia 2000), the contents of the red paeonia in 1g of the preparation are measured according to paeoniflorin C23H28O11 and can not be less than 2.0mg; second, the contents of pseudo-ginseng are determined according to HPLC (an appendix VID of subdivision one of Chinese pharmacopoeia 2000), and the contents of pseudo-ginseng in 1g of the preparation are measured according to ginsenoside Rg1C42H72O14 and can not be less than 1.2mg.

Description

A kind of method of quality control for the treatment of medicine for gynecopathy

Technical field

The present invention relates to a kind of method of quality control of botanical herbs compound preparation.

Background technology

World Health Organization (WHO) shows Chinese women's investigation: the women of China 41% suffers from gynaecopathia in various degree, and sickness rate is ascendant trend year by year, gynecopathy therapeutic pharmaceutical market scale enlarges rapidly, the medicine of market expectation good effect, gynaecopathia that toxic and side effects is low, and method of quality control is the essential condition for preparing medicine.

Summary of the invention

The object of the present invention is to provide the method for quality control of the strong treatment medicine for gynecopathy of a kind of specificity.

The method of quality control of treatment medicine for gynecopathy of the present invention is made up of following steps:

One, differentiates

1, differentiates tanshinone with thin layer chromatography;

2, differentiate tetrahydropalmatine with thin layer chromatography;

Two, check

Relative density should be not less than 1.30 (" appendix VIIA of Chinese pharmacopoeia).

Other should meet every regulation relevant under the soft extract item (" an appendix I of Chinese pharmacopoeia F).

Three, assay

1, measures Radix Paeoniae Rubra content according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000).

2, measure Radix Notoginseng content according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000).

The pharmaceutical preparation of treatment gynaecopathia of the present invention is the oral formulations of being made by the following materials based on weight medicine: Radix Salviae Miltiorrhizae 450, Rhizoma Curcumae 250, HERBA BUPLEURI 250, Radix Notoginseng 150, Radix Paeoniae Rubra 250, Radix Angelicae Sinensis 250, rhizoma sparganic 175, Rhizoma Cyperi 150, Rhizoma Corydalis 175, Radix Glycyrrhizae 100.

Its method for making is: above ten flavor medical materials, percolation under Radix Salviae Miltiorrhizae, Rhizoma Curcumae, Radix Notoginseng, Radix Angelicae Sinensis, Rhizoma Corydalis photograph fluid extract and the extractum item (" an appendix I of Chinese pharmacopoeia O), make solvent with 70% ethanol, flood and carry out percolation after 24 hours, the collection liquid of filtering, decompression recycling ethanol also is concentrated into the thick paste that relative density is about 1.30 (50 ℃); The five tastes such as medicinal residues and all the other HERBA BUPLEURI decoct with water secondary, decoct 2 hours at every turn, and collecting decoction filters, and filtrate is concentrated into the thick paste that relative density is 1.30 (50 ℃); Merge above-mentioned two kinds of thick pastes, add Mel (refining) 100, sugar refinery 165, potassium sorbate 3 (being parts by weight), mixing promptly gets the soft extracts preparation.

The effect of medicine of the present invention is a blood circulation promoting and blood stasis dispelling, depressed liver-energy dispersing and QI regulating, menstruction regulating and pain relieving, softening the hard mass eliminate indigestion.Be used for menoxenia due to women's stagnation of blood stasis, dysmenorrhea, menstrual period, uncomfortable, WEIJIA was gathered, endometriosis of pelvis sees above-mentioned symptom person.

Pharmaceutical preparation of the present invention is tested, is gathered experiment, the model experiment of rat endometrium dystopy, its experiment conclusion at the experiment of body uterus, external rabbit platelet through isolated uterine:

1, the present invention external (1.5,3.0mg crude drug/mL) significantly strengthens isolated rat uterus maximum shrinkage amplitude (P＜0.05); (3.0mg crude drug/mL) significantly reduces basic tension force (P＜0.01), and is significantly resisting neurohypophyseal hormone effect (P＜0.05) aspect basic tension force and the maximum shrinkage amplitude.2, the present invention (13.3,26.6g crude drug/kg) is to being dual regulation at body rat uterus contraction frequency, significantly resisting neurohypophyseal hormone effect (P＜0.05) aspect shrinkage factor and the shrinkage amplitude simultaneously.3, the present invention (10g/L) assembles external rabbit platelet inhibitory action, and suppression ratio reaches 60%.4, the present invention (5.32g crude drug/kg.d) similar with DALAZUO to rat endometrium dystopy disease model curative effect, it is better that the early treatment organizes curative effect, its curative effect realizes that by two-ways regulation cell and humoral immune function more single immunostimulant of the mechanism of action or immunosuppressant are better.

The specific embodiment

The method of quality control of treatment medicine for gynecopathy of the present invention is made up of following steps:

One, differentiates

1, differentiates tanshinone with thin layer chromatography

Get this product 5g, add water 20ml, shake up, add ammonia solution and regulate pH value to 9～10, extract 3 times with the ether jolting, each 20ml merges ether solution, adds an amount of anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution.Other gets the tanshinone reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 0.5mg, in contrast product solution.According to thin layer chromatography (" appendix VIB of Chinese pharmacopoeia) test, draw each 5～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with toluene-ethyl acetate (9.5:0.5), launch, taking-up is dried.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

2, differentiate tetrahydropalmatine with thin layer chromatography

Get the tetrahydropalmatine reference substance, add dehydrated alcohol and make the solution that every 1ml contains 0.5mg, in contrast product solution.According to thin layer chromatography (" an appendix VI of Chinese pharmacopoeia B) test, draw need testing solution and each 5～10 μ l of above-mentioned reference substance solution under the item of [discriminating] (1) respectively, put respectively on same silica gel g thin-layer plate, with normal hexane-chloroform-methanol (6:3.2:0.8) is developing solvent, launch, take out, dry, with iodine vapor smoked several minutes, take out, put wave the iodine that adsorbs on the most plate in the air after, put under the ultra-violet lamp (365nm) and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.

Two, check

Relative density should be not less than 1.30 (" appendix VIIA of Chinese pharmacopoeia).

Other should meet every regulation relevant under the soft extract item (" an appendix I of Chinese pharmacopoeia F).

Three, assay

1, Radix Paeoniae Rubra is measured according to high performance liquid chromatography (" appendix VID of Chinese pharmacopoeia).

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Acetonitrile-water-glacial acetic acid (12:88:0.2) is a mobile phase; The detection wavelength is 230nm.Number of theoretical plate calculates by the peoniflorin peak should be not less than 2000.

It is an amount of that the preparation precision of reference substance solution takes by weighing the peoniflorin reference substance, adds methanol and make the solution that every 1ml contains 25 μ g, promptly.

This product 5g is got in the preparation of need testing solution, and accurate the title decides, and adds water 5ml, stirs evenly, be transferred in the 100ml measuring bottle, an amount of methanol wash of container, washing liquid is incorporated in the measuring bottle, adds methanol 80ml, shake up, supersound process 10 minutes is put cold, add methanol and be diluted to scale, shake up, filter, precision is measured subsequent filtrate 20ml, puts in the tool plug conical flask, the accurate 5% lead acetate solution 20ml that adds, shake up, left standstill about 30 minutes, filter.Precision is measured subsequent filtrate 10ml, and evaporate to dryness, residue add water makes dissolving in right amount, be added on neutral alumina post (100 ~ 200 orders, 5g, internal diameter 10～15mm, water 30ml prewashing) on, the water eluting is collected eluent 22ml, put in the 25ml measuring bottle, add 2% metabisulfite solution 2ml, add water to scale, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

The every 1g of this product contains Radix Paeoniae Rubra with peoniflorin (C ₂₃H ₂₈O ₁₁) meter, must not be less than 2.0mg.

2, Radix Notoginseng is measured according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia D).

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; With mobile phase A: second cyanogen, B: water, according to the form below carries out gradient elution; Flow velocity is per minute 1.0ml; The detection wavelength is 203nm.Number of theoretical plate is by the ginsenoside Rg ₁The peak calculates should be not less than 5000.

The preparation precision of reference substance solution takes by weighing the ginsenoside Rg ₁Reference substance is an amount of, adds methanol and makes the solution that every 1ml contains 0.5mg, promptly.

This product 5g is got in the preparation of need testing solution, and accurate the title decides, and puts in the 100ml tool plug conical flask, the accurate methanol 50ml that adds claims to decide weight, supersound process 20 minutes, standing over night, supersound process 10 minutes is put cold, claim again to decide weight, supply the weight that subtracts mistake, filter with methanol, precision is measured subsequent filtrate 25ml, volatilizes, and adds water 25ml and makes dissolving, be transferred in the separatory funnel, with chloroform washing 2 times, each 25ml, discard chloroform solution, water liquid extracts 5 (30ml with water saturated butanol solution jolting, 25ml, 15ml, 15ml, 15ml), merge n-butyl alcohol liquid, be concentrated into 25ml, quantitatively be transferred in the separatory funnel, with ammonia solution washing 2 times, each 25ml discards ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue adds methanol makes dissolving, is transferred in the 10ml measuring bottle, and adds methanol and be diluted to scale, shake up, filter, get subsequent filtrate, promptly.

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly.

The every 1g of this product contains Radix Notoginseng with the ginsenoside Rg ₁(C ₄₂H ₇₂O ₁₄) meter, must not be less than 1.2mg.

Claims (1)

1, a kind of method of quality control for the treatment of medicine for gynecopathy is characterized in that being made up of following steps:

One, differentiates

(1), differentiates tanshinone with thin layer chromatography

Get this product 5g, add water 20ml, shake up, add ammonia solution and regulate pH value to 9～10, extract 3 times with the ether jolting, each 20ml merges ether solution, adds an amount of anhydrous sodium sulfate dehydration, filter, filtrate evaporate to dryness, residue add dehydrated alcohol 1ml makes dissolving, as need testing solution; Other gets the tanshinone reference substance, adds dehydrated alcohol and makes the solution that every 1ml contains 0.5mg, in contrast product solution; According to the thin layer chromatography test, draw each 5～10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with toluene-ethyl acetate 9.5:0.5, launch, take out, dry; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

(2), differentiate tetrahydropalmatine with thin layer chromatography

Get the tetrahydropalmatine reference substance, add dehydrated alcohol and make the solution that every 1ml contains 0.5mg, in contrast product solution; According to the thin layer chromatography test, draw need testing solution and each 5～10 μ l of above-mentioned reference substance solution of differentiating under (1) item respectively, put respectively on same silica gel g thin-layer plate, with normal hexane-chloroform-methanol 6:3.2:0.8 is developing solvent, launches, and takes out, dry, with iodine vapor smoked several minutes, take out, put wave the iodine that adsorbs on the most plate in the air after, put under the ultra-violet lamp 365nm and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;

Two, check

Relative density should be not less than 1.30;

Other should meet in the Chinese Pharmacopoeia every regulation relevant under the soft extract item;

Three, assay

(1), Radix Paeoniae Rubra is according to high effective liquid chromatography for measuring

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Acetonitrile-water-glacial acetic acid 12:88:0.2 is a mobile phase; The detection wavelength is 230nm; Number of theoretical plate calculates by the peoniflorin peak should be not less than 2000;

It is an amount of that the preparation precision of reference substance solution takes by weighing the peoniflorin reference substance, adds methanol and make the solution that every 1ml contains 25 μ g, promptly;

This product 5g is got in the preparation of need testing solution, and accurate the title decides, and adds water 5ml, stirs evenly, be transferred in the 100ml measuring bottle, an amount of methanol wash of container, washing liquid is incorporated in the measuring bottle, adds methanol 80ml, shake up, supersound process 10 minutes is put cold, add methanol and be diluted to scale, shake up, filter, precision is measured subsequent filtrate 20ml, puts in the tool plug conical flask, the accurate 5% lead acetate solution 20ml that adds, shake up, left standstill about 30 minutes, filter; Precision is measured subsequent filtrate 10ml, and evaporate to dryness, residue add water makes dissolving in right amount, is added on the neutral alumina post, and the neutral alumina post is 100～200 orders, 5g, internal diameter 10～15mm, water 30ml prewashing; The water eluting is collected eluent 22ml, puts in the 25ml measuring bottle, adds 2% metabisulfite solution 2ml, adds water to scale, shakes up, and filters, and gets subsequent filtrate, promptly;

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly;

The every 1g of this product contains Radix Paeoniae Rubra with peoniflorin C ₂₃H ₂₈O ₁₁Meter must not be less than 2.0mg;

(2), Radix Notoginseng is according to high effective liquid chromatography for measuring;

Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; With mobile phase A: second cyanogen, B: water, according to the form below carries out gradient elution; Flow velocity is per minute 1.0ml; The detection wavelength is 203nm; Number of theoretical plate is by the ginsenoside Rg ₁The peak calculates should be not less than 5000;

The preparation precision of reference substance solution takes by weighing the ginsenoside Rg ₁Reference substance is an amount of, adds methanol and makes the solution that every 1ml contains 0.5mg, promptly;

This product 5g is got in the preparation of need testing solution, and accurate the title decides, and puts in the 100ml tool plug conical flask, the accurate methanol 50ml that adds claims to decide weight, supersound process 20 minutes, standing over night, supersound process 10 minutes is put cold, claim again to decide weight, supply the weight that subtracts mistake, filter with methanol, precision is measured subsequent filtrate 25ml, volatilizes, and adds water 25ml and makes dissolving, be transferred in the separatory funnel, with chloroform washing 2 times, each 25ml, discard chloroform solution, water liquid extracts 5 30ml with water saturated butanol solution jolting, 25ml, 15ml, 15ml, 15ml merges n-butyl alcohol liquid, be concentrated into 25ml, quantitatively be transferred in the separatory funnel, with ammonia solution washing 2 times, each 25ml discards ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue adds methanol makes dissolving, is transferred in the 10ml measuring bottle, and adds methanol and be diluted to scale, shake up, filter, get subsequent filtrate, promptly;

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, promptly;

The every 1g of this product contains Radix Notoginseng with the ginsenoside Rg ₁C ₄₂H ₇₂O ₁₄Meter must not be less than 1.2mg.