CN102335333A - Method for detecting traditional Chinese medicine preparation of compound nutritions paste - Google Patents
Method for detecting traditional Chinese medicine preparation of compound nutritions paste Download PDFInfo
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- CN102335333A CN102335333A CN2010102341163A CN201010234116A CN102335333A CN 102335333 A CN102335333 A CN 102335333A CN 2010102341163 A CN2010102341163 A CN 2010102341163A CN 201010234116 A CN201010234116 A CN 201010234116A CN 102335333 A CN102335333 A CN 102335333A
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Abstract
The invention relates to a method for detecting a traditional Chinese medicine preparation of a compound nutritions paste. The preparation is prepared by processing 160g of Radix Codonopsis, 40g of prepared rehmannia root, 80g of polygonatum rhizome, 40g of wolfberry and 40g of fleece-flower root. The invention is characterized in that the detection method comprises an identification method and a content determination method.
Description
Invention field
The present invention relates to compound recipe nourishing power cream Chinese medicine preparation detection method, particularly the detection method of compound recipe nourishing power cream cream.
Background technology
Compound nourishing power cream cream records in the drug standards (WS promulgated by the ministries or commissions of the Central Government
3-B-2754-97), form by five kinds of Chinese medicine such as Radix Codonopsis, prepared rhizome of rehmannia, sealworts, be mainly used in insufficiency of vital energy and blood, to suffer from a deficiency of the kidney, muscle power is weak, soreness of the loins and weakness of the limbs, tinnitus dim eyesight.These article do not have discriminating, content assaying method in former ministry standard; Be the control product quality; Wherein main flavour of a drug Radix Codonopsis, Radix Rehmanniae Preparata have been carried out the thin layer chromatography discriminating, and in the Radix Rehmanniae Preparata in the compound recipe nourishing power cream cream that adopted high effective liquid chromatography for measuring the content of verbascoside with this quality control standard as product.
Summary of the invention
The object of the invention is in the detection method that a kind of compound recipe nourishing power paste formulation is provided.
The prescription of compound recipe nourishing power cream of the present invention:
Radix Codonopsis 160g Radix Rehmanniae Preparata 40g Rhizoma Polygonati 80g Fructus Lycii 40g Radix Polygoni Multiflori 40g
The method for preparing of compound recipe nourishing power cream of the present invention is:
The above five tastes, the decocte with water secondary, each 2 hours, collecting decoction, filtrating is condensed into thick paste.Other gets sucrose 600g and processes simple syrup, add sodium benzoate 3g stirring and dissolving after, add in the above-mentioned thick paste, mixing is processed 1000ml, filters, and promptly gets.
Detection method of the present invention comprises to be differentiated and content assaying method:
Wherein, discrimination method: comprise a kind of of following method and/or several kinds
A. get these article 3g, add water 50ml, measure 15ml, (internal diameter is 1.5cm through D101 type macroporous adsorptive resins; The post height is 15cm), water 100ml eluting is arranged, discard water liquid, reuse 50% ethanol 100ml eluting; Collect eluent, evaporate to dryness, residue add methanol 1ml makes dissolving; Get lobetyolin's reference substance in addition as need testing solution, add methanol and process the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography, draw need testing solution and each 2ul of reference substance solution and put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-glacial acetic acid-water (7; 1: 0.5) for developing solvent, launch, take out, to dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃, puts under the ultra-violet lamp (365nm) and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
B. get these article 2g, add water 40ml, measure 5ml, full each the n-butanol extraction jolting of water is extracted 4 times; Each 20ml merges n-butyl alcohol liquid, and evaporate to dryness, residue add methanol 2ml makes dissolving; As need testing solution, other gets the verbascoside reference substance, adds methanol alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography, draw need testing solution and each 5ul of reference substance solution and put respectively on same silica gel g thin-layer plate, be developing solvent with ethyl acetate-methanol formic acid (16: 0.5: 2), launch to take out and dry; With 0.1% 2, the bitter diazanyl ethanol solution dipping of 2-diphenyl-1-dries.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
Said content assaying method: according to HPLC (version appendix VI D in 2005) mensuration chromatographic condition and system suitability condition is filler with the octadecyl silane, and the glacial acetic acid of acetonitrile-0.1% (15: 85) is a mobile phase; The detection wavelength is 334nm; Number of theoretical plate calculates by the verbascoside peak should be not less than 5000
The preparation of reference substance solution: it is an amount of to get the verbascoside reference substance, and accurate the title decides, and adds mobile phase and processes the solution that every 1ml contains 20ug, promptly gets.
The preparation of need testing solution: get the about 1.0g of these article, the accurate title, decide, and puts in the tool plug conical flask, the accurate 50% methanol-methanol 50ml that adds; Claim decide weight, water-bath backflow-supersound process 10-70 minute is put coldly, and weight decided in title again; Add 50% methanol-methanol and supply weight, shake up, filter, precision is measured subsequent filtrate 2ml and is put in the 10ml volumetric flask; Add mobile phase to scale, shake up, filter, get subsequent filtrate and promptly get.The every g of these article contains Radix Rehmanniae Preparata must not be less than 0.06mg in verbascoside (C29H36O15).
Detection method of the present invention, precision is high, good reproducibility, good stability, the response rate is high, and reliable results has guaranteed the quality and the curative effect of product.
The specific embodiment
Below in conjunction with embodiment the present invention is described further, but not as restriction.
The research of experimental example 1, verbascoside content assaying method
(1) instrument and reagent
High performance liquid chromatograph comprises Tianjin, island LC-10AT pump, SPD-10AVP dual pathways UV-detector; Tianjin, island column oven; Zhejiang University's liquid chromatograph work station, verbascoside reference substance (lot number: 1530-200202 supplies assay usefulness, Nat'l Pharmaceutical & Biological Products Control Institute) acetonitrile is a U.S. Tedia chromatographically pure; Water is ultra-pure water, and other reagent is analytical pure.Drug sample to be measured is peace two appearance cream (lot number: 090101 Jiangxi Jimin Kexin Drug Industry Co., Ltd)
2) chromatographic condition is with reference to the Radix Rehmanniae Preparata assay
Chromatographic column: Penomenex Luna C18 (2) 4.6mm * 150mm column temperature: 30 ℃ of mobile phases: acetonitrile-0.1% acetum; The detection wavelength is 320nm; Flow velocity: 1.0ml/min
3) selection of detection wavelength
Get the verbascoside reference substance solution, in 200nm～400nm scope interscan, the result has absorption maximum at the 334nm place, so select 334nm as detecting wavelength.
4) extract choice of Solvent: respectively with 50% methanol, methanol as solvent, according to the operation down of assay item, (lot number: the content of verbascoside is measured 090101), and the result sees table 1 to sample.
Table 1 solvent is to measuring result's influence
Therefore, select for use methanol as solvent
5) extraction conditions selection: take by weighing testing sample respectively and carry out supersound process, water-bath reflow treatment, (lot number: the content of verbascoside 090101), the result sees table 2 to measure all article
Table 2 extraction conditions is to measuring result's influence
Therefore, select for use water-bath to reflux as extraction conditions.
6) selection of water-bath backflow: according to the operation down of assay item, the supersound process time was respectively 10,20,60,70 minutes, and (lot number is: the content of verbascoside 090101), the result sees table 3 to working sample
The table 3 supersound process time is to measuring result's influence
Therefore, the water-bath return time was selected for use 60 minutes.
7) blank assay
In the ratio of prescription taste of Chinese medicine, be furnished with the blank preparation that contains compound recipe nourishing power cream cream certainly, process blank solution as stated above, measure in accordance with the law, blank solution is not seen apparent chromatographic peak at the place of identical retention time with the verbascoside reference substance as a result, so think no interference.
8) linear relationship is investigated
Accurate above-mentioned reference substance solution (0.0216mg/ml) 2,4,6,8, the 10ul of drawing injects chromatograph of liquid, measures peak area, is horizontal vertical mark with the sample introduction (ug) of reference substance, is vertical coordinate with the peak area, the drawing standard curve.The result sees table 4
Table 4 linear relationship experimental data (n=5)
Y＝400000X+58596
R＝0.9996
The result shows: in 0.0432～0.216ug scope, be good linear relationship.
9) stability test
The accurate reference substance solution 10ul that draws, interval certain hour sample introduction, the peak area of mensuration verbascoside, BSD is: 183%, the result shows that the mensuration result is stable in 24 hours, the result sees table 5.
Table 5 stability test data (n=5)
10) precision test
The accurate need testing solution 10ul that draws injects chromatograph of liquid, measures the peak area of verbascoside, repeats sample introduction 5 times, and RSD is 0.69% as a result, sees table 6
Table 6 precision test data (n=5)
11) repeatability test
According to the operation down of assay item, (lot number: 090101) sample is measured for 5 parts, and trying to achieve relative standard deviation RSD is 0.28%, and the result sees table 7 to same lot number
Table 7 repeatability test data
12) recovery test
Reclaim with application of sample; Precision take by weighing known verbascoside content same lot number (lot number: the about 0.5g of test sample 0901010.076mg/g), it is a certain amount of that precision adds verbascoside reference substance solution (0.216mg/ml) respectively, according to the operation down of assay item; Be calculated as follows the response rate, the result sees table 8
Table 8 average recovery experimental data (n=6)
Average recovery rate is: 99.16% RSD is 0.52%
The assay of verbascoside in the experimental example 2 compound recipes nourishing power cream cream
According to the method operation of above-mentioned assay, surveyed the content of verbascoside in the 10 reply sides nourishing power cream cream, the result sees table 9
The assay of verbascoside in the table 9 compound recipe nourishing power cream cream
The content that records verbascoside in the compound recipe nourishing power cream paste formulation is all more than 0.88mg/g; Consider the loss error in the industry amplification; Therefore on the basis of minimum content; Float downward 20%, therefore obtaining examination criteria is to contain verbascoside content in the every gram of compound recipe nourishing power cream paste formulation should be not less than 0.06mg.
Embodiment 2, detection method
1, differentiates detection method
A. get these article 3g, add water 50ml, measure 15ml, (internal diameter is 1.5cm through D101 type macroporous adsorptive resins; The post height is 15cm), water 100ml eluting is arranged, discard water liquid, reuse 50% ethanol 100ml eluting; Collect eluent, evaporate to dryness, residue add methanol 1ml makes dissolving; Get lobetyolin's reference substance in addition as need testing solution, add methanol and process the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography, draw need testing solution and each 2ul of reference substance solution and put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-glacial acetic acid-water (7; 1: 0.5) for developing solvent, launch, take out, to dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃, puts under the ultra-violet lamp (365nm) and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
B. get these article 2g, add water 40ml, measure 5ml, full each the n-butanol extraction jolting of water is extracted 4 times; Each 20ml merges n-butyl alcohol liquid, and evaporate to dryness, residue add methanol 2ml makes dissolving; As need testing solution, other gets the verbascoside reference substance, adds methanol alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography, draw need testing solution and each 5ul of reference substance solution and put respectively on same silica gel g thin-layer plate, be developing solvent with ethyl acetate-methanol formic acid (16: 0.5: 2), launch to take out and dry; With 0.1% 2, the bitter diazanyl ethanol solution dipping of 2-diphenyl-1-dries.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
2, assay: measure according to HPLC (version appendix VID in 2005)
Chromatographic condition and system suitability condition are filler with the octadecyl silane, and the glacial acetic acid of acetonitrile-0.1% (15: 85) is a mobile phase; The detection wavelength is 334nm; Number of theoretical plate calculates by the verbascoside peak should be not less than 5000
The preparation of reference substance solution: it is an amount of to get the verbascoside reference substance, and accurate the title decides, and adds mobile phase and processes the solution that every 1ml contains 20ug, promptly gets.
The preparation of need testing solution: get the about 1.0g of these article, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 50ml that adds; Claim decide weight, water-bath refluxed 60 minutes, put coldly, and weight decided in title again; Add methanol and supply weight, shake up, filter, precision is measured subsequent filtrate 2ml and is put in the 10ml volumetric flask; Add mobile phase to scale, shake up, filter, get subsequent filtrate and promptly get.The every g of these article contains Radix Rehmanniae Preparata must not be less than 0.06mg in verbascoside (C29H36O15).
Claims (6)
1. the detection method of compound recipe nourishing power paste formulation, said preparation be by Radix Codonopsis 160g, Radix Rehmanniae Preparata 40g, and Rhizoma Polygonati 80g, Fructus Lycii 40g and Radix Polygoni Multiflori 40g are processed into, and it is characterized in that, and said detection method comprises discriminating and content assaying method.
2. detection method according to claim 1 is characterized in that, said discrimination method comprises:
Get preparation 3g, add water 50ml, measure 15ml, through D101 type macroporous adsorptive resins (, water 100ml eluting is arranged; Discard water liquid, reuse 50% ethanol 100ml eluting is collected eluent, evaporate to dryness; Residue adds methanol 1ml makes dissolving, gets lobetyolin's reference substance in addition as need testing solution, adds methanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography, draw need testing solution and each 2ul of reference substance solution and put respectively on same silica gel g thin-layer plate, be that 7: 1: 0.5 n-butyl alcohol-glacial acetic acid-water is developing solvent with volume ratio, launch; Take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; Put under the 365nm ultra-violet lamp and inspect, in the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
3. detection method according to claim 1 is characterized in that, said discrimination method comprises:
Get preparation 2g, add water 40ml, measure 5ml, full each the n-butanol extraction jolting of water is extracted 4 times; Each 20ml merges n-butyl alcohol liquid, and evaporate to dryness, residue add methanol 2ml makes dissolving; As need testing solution, other gets the verbascoside reference substance, adds methanol alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography, draw need testing solution and put respectively on same silica gel g thin-layer plate with each 5ul of reference substance solution, be that ethyl acetate-methanol formic acid of 16: 0.5: 2 is developing solvent with volume ratio, launch taking-up and dry; With 0.1% 2, the bitter diazanyl ethanol solution dipping of 2-diphenyl-1-dries; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
4. detection method according to claim 1 is characterized in that, said content assaying method may further comprise the steps:
According to version appendix VI D high effective liquid chromatography for measuring in 2005
Chromatographic condition and system suitability condition are filler with the octadecyl silane, are that the glacial acetic acid of 15: 85 acetonitrile-0.1% is a mobile phase with volume ratio; The detection wavelength is 334nm; Number of theoretical plate calculates by the verbascoside peak should be not less than 5000,
The preparation of reference substance solution: it is an amount of to get the verbascoside reference substance, and accurate the title decides, and adds mobile phase and processes the solution that every 1ml contains 20ug, promptly get,
The preparation of need testing solution: get preparation 1.0g, the accurate title, decide, and puts in the tool plug conical flask, the accurate 50% methanol-methanol 50ml that adds; Claim decide weight, water-bath backflow-supersound process 10-70 minute is put coldly, and weight decided in title again; Add 50% methanol-methanol and supply weight, shake up, filter, precision is measured subsequent filtrate 2ml and is put in the 10ml volumetric flask; Add mobile phase to scale, shake up, filter, get subsequent filtrate and promptly get.The every g of these article contains Radix Rehmanniae Preparata must not be less than 0.06mg in verbascoside (C29H36O15).
5. detection method according to claim 1 is characterized in that, said detection method may further comprise the steps: detection method is differentiated in (1)
A. get these article 3g, add water 50ml, measure 15ml, through D101 type macroporous adsorptive resins; Water 100ml eluting is arranged, discard water liquid, reuse 50% ethanol 100ml eluting; Collect eluent, evaporate to dryness, residue add methanol 1ml makes dissolving; Get lobetyolin's reference substance in addition as need testing solution, add methanol and process the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography, draw need testing solution and each 2ul of reference substance solution and put respectively on same silica gel g thin-layer plate, with n-butyl alcohol-glacial acetic acid-water (7; 1: 0.5) for developing solvent, launch, take out, to dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃, puts under the ultra-violet lamp (365nm) and inspects.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
B. get these article 2g, add water 40ml, measure 5ml, full each the n-butanol extraction jolting of water is extracted 4 times; Each 20ml merges n-butyl alcohol liquid, and evaporate to dryness, residue add methanol 2ml makes dissolving; As need testing solution, other gets the verbascoside reference substance, adds methanol alcohol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography, draw need testing solution and each 5ul of reference substance solution and put respectively on same silica gel g thin-layer plate, be developing solvent with ethyl acetate-methanol formic acid (16: 0.5: 2), launch to take out and dry; With 0.1% 2, the bitter diazanyl ethanol solution dipping of 2-diphenyl-1-dries.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.
(2) assay: version appendix VID was according to high effective liquid chromatography for measuring in 2005
Chromatographic condition and system suitability condition are filler with the octadecyl silane, and the glacial acetic acid of acetonitrile-0.1% (15: 85) is a mobile phase; The detection wavelength is 334nm; Number of theoretical plate calculates by the verbascoside peak should be not less than 5000
The preparation of reference substance solution: it is an amount of to get the verbascoside reference substance, and accurate the title decides, and adds mobile phase and processes the solution that every 1ml contains 20ug, promptly gets.
The preparation of need testing solution: get the about 1.0g of these article, the accurate title, decide, and puts in the tool plug conical flask, the accurate methanol 50ml that adds; Claim decide weight, water-bath refluxed 60 minutes, put coldly, and weight decided in title again; Add methanol and supply weight, shake up, filter, precision is measured subsequent filtrate 2ml and is put in the 10ml volumetric flask; Add mobile phase to scale, shake up, filter, get subsequent filtrate and promptly get.The every g of these article contains Radix Rehmanniae Preparata must not be less than 0.06mg in verbascoside (C29H36O15).
6. detection method according to claim 1 is characterized in that, the method for preparing of said preparation may further comprise the steps:
Get Radix Codonopsis 160g, Radix Rehmanniae Preparata 40g, Rhizoma Polygonati 80g, Fructus Lycii 40g, Radix Polygoni Multiflori 40g; The above five tastes, decocte with water 2 times, each 2 hours, collecting decoction; Filtrating be condensed into thick paste, other gets sucrose 600g and processes simple syrup, add sodium benzoate 3g stirring and dissolving after, add in the above-mentioned thick paste; Mixing is processed 1000ml, filters, and promptly gets.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103622989A (en) * | 2013-11-21 | 2014-03-12 | 无限极（中国）有限公司 | Compound polysaccharide composition as well as preparation method and application of compound polysaccharide composition |
| CN109541044A (en) * | 2017-09-22 | 2019-03-29 | 广州白云山中药业有限公司 | The quality determining method of glutinous rehmannia medicinal material |
| CN110448631A (en) * | 2019-09-24 | 2019-11-15 | 慈幼(上海)健康管理咨询有限公司 | A kind of children massage with kidney tonifying cream and preparation method thereof |
| CN112569306A (en) * | 2019-09-27 | 2021-03-30 | 王艳萍 | Life-prolonging health-care liquid and preparation method thereof |
-
2010
- 2010-07-22 CN CN2010102341163A patent/CN102335333A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103622989A (en) * | 2013-11-21 | 2014-03-12 | 无限极（中国）有限公司 | Compound polysaccharide composition as well as preparation method and application of compound polysaccharide composition |
| CN103622989B (en) * | 2013-11-21 | 2016-01-20 | 无限极（中国）有限公司 | A kind of complex polysaccharide composition and method of making the same and application |
| CN109541044A (en) * | 2017-09-22 | 2019-03-29 | 广州白云山中药业有限公司 | The quality determining method of glutinous rehmannia medicinal material |
| CN110448631A (en) * | 2019-09-24 | 2019-11-15 | 慈幼(上海)健康管理咨询有限公司 | A kind of children massage with kidney tonifying cream and preparation method thereof |
| CN112569306A (en) * | 2019-09-27 | 2021-03-30 | 王艳萍 | Life-prolonging health-care liquid and preparation method thereof |
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