CN102078503A - Detection method for pulse-activating decoction traditional Chinese medicine preparation - Google Patents
Detection method for pulse-activating decoction traditional Chinese medicine preparation Download PDFInfo
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- CN102078503A CN102078503A CN2009101865883A CN200910186588A CN102078503A CN 102078503 A CN102078503 A CN 102078503A CN 2009101865883 A CN2009101865883 A CN 2009101865883A CN 200910186588 A CN200910186588 A CN 200910186588A CN 102078503 A CN102078503 A CN 102078503A
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Abstract
The invention relates to a detection method for a pulse-activating decoction traditional Chinese medicine preparation, in particular relating to a detection method for pulse-activating decoction. The detection method for a pulse-activating decoction oral liquid comprises the steps of identification and content determination, wherein during the identification, thin-layer chromatography is adopted, and Chinese magnoliavine fruit and dwarf lilyturf tuber are taken as control for identifying pharmaceutical ingredients in the pulse-activating decoction oral liquid; and during the content determination, high performance liquid chromatography is adopted for determining the content of schizandrin in the pulse-activating decoction oral liquid.
Description
Invention field
The present invention relates to the detection method of SHENGMAI YIN Chinese medicine preparation, particularly the detection method of SHENGMAI YIN (Radix Codonopsis side).
Background technology
SHENGMAI YIN KOUFUYE, its function cure mainly and are Yiqi and vein recovery, YIN nourishing and the production of body fluid promoting.Be used for QIYINLIANGXU, shortness of breath and palpitation, faint pulse spontaneous perspiration.Determined curative effect, the little welcome that is subjected to numerous doctors and patient deeply of side effect always, good market prospects have been hauled oneself willingly into since the market.
SHENGMAI YIN KOUFUYE is by the method for making of ministry standard Chinese traditional patent formulation preparation, and Radix Codonopsis 300g, Radix Ophiopogonis, 200g, above three flavors of Fructus Schisandrae Chinensis 100g decocted with water secondary, 2 hours for the first time, 1.5 hours for the second time, collecting decoction filtered, and filtrate is concentrated into about 300ml, put coldly, add ethanol 600ml, placed 24 hours, filter, filtrate decompression is condensed into the thick paste shape, adds water and dilutes in right amount, filter, other add simple syrup 300ml and antiseptic an amount of, add water to 1000ml, stir evenly, leave standstill, filter, embedding, sterilization, promptly.
Existing quality standard only its relative density (should be not less than 1.08) is controlled, and is unfavorable for controlling aborning the quality with monitor drug.
This just requires us to produce high-quality product, but the initial quality controlling party is difficult to the product of different dosage form is not carried out effective quality control.
The influence factor is more for the existing product detection method, and repeatability is relatively poor as a result, can not the better controlled drug quality, thus can not guarantee the stable uniform of product quality well.
For guaranteeing its steady quality homogeneous, this just requires have a good method of quality control that it is carried out effective quality control to product, makes product accomplish " safe, effective, quality controllable ".
The present invention adopts high performance liquid chromatography to carry out assay, adds to cooperate thin layer to differentiate, has more characteristics accurate, favorable reproducibility, and more former method precision, sensitivity, stability are all higher.Thereby can accurately strictly control the quality and the homogeneity of product,
Summary of the invention
The detection method of SHENGMAI YIN KOUFUYE of the present invention comprises and differentiating and the step of assay:
Thin layer chromatography is adopted in described discriminating, and with Fructus Schisandrae Chinensis, be contrast Radix Ophiopogonis, differentiates the ingredient in the SHENGMAI YIN KOUFUYE;
Described assay adopts the content of schizandrin in the high effective liquid chromatography for measuring SHENGMAI YIN KOUFUYE.
The detection method of SHENGMAI YIN KOUFUYE of the present invention, wherein discrimination method may further comprise the steps:
A. get this product 20ml and add chloroform 20ml, jolting is extracted, getting chloroform liquid puts in the water-bath and to steam residue and add methanol 0.5ml and make dissolving, as need testing solution. other gets Fructus Schisandrae Chinensis control medicinal material 0.5g, shine drug solns in pairs with legal system. get the schisandrin B reference substance again, add methanol and make the solution that every 1ml contains 1mg, product solution in contrast. according to thin layer chromatography (appendix IVB) test, draw each 2～5ul of above-mentioned three kinds of solution, the cross point is on same silica GF254 lamellae respectively, with toluene-ethyl acetate (9: 1) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (254nm) and inspect. in the test sample chromatograph, with the corresponding position of reference substance chromatograph of control medicinal material on, show the speckle of same color.
B. get this product 50ml, add hydrochloric acid 2ml, heated and boiled 5 minutes is put coldly, extracts with chloroform 40ml jolting, divides and gets chloroform liquid, and evaporate to dryness, residue add chloroform liquid 1ml makes dissolving, as need testing solution.Other gets control medicinal material 1g Radix Ophiopogonis, adds water 20ml, decocts 10 minutes, filters, and filtrate adds hydrochloric acid 0.5ml, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix IVB) test, draw above-mentioned two each 5ul of clock solution, put on same silica gel g thin-layer plate respectively.With chloroform: acetone (5: 1) is developing solvent, launches, and takes out, and dries, and spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
The detection method of SHENGMAI YIN KOUFUYE of the present invention, wherein content assaying method may further comprise the steps:
Measure according to high performance liquid chromatography (version appendix VID in 2005)
Chromatographic condition and system suitability condition: with the octadecyl silane is filler, methanol: water (30: 70) is mobile phase; The detection wavelength is 250nm; Number of theoretical plate calculates the preparation that should be not less than 2000 reference substance solution by the schisandrin peak: it is an amount of to get the schisandrin reference substance, the accurate title, decide, add methanol and make the solution that every 1ml contains 10ug, promptly. the preparation of need testing solution: precision is measured this product 2ml, put in 25～50ml measuring bottle, add methanol and be diluted to scale, shake up, filter, promptly. algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure. promptly get the every 1ml of this product and contain Fructus Schisandrae Chinensis, must not be less than 0.30mg in schizandrin (C24H32O7)
Thereby the SHENGMAI YIN KOUFUYE quality has been carried out effective control.
Monitoring method of the present invention is obtaining of process experiment sieving, with experimental example the present invention is further specified below.
Experimental example 1The research of schisandrin content assaying method
(1) instrument and reagent
High performance liquid chromatograph, comprise the Waters1525 pump, 2487 dual pathways UV-detector, the Waters column oven, Beerze liquid chromatograph work station, schisandrin reference substance (lot number: for assay usefulness, Nat'l Pharmaceutical ﹠ Biological Products Control Institute) methanol is U.S. Tedia chromatographically pure, water is ultra-pure water, and other reagent is analytical pure.Drug sample to be measured is SHENGMAI YIN (Radix Codonopsis side) (lot number: 080101 Jiangxi Jimin Kexin Drug Industry Co., Ltd)
2) chromatographic condition is with reference to the schisandra chinensis medicinal material assay
Chromatographic column: Penomenex Luna C18 (2) 4.6mm * 150mm column temperature: 30 ℃ of mobile phases: methanol-water (30: 70); The detection wavelength is 250nm; Flow velocity: 1.0ml/min
3) selection of detection wavelength
Get the schisandrin reference substance solution, in 200nm～400nm scope interscan, the result has absorption maximum at the 250nm place, so select 250nm as detecting wavelength.
4) choice of Solvent: respectively with 50% methanol, methanol as solvent, according to the operation down of assay item, (lot number: the content of schisandrin is measured 080101), the results are shown in Table 1 to sample.
Table 1 solvent is to the influence of measurement result
Therefore, select for use methanol as solvent
5) selection of solvent load: accurate respectively absorption test sample 2ml is settled to 25ml, 50ml.Filter then, (lot number: the content of the schisandrin 081001) is measured, and the results are shown in Table 2 to sample.
Table 2 solvent load is to the influence of measurement result
Therefore, select for use and be settled to 50ml as need testing solution.
6) blank assay
In the flavour of a drug ratio in the prescription, autogamy does not contain the blank preparation of Fructus Schisandrae Chinensis, makes blank solution as stated above, measures in accordance with the law, and blank solution is not seen apparent chromatographic peak at the place of identical retention time with the schisandrin reference substance as a result, so think noiseless.
7) linear relationship is investigated
Accurate above-mentioned reference substance solution (0.01223mg/ml) 2,4,6,8, the 10ul of drawing injects chromatograph of liquid, measures peak area, is abscissa with the sample size (ug) of reference substance, is vertical coordinate with the peak area, the drawing standard curve.The results are shown in Table 3.
Table 3 linear relationship experimental data (n=5)
Y＝500000X+11189
r＝0.9999
The result shows: be good linear relationship in .02446～0.1223ug scope.
8) stability test
The accurate need testing solution 10ul that draws, interval certain hour sample introduction, the peak area of mensuration schisandrin, RSD is 0.78%, the result shows that measurement result is stable in 24 hours, the results are shown in Table 4
Table 4 stability test data (n=5)
9) precision test
The accurate need testing solution 10ul that draws injects chromatograph of liquid, measures the peak area of schisandrin, repeats sample introduction 5 times, and RSD is 0.59% to be to see Table 5 as a result
Table 5 precision test data (n=5)
10) repeatability test
According to the operation down of assay item, (lot number: 081001) sample is surveyed for 5 parts, tries to achieve relative standard deviation RSD to be, the results are shown in Table 6 to same lot number
Table 6 repeatability test data (n=5)
11) recovery test
Reclaim with application of sample, and the same lot number of the known schisandrin content of accurate absorption (lot number: 081001, sample 1ml 0.36mg/ml), the accurate respectively schisandrin reference substance solution (0.2mg/ml) that adds is a certain amount of, according to the operation down of assay item, be calculated as follows the response rate, the results are shown in Table 7
Table 9 average recovery experimental data (n=6)
Average recovery rate is: 98.00%RSD is 0.65%
The assay of schisandrin in experimental example 1 SHENGMAI YIN
According to the method operation of above-mentioned assay, survey and know clearly 10 batches
The content that records the schisandrin in the SHENGMAI YIN preparation is more than 0.36mg/ml, consider the loss error in the industry amplification, therefore on the basis of minimum content, float downward 15%, therefore obtaining examination criteria is to contain schisandrin content among the every 1ml of SHENGMAI YIN should be not less than 0.30mg.
The specific embodiment
Embodiment 1
Give birth to the method for hydroxylamine drink quality testing
1, differentiates detection method
A; get this product 20ml and add chloroform 20ml; jolting is extracted; get chloroform liquid and put and steam residue in the water-bath and add methanol 0.5ml and make dissolving; as need testing solution. other gets Fructus Schisandrae Chinensis control medicinal material 0.5g; shine drug solns in pairs with legal system. get the schisandrin B reference substance again; add methanol and make the solution that every 1ml contains 1mg; product solution in contrast. according to thin layer chromatography (appendix IVB) test; draw each 2～5ul of above-mentioned three kinds of solution; the cross point is on same silica GF254 lamellae respectively; with toluene-ethyl acetate (9: 1) be developing solvent, launch, take out; dry; put under the ultra-violet lamp (254nm) and inspect. in the test sample chromatograph, with the corresponding position of reference substance chromatograph of control medicinal material on, the speckle of apparent same color.
B, get this product 50ml, add hydrochloric acid 2ml, heated and boiled 5 minutes is put coldly, extracts with chloroform 40ml jolting, divides and gets chloroform liquid, and evaporate to dryness, residue add chloroform liquid 1ml makes dissolving, as need testing solution.Other gets control medicinal material 1g Radix Ophiopogonis, adds water 20ml, decocts 10 minutes, filters, and filtrate adds hydrochloric acid 0.5ml, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix IVB) test, draw above-mentioned two each 5ul of clock solution, put on same silica gel g thin-layer plate respectively.With chloroform: acetone (5: 1) is developing solvent, launches, and takes out, and dries, and spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
2, assay: measure according to high performance liquid chromatography (version appendix VID in 2005)
Chromatographic condition and system suitability condition are filler with the octadecyl silane, methanol: water (30: 70) is mobile phase; The detection wavelength is 250nm; Number of theoretical plate calculates the preparation that should be not less than 2000 reference substance solution by the schisandrin peak: it is an amount of to get the schisandrin reference substance, the accurate title, decide, add methanol and make the solution that every 1ml contains 10ug, promptly. the preparation of need testing solution: precision is measured this product 2ml, put in the 50ml measuring bottle, add methanol and be diluted to scale, shake up, filter, promptly. algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure. promptly get the every 1ml of this product and contain Fructus Schisandrae Chinensis, must not be less than 0.30mg in schizandrin (C24H32O7).
Claims (7)
1. the detection method of SHENGMAI YIN Chinese medicine preparation comprises and differentiating and the step of assay:
2. the detection method of claim 1 is characterized in that, thin layer chromatography is adopted in described discriminating, and with Fructus Schisandrae Chinensis, be contrast Radix Ophiopogonis, differentiates the ingredient in the SHENGMAI YIN KOUFUYE;
3. the detection method of claim 1 is characterized in that, described assay adopts the content of schizandrin in the high effective liquid chromatography for measuring SHENGMAI YIN KOUFUYE.
4. the detection method of claim 1 is characterized in that, wherein discrimination method may further comprise the steps:
A. get this product 20ml and add chloroform 20ml, jolting is extracted, getting chloroform liquid puts in the water-bath and to steam residue and add methanol 0.5ml and make dissolving, as need testing solution. other gets Fructus Schisandrae Chinensis control medicinal material 0.5g, shine drug solns in pairs with legal system. get the schisandrin B reference substance again, add methanol and make the solution that every 1ml contains 1mg, product solution in contrast. according to thin layer chromatography (appendix IVB) test, draw each 2～5ul of above-mentioned three kinds of solution, the cross point is on same silica GF254 lamellae respectively, with toluene-ethyl acetate (9: 1) is developing solvent, launches, and takes out, dry, put under the ultra-violet lamp (254nm) and inspect. in the test sample chromatograph, with the corresponding position of reference substance chromatograph of control medicinal material on, show the speckle of same color.
5. the detection method of claim 1 is characterized in that, wherein discrimination method may further comprise the steps:
B. get this product 50ml, add hydrochloric acid 2ml, heated and boiled 5 minutes is put coldly, extracts with chloroform 40ml jolting, divides and gets chloroform liquid, and evaporate to dryness, residue add chloroform liquid 1ml makes dissolving, as need testing solution.Other gets control medicinal material 1g Radix Ophiopogonis, adds water 20ml, decocts 10 minutes, filters, and filtrate adds hydrochloric acid 0.5ml, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix IVB) test, draw above-mentioned two each 5ul of clock solution, put on same silica gel g thin-layer plate respectively.With chloroform: acetone (5: 1) is developing solvent, launches, and takes out, and dries, and spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
6. the detection method of claim 1, content assaying method wherein, step is as follows:
Measure according to high performance liquid chromatography (version appendix VID in 2005)
Chromatographic condition and system suitability condition: with the octadecyl silane is filler, methanol: water (30: 70) is mobile phase; The detection wavelength is 250nm; Number of theoretical plate calculates the preparation that should be not less than 2000 reference substance solution by the schisandrin peak: it is an amount of to get the schisandrin reference substance, the accurate title, decide, add methanol and make the solution that every 1ml contains 10ug, promptly. the preparation of need testing solution: precision is measured this product 2ml, put in 25～50ml measuring bottle, add methanol and be diluted to scale, shake up, filter, promptly. algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure. promptly get the every 1ml of this product and contain Fructus Schisandrae Chinensis, must not be less than 0.30mg in schizandrin (C24H32O7)
7. the detection method of claim 1, step is as follows:
1, differentiates detection method
A; get this product 20ml and add chloroform 20ml; jolting is extracted; get chloroform liquid and put and steam residue in the water-bath and add methanol 0.5ml and make dissolving; as need testing solution. other gets Fructus Schisandrae Chinensis control medicinal material 0.5g; shine drug solns in pairs with legal system. get the schisandrin B reference substance again; add methanol and make the solution that every 1ml contains 1mg; product solution in contrast. according to thin layer chromatography (appendix IVB) test; draw each 2～5ul of above-mentioned three kinds of solution; the cross point is on same silica GF254 lamellae respectively; with toluene-ethyl acetate (9: 1) be developing solvent, launch, take out; dry; put under the ultra-violet lamp (254nm) and inspect. in the test sample chromatograph, with the corresponding position of reference substance chromatograph of control medicinal material on, the speckle of apparent same color.
B, get this product 50ml, add hydrochloric acid 2ml, heated and boiled 5 minutes is put coldly, extracts with chloroform 40ml jolting, divides and gets chloroform liquid, and evaporate to dryness, residue add chloroform liquid 1ml makes dissolving, as need testing solution.Other gets control medicinal material 1g Radix Ophiopogonis, adds water 20ml, decocts 10 minutes, filters, and filtrate adds hydrochloric acid 0.5ml, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix IVB) test, draw above-mentioned two each 5ul of clock solution, put on same silica gel g thin-layer plate respectively.With chloroform: acetone (5: 1) is developing solvent, launches, and takes out, and dries, and spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the speckle of same color.
2, assay: according to high performance liquid chromatography (version appendix VID in 2005) mensuration chromatographic condition and system suitability condition is filler with the octadecyl silane, and methanol: water (30: 70) is mobile phase; The detection wavelength is 250nm; Number of theoretical plate calculates the preparation that should be not less than 2000 reference substance solution by the schisandrin peak: it is an amount of to get the schisandrin reference substance, the accurate title, decide, add methanol and make the solution that every 1ml contains 10ug, promptly. the preparation of need testing solution: precision is measured this product 2ml, put in the 50ml measuring bottle, add methanol and be diluted to scale, shake up, filter, promptly. algoscopy: accurate respectively reference substance solution and each 10ul of need testing solution of drawing, inject chromatograph of liquid, measure. promptly get the every 1ml of this product and contain Fructus Schisandrae Chinensis, must not be less than 0.30mg in schizandrin (C24H32O7).
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102879514A (en) * | 2012-08-07 | 2013-01-16 | 何述金 | Mass control method for medicinal composition for treating hepatitis |
| CN103495018A (en) * | 2013-09-29 | 2014-01-08 | 黑龙江大学 | Method for preparing refreshing Shengmai oral liquid |
| CN103822975A (en) * | 2013-08-07 | 2014-05-28 | 浙江工业大学 | Test method for pulse-activating preparation adopting pilose asiabell root |
| CN113777205A (en) * | 2021-08-31 | 2021-12-10 | 湖南省药品检验研究院(湖南药用辅料检验检测中心) | Method for detecting adulteration of radix ophiopogonis in pulse-activating decoction |
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2009
- 2009-11-30 CN CN2009101865883A patent/CN102078503A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102879514A (en) * | 2012-08-07 | 2013-01-16 | 何述金 | Mass control method for medicinal composition for treating hepatitis |
| CN103822975A (en) * | 2013-08-07 | 2014-05-28 | 浙江工业大学 | Test method for pulse-activating preparation adopting pilose asiabell root |
| CN103822975B (en) * | 2013-08-07 | 2015-10-28 | 浙江工业大学 | A kind of detection method of formulation of ' Sheng Mai ' of Radix Codonopsis side |
| CN103495018A (en) * | 2013-09-29 | 2014-01-08 | 黑龙江大学 | Method for preparing refreshing Shengmai oral liquid |
| CN103495018B (en) * | 2013-09-29 | 2016-06-01 | 黑龙江大学 | The preparation method of the raw arteries and veins oral liquid of a kind of restoring consciouness |
| CN113777205A (en) * | 2021-08-31 | 2021-12-10 | 湖南省药品检验研究院(湖南药用辅料检验检测中心) | Method for detecting adulteration of radix ophiopogonis in pulse-activating decoction |
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Application publication date: 20110601 |