CN100536870C - Qulity control method for new compound isatis leaf preparation - Google Patents
Qulity control method for new compound isatis leaf preparation Download PDFInfo
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- CN100536870C CN100536870C CNB2006100729977A CN200610072997A CN100536870C CN 100536870 C CN100536870 C CN 100536870C CN B2006100729977 A CNB2006100729977 A CN B2006100729977A CN 200610072997 A CN200610072997 A CN 200610072997A CN 100536870 C CN100536870 C CN 100536870C
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Abstract
The present invention relates to the quality control method for new compound isatis leaf preparation. The new compound isatis leaf preparation has the recipe comprising compound isatis leaf extract, paracetamol, coffeine, amobarbital and vitamin C in certain weight proportion. The quality control method includes the following steps: identification of content and the content measurement of effective components.
Description
Technical field:
The present invention relates to a kind of method of quality control of pharmaceutical preparation, specifically relate to the method for quality control of new compound indigowoad leaf preparation.
Background technology:
New compound Folium Isatidis blade is a kind of Chinese medicine and western medicine compound preparation that has gone on the market, and has pestilence clearly, antiinflammatory, refrigeration function.But because of its production technology limits, this product is full extractum film-making, and tablet hardness is big, the phenomenon that disintegrate does not meet national standard often occurs, has limited the performance of pharmaceutical effectiveness.Do adhesive, filler, disintegrating agent, covering agent, increased technical process, waste energy, increased cost owing to needing tabletting, coating, granulation, tabletting, coating to add adjuvant again simultaneously.On the other hand, existing new compound Folium Isatidis blade exists method of quality control to fall behind the uppity shortcoming of product quality.Its composition is not differentiated in the existing method of quality control, or its composition is carried out the content of assay.So existing method of quality control can not effectively be controlled the quality of new compound Folium Isatidis blade, thereby will influence the production of product and ensure the quality of products.
For effectively controlling product quality, we have set up the new method of quality control of new compound indigowoad leaf preparation, and this method adopts thin layer chromatography that composition is wherein differentiated, the employing high performance liquid chromatography is carried out assay to composition wherein.Up to the present report is not studied this target level of product quality comprehensively, only there are indivedual researcheres that wherein chlorogenic acid, acetaminophen were done separately research, resemble high performance liquid chromatogram interlock that we adopt and measure the report of three active component simultaneously and more do not have.This method of quality control precision, sensitivity, stability are all good, and method is simpler, can farthest guarantee " safety, the homogeneous, stable, effective, controlled " of product quality.
Summary of the invention:
The object of the invention is to provide the preparation method and the method for quality control of a kind of taking convenience, better efficacy, quality controllable new compound Folium Isatidis capsule preparations.
One, this kind capsule has than the advantage of tablet:
1, reduces production tabletting, coating operation, save the process time, reduce power consumption;
2, reduce the adjuvants such as disintegrating agent that tabletting increased, economize on resources;
3, reduce the required a large amount of adjuvants of tablet coating, economize on resources, reduce the absorption of patient invalid adjuvant;
4, shorten disintegrating procedue, the dissolving that is beneficial to medicine absorbs.
Because tablet need be pulverized dried cream, add adjuvant (starch, ethanol) and granulate, with disintegrating agent, lubricant granulate, tabletting, with sucrose, Pulvis Talci coating, packing.And capsule only need be broken with dried cream powder, adds lubricant, and fill promptly.Save three process, reduce adjuvant account for sheet heavy about 50%.
Former label content of dispersion .0.3g, add adjuvant granulation, tabletting, coating after, the heavy 0.56-0.60g of sheet
Tablet and capsule contrast
| Project | Tablet | Capsule |
| Granulate | Need | Do not need |
| Mix granulate | Need | Need |
| Tabletting | Need | Do not need |
| Coating | Need | Do not need |
| Adjuvant | Need | Do not need |
| Gross weight | 0.58-0.63g | 0.3g |
| Disintegration | 45-65 minute | 25-35 minute |
| Cycle in man-hour | 0.4/ ten thousand | 0.08/ ten thousand |
| The adjuvant expense | 13.65 unit/ten thousand | 0.345/ ten thousand |
New compound Folium Isatidis capsule prescription consists of:
[prescription] compound Folium Isatidis extract 200g acetaminophen 75g
Caffeine 7.5g amobarbital 7.5g vitamin C 10g
[method for making] above five tastes pulverize separately, mixing adds right amount of auxiliary materials, makes granule, dresses up 1000 of capsules, promptly.
The present invention be directed to new compound Folium Isatidis capsule preparations and propose method of quality control, but also do not limit and new compound Folium Isatidis capsule, also can comprise other oral formulations such as tablet, granule.
Its prescription of other dosage forms of new compound Folium Isatidis that the present invention includes is identical with capsule preparations.
Wherein said compound Folium Isatidis extract is by Chinese medicine of the five flavours: Folium Isatidis, Flos Lonicerae, Radix Et Rhizoma Rhei, Rhizoma Et Radix Notopterygii, Rhizoma Bistortae, Folium Isatidis by weight proportion: Flos Lonicerae: Radix Et Rhizoma Rhei: Rhizoma Et Radix Notopterygii: after Rhizoma Bistortae=4:2:1:1:1 proportioning through extracting processing and preparing, described extraction processing method can be according to prior art such as ministry standard, in patent and the non-patent literature in the preparation of 'Compound satis Leaf ' relevant extracting method extract.
'Compound satis Leaf ' is a kind of known drug, has listed the 5th of ministry standard in.
Above new compound Folium Isatidis prescription is formed and is by weight as proportioning, can increase or reduce according to corresponding proportion when producing, and be no more than 100% at most.
Large-scale production can be unit with the kilogram, or is unit with the ton.More than composition can be made into 1000 doses of pharmaceutical preparatioies, described 1000 doses of fingers, and the final drug preparation of making, as make 1000 of capsule preparations, and 1000 in tablet, granule 1000g etc.,
New compound indigowoad leaf preparation of the present invention, when making preparation, can add the medicine acceptable carrier as required, these carriers can be any carriers that is fit to make preparation of the present invention, as: starch, sucrose, lactose, cellulose and derivant thereof, Pulvis Talci, titanium dioxide, gelatin, polyvinylpyrrolidone, ethanol, β-cyclodextrin etc.
New compound indigowoad leaf preparation, particularly capsule preparations of the present invention, it is as follows that its function cures mainly usage and dosage: [function cures mainly] clear pestilence, antiinflammatory, analgesic.Be used for Common Cold, fever and headache, rhinorrhea with clear discharge, joint is ached.
[usage and dosage] is oral, a 3-4 grain, 2 times on the one.
The present invention is to provide the method for quality control at above new compound indigowoad leaf preparation, particularly capsule preparations, be the quality of control new compound indigowoad leaf preparation.At its characteristics and prescription of the present invention, we provide following method of quality control:
Method of quality control of the present invention may further comprise the steps:
The discriminating of content is carried out assay to the composition that contains, and therefore, the main step of method of quality control of the present invention is:
The discriminating of content, step is:
The discriminating of [discriminating] (1) Folium Isatidis: get the about 5g of this product content, add water 80ml water-bath warm macerating (80 ℃) and made dissolving in 30 minutes, leave standstill put cold, filter, filtrate is used ethyl acetate extraction 3 times, each 30ml, combined ethyl acetate liquid, with 1% sodium hydroxide solution washing 3 times, each 20ml, combined ethyl acetate liquid, water bath method, residue adds the 0.5ml ethyl acetate makes dissolving, as need testing solution.It is an amount of that other gets the indirubin reference substance, adds ethyl acetate and make the solution that every 1ml contains 0.1mg, in contrast product solution.Test according to thin layer chromatography (2005 editions appendix VIB of Chinese Pharmacopoeia), draw need testing solution 5～10 μ 1, reference substance solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethyl starch sodium, with toluene-ethyl acetate-acetone (5:4:1) is developing solvent, launch, take out, dry.In the test sample chromatograph, with contrast chromatograph corresponding position, show the lavender speckle of same color.
(2) discriminating of Radix Et Rhizoma Rhei: get the about 1.5g of this product content, add methanol 30ml supersound process 15 minutes, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, adds hydrochloric acid 1ml again, put warm macerating 30min in the water-bath (80 ℃), cooling immediately extracts twice with the ethyl acetate jolting, each 10ml, combined ethyl acetate liquid, water bath method, residue add methanol 1ml makes dissolving, as need testing solution.Get Radix Et Rhizoma Rhei control medicinal material 0.5g, add methanol 20ml, prepare control medicinal material solution according to the test sample preparation method.Other gets the chrysophanol reference substance, adds methanol and makes the solution that 1ml contains 0.5mg, in contrast product solution.Test according to thin layer chromatography (2005 editions appendix VIB of Chinese Pharmacopoeia), draw control medicinal material solution and reference substance solution each 2 μ l, need testing solution 5 μ 1, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethyl starch sodium, upper solution with petroleum ether (30-60 ℃)-Ethyl formate-formic acid (15:5:1) is developing solvent, launch, take out, dry, put under the uviol lamp (365nm) and inspect, in the test sample chromatograph, with control medicinal material, the corresponding position of reference substance chromatograph on show the fluorescence speckle of same color.
(3) ascorbic discriminating: get this product content 3g, add 0.1mol/l hydrochloric acid solution 50ml, jolting 30 minutes filters, and gets subsequent filtrate 10ml, adds phenylhydrazine hydrochloride 0.1g, and 60 ℃ of water-bath warm macerating 1 hour (jolting constantly) are put coldly, filter, as need testing solution.Other gets vitamin C reference substance 50mg, adds 0.1mol/l hydrochloric acid solution 10ml and phenylhydrazine hydrochloride 0.1g, makes with method, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw each 10 μ l of need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene-ethyl acetate-acetone (5:5:1) is developing solvent, launch, take out, dry.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the glassy yellow speckle of same color.
(4) discriminating of Rhizoma Et Radix Notopterygii: get this product content 3g, use ethyl acetate extraction 4 times, each 10ml, combined ethyl acetate liquid, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Make negative need testing solution with method.Other gets Rhizoma Et Radix Notopterygii control medicinal material powder 1.0g, adds ethanol 30ml, floods 1 hour, filters, and filtrate evaporate to dryness, residue add ethanol 1ml and dissolve medical material solution in contrast.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), draw each 5 μ l of need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, upper solution with ethyl acetate-methanol-water (10:2:3) is developing solvent, launch, take out, dry.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
The composition that contains is carried out assay, and step is:
[assay]
1, chlorogenic acid is measured chromatographic condition and system suitability test according to high performance liquid chromatography (2005 editions appendix VID of Chinese Pharmacopoeia): with octadecylsilane chemically bonded silica is filler; Acetonitrile-0.4% phosphoric acid solution (13:87) is a mobile phase; Detect wavelength 327nm.Number of theoretical plate calculates by the chlorogenic acid peak should be lower than 2000.
The preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, puts in the brown measuring bottle, adds 50% methanol and make the solution that every 1ml contains 20 μ g.
The preparation of need testing solution: precision takes by weighing this product powder 0.3g, puts in the tool plug conical flask, and precision adds 50% methanol 50ml, claims decide weight, and supersound process 30 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, promptly.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
The every gram of this product contains Flos Lonicerae and Rhizoma Bistortae must not be less than 3.0mg by chlorogenic acid (C16H18O9).
2, acetaminophen, caffeine and amobarbital are measured according to high performance liquid chromatography (2005 editions appendix VID of Chinese Pharmacopoeia)
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Methanol-water (50:50) is a mobile phase; Detect wavelength 215nm.Number of theoretical plate calculates by acetaminophen peak, caffeine peak-to-peak and amobarbital peak all should be not less than 3000.
The preparation of reference substance solution: it is an amount of that precision takes by weighing acetaminophen, caffeine and amobarbital reference substance, add 50% methanol and make every 1ml and contain acetaminophen 500 μ g and contain the respectively reference substance mixed solution of 50 μ g of caffeine and amobarbital, promptly.
The preparation of need testing solution: precision takes by weighing this product powder 0.5g, puts in the tool plug conical flask, and precision adds 50% methanol 50ml, claims decide weight, and supersound process 20 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, promptly.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
This product contains 85%～115%. pair of composition that contains that acetaminophen, caffeine and amobarbital all should be labelled amount and carries out assay, also can adopt following steps:
[assay]
1, chlorogenic acid is measured chromatographic condition and system suitability test according to high performance liquid chromatography (2005 editions appendix VID of Chinese Pharmacopoeia): with octadecylsilane chemically bonded silica is filler; Acetonitrile-0.4% phosphoric acid solution (13:87) is a mobile phase; Detect wavelength 327nm.Number of theoretical plate calculates by the chlorogenic acid peak should be lower than 2000.
The preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, puts in the brown measuring bottle, adds 50% methanol and make the solution that every 1ml contains 20 μ g.
The preparation of need testing solution: precision takes by weighing this product powder 0.3g, puts in the tool plug conical flask, and precision adds 50% methanol 50ml, claims decide weight, and supersound process 30 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, promptly.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
The every gram of this product contains Flos Lonicerae and Rhizoma Bistortae must not be less than 3.0mg by chlorogenic acid (C16H18O9).
Caffeine and amobarbital are measured according to high performance liquid chromatography (2005 editions appendix VID of Chinese Pharmacopoeia).
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Methanol-water (50:50) is a mobile phase; Detect wavelength 215nm.Number of theoretical plate calculates by caffeine peak and amobarbital peak all should be not less than 3000.
The preparation of reference substance solution: precision takes by weighing caffeine and the amobarbital reference substance is an amount of, adds 50% methanol solution and makes the solution that every 1ml contains 50 μ g, promptly.
The preparation of need testing solution: precision takes by weighing this product powder 0.1g, puts in the tool plug conical flask, and precision adds 50% methanol solution 50ml, claims decide weight, and supersound process 20 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, promptly.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
This product contains caffeine and amobarbital all should be 85%～115%. of labelled amount
Vitamin C and acetaminophen are measured according to high performance liquid chromatography (2005 editions appendix VID of Chinese Pharmacopoeia)
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile-0.4 phosphoric acid solution (13:87) is a mobile phase; Detect wavelength 242nm.Number of theoretical plate calculates by vitamin C peak and acetaminophen peak all should be not less than 3000.
The preparation of reference substance solution: precision takes by weighing vitamin C and the acetaminophen reference substance is an amount of, adds mobile phase and makes the mixed solution that every 1ml contains vitamin C 30 μ g and contains acetaminophen 250 μ g, promptly.The preparation of need testing solution: precision takes by weighing this product powder 0.5g, puts in the tool plug conical flask, and precision adds mobile phase 50ml, claims decide weight, and supersound process 20 minutes is put coldly, claims to decide weight again, supplies with mobile phase to subtract weight loss, shake up, and filtration, promptly.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
This product contains vitamin C should be not less than 70% of labelled amount, and containing acetaminophen is 85%～115% of labelled amount.
Method of the present invention obtains through screening, and screening process is described as follows:
[discriminating]
(1) TLC of Folium Isatidis differentiates: the discriminating of the 615th page of " GANMAO TUIRE KELI " middle Folium Isatidis of test reference " one one of Chinese Pharmacopoeia " once version in 2000, with the indirubin is contrast, found that there is a yellow spotting serious interference at indirubin speckle place in sample, medicine according to this product is formed, select for use 1% sodium hydroxide solution that extract is carried out preparing sample after the carrying out washing treatment, the yellow interference disappears in the chromatograph as a result, and chromatograph is clear.Show that through many batch sample tests this method is stablized feasible, repeatability is good, the discriminating of Folium Isatidis in the side of can be used as.
(2) TLC of Radix Et Rhizoma Rhei differentiates: with reference to the discriminating under the 18th page of " Radix Et Rhizoma Rhei " item of " one one of Chinese Pharmacopoeia " version in 2000, with rhubarb medicinal material and chrysophanol is contrast, test according to thin layer chromatography (2005 editions appendix VIB of Chinese Pharmacopoeia), draw control medicinal material and reference substance solution 2 μ l, need testing solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethyl starch sodium, upper solution with petroleum ether (30-60 ℃)-Ethyl formate-formic acid (15:5:1) is developing solvent, launch, take out, dry, put under the uviol lamp (365nm) and inspect, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on show the fluorescence speckle of same color.
(3) ascorbic thin layer is differentiated: test is carried out physicochemical identification with the sodium dichlorophenol indophenolate test solution to vitamin C once with reference to official method, and feminine gender has interference as a result.By screening vitamin C has been carried out the thin layer discriminating, test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2005), with test sample and reference substance put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene-ethyl acetate-acetone (5:5:1) is developing solvent, launch, take out, dry.In the test sample chromatograph, with contrast chromatograph corresponding position on, show the glassy yellow speckle of same color.Show that through many batch sample tests this method is stablized feasible, repeatability is good, ascorbic discriminating in the side of can be used as.
(4) TLC of Rhizoma Et Radix Notopterygii differentiates: test is once with reference to the thin layer discrimination method of controlling Rhizoma Et Radix Notopterygii and Herba Andrographis water soluble ingredient in the good capsule of sense, use n-butanol extraction, with chloroform-ethyl acetate-methanol (4:5:2) is that developing solvent is differentiated, corresponding chromatograph speckle was not obvious during the result supplied examination and contrasts.Using ethyl acetate extraction afterwards, is developing solvent with ethyl acetate-methanol-water (10:2:3), in the test sample chromatograph, with contrast chromatograph corresponding position on, show the fluorescence speckle of same color.
[assay]
One, the assay of chlorogenic acid in Flos Lonicerae and the Rhizoma Bistortae
Chlorogenic acid is the main component of Flos Lonicerae, also is the main component of new compound Folium Isatidis blade anti-inflammation, and it is necessary to set up its assay.But the Rhizoma Bistortae in the compound indigowoad leaf extractum side contains a small amount of chlorogenic acid too, for this reason, adopts chlorogenic acid contents in hplc simultaneous determination Flos Lonicerae and the Rhizoma Bistortae.With reference to content assaying method and the relevant document under the 21st page of Flos Lonicerae item of Chinese Pharmacopoeia version in 2005, determine that method is simple, the accuracy height with chlorogenic acid contents in high effective liquid chromatography for measuring Flos Lonicerae and the Rhizoma Bistortae.
1, chromatographic condition and system suitability test
With octadecylsilane chemically bonded silica is filler; Acetonitrile-0.4% phosphoric acid solution (13:87) is a mobile phase; Detect wavelength 327nm, flow velocity 1/min.Number of theoretical plate calculates by the chlorogenic acid peak should be lower than 2000.The mobile phase of once screening in the test has:
Methanol-0.4% phosphoric acid solution (15:85)
Result: T
R=45min, reference substance and sample peak shape are good, but retention time is oversize;
Acetonitrile-0.4% phosphoric acid solution (13:87)
Result: T
R=10min, reference substance and sample evaluation are good.
So select mobile phase to be: acetonitrile-0.4% phosphoric acid solution (13:87).
2 ranges of linearity are investigated: it is an amount of to get the chlorogenic acid reference substance, and accurate the title decides, and adds 50% methanol and makes the solution that every 1ml contains 37 μ g, and accurate respectively 20 μ l, 15 μ l, 10 μ l, 8 μ l, the 5 μ l sample introductions drawn are measured its peak area.The results are shown in Table 15-1.With peak area (Y) sample size (X) is returned, get the standard curve equation.Y＝57.659X-0.1899(r＝0.9998)。Above result shows that chlorogenic acid peak area value and sample size have good linear relationship in 0.185 μ g～0.740 μ g scope.
Table 1 chlorogenic acid reference substance measurement result
| Sampling volume (μ l) | 20 | 15 | 10 | 8 | 5 |
| Sample size (μ g) | 0.740 | 0.555 | 0.370 | 0.296 | 0.185 |
| Peak area | 42.647 | 31.450 | 21.332 | 16.918 | 10.439 |
3 precision tests: draw reference substance solution (20 μ g/ml) and same duplicate samples solution and repeat sample introduction respectively 6 times, 20 μ l respectively at every turn calculate the RSD of both peak areas respectively, the results are shown in Table 2.
Table 2 Precision test result
The result shows: reference substance peak area RSD is 0.25%, and test sample peak area RSD is 0.56%, and precision is good.
4 repeatability test: get 041001 batch 6 parts in sample, press preparation under the test sample preparation, mensuration respectively, the results are shown in Table 3.
Table 3 reproducible test results
The result shows: the average content of 6 parts of test samples is 4.23mg/g, and RSD is 0.93%, and the repeatability of test method is good.
5 recovery tests: (adopting the application of sample absorption method) precision takes by weighing 041001 batch, and (known content is
4.21mg/g) fine powder 0.15g, nominal is got 5 parts, adds identical chlorogenic acid reference substance respectively, measures total content, calculate recovery rate.The response rate=(measuring in total amount-sample)/pure product amount of adding.The results are shown in Table 4.
Table 4 recovery test result
The result shows: average recovery rate is 97.5%, and RSD is 0.76%, and average recovery is good.
6 sample determinations: get ten batch samples, measure by the method for working out, chlorogenic acid content sees Table 5.
Chlorogenic acid content measurement result table in the table 5 new compound Folium Isatidis capsule
According to above-mentioned result of the test, more than ten batch sample content minimum be 041103 batch, content is 3.81mg/g, considers the source of medical material, and the influence of factors such as preparation production, storage, tentative this product chlorogenic acid content must not be and is less than 3.0mg/g.
Two, caffeine and amobarbital Determination on content
This side is the Chinese medicine and western medicine compound preparation, and the quantitative criterion of setting up Western medicine is very necessary.We carry out purification treating method by screening chromatographic condition, sample with reference to relevant document, have set up the translocation method of caffeine, amobarbital.
1, the selection of chromatographic condition
With octadecylsilane chemically bonded silica is filler; Detect wavelength 215nm.Number of theoretical plate calculates by caffeine peak and amobarbital peak all should be not less than 3000.Flow velocity 1ml/min, following mobile phase once on probation and wavelength:
Methanol-water (75:25), the result: the caffeine retention time is: 3.17min, the amobarbital retention time is: 3.95min, reference substance and sample peak all trail.
Methanol-water (50:50), the result: the caffeine retention time is: 4.45min, the amobarbital retention time is: 15.26min, sample and reference substance are under three wavelength, peak shape is all good, under each wavelength in the sample labelled amount be more or less the same, caffeine and amobarbital all have bigger absorption under 215nm.
So mobile phase is defined as methanol: water (50:50), detect wavelength and be defined as 215nm.
2, the range of linearity is investigated: get caffeine and the amobarbital reference substance is an amount of, the accurate title, decide, add 50% methanol and make the mixed solution that every 1ml contains caffeine 102 μ g and contains amobarbital 97 μ g, accurate respectively 5 μ l, 8 μ l, 10 μ l, 15 μ l, the 20 μ l sample introductions drawn are measured its peak area.The results are shown in Table 6,7.With peak area (Y) sample size (X) is returned, get the standard curve equation.Caffeine: Y=72.828X+3.1176 (r=0.9997).Above result shows caffeine in 0.510 μ g～2.040 μ g scopes, and caffeine peak area value and sample size have good linear relationship; Amobarbital: Y=33.92X-0.7237 (r=0.9999) is in 0.458 μ g～1.940 μ g scopes, and amobarbital peak area value and sample size have good linear relationship.
Table 6 caffeine reference substance measurement result
| Sampling volume (μ l) | 20 | 15 | 10 | 8 | 5 |
| Sample size (μ g) | 2.040 | 1.530 | 1.020 | 0.816 | 0.510 |
| Peak area | 150.576 | 116.02 | 77.509 | 62.998 | 39.334 |
Table 7 amobarbital reference substance measurement result
| Sampling volume (μ l) | 20 | 15 | 10 | 8 | 5 |
| Sample size (μ g) | 1.940 | 1.455 | 0.970 | 0.776 | 0.458 |
| Peak area | 65.248 | 48.580 | 31.820 | 25.550 | 15.100 |
3, precision test: draw reference substance solution and same duplicate samples solution and repeat sample introduction respectively 6 times, each each 20 μ l calculate the RSD of both peak areas respectively, the results are shown in Table 8 and 9.
Table 8 caffeine Precision test result
The result shows: reference substance peak area RSD is 0.11%, and test sample peak area RSD is 0.26%, and precision is good.
Table 9 amobarbital Precision test result
The result shows: reference substance peak area RSD is 0.37%, and test sample peak area RSD is 0.26%, and precision is good.
4, repeatability test: get 050105 batch 6 parts in sample, press respectively under the test sample preparation prepare, measurement result, caffeine and amobarbital content see Table 10 and 11.
Table 10 caffeine reproducible test results
The result shows: the caffeine average content of 6 parts of test samples is 25.8mg/g, and RSD is 0.20%, and the repeatability of test method is good.
Table 11 amobarbital reproducible test results
The result shows: the amobarbital average content of 6 parts of test samples is 22.2mg/g, and RSD is 0.23%, and the repeatability of test method is good.
5, recovery test: (adopting the application of sample absorption method) precision takes by weighing 041002 batch, and (known content of caffeine is 25.7mg/g, amobarbital content is 22.2mg/g) new compound Folium Isatidis capsule fine powder 0.05g, nominal is got 5 parts, add identical caffeine and amobarbital reference substance respectively, measure total content, calculate recovery rate.The response rate=(measuring in total amount-sample)/pure product amount of adding.The results are shown in Table 12 and 13.
Table 12 caffeine recovery test result
The result shows: average recovery rate is 99%, and RSD is 0.65%, and average recovery is good.
Table 13 amobarbital recovery test result
The result shows: average recovery rate is 100.6%, and RSD is 2.1%, and average recovery is good
6, sample determination: get ten batch samples, measure by the method for working out, caffeine and amobarbital content see Table 14 and table 15.
Table 14 ten batch sample content of caffeine measurement result tables
Table 15 ten batch sample amobarbital assays are table as a result
According to above-mentioned result of the test, more than ten batch sample content of caffeine minimum be 041103 batch, content is 92.8%, what amobarbital content was minimum is 041103 batch, content is 85.3%.For guaranteeing the safe and effective of preparation, tentative caffeine and amobarbital content are 85%～115% of labelled amount.
Three, acetaminophen, caffeine, amobarbital Determination on content
Because four flavor Western medicine are arranged in this product, we find in the test, and also available Same Way is measured the content of three compositions simultaneously, can reduce the mensuration program like this, economizes on resources and the time, makes method simpler.Therefore we screen the treatment conditions of chromatographic condition, sample, have set up feasible method.
1, chromatographic condition and system suitability test
With octadecylsilane chemically bonded silica is filler; Methanol-water (50:50) is a mobile phase; Detect wavelength 215nm.Flow velocity 1ml/min; Number of theoretical plate calculates by acetaminophen peak, caffeine peak-to-peak and amobarbital peak all should be not less than 3000.
Once following mobile phase on probation and wavelength: methanol-water (25:75)
The result: the acetaminophen retention time is: 5.9min, under wavelength 215nm, 218nm, 248nm, 257nm, contrast is good with the evaluation of sample collection of illustrative plates.
The caffeine retention time is: 15.7min, and under wavelength 215nm and 218nm, contrast is good with the evaluation of sample collection of illustrative plates; Under 248nm and 257nm, contrast is general with the sample evaluation.
Amobarbital contrast and sample do not see that yet collection of illustrative plates occurs behind operation 63min.This scheme is infeasible.Methanol-water (50:50)
The result: the acetaminophen retention time is: 3.1min, and under wavelength 215nm and 218nm, contrast is good with the evaluation of sample collection of illustrative plates; Under 248nm and 257nm, the chromatographic column overload appears owing to sample concentration is excessive, and collection of illustrative plates is estimated bad;
The caffeine retention time is: 4.2min, and under wavelength 215nm and 218nm, the collection of illustrative plates evaluation is good; Under 248nm and 257nm, the collection of illustrative plates evaluation is general.
The amobarbital retention time is: 14.1min, and under wavelength 215nm and 218nm, contrast is good with the evaluation of sample collection of illustrative plates; Under 248nm and 257nm, contrast does not all have absorption with sample.
From test as can be known, when being mobile phase with methanol-water (50:50), sample and reference substance are under 215nm and 218nm wavelength, the collection of illustrative plates evaluation is good, caffeine and amobarbital all have bigger absorption under 215nm, so mobile phase is defined as methanol: water (50:50), detect wavelength and be defined as 215nm.
2, the range of linearity is investigated: it is an amount of to get acetaminophen, caffeine, amobarbital reference substance, the accurate title, decide, adding 50% methanol makes every 1ml and contains acetaminophen 779 μ g, contain caffeine 82 μ g, the mixing reference substance solution that contains amobarbital 75 μ g, accurate respectively 20 μ l, 15 μ l, 10 μ l, 8 μ l, the 5 μ l sample introductions drawn are measured its peak area.With peak area (y) sample size (x) is returned, getting acetaminophen standard curve equation is y=24.152x+0.9007 (r=0.9998), getting caffeine standard curve equation is y=67.028x-0.2375 (r=0.9999), and getting amobarbital standard curve equation is y=32.14x-0.3509 (r=0.9998).Above result shows acetaminophen in 3.895 μ g～15.580 μ g scopes, and caffeine is in 0.410 μ g～1.640 μ g scopes, and amobarbital is in 0.375 μ g～1.500 μ g scopes, and peak area value and sample size have good linear relationship.The results are shown in following table.
Table 16 acetaminophen standard curve determination result
| Sampling volume (μ l) | 20 | 15 | 10 | 8 | 5 |
| Sample size (μ g) | 15.580 | 11.685 | 7.790 | 6.232 | 3.895 |
| Peak area | 379.468 | 279.993 | 188.031 | 152.077 | 96.155 |
Table 17 caffeine standard curve determination result
| Sampling volume (μ l) | 20 | 15 | 10 | 8 | 5 |
| Sample size (μ g) | 1.640 | 1.230 | 0.820 | 0.656 | 0.410 |
| Peak area | 110.115 | 81.490 | 55.002 | 43.524 | 27.465 |
Table 18 amobarbital standard curve determination result
| Sampling volume (μ l) | 20 | 15 | 10 | 8 | 5 |
| Sample size (μ g) | 1.500 | 1.125 | 0.750 | 0.600 | 0.375 |
| Peak area | 48.117 | 35.394 | 23.851 | 18.862 | 11.832 |
3, precision test: draw reference substance solution and same duplicate samples solution and repeat sample introduction respectively 6 times, each each 20 μ l calculate the RSD of both peak areas respectively, the results are shown in following table.
Table 19 acetaminophen Precision test result
The result shows: reference substance peak area RSD is 0.26%, and test sample peak area RSD is 0.38%, and precision is good.
Table 20 caffeine Precision test result
The result shows: reference substance peak area RSD is 0.31%, and test sample peak area RSD is 0.29%, and precision is good.
Table 21 amobarbital Precision test result
The result shows: reference substance peak area RSD is 0.25%, and test sample peak area RSD is 0.22%, and precision is good.
4 repeatability tests: get 041002 batch 6 parts in sample, press respectively under the test sample preparation prepare, measurement result, acetaminophen, caffeine, amobarbital content see the following form.
Table 22 acetaminophen reproducible test results
The result shows: the acetaminophen average content of 6 parts of test samples is 227.5mg/g, and RSD is 0.30%, and the repeatability of test method is good.
Table 23 caffeine reproducible test results
The result shows: the caffeine average content of 6 parts of test samples is 24.6mg/g, and RSD is 1.05%, and the repeatability of test method is good.
Table 24 amobarbital reproducible test results
The result shows: the different not barbital average content of 6 parts of test samples is 24.3mg/g, and RSD is 1.20%, and the repeatability of test method is good.
5 recovery tests: (adopting the application of sample absorption method) precision takes by weighing 041002 crowd of new compound Folium Isatidis capsule fine powder 0.05g, and nominal is got 5 parts, adds identical reference substance respectively, measures total content, calculate recovery rate.The response rate=(measuring in total amount-sample)/pure product amount of adding.The results are shown in following table.
Table 25 acetaminophen recovery test result
The result shows: average recovery rate is 98.6%, and RSD is 0.38%, and average recovery is good.
Table 26 recovery test result
The result shows: average recovery rate is 98.2%, and RSD is 0.53%, and average recovery is good.
Table 27 recovery test result
The result shows: average recovery rate is 99.1%, and RSD is 0.58%, and average recovery is good.
6 sample determinations: get ten batch samples, measure by the method for drafting, measurement result sees the following form.
Table 28 ten batch sample acetaminophen assays are table as a result
Table 29 ten batch sample content of caffeine measurement result tables
Table 30 ten batch sample amobarbital assays are table as a result
According to above-mentioned result of the test, more than ten batch sample acetaminophen content minimum be 041106 batch, content is 91.7%; What content of caffeine was minimum is 041106 batch, and content is 96.6%, and what amobarbital content was minimum is 041106 batch, and content is 96.9%.For guaranteeing the safe and effective of preparation, consider the influence of each side such as production simultaneously, tentative acetaminophen, caffeine, amobarbital content are 85%～115% of labelled amount.
More than this product described in the method for quality control of the present invention, be meant new compound Folium Isatidis capsule according to prepared of the present invention, when other dosage forms are measured, refer to corresponding other dosage forms.
The specific embodiment:
Further specify the present invention by the following examples, but not as limitation of the present invention.
The quality control of embodiment 1 new compound Folium Isatidis capsule,
May further comprise the steps: the discriminating of content, step is:
The discriminating of [discriminating] (1) Folium Isatidis: get the about 5g of this product content, add water 80ml water-bath warm macerating (80 ℃) and made dissolving in 30 minutes, leave standstill put cold, filter, filtrate is used ethyl acetate extraction 3 times, each 30ml, combined ethyl acetate liquid, with 1% sodium hydroxide solution washing 3 times, each 20ml, combined ethyl acetate liquid, water bath method, residue adds the 0.5ml ethyl acetate makes dissolving, as need testing solution.It is an amount of that other gets the indirubin reference substance, adds ethyl acetate and make the solution that every 1ml contains 0.1mg, in contrast product solution.Test according to thin layer chromatography (2005 editions appendix VIB of Chinese Pharmacopoeia), draw need testing solution 5～10 μ 1, reference substance solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethyl starch sodium, with toluene-ethyl acetate-acetone (5:4:1) is developing solvent, launch, take out, dry.In the test sample chromatograph, with contrast chromatograph corresponding position, show the lavender speckle of same color.
(2) discriminating of Radix Et Rhizoma Rhei: get the about 1.5g of this product content, add methanol 30ml supersound process 15 minutes, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, adds hydrochloric acid 1ml again, put warm macerating 30min in the water-bath (80 ℃), cooling immediately extracts twice with the ethyl acetate jolting, each 10ml, combined ethyl acetate liquid, water bath method, residue add methanol 1ml makes dissolving, as need testing solution.Get Radix Et Rhizoma Rhei control medicinal material 0.5g, add methanol 20ml, prepare control medicinal material solution according to the test sample preparation method.Other gets the chrysophanol reference substance, adds methanol and makes the solution that 1ml contains 0.5mg, in contrast product solution.Test according to thin layer chromatography (2005 editions appendix VIB of Chinese Pharmacopoeia), draw control medicinal material solution and reference substance solution each 2 μ l, need testing solution 5 μ 1, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethyl starch sodium, upper solution with petroleum ether (30-60 ℃)-Ethyl formate-formic acid (15:5:1) is developing solvent, launch, take out, dry, put under the uviol lamp (365nm) and inspect, in the test sample chromatograph, with control medicinal material, the corresponding position of reference substance chromatograph on show the fluorescence speckle of same color.
(3) ascorbic discriminating: get this product content 3g, add 0.1mol/l hydrochloric acid solution 50ml, jolting 30 minutes filters, and gets subsequent filtrate 10ml, adds phenylhydrazine hydrochloride 0.1g, and 60 ℃ of water-bath warm macerating 1 hour (jolting constantly) are put coldly, filter, as need testing solution.Other gets vitamin C reference substance 50mg, adds 0.1mol/l hydrochloric acid solution 10ml and phenylhydrazine hydrochloride 0.1g, makes with method, in contrast product solution.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw each 10 μ l of need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene-ethyl acetate-acetone (5:5:1) is developing solvent, launch, take out, dry.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the glassy yellow speckle of same color.
(4) discriminating of Rhizoma Et Radix Notopterygii: get this product content 3g, use ethyl acetate extraction 4 times, each 10ml, combined ethyl acetate liquid, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution.Make negative need testing solution with method.Other gets Rhizoma Et Radix Notopterygii control medicinal material powder 1.0g, adds ethanol 30ml, floods 1 hour, filters, and filtrate evaporate to dryness, residue add ethanol 1ml and dissolve medical material solution in contrast.Test according to thin layer chromatography (appendix VIB of Chinese Pharmacopoeia version in 2000), draw each 5 μ l of need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, upper solution with ethyl acetate-methanol-water (10:2:3) is developing solvent, launch, take out, dry.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.
The composition that contains is carried out assay, and step is:
[assay]
1, chlorogenic acid is measured chromatographic condition and system suitability test according to high performance liquid chromatography (2005 editions appendix VID of Chinese Pharmacopoeia): with octadecylsilane chemically bonded silica is filler; Acetonitrile-0.4% phosphoric acid solution (13:87) is a mobile phase; Detect wavelength 327nm.Number of theoretical plate calculates by the chlorogenic acid peak should be lower than 2000.
The preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, puts in the brown measuring bottle, adds 50% methanol and make the solution that every 1ml contains 20 μ g.
The preparation of need testing solution: precision takes by weighing this product powder 0.3g, puts in the tool plug conical flask, and precision adds 50% methanol 50ml, claims decide weight, and supersound process 30 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, promptly.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
The every gram of this product contains Flos Lonicerae and Rhizoma Bistortae by chlorogenic acid (C
16H
18O
9) must not count and be less than 3.0mg.
2, acetaminophen, caffeine and amobarbital are measured according to high performance liquid chromatography (2005 editions appendix VID of Chinese Pharmacopoeia)
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Methanol-water (50:50) is a mobile phase; Detect wavelength 215nm.Number of theoretical plate calculates by acetaminophen peak, caffeine peak-to-peak and amobarbital peak all should be not less than 3000.
The preparation of reference substance solution: it is an amount of that precision takes by weighing acetaminophen, caffeine and amobarbital reference substance, add 50% methanol and make every 1ml and contain acetaminophen 500 μ g and contain the respectively reference substance mixed solution of 50 μ g of caffeine and amobarbital, promptly.
The preparation of need testing solution: precision takes by weighing this product powder 0.5g, puts in the tool plug conical flask, and precision adds 50% methanol 50ml, claims decide weight, and supersound process 20 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, promptly.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
This product contains acetaminophen, caffeine and amobarbital and all should be 85%～115%. of labelled amount
Embodiment 2,
The capsular another kind of content assaying method of new compound Folium Isatidis is as follows: the composition that contains is carried out assay, also can adopt following steps:
[assay]
1, chlorogenic acid is measured chromatographic condition and system suitability test according to high performance liquid chromatography (2005 editions appendix VID of Chinese Pharmacopoeia): with octadecylsilane chemically bonded silica is filler; Acetonitrile-0.4% phosphoric acid solution (13:87) is a mobile phase; Detect wavelength 327nm.Number of theoretical plate calculates by the chlorogenic acid peak should be lower than 2000.
The preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, puts in the brown measuring bottle, adds 50% methanol and make the solution that every 1ml contains 20 μ g.
The preparation of need testing solution: precision takes by weighing this product powder 0.3g, puts in the tool plug conical flask, and precision adds 50% methanol 50ml, claims decide weight, and supersound process 30 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, promptly.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
The every gram of this product contains Flos Lonicerae and Rhizoma Bistortae must not be less than 3.0mg by chlorogenic acid (C16H18O9).
Caffeine and amobarbital are measured according to high performance liquid chromatography (2005 appendix VID of Chinese Pharmacopoeia).Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Methanol-water (50:50) is a mobile phase; Detect wavelength 215nm.Number of theoretical plate calculates by caffeine peak and amobarbital peak all should be not less than 3000.
The preparation of reference substance solution: precision takes by weighing caffeine and the amobarbital reference substance is an amount of, adds 50% methanol solution and makes the solution that every 1ml contains 50 μ g, promptly.
The preparation of need testing solution: precision takes by weighing this product powder 0.1g, puts in the tool plug conical flask, and precision adds 50% methanol solution 50ml, claims decide weight, and supersound process 20 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, promptly.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
This product contains caffeine and amobarbital all should be 85%～115%. of labelled amount
Vitamin C and acetaminophen are measured according to high performance liquid chromatography (2005 editions appendix VID of Chinese Pharmacopoeia)
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; Acetonitrile-0.4 phosphoric acid solution (13:87) is a mobile phase; Detect wavelength 242nm.Number of theoretical plate calculates by vitamin C peak and acetaminophen peak all should be not less than 3000.
The preparation of reference substance solution: precision takes by weighing vitamin C and the acetaminophen reference substance is an amount of, adds mobile phase and makes the mixed solution that every 1ml contains vitamin C 30 μ g and contains acetaminophen 250 μ g, promptly.The preparation of need testing solution: precision takes by weighing this product powder 0.5g, puts in the tool plug conical flask, and precision adds mobile phase 50ml, claims decide weight, and supersound process 20 minutes is put coldly, claims to decide weight again, supplies with mobile phase to subtract weight loss, shake up, and filtration, promptly.
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly.
This product contains vitamin C should be not less than 70% of labelled amount, and containing acetaminophen is 85%～115% of labelled amount.
Embodiment 3, the quality control of new compound Folium Isatidis tablet, and method changes the detected object capsule into tablet with embodiment 1, and detection method is constant.
Embodiment 4, the quality control of new compound Folium Isatidis tablet, and method changes the detected object capsule into tablet with embodiment 2, and detection method is constant.
Embodiment 5, the quality control of new compound Folium Isatidis granule, and method changes the detected object capsule into granule with embodiment 1, and detection method is constant.
Claims (4)
1, a kind of detection method of new compound Folium Isatidis capsule is characterized in that, comprises the discriminating of content, and the composition that contains is carried out the step of assay, and described method step is as follows:
The discriminating of content, step is:
The discriminating of [discriminating] (1) Folium Isatidis: get this product content 5g, add 80 ℃ of warm macerating of water 80ml water-bath and made dissolving in 30 minutes, leave standstill put cold, filter, filtrate is used ethyl acetate extraction 3 times, each 30ml, combined ethyl acetate liquid, with 1% sodium hydroxide solution washing 3 times, each 20ml, combined ethyl acetate liquid, water bath method, residue adds the 0.5ml ethyl acetate makes dissolving, as need testing solution; It is an amount of that other gets the indirubin reference substance, adds ethyl acetate and make the solution that every 1ml contains 0.1mg, in contrast product solution; 2005 editions appendix VIB of Chinese Pharmacopoeia are according to the thin layer chromatography test, draw need testing solution 5-10 μ l, reference substance solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethyl starch sodium, with toluene-ethyl acetate-acetone, ratio is that 5:4:1 is developing solvent, launches, take out, dry; In the test sample chromatograph, with contrast chromatograph corresponding position, show the lavender speckle of same color;
(2) discriminating of Radix Et Rhizoma Rhei: get this product content 1.5g, add methanol 30ml supersound process 15 minutes, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, adds hydrochloric acid 1ml again, put warm macerating 30min in 80 ℃ of water-baths, cooling immediately extracts twice with the ethyl acetate jolting, each 10ml, combined ethyl acetate liquid, water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Get Radix Et Rhizoma Rhei control medicinal material 0.5g, add methanol 20ml, prepare control medicinal material solution according to the test sample preparation method; Other gets the chrysophanol reference substance, adds methanol and makes the solution that 1ml contains 0.5mg, in contrast product solution; 2005 editions appendix VIB of Chinese Pharmacopoeia are according to the thin layer chromatography test, draw control medicinal material solution and reference substance solution each 2 μ l, need testing solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethyl starch sodium, with 30-60 ℃ of petroleum ether-Ethyl formate-formic acid ratio is that the upper solution of 15:5:1 is developing solvent, launch, take out, dry, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with control medicinal material, the corresponding position of reference substance chromatograph on show the fluorescence speckle of same color;
(3) ascorbic discriminating: get this product content 3g, add 0.1mol/l hydrochloric acid solution 50ml, jolting 30 minutes filters, and gets subsequent filtrate 10ml, adds phenylhydrazine hydrochloride 0.1g, 60 ℃ of water-bath warm macerating 1 hour, and jolting is constantly put coldly, filters, as need testing solution; Other gets vitamin C reference substance 50mg, adds 0.1mol/l hydrochloric acid solution 10ml and phenylhydrazine hydrochloride 0.1g, makes with method, in contrast product solution; An appendix VIB is according to the thin layer chromatography test for Chinese Pharmacopoeia version in 2005, draw each 10 μ l of need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, ratio with toluene-ethyl acetate-acetone is that 5:5:1 is developing solvent, launch, take out, dry; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the glassy yellow speckle of same color;
(4) discriminating of Rhizoma Et Radix Notopterygii: get this product content 3g, use ethyl acetate extraction 4 times, each 10ml, combined ethyl acetate liquid, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Make negative need testing solution with method; Other gets Rhizoma Et Radix Notopterygii control medicinal material powder 1.0g, adds ethanol 30ml, floods 1 hour, filters, and filtrate evaporate to dryness, residue add ethanol 1ml and dissolve medical material solution in contrast; An appendix VIB is according to the thin layer chromatography test for Chinese Pharmacopoeia version in 2005, draw each 5 μ l of need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with ethyl acetate-methanol-water ratio is that the upper solution of 10:2:3 is developing solvent, launch, take out, dry; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
The composition that contains is carried out assay, and step is:
[assay]
1), 2005 editions appendix VID of chlorogenic acid Chinese Pharmacopoeia test according to high effective liquid chromatography for measuring chromatographic condition and system suitability: with octadecylsilane chemically bonded silica is filler; Acetonitrile-0.4% phosphoric acid solution ratio is that 13:87 is a mobile phase; Detect wavelength 327nm, number of theoretical plate must not calculate by the chlorogenic acid peak and is lower than 2000,
The preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, puts in the brown measuring bottle, and add 50% methanol and make the solution that every 1ml contains 20 μ g,
The preparation of need testing solution: precision takes by weighing this product powder 0.3g, puts in the tool plug conical flask, and precision adds 50% methanol 50ml, claims decide weight, and supersound process 30 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, that is,
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, that is,
The every gram of this product contains Flos Lonicerae and Rhizoma Bistortae by chlorogenic acid C
16H
18O
9Meter must not be less than 3.0mg,
2), acetaminophen, caffeine and 2005 editions appendix VID of amobarbital Chinese Pharmacopoeia are according to high effective liquid chromatography for measuring
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; The ratio of methanol-water is that 50:50 is a mobile phase; Detect wavelength 215nm, number of theoretical plate calculates by acetaminophen peak, caffeine peak-to-peak and amobarbital peak and all is not less than 3000,
The preparation of reference substance solution: it is an amount of that precision takes by weighing acetaminophen, caffeine and amobarbital reference substance, and add 50% methanol and make every 1ml and contain acetaminophen 500 μ g and contain the respectively reference substance mixed solution of 50 μ g of caffeine and amobarbital, that is,
The preparation of need testing solution: precision takes by weighing this product powder 0.5g, puts in the tool plug conical flask, and precision adds 50% methanol 50ml, claims decide weight, and supersound process 20 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, that is,
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, that is,
This product contains acetaminophen, caffeine and amobarbital and is 85%～115% of labelled amount.
2, a kind of detection method of new compound Folium Isatidis capsule is characterized in that, comprises the discriminating of content, and the composition that contains is carried out the step of assay, and described method step is as follows:
The discriminating of content, step is:
The discriminating of [discriminating] (1) Folium Isatidis: get this product content 5g, add water 80ml, 80 ℃ of water-bath warm macerating, made dissolving in 30 minutes, and left standstill and put coldly, filter, filtrate is used ethyl acetate extraction 3 times, each 30ml, combined ethyl acetate liquid, with 1% sodium hydroxide solution washing 3 times, each 20ml, combined ethyl acetate liquid, water bath method, residue adds the 0.5ml ethyl acetate makes dissolving, as need testing solution; It is an amount of that other gets the indirubin reference substance, adds ethyl acetate and make the solution that every 1ml contains 0.1mg, in contrast product solution; 2005 editions appendix VIB of Chinese Pharmacopoeia are according to the thin layer chromatography test, draw need testing solution 5-10 μ l, reference substance solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethyl starch sodium, with toluene-ethyl acetate-acetone ratio is that 5:4:1 is developing solvent, launch, take out, dry; In the test sample chromatograph, with contrast chromatograph corresponding position, show the lavender speckle of same color;
(2) discriminating of Radix Et Rhizoma Rhei: get this product content 1.5g, add methanol 30ml supersound process 15 minutes, filter, filtrate evaporate to dryness, residue add water 10ml makes dissolving, adds hydrochloric acid 1ml again, put warm macerating 30min in 80 ℃ of water-baths, cooling immediately extracts twice with the ethyl acetate jolting, each 10ml, combined ethyl acetate liquid, water bath method, residue add methanol 1ml makes dissolving, as need testing solution; Get Radix Et Rhizoma Rhei control medicinal material 0.5g, add methanol 20ml, prepare control medicinal material solution according to the test sample preparation method; Other gets the chrysophanol reference substance, adds methanol and makes the solution that 1ml contains 0.5mg, in contrast product solution; 2005 editions appendix VIB of Chinese Pharmacopoeia are according to the thin layer chromatography test, draw control medicinal material solution and reference substance solution each 2 μ l, need testing solution 5 μ l, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethyl starch sodium, with 30-60 ℃ of petroleum ether-Ethyl formate-formic acid ratio is that the upper solution of 15:5:1 is developing solvent, launch, take out, dry, put under the 365nm uviol lamp and inspect, in the test sample chromatograph, with control medicinal material, the corresponding position of reference substance chromatograph on show the fluorescence speckle of same color;
(3) ascorbic discriminating: get this product content 3g, add 0.1mol/l hydrochloric acid solution 50ml, jolting 30 minutes filters, and gets subsequent filtrate 10ml, adds phenylhydrazine hydrochloride 0.1g, 60 ℃ of water-bath warm macerating 1 hour, and jolting is constantly put coldly, filters, as need testing solution; Other gets vitamin C reference substance 50mg, adds 0.1mol/l hydrochloric acid solution 10ml and phenylhydrazine hydrochloride 0.1g, makes with method, in contrast product solution; An appendix VIB is according to the thin layer chromatography test for Chinese Pharmacopoeia version in 2005, draw each 10 μ l of need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with toluene-ethyl acetate acetone ratio is that 5:5:1 is developing solvent, launch, take out, dry; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the glassy yellow speckle of same color;
(4) discriminating of Rhizoma Et Radix Notopterygii: get this product content 3g, use ethyl acetate extraction 4 times, each 10ml, combined ethyl acetate liquid, evaporate to dryness, residue add ethanol 1ml makes dissolving, as need testing solution; Make negative need testing solution with method; Other gets Rhizoma Et Radix Notopterygii control medicinal material powder 1.0g, adds ethanol 30ml, floods 1 hour, filters, and filtrate evaporate to dryness, residue add ethanol 1ml and dissolve medical material solution in contrast; An appendix VIB is according to the thin layer chromatography test for Chinese Pharmacopoeia version in 2005, draw each 5 μ l of need testing solution and reference substance solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with the ratio of ethyl acetate-methanol-water is that the upper solution of 10:2:3 is developing solvent, launch, take out, dry; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color;
The composition that contains is carried out assay, and step is:
[assay]
1), 2005 editions appendix VID of chlorogenic acid Chinese Pharmacopoeia are according to high effective liquid chromatography for measuring
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; The ratio of acetonitrile-0.4% phosphoric acid solution is that 13:87 is a mobile phase; Detect wavelength 327nm; Number of theoretical plate must not calculate by the chlorogenic acid peak and is lower than 2000;
The preparation of reference substance solution: it is an amount of that precision takes by weighing the chlorogenic acid reference substance, puts in the brown measuring bottle, adds 50% methanol and make the solution that every 1ml contains 20 μ g;
The preparation of need testing solution: precision takes by weighing this product powder 0.3g, puts in the tool plug conical flask, and precision adds 50% methanol 50ml, claims decide weight, and supersound process 30 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, promptly;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
The every gram of this product contains Flos Lonicerae and Rhizoma Bistortae by chlorogenic acid C
16H
18O
9Meter must not be less than 3.0mg;
2) 2005 editions appendix VID of caffeine and amobarbital Chinese Pharmacopoeia are according to high effective liquid chromatography for measuring;
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; The ratio of methanol-water is that 50:50 is a mobile phase; Detect wavelength 215nm; Number of theoretical plate calculates by caffeine peak and amobarbital peak and all is not less than 3000;
The preparation of reference substance solution: precision takes by weighing caffeine and the amobarbital reference substance is an amount of, adds 50% methanol solution and makes the solution that every 1ml contains 50 μ g, promptly;
The preparation of need testing solution: precision takes by weighing this product powder 0.1g, puts in the tool plug conical flask, and precision adds 50% methanol solution 50ml, claims decide weight, and supersound process 20 minutes is put coldly, claims to decide weight again, supplies with 50% methanol to subtract weight loss, shake up, and filtration, promptly;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
This product contains caffeine and amobarbital is 85%～115%. of labelled amount
3) 2005 editions appendix VID of vitamin C and acetaminophen Chinese Pharmacopoeia are according to high effective liquid chromatography for measuring
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; The ratio of acetonitrile-0.4 phosphoric acid solution is that 13:87 is a mobile phase; Detect wavelength 242nm; Number of theoretical plate calculates by vitamin C peak and acetaminophen peak and all is not less than 3000;
The preparation of reference substance solution: precision takes by weighing vitamin C and the acetaminophen reference substance is an amount of, adds mobile phase and makes the mixed solution that every 1ml contains vitamin C 30 μ g and contains acetaminophen 250 μ g, promptly;
The preparation of need testing solution: precision takes by weighing this product powder 0.5g, puts in the tool plug conical flask, and precision adds mobile phase 50ml, claims decide weight, and supersound process 20 minutes is put coldly, claims to decide weight again, supplies with mobile phase to subtract weight loss, shake up, and filtration, promptly;
Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;
This product contains vitamin C and is not less than 70% of labelled amount, and containing acetaminophen is 85%～115% of labelled amount.
According to the method for claim 1, it is characterized in that 3, described new compound Folium Isatidis capsule is to be made by the raw material of following weight: compound Folium Isatidis extract 200g, acetaminophen 75g, caffeine 7.5g, amobarbital 7.5g, vitamin C 10g.
According to the method for claim 1, it is characterized in that 4, described new compound Folium Isatidis capsule prepares by the following method: compound Folium Isatidis extract 200g, acetaminophen 75g, caffeine 7.5g, amobarbital 7.5g, vitamin C 10g, pulverize separately, mixing adds right amount of auxiliary materials, makes granule, dress up 1000 of capsules, promptly.
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| CN101698087B (en) * | 2009-11-06 | 2012-10-31 | 合肥神鹿双鹤药业有限责任公司 | Detection method of Yidan tablets |
| CN104345108B (en) * | 2013-07-29 | 2017-02-22 | 九芝堂股份有限公司 | Qualitative quantitative determination method for liver-heat-clearing tablet |
| CN103512983B (en) * | 2013-10-12 | 2015-09-09 | 中国热带农业科学院分析测试中心 | The high-performance liquid chromatogram determination method of caffeine in a kind of coffee and goods |
| CN103926369B (en) * | 2014-05-06 | 2015-06-17 | 山东师范大学 | Extraction method of biotin in molasses and thin layer chromatography (TLC) scanning detection method thereof |
| CN105738536A (en) * | 2016-04-22 | 2016-07-06 | 广西壮族自治区梧州食品药品检验所 | Extraction method for indirubin in folium isatidis |
| CN106248823A (en) * | 2016-07-21 | 2016-12-21 | 李时珍医药集团有限公司 | A kind of moxa roll Content of Chlorogenic Acid detection method |
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| CN107655993A (en) * | 2017-09-28 | 2018-02-02 | 广东省药品检验所（广东省药品质量研究所、广东省口岸药品检验所） | A kind of method of paracetamol and other chemicalses in detection herbal beverage |
| CN108918734B (en) * | 2018-07-31 | 2021-06-22 | 四川新绿色药业科技发展有限公司 | High performance liquid chromatography method for detecting characteristic spectrums of folium isatidis medicinal materials, decoction pieces, standard decoction and formula granules |
| CN108918721A (en) * | 2018-08-01 | 2018-11-30 | 山西大学 | A kind of measuring method of tussilago Content of Chlorogenic Acid |
| CN108982720B (en) * | 2018-10-08 | 2021-10-08 | 湖南省中医药研究院 | Construction method and application of HPLC standard fingerprint of compound Ganmaoling granules |
| CN111103362A (en) * | 2018-10-26 | 2020-05-05 | 荣昌制药(淄博)有限公司 | Detection method of compound folium isatidis composition |
| CN109674870A (en) * | 2019-01-08 | 2019-04-26 | 成都威迪特生物科技有限公司 | A kind of antibacterial growth promotion Chinese medicine composition and preparation method thereof and detection method |
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