CN101698034B - Method for controlling quality of modified ageratum pills - Google Patents
Method for controlling quality of modified ageratum pills Download PDFInfo
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- CN101698034B CN101698034B CN2009101936431A CN200910193643A CN101698034B CN 101698034 B CN101698034 B CN 101698034B CN 2009101936431 A CN2009101936431 A CN 2009101936431A CN 200910193643 A CN200910193643 A CN 200910193643A CN 101698034 B CN101698034 B CN 101698034B
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Abstract
The invention discloses a method for detecting the modified inspirex extract, which distinguishes the thin-layer chromatography of officinal magnolia bark, pogostemon cablin and radix angelicae and detects the content of hesperidin in dried tangerine or orange peel. The method improves the quality standard of the modified inspirex extract, has higher specificity and good reproducibility, and is good for effectively controlling the content of the finished product of the modified inspirex extract.
Description
Technical field
The present invention relates to a kind of detection method of JIAWEI HUOXIANG ZHENGQI WAN.
Background technology
JIAWEI HUOXIANG ZHENGQI WAN (HUOXIANG ZHENGQI WAN) records in the 14 in " Drug Standard of Ministry of Public Health of the Peoples Republic of China " Chinese traditional patent formulation preparation, and standard No. is WS
3-B-2682-97.JIAWEI HUOXIANG ZHENGQI WAN is processed by ten Herba indigoferae Pseudotinctoriae such as Herba Pogostemonis, Folium Perillaes, has the removing dampness of inducing sweat, regulate the flow of vital energy with in effect, be a kind of affection of exogenous wind-cold that is used to treat, the internal injury humidity hysteresis, headache dusk is heavy, the chest and diaphragm painful abdominal mass is vexed, abdominal distention, the Chinese medicine that vomiting is had loose bowels.The JIAWEI HUOXIANG ZHENGQI WAN primary standard has only microscopical identification, is difficult to effectively control the quality of finished product.
Summary of the invention
The objective of the invention is for a kind of detection method of the JIAWEI HUOXIANG ZHENGQI WAN that can control effectively to the quality of JIAWEI HUOXIANG ZHENGQI WAN finished product is provided.
For realizing above-mentioned purpose, the technical scheme below the present invention has taked:
A kind of detection method of JIAWEI HUOXIANG ZHENGQI WAN is characterized in that: one or multinomial detection the below adopting in discriminating and the assay project:
(1) thin layer of Cortex Magnoliae Officinalis and Herba Pogostemonis is differentiated: get JIAWEI HUOXIANG ZHENGQI WAN finished product 4-8g, and porphyrize, the 20-30ml that adds diethyl ether, jolting, dipping spends the night, and filters, and filtrating volatilizes, and residue adds ethanol 1-3ml makes dissolving, as need testing solution; Other gets Cortex Magnoliae Officinalis control medicinal material 0.5-1.5g, shines medical material solution in pairs with legal system; Get the patchouli alcohol reference substance again, add ethyl acetate and process the solution that every 1ml contains 1.5-2.5mg, as reference substance solution; According to the test of 2005 editions one appendix VI B of Chinese Pharmacopoeia thin layer chromatography, draw each 10 μ l of need testing solution and control medicinal material solution, reference substance solution 5 μ l, put respectively on same silica gel thin-layer plate; 60～90 ℃ of petroleum ether-ethyl acetate-formic acid that with the volume ratio are 80-90: 10-20: 1-3 are developing solvent; Exhibition is taken out to about 15cm, dries; Putting wavelength is to inspect under the 254nm ultra-violet lamp; In the test sample chromatograph, with the corresponding position of Cortex Magnoliae Officinalis control medicinal material chromatograph on, show the fluorescent quenching speckle of same color; Spray is with 5% vanillin sulfuric acid solution, and 105 ℃ are heated to clear spot; In the test sample chromatograph, with the corresponding position of patchouli alcohol reference substance chromatograph on, show the speckle of same color;
(2) thin layer of the Radix Angelicae Dahuricae is differentiated: get JIAWEI HUOXIANG ZHENGQI WAN finished product 15-20g, porphyrize adds water 100-200ml and decocts; Put coldly, the centrifuging and taking supernatant is transferred pH to 1～2 with hydrochloric acid; With chloroform extraction 2-3 time, each 15-25ml merges chloroform liquid; Evaporate to dryness, residue add chloroform 1-2ml makes dissolving, as need testing solution; Other gets Radix Angelicae Dahuricae control medicinal material 0.5-1.5g, shines medical material solution in pairs with legal system; According to 2005 editions one appendix VIB thin layer chromatography test of Chinese Pharmacopoeia, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate; Chloroform-the methanol that with the volume ratio is 8-12: 0.5-15 is developing solvent; Put in the expansion cylinder presaturation 15-25 minute, exhibition is to about 15cm, taking-up; Dry, putting wavelength is to inspect under the 365nm ultra-violet lamp; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
(3) Determination of Hesperidin Content in the high effective liquid chromatography for measuring Pericarpium Citri Reticulatae may further comprise the steps:
1) chromatographic condition: with the octadecylsilane chemically bonded silica is filler, is that acetonitrile-0.2% phosphoric acid solution of 20-24: 76-80 is a mobile phase with the volume ratio, and the detection wavelength is 283nm, and number of theoretical plate calculates by the Hesperidin peak should be not less than 2000;
2) preparation of reference substance solution: it is an amount of to get the Hesperidin reference substance, and accurate the title decides, and adds methanol and processes the solution that every 1ml contains 0.1-0.2mg, promptly gets reference substance solution;
3) preparation of need testing solution: it is an amount of to get the JIAWEI HUOXIANG ZHENGQI WAN finished product, and porphyrize is got about 1g, and accurate the title decides; Put in the tool plug conical flask, the accurate methanol 50ml that adds claims to decide weight, and heating in water bath refluxes; Put coldly, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol; Filter, get subsequent filtrate, promptly get need testing solution;
4) algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly get; Calculate according to the JIAWEI HUOXIANG ZHENGQI WAN finished product, every gram JIAWEI HUOXIANG ZHENGQI WAN contains Pericarpium Citri Reticulatae in Hesperidin, and it is qualified being no less than setting value.
Every gram JIAWEI HUOXIANG ZHENGQI WAN contains Pericarpium Citri Reticulatae in Hesperidin, and setting value is 3.2mg.
Above-mentioned quality determining method need not carry out according to sequencing; And can also carry out conventional sense simultaneously; Observation and microscopical identification like shape; That is: get the JIAWEI HUOXIANG ZHENGQI WAN finished product, put microscopically and observe: irregular particle shape agglomerate and divide dendritic agglomerate colourless, meet chloral hydrate liquid and gradually dissolve; Hyphae colorless or light brown, elongated, crooked slightly, branch is arranged, diameter 3～8 μ m (Poria).Prism of calcium oxalate is present in (Pericarpium Citri Reticulatae) in the parenchyma cell in flakes.Needle-like calcium oxalate crystal is tiny, and long 10～32 μ m are present in (Rhizoma Atractylodis Macrocephalae) in the parenchyma cell.The needle-like calcium oxalate crystal bundle is present in the mucilage cell, or is dispersed in, long 20～144 μ m (Rhizoma Pinelliae) of needle.The fiber bunchy, parenchyma cell contains prism of calcium oxalate on every side, forms crystalline cellulose (Radix Glycyrrhizae).The stone cell class is square, oval, oval or irregular branch are dendritic, visible sometimes laminated striation (Cortex Magnoliae Officinalis).This finished product also should meet each item regulation relevant under the Chinese Pharmacopoeia pill item.
The JIAWEI HUOXIANG ZHENGQI WAN finished product that meets above condition is qualified.
The present invention has revised and enlarged Cortex Magnoliae Officinalis and Herba Pogostemonis, the Radix Angelicae Dahuricae on the basis of initial quality standard thin layer chromatography differentiate and Pericarpium Citri Reticulatae in Determination of Hesperidin Content measure; The present invention has improved the quality standard of JIAWEI HUOXIANG ZHENGQI WAN; Have stronger specificity and good repeatability, help realizing effective control the quality of JIAWEI HUOXIANG ZHENGQI WAN finished product.
The specific embodiment
Specify the present invention below in conjunction with embodiment.
Embodiment 1:
The JIAWEI HUOXIANG ZHENGQI WAN that the present embodiment detection method selects for use Guangzhou Zhongyi Medicine Industry Co., Ltd to produce is a test sample, and these article are bluish yellow color to brown xanchromatic concentrated watered pill; Gas fragrance, sweet in the mouth, little hardship.
The detection method of this JIAWEI HUOXIANG ZHENGQI WAN comprises following discriminating and assay:
A, discriminating
(1) discriminating of Cortex Magnoliae Officinalis and Herba Pogostemonis: get JIAWEI HUOXIANG ZHENGQI WAN finished product 6g, porphyrize, the 25ml that adds diethyl ether, jolting, dipping spends the night, and filters, and filtrating volatilizes, and residue adds ethanol 2ml makes dissolving, as need testing solution.Other gets Cortex Magnoliae Officinalis control medicinal material 1g, shines medical material solution in pairs with legal system.Get the patchouli alcohol reference substance again, add ethyl acetate and process the solution that every 1ml contains 2mg, as reference substance solution.According to thin layer chromatography (2005 editions one appendix VI B of Chinese Pharmacopoeia) test, draw each 10 μ l of need testing solution and control medicinal material solution, reference substance solution 5 μ l, put in same silica gel G F respectively
254On the lamellae, be developing solvent with petroleum ether (60～90 ℃)-ethyl acetate-formic acid (85: 15: 2), exhibition is to about 15cm; Take out, dry, put under the ultra-violet lamp (254nm) and inspect; In the test sample chromatograph, with the corresponding position of Cortex Magnoliae Officinalis control medicinal material chromatograph on, show the fluorescent quenching speckle of same color; Spray is with 5% vanillin sulfuric acid solution, and 105 ℃ are heated to clear spot.In the test sample chromatograph, with the corresponding position of patchouli alcohol reference substance chromatograph on, show the speckle of same color.
(2) discriminating of the Radix Angelicae Dahuricae: get JIAWEI HUOXIANG ZHENGQI WAN finished product 18g, porphyrize adds water 150ml and decocted 30 minutes, puts cold; Centrifugal (1500 rev/mins) 10 minutes are got supernatant, transfer pH to 1～2 with hydrochloric acid; With chloroform extraction 2 times, each 20ml merges chloroform liquid; Evaporate to dryness, residue add chloroform 1ml makes dissolving, as need testing solution.Other gets Radix Angelicae Dahuricae control medicinal material 1g, shines medical material solution in pairs with legal system.According to thin layer chromatography (2005 editions appendix VIB of Chinese Pharmacopoeia) test, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate; With chloroform-methanol (10: 1) is developing solvent; Put in the expansion cylinder presaturation 20 minutes, exhibition is to about 15cm, taking-up; Dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
B, assay
According to Determination of Hesperidin Content in an appendix VI of Chinese Pharmacopoeia version in 2005 the D high effective liquid chromatography for measuring Pericarpium Citri Reticulatae.Determination step comprises:
(1) test of chromatographic condition and system suitability is a filler with the octadecylsilane chemically bonded silica; With acetonitrile-0.2% phosphoric acid solution (22: 78) is mobile phase; The detection wavelength is 283nm.Number of theoretical plate calculates by the Hesperidin peak should be not less than 2000.
(2) it is an amount of that the Hesperidin reference substance is got in the preparation of reference substance solution, and accurate the title decides, and adds methanol and processes the solution that every 1ml contains 0.15mg, promptly gets.
(3) preparation of need testing solution is got these article in right amount, and porphyrize is got about 1g, and accurate the title decides, and puts in the tool plug conical flask; The accurate methanol 50ml that adds claims decide weight, and heating in water bath refluxed 30 minutes, puts coldly, and weight decided in title again; Supply the weight that subtracts mistake with methanol, shake up, filter, get subsequent filtrate, promptly get.
(4) accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of algoscopy inject chromatograph of liquid, measure, and promptly get.
The every 1g of these article contains Pericarpium Citri Reticulatae with Hesperidin (C
28H
34O
15) meter, must not be less than 3.2mg.
Above-mentioned quality determining method need not carry out according to sequencing, and can also carry out conventional sense simultaneously, and like the observation and the microscopical identification of shape, this finished product also should meet each item regulation relevant under an appendix I of Chinese Pharmacopoeia version in 2005 the A pill item.
The JIAWEI HUOXIANG ZHENGQI WAN finished product that meets above condition is qualified.
Below the methodology of quality determining method of the present invention is investigated:
The methodology checking that A, Cortex Magnoliae Officinalis and Herba Pogostemonis are differentiated
1. specificity is investigated
Draw need testing solution, (Nat'l Pharmaceutical & Biological Products Control Institute provides the Cortex Magnoliae Officinalis control medicinal material; Lot number: 121285-200301) solution 10 μ l, Cortex Magnoliae Officinalis negative control solution 10 μ l, (Nat'l Pharmaceutical & Biological Products Control Institute provides the patchouli alcohol reference substance; Lot number: 110772-200404) solution 5 μ l, Herba Pogostemonis negative control solution 10 μ l; Put respectively on same silica GF254 lamellae, make an experiment by discrimination method of the present invention.The result does not see that the Cortex Magnoliae Officinalis negative control differentiates that to the thin layer chromatography of patchouli alcohol interference is arranged to the thin layer chromatography of Cortex Magnoliae Officinalis and Herba Pogostemonis negative control.
2. ruggedness is investigated
2.1 the comparison of different lamellaes
Get the silica gel G F of hands shop
254Lamellae and prefabricated silica gel G F
254Lamellae makes an experiment by discrimination method of the present invention respectively, and the result shows the separating effect basically identical of manual bed board of self-control and precoated plate.
2.2 temperature is investigated
Get the lamellae behind the point sample, by by discrimination method of the present invention expansion in 4 ℃ and 35 ℃ respectively.The result is presented at test in 4 ℃～35 ℃ temperature ranges, and temperature differentiates not have obviously influence to this.
2.3 humidity is investigated
Get the lamellae behind the point sample, regulate in the relative humidity 20% in normal humidity 65% with sulphuric acid respectively by discrimination method of the present invention and launch.The result is presented to test in 20%～65% relative humidity scope does not have significantly influence to differentiating.
Learn demonstration test through said method, it is little that the result shows that lamellae, humiture change launching influential effect, therefore, thinks the good tolerance of Cortex Magnoliae Officinalis and Herba Pogostemonis discrimination method.
The methodology checking that B, the Radix Angelicae Dahuricae are differentiated
1. specificity is investigated
Draw each 10 μ l of need testing solution and negative control solution, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate, by discrimination method test of the present invention.The result does not see that Radix Angelicae Dahuricae negative control has interference to the thin layer chromatography discriminating of the Radix Angelicae Dahuricae.
2. ruggedness is investigated
2.1 the comparison of different lamellaes
Get the silica gel g thin-layer plate and the prefabricated silica gel g thin-layer plate of hands shop, respectively by discrimination method experiment of the present invention.As a result, the separating effect of precoated plate is superior to manual bed board.
2.2 temperature is investigated
Get the lamellae behind the point sample, by discrimination method of the present invention expansion in 35 ℃ and 4 ℃ respectively.The result is presented at test in 4 ℃～35 ℃ temperature ranges, to not significantly influence of result.
2.3 humidity is investigated
Get the lamellae behind the point sample, regulate in normal humidity (65%) with sulphuric acid respectively in the relative humidity (20%) by discrimination method of the present invention and launch.The result is presented to test in 20%～65% relative humidity scope does not have significantly influence to differentiating.
Learn demonstration test through said method, the result shows the good tolerance of Radix Angelicae Dahuricae discrimination method.
Content of hesperidin method for measuring in C, the Pericarpium Citri Reticulatae
Used instrument is a SHIMADZU LC-2010A high performance liquid chromatograph; Data processing software system: SHIMADZU LCsolution; Ultrasound Instrument: KQ-300DA: ultraviolet spectrometer: UV-2450.
1 chromatographic condition and system suitability test
1.1 the selection of mobile phase
Mobile phase list of references " content of hesperidin in the high effective liquid chromatography for measuring HUOXIANGZHENGQI KOUFUYE " is also changed slightly, selects for use acetonitrile-0.2% phosphoric acid solution (22: 78) to be mobile phase.
1.2 measure the selection of wavelength
Precision takes by weighing Hesperidin reference substance (lot number: 110721-200612, assay usefulness, Nat'l Pharmaceutical & Biological Products Control Institute) 10.36mg; Put in the 250ml measuring bottle; Add dissolve with methanol and be diluted to scale, in the interscan of 200～380nm wave-length coverage, the result shows; Hesperidin has absorption maximum at the 283nm place, and is consistent with the detection wavelength (283nm) that Chinese Pharmacopoeia was adopted under a medical material Pericarpium Citri Reticulatae item in 2005.So selected 283nm is as the mensuration wavelength of these article.
1.3 chromatographic column
With reference to " method of 2005 editions one " Pericarpium Citri Reticulatae " [assay] of Chinese pharmacopoeia selects for use reverse HPLC-UV-detector method to measure, and selecting for use with the octadecylsilane chemically bonded silica is the chromatographic column of filler.
The checking of 2 methodologies
2.1 accuracy
2.1.1 application of sample reclaims the reference substance stock solution preparation of usefulness
Precision takes by weighing Hesperidin reference substance 10.50mg, puts in the 50ml measuring bottle, adds methanol and makes dissolving in right amount, and be diluted to scale, shakes up, as stock solution (0.2100mg/ml).
2.1.2 application of sample reclaims the preparation of need testing solution
Get totally 6 parts in concentrated pill (Guangzhou Zhongyi Medicine Industry Co., Ltd produce, lot number K00123 the contains Hesperidin 8.46mg/g) powder of same lot number, get about 0.5g, the accurate title calmly; Put in the tool plug conical flask, the accurate respectively reference substance stock solution 20ml that adds above-mentioned application of sample recovery usefulness, the accurate again methanol 30ml that adds claims to decide weight; Heating in water bath refluxed 30 minutes, put coldly, claimed to decide weight again, supplied the weight that subtracts mistake with methanol; Shake up, filter, get subsequent filtrate, promptly get by (table 1).
Table 1 accuracy test result
2.2 precision (replica test)
Get totally 6 parts in concentrated pill (Guangzhou Zhongyi Medicine Industry Co., Ltd produce, the lot number K00123) powder of same lot number, get about 1g, accurately claim surely, press the need testing solution method for preparing and prepare, mensuration promptly gets.Result's repeatability good (table 2).
Table 2 replica test result
2.3 specificity
Accurate reference substance solution, need testing solution, each 10 μ l of negative control solution of drawing; Inject high performance liquid chromatograph respectively; Press content assaying method and measure, as a result negative control solution with the corresponding position of Hesperidin reference substance chromatographic retention on noiseless.
2.4 it is linear
Precision takes by weighing Hesperidin reference substance 19.32mg, puts in the 20ml measuring bottle, adds methanol and makes dissolving, is diluted to scale; Shake up, as stock solution (0.9660mg/ml), the accurate stock solution 1ml that draws puts in the 10ml measuring bottle; Add methanol and be diluted to scale, shake up, as stock solution A (0.09660mg/ml); The accurate stock solution 2ml that draws puts in the 10ml measuring bottle, adds methanol and is diluted to scale, shakes up, as stock solution B (0.1932mg/ml); The accurate stock solution 3ml that draws puts in the 10ml measuring bottle, adds methanol and is diluted to scale, shakes up, as stock solution C (0.2898mg/ml).Accurate stock solution A (0.09660mg/ml) 3 μ l, the 10 μ l of drawing, stock solution B (0.1932mg/ml) 10 μ l, 12 μ l, stock solution C (0.2898mg/ml) 10 μ l, 15 μ l inject chromatograph of liquid respectively.With the peak area is that vertical coordinate, content of hesperidin (μ g) are abscissa, obtains regression equation and is: y=1757874.6832x+73713.5567, r=0.9999.Experimental result shows that Hesperidin is good linear relationship (table 3) with peak area in the scope of 0.2898 μ g～4.347 μ g.
The linear result that investigates of table 3
2.5 scope
2.5.1 scope-scope experimental design
Draft [assay] following every 1g and contain Pericarpium Citri Reticulatae with Hesperidin (C
28H
34O
15) meter, must not be less than 3mg.The investigation of scope can be by (table 4) contrived experiment.
Table 4 scope experimental design
2.5.2 scope-accuracy prepares with the reference substance stock solution
Precision takes by weighing Hesperidin reference substance 10.90mg, puts in the 50ml measuring bottle, adds methanol and makes dissolving in right amount, and be diluted to scale, shakes up, as stock solution (0.2180mg/ml).
2.5.3 scope-accuracy prepares with the test sample stock solution
Get totally 6 parts in concentrated pill (Guangzhou Zhongyi Medicine Industry Co., Ltd produce, lot number K00123 the contains Hesperidin 8.46mg/g) powder of same lot number; Get about 0.12g; The accurate title, decide, and puts respectively in the tool plug conical flask, the accurate reference substance stock solution 5ml that adds; The accurate again methanol 45ml that adds presses content assaying method and measures.Experiment shows: the investigation scope of this analytical method is that 2mg/g～8.46mg/g all has good accuracy (seeing table 5) at test sample content.
Table 5 scope-accuracy test is measured the result
2.6 ruggedness
2.6.1 stability test
Get same need testing solution, after the preparation, every certainly at a distance from 2.5 hours, press content assaying method and measure.Need testing solution is stable in 19 hours as a result.The result sees table 6.
Table 6 stability test result
2.6.2 the investigation of method for distilling
Get same lot number (Guangzhou Zhongyi Medicine Industry Co., Ltd produces, and K00123) sample powder is two parts, every part of about 1g, the accurate title, put in the tool plug conical flask calmly; The accurate methanol 50ml that adds claims to decide weight, the mode that adopts supersound process (power 300W, frequency 40kHz) and heating in water bath to reflux respectively; Extraction time is 30 minutes, puts coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol; Shake up, filter, get subsequent filtrate, Determination of Hesperidin Content in the working sample.Test shows that heating in water bath backflow effect is obviously good than supersound process, so select for use heating in water bath to reflux as method for distilling, the result sees table 7.
The comparison of the different extracting modes of table 7
2.6.3 whether adopt the selection of petroleum ether roguing
Get same lot number (Guangzhou Zhongyi Medicine Industry Co., Ltd produces, and K00123) sample powder is two parts, every part of about 1g, the accurate title, put in the tool plug conical flask calmly; Adopt (1) accurate petroleum ether (60～90 ℃) 50ml that adds respectively, heating in water bath refluxed 30 minutes, discarded petroleum ether liquid, and medicinal residues volatilize petroleum ether, the accurate again methanol 50ml that adds; Claim decide weight, heating in water bath refluxed 30 minutes, put coldly, and weight decided in title again, supplies the weight that subtracts mistake with methanol; Shake up, filter, get subsequent filtrate, promptly get; (2) the directly accurate methanol 50ml that adds claims decide weight, and heating in water bath refluxed 30 minutes, puts coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol, shakes up, and filtration is got subsequent filtrate, content of hesperidin in the working sample.The result shows that the impurity peaks quantity after two kinds of methods are handled do not have significant difference, directly adds methanol water-bath reflux (table 8) so select for use.
Whether table 8 adopts the result of petroleum ether roguing
2.6.4 the selection of reflux extracting time
(Guangzhou Zhongyi Medicine Industry Co., Ltd produces, and K00123) sample powder is three parts, every part of about 1g, the accurate title calmly to get same lot number; Put in the tool plug conical flask, the accurate methanol 50ml that adds claims to decide weight, and reflux is 30,45,60 minutes respectively; Put coldly, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol; Filter, get subsequent filtrate, content of hesperidin in the working sample.Test shows that different extraction times are measured the result and are more or less the same, and will be decided to be 30 minutes (seeing table 9) extraction time.
The comparison of table 9 different extraction times
2.6.5 extract the investigation of solvent load
(Guangzhou Zhongyi Medicine Industry Co., Ltd produces, and K00123) sample powder is totally three parts, every part of about 1g, the accurate title calmly to get same lot number; Put in the tool plug conical flask, accurate respectively methanol 25,50, the 75ml of adding claims to decide weight, and heating in water bath refluxed 30 minutes; Put coldly, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol; Filter, get subsequent filtrate, content of hesperidin in the working sample.Test shows that the mensuration result of consumption 25ml is low than the mensuration result of 50ml, 75ml, and the result of 50ml, 75ml mensuration is more or less the same, and is chosen as 50ml (table 10) so extract the consumption of solvent.
The different comparisons of extracting quantity of solvent of table 10
2.6.6 the comparison of different size chromatographic column
Get same reference substance solution, need testing solution (Guangzhou Zhongyi Medicine Industry Co., Ltd's production; Lot number: K00009); Press the content assaying method operation, investigate the influence degree of different chromatographic columns to content of hesperidin mensuration result, test shows; Use different manufacturers' chromatographic column to analyze, the gained result does not have significance difference (table 11).
The mensuration result of the different chromatographic columns of table 11
Remarks: A.Agilent Eclipse XDB-C18 (4.6*150mm*5 μ m); B.Diamonsil diamond C18 (4.6*200mm*5 μ m);
C.Waters?SymmetryShield?RP-C18(4.6*150mm*5μm)
3 test product assays
Get the concentrated pill of different product batch numbers (Guangzhou Zhongyi Medicine Industry Co., Ltd's production), press the content assaying method operation, measure, the result sees table 12.
Ten crowdes of assay results of table 12
Confirming of 4 content limits
Above-mentioned 10 batches of average contents are 8.86mg/g.Because in these 10 lot sample article, the feed intake content of hesperidin of the Pericarpium Citri Reticulatae of selecting for use of producer is high, so improper with this limit foundation as assay.Consider that the sample that acceptable material, reasonable technology are produced is certified products, so it is more reasonable to establish the limits with the minimum content of Hesperidin in the Pericarpium Citri Reticulatae of pharmacopeia regulation.
Calculate with dry product by pharmacopeia regulation Pericarpium Citri Reticulatae; Content of hesperidin must not be lower than 3.5% (moisture must not cross 13%, and promptly Pericarpium Citri Reticulatae contains Hesperidin and should not be lower than 3.045%), and the powder pill rate of transform that feeds intake should not be lower than 90% and calculates; Conversion stipulates that with pharmacopeia the Pericarpium Citri Reticulatae of minimum content of hesperidin feeds intake; Content of hesperidin is 3.2mg/g in the JIAWEI HUOXIANG ZHENGQI WAN, as certificate, drafts these article and contains Pericarpium Citri Reticulatae with Hesperidin (C
28H
34O
15) meter, must not be less than 3.2mg/g.
Claims (2)
1. the detection method of a JIAWEI HUOXIANG ZHENGQI WAN is characterized in that: differentiate below said detection method adopts and the assay project in one or multinomial:
(1) thin layer of Cortex Magnoliae Officinalis and Herba Pogostemonis is differentiated: get JIAWEI HUOXIANG ZHENGQI WAN finished product 4-8g, and porphyrize, the 20-30ml that adds diethyl ether, jolting, dipping spends the night, and filters, and filtrating volatilizes, and residue adds ethanol 1-3ml makes dissolving, as need testing solution; Other gets Cortex Magnoliae Officinalis control medicinal material 0.5-1.5g, shines medical material solution in pairs with legal system; Get the patchouli alcohol reference substance again, add ethyl acetate and process the solution that every 1ml contains 1.5-2.5mg, as reference substance solution; According to the test of 2005 editions one appendix VI B of Chinese Pharmacopoeia thin layer chromatography, draw each 10 μ l of need testing solution and control medicinal material solution, reference substance solution 5 μ l, put respectively on same silica gel thin-layer plate; 60～90 ℃ of petroleum ether-ethyl acetate-formic acid that with the volume ratio are 80-90: 10-20: 1-3 are developing solvent; Exhibition is taken out to about 15cm, dries; Putting wavelength is to inspect under the 254nm ultra-violet lamp; In the test sample chromatograph, with the corresponding position of Cortex Magnoliae Officinalis control medicinal material chromatograph on, show the fluorescent quenching speckle of same color; Spray is with 5% vanillin sulfuric acid solution, and 105 ℃ are heated to clear spot; In the test sample chromatograph, with the corresponding position of patchouli alcohol reference substance chromatograph on, show the speckle of same color;
(2) thin layer of the Radix Angelicae Dahuricae is differentiated: get JIAWEI HUOXIANG ZHENGQI WAN finished product 15-20g, porphyrize adds water 100-200ml and decocts; Put coldly, the centrifuging and taking supernatant is transferred pH to 1～2 with hydrochloric acid; With chloroform extraction 2-3 time, each 15-25ml merges chloroform liquid; Evaporate to dryness, residue add chloroform 1-2ml makes dissolving, as need testing solution; Other gets Radix Angelicae Dahuricae control medicinal material 0.5-1.5g, shines medical material solution in pairs with legal system; According to the test of 2005 editions one appendix VI B of Chinese Pharmacopoeia thin layer chromatography, draw need testing solution 10 μ l, control medicinal material solution 5 μ l, put respectively on same silica gel g thin-layer plate; Chloroform-the methanol that with the volume ratio is 8-12: 0.5-1.5 is developing solvent; Put in the expansion cylinder presaturation 15-25 minute, exhibition is to about 15cm, taking-up; Dry, putting wavelength is to inspect under the 365nm ultra-violet lamp; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;
(3) Determination of Hesperidin Content in the high effective liquid chromatography for measuring Pericarpium Citri Reticulatae: may further comprise the steps: 1) chromatographic condition: be filler with the octadecylsilane chemically bonded silica; Acetonitrile-0.2% phosphoric acid solution that with the volume ratio is 20-24: 76-80 is a mobile phase; The detection wavelength is 283nm, and number of theoretical plate calculates by the Hesperidin peak should be not less than 2000; 2) preparation of reference substance solution: it is an amount of to get the Hesperidin reference substance, and accurate the title decides, and adds methanol and processes the solution that every 1ml contains 0.1-0.2mg, promptly gets reference substance solution; 3) preparation of need testing solution: it is an amount of to get the JIAWEI HUOXIANG ZHENGQI WAN finished product, and porphyrize is got about 1g, and accurate the title decides; Put in the tool plug conical flask, the accurate methanol 50ml that adds claims to decide weight, and heating in water bath refluxes; Put coldly, claim again to decide weight, supply the weight that subtracts mistake, shake up with methanol; Filter, get subsequent filtrate, promptly get need testing solution; 4) algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly get.
2. the detection method of JIAWEI HUOXIANG ZHENGQI WAN according to claim 1 is characterized in that: calculate according to the JIAWEI HUOXIANG ZHENGQI WAN finished product, every gram JIAWEI HUOXIANG ZHENGQI WAN contains Pericarpium Citri Reticulatae in Hesperidin, and content is more than or equal to 3.2mg.
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| CN1843442A (en) * | 2006-02-13 | 2006-10-11 | 浙江大德药业集团有限公司 | Oral disintegration tablet of 'Huo Xiang Zheng Qi' and preparation method and quality control method |
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| CN1843442A (en) * | 2006-02-13 | 2006-10-11 | 浙江大德药业集团有限公司 | Oral disintegration tablet of 'Huo Xiang Zheng Qi' and preparation method and quality control method |
Non-Patent Citations (2)
| Title |
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| 刘英敏等.高效液相色谱测定加味藿香正气丸中橙皮苷的含量.《临床和实验医学杂志》.2006,第5卷(第11期),1838. * |
| 张小兵等.加味藿香正气丸的质量标准研究.《解放军药学学报》.2007,第23卷(第4期),291-293. * |
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Owner name: GUANGZHOU BAIYUNSHAN ZHONGYI PHARMACEUTICAL CO., L Free format text: FORMER NAME: GUANGZHOU ZHONGYI PHARMACEUTICAL CO., LTD. |
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Address after: 510530, No. 32, Po pan Road, Guangzhou, Guangdong, Luogang District Patentee after: Guangzhou Baiyunshan Zhongyi Pharmaceutical Co., Ltd. Address before: 510530, No. 32, Po Po Road, Yun Po Industrial Area, Guangzhou, Guangdong, Luogang District Patentee before: Guangzhou Zhongyi Pharmaceutical Co., Ltd. |