CN110231415A - Gallic acid content measuring method in one seed ginseng times Guchang capsule - Google Patents
Gallic acid content measuring method in one seed ginseng times Guchang capsule Download PDFInfo
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Abstract
The invention discloses gallic acid content measuring methods in ginseng times Guchang capsule, include following steps: preparing reference substance solution by solvent of methanol solution: taking the content under this product content uniformity item is finely ground to obtain to test sample；Test solution is prepared by solvent of methanol；Using Determination of Gallic content；Chromatographic test strip part are as follows: using octadecylsilane chemically bonded silica as filler；Mobile phase is 2% methanol-acetonitrile-water-phosphoric acid-triethylamine (3:97:0.1:0.1)；Detection wavelength is 216nm；Sample volume 10uL；The technical solution used in the present invention is solved since other Chinese medicinal ingredients in ginseng times Guchang capsule are easy to cause biggish interference to the detection of gallic acid, leads to be difficult to the problem of rapidly and accurately measuring the content of gallic acid in ginseng times Guchang capsule.
Description
Technical field
The present invention relates to drug ingredient detection technique fields, and in particular to gallic acid content in a seed ginseng times Guchang capsule
Measuring method.
Background technique
A ginseng times Guchang capsule is a kind of for incoordination between the liver and the spleen, diarrhea dysentery abdominal pain, in chronic nonspecific ulcerative colitis
Patent medicine, composition mainly include Chinese gall, nutmeg (stewing), terminalia flesh (stewing), dark plum, radix aucklandiae, rhizoma atractylodis, Poria cocos, cornu cerve degelatinatum,
Red ginseng etc.；Wherein in Chinese gall and terminalia flesh rich in there is gallic acid；Gallic acid is a kind of organic acid, has antibacterial, disease-resistant
Poison, antineoplastic action；Therefore in ginseng times Guchang capsule, the content of gallic acid plays large effect to drug effect；Though at present
So there is the measuring method of gallic acid content in various detection plants, but since other Chinese medicinal ingredients in ginseng times Guchang capsule are held
Biggish interference easily is caused to the detection of gallic acid, causes to be difficult to rapidly and accurately to measure gallic acid in ginseng times Guchang capsule
Content the problem of.
Summary of the invention
The purpose of the present invention is to provide gallic acid content measuring methods in a seed ginseng times Guchang capsule, solve by because of ginseng
Other Chinese medicinal ingredients in times Guchang capsule cause biggish interference to the detection of gallic acid, cause to be difficult to quickly and accurately
The problem of measuring the content of gallic acid in ginseng times Guchang capsule.
In order to solve the above technical problems, the invention adopts the following technical scheme: galla turcica in a seed ginseng times Guchang capsule
Acid content measuring method, includes following steps:
1) reference substance solution is prepared by solvent of methanol solution:
2) take the content under this product content uniformity item is finely ground to obtain to test sample；
3) test solution is prepared by solvent of methanol；
4) Determination of Gallic content is used；
Chromatographic test strip part are as follows: HPLC chromatogram column is Agilent C18, and octadecylsilane chemically bonded silica is filler；Flowing
It is mutually 2% methanol-acetonitrile-water-phosphoric acid-triethylamine (3:97:0.1:0.1)；Detection wavelength is 220nm；Sample volume 20uL；
Further, the concrete operations of reference substance solution are prepared in step 1) are as follows:
Step S1: precision weighs gallic acid reference substance 12.5mg, is placed in 50mL volumetric flask, adds 50% methanol to dissolve and dilutes
It to scale, shakes up, obtains the intermediate solution that concentration is 250ug/mL；
Step S2: precision measures 2mL intermediate solution, is placed in 25mL volumetric flask, adds 50% methanol dilution to scale, shake up
Obtain the reference substance solution that concentration is 20ug/mL；
Further, the concrete operations of test solution are prepared in step 3) are as follows:
Step A1: precision weighs 0.2g and waits for test sample, is placed in 50mL volumetric flask, adds 50% methanol 40mL, is ultrasonically treated 15min, puts
It is cold, add 50% methanol dilution to scale, shakes up, filtrate is obtained after filtering；
Step A2: precision measures 1mL filtrate, is placed in 10mL volumetric flask, adds 50% methanol dilution to scale, shake up, can be obtained
Test sample；
Further, in step A1, ultrasonic hertz is 40KHZ；It is let cool after ultrasound to 20 DEG C~30 DEG C；
Further, the column temperature that chromatography detects in step 1) is 20 DEG C~40 DEG C；
Further, chromatographic column number of theoretical plate presses gallic acid peak and calculates >=3000 in step 1)；
Further technical solution is that the type of elution that chromatography detects in step 1) is isocratic elution.
Compared with prior art, the beneficial effects of the present invention are:
1, the present invention can accurately measure the content of gallic acid in the ginseng times Guchang capsule with more interfering components；
2, by selecting suitable testing conditions, so that the separation of gallic acid content in times Guchang capsule is joined in HPLC measurement
Degree, accuracy, reproducibility, stability etc. all meet standard requirements；
3, the intermediate transfer amount of test solution preparation is increased, more convenient operation, accuracy is higher；
4, the sample volume for reducing reference substance solution and test solution, effectively prevent chromatography column overload, and peak type is more preferable.
Detailed description of the invention
Fig. 1 is reference substance solution chromatogram.
Fig. 2 is test solution chromatogram.
Fig. 3 is gallic acid canonical plotting.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
Gallic acid content measuring method in 1: one seed ginseng times Guchang capsule of embodiment, includes following steps:
1) reference substance solution is prepared by solvent of 50% methanol solution；
2) take the content under this product content uniformity item is finely ground to obtain to test sample；
3) test solution is prepared using 50% methanol as solvent；
4) using the gallic acid content in high effective liquid chromatography for measuring reference substance solution and test sample solution；
Chromatographic test strip part are as follows: using octadecylsilane chemically bonded silica as filler；Mobile phase is 2% methanol-acetonitrile-water-phosphorus
Acid-triethylamine (3:97:0.1:0.1)；Detection wavelength is 216nm；Sample volume 10uL.
Wherein, it selects 2% methanol-acetonitrile-water-phosphoric acid-triethylamine (3:97:0.1:0.1) as mobile phase, can be improved
Separative efficiency reduces interference of other Chinese medicinal ingredients to assay in ginseng times Guchang capsule.
The concrete operations of reference substance solution are prepared in step 1) are as follows:
The concrete operations of reference substance solution are prepared in step 1) are as follows: take gallic acid reference substance appropriate, it is accurately weighed, add 50% first
Alcohol makes solution of the melt into 1mL containing 0.02mg to get reference substance solution.
Methanol source are as follows: the import of the U.S. world, lot number: MS1922-801；Gallic acid reference substance source are as follows: Chinese food
Drug assay research institute, lot number: 110831-201204.
Further, the concrete operations of test solution are prepared in step 3) are as follows:
The concrete operations of test solution are prepared in step 3) are as follows:
Step A1: precision weighs 0.2g and waits for test sample, is placed in 100mL stuffed conical flask, adds 50% methanol 50mL, ultrasonic treatment
15min is let cool, then weighed weight, and the weight of less loss is supplied with 50% methanol, is shaken up, and is abandoned primary filtrate, is taken subsequent filtrate；
Step A2: precision measures 5mL subsequent filtrate, is placed in 50mL volumetric flask, adds 50% methanol dilution to scale, shake up, can obtain
To test sample.
Wherein ultrasonic treatment can speed up medium particle movement, and cavitation, the vibration of ultrasonic wave, which homogenizes, makes sample media
The effect that interior each point is subject to is consistent, makes entire sample extraction more evenly.
It does not need to heat when ultrasonic wave extraction, avoids conventional decocting method, circumfluence method long-time heating to effective component
Adverse effect.Ultrasonic wave extraction improves the recovery rate of effective ingredient, is conducive to making full use of for resource, improves economy
Benefit.Solvent usage is few, has saved solvent.Ultrasonic wave extraction is a physical process, without chemical anti-in entire leaching process
It should occur, not influence the physiological activity of most drugs effective component.Extract active constituent content is high, advantageous dry further essence
System.
Further, in step A1, ultrasonic hertz is 50KHZ；It is let cool after ultrasound to 25 DEG C~35 DEG C.
Further, the column temperature that chromatography detects in step 1) is 25 DEG C~35 DEG C.
Further, chromatographic column number of theoretical plate presses gallic acid peak and calculates >=2500 in step 1).
Further technical solution is that the type of elution that chromatography detects in step 1) is isocratic elution.
Testing result is as depicted in figs. 1 and 2.Wherein, the peak area in spectrogram measured by reference substance solution is
3338.63452 peak height 116.30012, peak width 0.4339；Peak area in spectrogram measured by test solution is
3329.25610 peak height 115.39370, peak width 0.4397；The number of plates 3509.
This method verification result is as follows:
(1) linear to investigate
Precision draws gallic acid reference substance solution (0.023797mg/ml) 2,6,10,14,18,22 μ l sample introduction, records chromatography,
Using peak area as abscissa, it is ordinate mapping with sample volume (μ g), calculates regression equation are as follows: A=7543182.92C-
5794.78 (r=0.9999), regression equation were fitted the linear equation of origin are as follows: A=7528876.953C；One test sample is molten
Sample introduction is analyzed for liquid, and gained peak area substitutes into upper two formula respectively and calculates, relative deviation 0.29%, therefore is believed that standard curve is excessively former
Point, regression equation no intercept.Thus one point external standard method calculating can be used in assay.Between 0.0476 μ of μ g~0.5235 g
Linear relationship is good.
1 gallic acid linear relationship of table is investigated
(2) precision test
Reference substance solution 0.023797mg/ml is taken, is repeated sample introduction 6 times, chromatography is recorded, the results are shown in Table 2 and precision map, RSD
It is 0.19%, illustrates there is good precision.
2 precision test of table
(3) repetitive test
Take a sample by 6 parts of test solutions are prepared described in text, sample introduction, records chromatography respectively, calculates, and the results are shown in Table 3, repeats
Property map, RSD 0.58% illustrate there is good repeatability.
3 repetitive test of table
(4) recovery test
Recycled using sample-adding, precision draws the sample 0.1g(61.82mg/g for having measured content), add gallic acid reference substance
In right amount, it is measured by chromatographic condition described in text, calculates the rate of recovery, the results are shown in Table 4, rate of recovery map, the rate of recovery of gallic acid
For 100.35%, RSD 0.96%, illustrate that this law has the good rate of recovery.
4 recovery test of table
(5) specificity
It takes ginseng times Guchang capsule and negative sample to prepare sample solution and negative sample solution by sample solution preparation method, presses
Above-mentioned chromatographic condition takes reference substance solution, sample solution, negative sample solution liquid to enter liquid chromatograph, sample solution color respectively
Spectrum is equipped with corresponding chromatographic peak in reference substance solution chromatography corresponding positions, and negative noiseless.
(6) durability
The selection of sample extraction solvent: taking the content under content uniformity item, takes 5 parts, every part of about 0.2g, accurately weighed, sets tool
It fills in conical flask, it is accurate respectively to be added 20% methanol, 30% methanol, 40% methanol, 50% methanol, 60% methanol each 50ml, it is weighed heavy
Amount is ultrasonically treated 15 minutes, lets cool, then weighed weight, respectively with 20% methanol, 30% methanol, 40% methanol, 50% methanol, 60% first
Alcohol supplies less loss weight, shakes up, filtration, and the accurate 1ml that draws is set in 10ml measuring bottle respectively, adds 50% methanol to scale, shakes up, use
0.45 μm of miillpore filter filtration, takes filtrate as test solution.Each 10 μ l injection liquid chromatograph of above-mentioned solution is taken respectively,
It calculates, the results are shown in Table 5, illustrate that this product uses 50% methanol as Extraction solvent, content is extracted more complete.
The selection of 5 Extraction solvent of table
Sample extraction time effects: taking the content under content uniformity item, finely ground, takes 3 parts, every part of about 0.2g, accurately weighed, point
It does not set in stuffed conical flask, 50% methanol 50ml is added in precision, ultrasonic extraction 10,15,30 minutes respectively, takes out, lets cool, with 50%
Methanol supplies less loss weight, shakes up, filtration, and the accurate 1ml that draws is set in 10ml measuring bottle respectively, and 50% methanol is added to shake up to scale,
It is filtered with 0.45 μm of miillpore filter, takes filtrate as test solution.10 μ l subsequent filtrates are taken to inject liquid chromatograph, meter respectively
Content is calculated, 7 is the results are shown in Table, just extracts within ultrasonic extraction 15 minutes gallic acid completely, therefore the selective extraction time is 15 minutes.
6 extraction time of table influences
Sample stability test: ginseng times Guchang capsule is taken, is prepared by sample solution preparation method, is measured by above-mentioned chromatographic condition
Gallic acid peak area, the results are shown in Table 7.
The test of 7 sample stability of table
(7) ten batches of assays: 8 be the results are shown in Table.
According to 10 batches of sample measurements, this product every is fixed tentatively containing Chinese gall, terminalia flesh in terms of gallic acid (C7H6O5), no
Less than 14.0mg.
Although reference be made herein to invention has been described for multiple explanatory embodiments of the invention, however, it is to be understood that
Those skilled in the art can be designed that a lot of other modification and implementations, these modifications and implementations will fall in this Shen
It please be within disclosed scope and spirit.It more specifically, within the scope of the present disclosure and claims, can be to master
The building block and/or layout for inscribing composite configuration carry out a variety of variations and modifications.In addition to what is carried out to building block and/or layout
Outside modification and improvement, to those skilled in the art, other purposes also be will be apparent.
Claims (7)
1. gallic acid content measuring method in a seed ginseng times Guchang capsule, which is characterized in that include following steps:
1) reference substance solution is prepared by solvent of 50% methanol solution；
2) take the content under this product content uniformity item is finely ground to obtain to test sample；
3) test solution is prepared using 50% methanol as solvent；
4) using the gallic acid content in high effective liquid chromatography for measuring reference substance solution and test sample solution；
Chromatographic test strip part are as follows: using octadecylsilane chemically bonded silica as filler；Mobile phase is 2% methanol-acetonitrile-water-phosphorus
Acid-triethylamine (3:97:0.1:0.1)；Detection wavelength is 216nm；Sample volume 10uL.
2. gallic acid content measuring method in a seed ginseng times Guchang capsule according to claim 1, it is characterised in that: step
The concrete operations of rapid 1) middle preparation reference substance solution are as follows: take gallic acid reference substance appropriate, it is accurately weighed, add 50% methanol to make molten
Solution at 1mL containing 0.02mg is to get reference substance solution.
3. gallic acid content measuring method in a seed ginseng times Guchang capsule according to claim 1, it is characterised in that: step
It is rapid 3) in prepare the concrete operations of test solution are as follows:
Step A1: precision weighs 0.2g and waits for test sample, is placed in 100mL stuffed conical flask, adds 50% methanol 50mL, ultrasonic treatment
15min is let cool, then weighed weight, and the weight of less loss is supplied with 50% methanol, is shaken up, and is abandoned primary filtrate, is taken subsequent filtrate；
Step A2: precision measures 5mL subsequent filtrate, is placed in 50mL volumetric flask, adds 50% methanol dilution to scale, shake up, can obtain
To test sample.
4. gallic acid content measuring method in a seed ginseng times Guchang capsule according to claim 3, it is characterised in that: step
In rapid A1, ultrasonic hertz is 50KHZ；It is let cool after ultrasound to 25 DEG C~35 DEG C.
5. gallic acid content measuring method in a seed ginseng times Guchang capsule according to claim 1, it is characterised in that: step
It is rapid 3) in chromatography detection column temperature be 25 DEG C~35 DEG C.
6. gallic acid content measuring method in a seed ginseng times Guchang capsule according to claim 1, it is characterised in that: step
It is rapid 3) in chromatographic column number of theoretical plate by gallic acid peak calculate >=2500.
7. gallic acid content measuring method in a seed ginseng times Guchang capsule according to claim 1, it is characterised in that: step
It is rapid 3) in chromatography detection type of elution be isocratic elution.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113504325A (en) * | 2021-07-07 | 2021-10-15 | 江西康恩贝天施康药业有限公司 | Detection method of changyanning preparation |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1712051A (en) * | 2005-06-08 | 2005-12-28 | 叶耀良 | Quality control of Gongyanping preparation |
| CN1994340A (en) * | 2006-11-16 | 2007-07-11 | 胡军 | Quality control method of turpinia indochinensis leaf and its formulation |
| CN102106972A (en) * | 2009-12-25 | 2011-06-29 | 庞来祥 | Medicament for treating empyrosis and scalding and acute skin bruise and contusion, preparation method and detection method |
| CN102784275A (en) * | 2012-09-03 | 2012-11-21 | 武汉市健恒药业有限公司 | Tongzhisurunjiang tablet for activating retardation and softening and preparation and quality control method thereof |
| KR20130027813A (en) * | 2011-09-08 | 2013-03-18 | (주)바이오메디앙 | Method for analyzing vitis vinifera extract |
-
2019
- 2019-05-21 CN CN201910425997.8A patent/CN110231415A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1712051A (en) * | 2005-06-08 | 2005-12-28 | 叶耀良 | Quality control of Gongyanping preparation |
| CN1994340A (en) * | 2006-11-16 | 2007-07-11 | 胡军 | Quality control method of turpinia indochinensis leaf and its formulation |
| CN102106972A (en) * | 2009-12-25 | 2011-06-29 | 庞来祥 | Medicament for treating empyrosis and scalding and acute skin bruise and contusion, preparation method and detection method |
| KR20130027813A (en) * | 2011-09-08 | 2013-03-18 | (주)바이오메디앙 | Method for analyzing vitis vinifera extract |
| CN102784275A (en) * | 2012-09-03 | 2012-11-21 | 武汉市健恒药业有限公司 | Tongzhisurunjiang tablet for activating retardation and softening and preparation and quality control method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 黄毅等: "高效液相色谱法测定妇康阴道泡腾片中没食子酸和盐酸小檗碱的含量", 《时珍国医国药》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113504325A (en) * | 2021-07-07 | 2021-10-15 | 江西康恩贝天施康药业有限公司 | Detection method of changyanning preparation |
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Inventor after: Luo Yujiang Inventor after: Zhou Ying Inventor after: Huang Guofang Inventor before: Luo Yuhong Inventor before: Zhou Ying Inventor before: Huang Guofang |
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