CN111443137B - Method for detecting content of gan Jiang Ling Zhu soup - Google Patents
Method for detecting content of gan Jiang Ling Zhu soup Download PDFInfo
- Publication number
- CN111443137B CN111443137B CN202010171766.1A CN202010171766A CN111443137B CN 111443137 B CN111443137 B CN 111443137B CN 202010171766 A CN202010171766 A CN 202010171766A CN 111443137 B CN111443137 B CN 111443137B
- Authority
- CN
- China
- Prior art keywords
- chromatography column
- mobile phase
- gingerol
- gan jiang
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/047—Standards external
Abstract
The invention discloses a method for detecting the content of gan Jiang Ling Zhu Tang, which is characterized in that a sample solution of the gan Jiang Ling Zhu Tang is taken for HPLC detection, and the chromatographic conditions of the HPLC detection comprise: a C18 chromatographic column is adopted, a phosphoric acid aqueous solution with the concentration of 0.1% is used as a mobile phase A, acetonitrile is used as a mobile phase B, the flow rate is 0.8-1.2 mL/min, the column temperature is 25-35 ℃, and the detection wavelength is 228-232 nm. The detection method can accurately detect the component information or the component content information in the substance standard of the gan Jiang Zhi Zhu decoction, and systematic methodology investigation and verification show that the method has strong specificity, good durability, repeatability and accuracy, and can accurately detect the component content in the substance standard of the gan Jiang Zhi Zhu decoction, thereby objectively evaluating the quality of the preparation.
Description
Technical Field
The invention relates to the technical field of analytical chemistry, in particular to a method for detecting the content of gan Jiang Ling Zhu decoction.
Background
Gan Jiang Ling Zhu Tang is from the golden Kui Yao L ü e of Han Zhong Jing, and the prescription is composed of rhizoma Zingiberis, rhizoma Atractylodis Macrocephalae, Poria, and radix Glycyrrhizae. The dried ginger is a monarch drug for warming the middle-jiao and dispelling cold, the tuckahoe and the atractylodes macrocephala are ministerial drugs for strengthening the spleen and eliminating dampness, and the liquorice is a conductant drug for harmonizing the other drugs, so that the whole formula has the effects of warming yang and dispelling cold and eliminating dampness.
The traditional Chinese medicine and the preparation thereof are all multi-component complex systems, so that the quality of the traditional Chinese medicine and the preparation thereof is evaluated by adopting a method suitable for the traditional Chinese medicine. The traditional Chinese medicine fingerprint is a comprehensive quantifiable identification means and is mainly used for evaluating the authenticity, the excellence and the stability of the quality of the traditional Chinese medicine and the preparation thereof. The method can provide a detection method with abundant identification information, and the establishment of the traditional Chinese medicine fingerprint spectrum can comprehensively reflect the types and the quantities of the traditional Chinese medicine and chemical components contained in the preparation of the traditional Chinese medicine, so as to integrally describe and evaluate the quality of the preparation. In order to better control the quality of the preparation and ensure the clinical curative effect, a component detection method is established to comprehensively evaluate the quality of the preparation.
The gan Jiang Ling Zhu Tang is a compound preparation with complex chemical components, and the effective components comprise flavonoids, terpenoids and the like. It is necessary to detect the chemical components in the compound preparation so as to better realize the quality control and evaluation of the compound preparation.
Disclosure of Invention
In order to achieve the above object, the present invention provides the following technical solutions:
a method for detecting the content of the gan Jiang Ling Zhu Tang is characterized in that a sample solution of the gan Jiang Ling Zhu Tang is taken for HPLC detection, and the chromatographic conditions of the HPLC detection comprise:
a C18 chromatographic column is adopted, a phosphoric acid aqueous solution with the concentration of 0.1% is used as a mobile phase A, acetonitrile is used as a mobile phase B, the flow rate is 0.8-1.2 mL/min, the column temperature is 25-35 ℃, and the detection wavelength is 228-232 nm. .
Optionally, the sample solution of gan Jiang Ling Zhu Tang is prepared by collecting about 0.3g of gan Jiang Zhu Tang substance standard powder, precisely adding 50% methanol, heating and refluxing for 45min, cooling, and supplementing weight.
Optionally, the preparation method of the gan Jiang Ling Zhu Tang substance reference powder comprises the following steps: taking four medicinal materials of 12g of dried ginger, 12g of tuckahoe, 6g of atractylodes macrocephala and 6g of liquorice, putting the medicinal materials into a decoction pot, adding a proper amount of purified water for decoction, filtering, cooling the filtrate to room temperature, carrying out rotary steaming and concentration on the filtrate, and carrying out program freeze drying on the concentrated liquid medicine to obtain the gan Jiang Ling Shu soup material reference powder.
Further, the chromatographic conditions include: the flow rate is 1.0mL/min, the column temperature is 30 ℃, and the detection wavelength is 230 nm.
Specifically, the C18 chromatographic column is selected from Agilent Zorbax SB-C18 chromatographic column, Phenomenex Gemini C18 chromatographic column, Thermo syncronics C18 chromatographic column, Agilent5TC-C18 chromatographic column, Kromasil 100-5-C18 chromatographic column, Welch Xitinate C18 chromatographic column, Waters symmetry C18 chromatographic column or Phenomenex Luna C18 chromatographic column.
Further, the detection method also comprises the detection of a reference substance, wherein the reference substance comprises one or more of 6-gingerol, liquiritin, ammonium glycyrrhizinate, pachymic acid, atractylenolide I, atractylenolide II and atractylenolide III standard substances.
Specifically, the reference substance is prepared into a solution for detection, and the method comprises the following steps: taking 6-gingerol, liquiritin, ammonium glycyrrhizinate, pachymic acid, atractylenolide I, atractylenolide II and atractylenolide III standard substances, and adding methanol to prepare single-standard or mixed-standard solution containing 20-100 mu g of methanol per 1 mL.
Further, the elution in the chromatographic conditions of the HPLC detection is a gradient elution, and the gradient elution procedure may be: 0-9 min, wherein the mobile phase is 81% A; 9-30 min, wherein the mobile phase is 81-60% A; 30-45 min, and the mobile phase is 60-15% A.
And finally obtaining the standard component information or component content information of the gan-ginger-poria-surgery decoction substance by establishing a standard curve of one or more reference substances.
The invention provides a method for detecting the component content of Ganjing Lingzhu decoction, which can accurately detect the component information or the component content information in the substance standard of the Ganjing Lingzhu decoction by using the detection method, and the systematic methodological investigation and verification shows that the method has strong specificity, good durability, repeatability and accuracy, and can accurately detect the component content in the substance standard of the Ganjing Lingzhu decoction, thereby objectively evaluating the quality of a preparation.
Drawings
FIG. 1 is the absorption spectrum of the liquiritin, ammonium glycyrrhizinate and 6-gingerol reference substance;
FIG. 2 shows different mobile phase investigation spectra of the content detection of the reference ingredients of the gan Jiang Ling Shu decoction;
FIG. 3 chromatogram obtained by gradient elution procedure for content detection of reference components of Ganjianlingzhu decoction;
FIG. 4 is a chromatogram for examining durability by the method for detecting the content of the reference component of Ganjianlingzhu decoction;
FIG. 5 chromatogram for examining the specificity of the content detection of the reference substance of Ganjianlingzhu decoction;
FIG. 6 reports the peak purities of glycyrrhizin, glycyrrhizic acid and 6-gingerol in the chromatogram for detecting the content of the reference components of gan Jiang Ling Zhu Tang.
Detailed Description
The invention discloses a method for detecting the reference components of a gan Jiang Ling Zhu soup substance, which can be realized by appropriately improving process parameters by referring to the contents in the text by the technical personnel in the field. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
If not indicated, the rhizoma zingiberis glabrae decoction provided by the invention is a basic ingredient of the material.
Instrument and reagent
Mettler Toledo AL204 electronic analytical balance (Mettler-Torledo, Switzerland)
Mettler Toledo XP6 electronic analytical balance (Mettler-Torledo, Switzerland)
KQ-250DB digital control ultrasonic cleaning instrument (Kunshan ultrasonic instrument Co., Ltd.)
DHG-9070A electric heating constant temperature air-blast drying oven (Shanghai Xinmiao medical appliance manufacturing Co., Ltd.)
Agilent1260 high performance liquid chromatograph (Agilent technologies, Inc.)
Milli-Q (IQ-7000) Water purifier (Mirabbo, USA)
Sample preparation: preparing rhizoma Zingiberis recens-LINGZHU reference powder (Jiangsu Kangyuan pharmaceutical industry GmbH, lot number: Z181104), decocting rhizoma Zingiberis 12g, Poria 12g, Atractylodis rhizoma 6g, and Glycyrrhrizae radix 6g in water, adding purified water, filtering, cooling the filtrate to room temperature, steaming the filtrate, concentrating, and freeze drying the concentrated solution to obtain rhizoma Zingiberis recens-LINGZHU decoction reference powder
Comparison products: liquiritin reference substance (China food and drug testing research institute, batch No.: 111610-201106 for content determination, purity 93.7%)
Ammonium glycyrrhizinate reference (Chinese food and drug testing institute, batch No. 110731 one 201619 for content determination, purity 93.0%)
6-gingerol (China institute for food and drug identification, batch No. 111833-
Atractylenolide I: purchased from China institute for testing and testing food and drug, lot No. 111975-201501, for content determination; the content is calculated by 99.9%
Atractylenolide II: purchased from China institute for testing and testing food and drug, lot No. 111976-201501, for content determination; the content is calculated by 99.9%
Atractylenolide III: purchased from China institute for testing and testing food and drug, lot No. 111978-201501, for content determination; the content is calculated by 99.9%
Pachymic acid: purchased from Dowmastt, Lot MUST-18072910, for use in content determination; the content is 98.1%
Reagent: acetonitrile is chromatographically pure (Tiandi, USA); the water is ultrapure water; other reagents are analytically pure
Example 1
1 chromatographic conditions
1.1 selection of detection wavelength
The detection wavelength of the content determination of 6-gingerol, liquiritin and ammonium glycyrrhizinate can be selected to be 230nm by performing full-wavelength scanning on the 6-gingerol, liquiritin and ammonium glycyrrhizinate reference substance solution and comprehensively considering. The results are shown in FIG. 1.
1.2 selection of the Mobile phase
Acetonitrile-0.1% phosphoric acid and acetonitrile-water are used as an elution system, and the 6-gingerol, the liquiritin and the ammonium glycyrrhizinate are detected by performing tests through the acetonitrile-0.1% phosphoric acid and the acetonitrile-water. Ammonium glycyrrhetate cannot be detected in an acetonitrile-water system; 6-gingerol, liquiritin and ammonium glycyrrhetate are detected under an acetonitrile-0.1% phosphoric acid system, and the separation effect of each chromatographic peak is good, so that the acetonitrile-0.1% phosphoric acid is selected as an elution system. The results are shown in Table 1 and FIG. 2.
TABLE 1 gradient elution procedure
By inspecting different elution gradients, the results of different elution procedures of gradient elution by using acetonitrile-0.1% phosphoric acid solution as a mobile phase are shown in tables 2-4 and figure 3.
Table 2 gradient elution procedure 1
Table 3 gradient elution procedure 2
Table 4 gradient elution procedure 3
By examining different elution gradients, under the elution conditions of the acetonitrile-0.1% phosphoric acid gradient elution program in the table 4, the elution program in the table 4 has the best effect, the separation degrees of liquiritin, glycyrrhizic acid, 6-gingerol and adjacent chromatographic peaks in the chromatogram of the test sample are all larger than 1.5, the baseline separation is achieved, and the symmetry factor is 1.0, so the acetonitrile-0.1% phosphoric acid gradient elution program 3 in the table 4 is selected.
The detection conditions finally established were:
the elution in the chromatographic conditions of the HPLC detection is a gradient elution with the following gradient elution procedure: 0-9 min, wherein the mobile phase is 81% A; 9-30 min, wherein the mobile phase is 81-60% A; 30-45 min, and the mobile phase is 60-15% A; the flow rate of HPLC detection is 1.0mL/min, the column temperature is 30 ℃, the sample injection amount is 10 mu l, and the detection wavelength is 230 nm.
1.3 System suitability test
1.3.1 preparation of control solutions
Taking appropriate amount of liquiritin and ammonium glycyrrhizinate as reference substances, precisely weighing, and adding methanol to obtain solution containing liquiritin, glycyrrhizic acid 60 μ g and glycyrrhizic acid 100 μ g per 1ml (weight of glycyrrhizic acid is ammonium glycyrrhizinate/1.0207). Taking another appropriate amount of 6-gingerol control, precisely weighing, and adding methanol to obtain a solution containing 6-gingerol 50 μ g per 1 ml.
1.3.2 preparation of test solutions
Accurately weighing about 0.3g of rhizoma Zingiberis recens rhizoma Atractylodis substance, placing into a conical flask with a plug, accurately adding 25ml of 50% methanol, sealing the plug, weighing, heating and refluxing for 45min, cooling, weighing again, supplementing 50% methanol reagent to the reduced weight, shaking, filtering, and collecting the filtrate.
1.3.3 determination
Precisely absorbing 10 μ l of 6-gingerol, liquiritin and ammonium glycyrrhizinate reference solution, injecting into high performance liquid chromatograph, continuously and repeatedly injecting sample for 6 times, and calculating relative standard deviation of peak area measurement value; precisely absorbing 10 μ l of sample solution, injecting into high performance liquid chromatograph, detecting according to the above screened chromatographic conditions, continuously repeating sample introduction for 6 times, and calculating theoretical plate number, separation degree and tailing factor of the component to be detected.
1.3.4 results
Under the chromatographic conditions, the separation degrees of liquiritin and glycyrrhizic acid from adjacent chromatographic peaks in the chromatogram of the test sample are all larger than 1.5, the baseline separation is achieved, the symmetry factor is 1.0, the relative standard deviation of the repeatability of the chromatographic system is 6-gingerol 0.11%, liquiritin 0.12% and glycyrrhizic acid 0.20%, the number of theoretical plates calculated by the liquiritin peaks is larger than 10000, and the number of the theoretical plates is not lower than 5000 calculated according to the liquiritin peaks in order to ensure the accuracy of quantitative analysis.
1.4 durability test
According to the selected conditions, factors such as detection wavelength, flow velocity, column temperature and the like in the chromatographic conditions are investigated, the results show that the changes of the detection wavelength, the flow velocity and the column temperature do not have obvious influence on the analysis results of the 6-gingerol, the liquiritin and the glycyrrhizic acid, and the test results show that the content determination method has good durability. The results are shown in Table 5 and FIG. 4.
TABLE 5 durability test results for chromatographic conditions
And (3) measuring and calculating the same batch of rhizoma Zingiberis recens rhizoma Atractylodis substance reference by adopting three chromatographic columns. The result shows that chromatographic columns of different brands have no significant influence on the determination results of 6-gingerol, liquiritin and glycyrrhizic acid. The results are shown in Table 6 and FIG. 4.
TABLE 6 test results for durability test of chromatographic columns
Three types of instruments are adopted to respectively analyze and measure the same batch of rhizoma zingiberis glabrae substance standard. The result shows that the high performance liquid chromatographs of different types have no obvious influence on the determination results of the 6-gingerol, the liquiritin and the glycyrrhizic acid. The results are shown in Table 7 and FIG. 4.
TABLE 7 test results for durability examination of instruments
1.5 examination of the preparation method of the test solution
The method is a reasonable and simple method for preparing the test solution, and the extraction solvent, the extraction mode, the solvent dosage and the extraction time are respectively considered. The results are shown in tables 8 to 10.
TABLE 8 test results of extraction solvent and extraction method
TABLE 9 extraction time test results
TABLE 10 extraction time test results
According to the test result, the preparation method of the test solution is determined as follows: accurately weighing about 0.3g of rhizoma Zingiberis recens rhizoma Atractylodis substance, placing into a conical flask with a plug, accurately adding 25ml of 50% methanol, sealing the plug, weighing, heating and refluxing for 45min, cooling, weighing again, supplementing 50% methanol reagent to the reduced weight, shaking, filtering, and collecting the filtrate.
1.6 specificity test
1.6.1 preparation of control solutions
Taking appropriate amount of liquiritin and ammonium glycyrrhizinate as reference substances, precisely weighing, and adding methanol to obtain solution containing liquiritin, glycyrrhizic acid 60 μ g and glycyrrhizic acid 100 μ g per 1ml (weight of glycyrrhizic acid is ammonium glycyrrhizinate/1.0207). Taking another appropriate amount of 6-gingerol control, precisely weighing, and adding methanol to obtain a solution containing 6-gingerol 50 μ g per 1 ml.
1.6.2 preparation of test solutions
Accurately weighing about 0.3g of rhizoma Zingiberis recens rhizoma Atractylodis substance, placing into a conical flask with a plug, accurately adding 25ml of 50% methanol, sealing the plug, weighing, heating and refluxing for 45min, cooling, weighing again, supplementing 50% methanol reagent to the reduced weight, shaking, filtering, and collecting the filtrate. 1.6.3 preparation of negative test solutions
1.6.3.1 preparation of dried ginger negative test solution
Poria cocos, bighead atractylodes rhizome and liquorice are taken and prepared into a negative preparation according to a substance standard preparation process of rhizoma zingiberis glabrae and then a rhizoma zingiberis negative test solution is prepared according to the same method as the preparation method of the test solution.
1.6.3.2 preparation of Licorice root negative sample solution
Taking rhizoma Zingiberis, Poria and Atractylodis rhizoma, making into negative preparation according to rhizoma Zingiberis-poria reference preparation process, and preparing Glycyrrhrizae radix negative test solution according to the same method as the test solution.
1.6.4 measurement
Precisely sucking 10 μ l of each of the reference solution, the test solution and the negative test solution, injecting into a high performance liquid chromatograph, and measuring, wherein the result shows that the negative test solution has no interference to the measurement of 6-gingerol, liquiritin and glycyrrhizic acid. The results are shown in FIG. 5.
1.7 Peak purity check
And (3) using a diode array detector to carry out purity inspection on chromatographic peaks of 6-gingerol, liquiritin and glycyrrhizic acid in the chromatogram of the test sample, wherein the chromatographic peak purity of the liquiritin and the glycyrrhizic acid in the chromatogram of the test sample reaches 99.9 percent and meets the requirement. The results are shown in FIG. 6.
1.8 Linear relationship investigation
Taking appropriate amount of 6-gingerol, liquiritin and ammonium glycyrrhizinate as reference substances, precisely weighing, and adding methanol to obtain solutions with concentration of 6-gingerol 203.30 μ g/ml, liquiritin 156.89 μ g/ml and glycyrrhizic acid (weight of glycyrrhizic acid is equal to weight of ammonium glycyrrhizinate/1.0207) 466.28 μ g/ml, i.e. 6-gingerol, liquiritin and glycyrrhizic acid reference substance stock solutions. Diluting the stock solution to obtain 6-gingerol with concentration of 203.30 μ g/ml, 162.64 μ g/ml, 121.98 μ g/ml, 81.32 μ g/ml, 40.66 μ g/ml, 20.33 μ g/ml, 10.16 μ g/ml; the liquorice and licorice concentration is 156.89 mug/ml, 125.51 mug/ml, 94.14 mug/ml, 62.76 mug/ml, 31.38 mug/ml, 15.69 mug/ml and 7.84 mug/ml; the glycyrrhizic acid concentration is 466.28 μ g/ml, 373.02 μ g/ml, 279.77 μ g/ml, 186.51 μ g/ml, 93.26 μ g/ml, 46.63 μ g/ml and 23.31 μ g/ml. Precisely sucking 10 μ l of the above solutions, respectively, injecting into high performance liquid chromatograph, and recording chromatographic peak area. Taking the concentration (mu g/ml) as an abscissa (X) and the peak area as an ordinate (Y), drawing a standard curve, and calculating a regression equation and a correlation coefficient as follows: 6-gingerol: 10316x + 5374.6; r2 ═ 0.9999; liquiritin: y 23537x + 2312.6; r2 ═ 1; glycyrrhizic acid: y 4351.9x + 2188.2; r2 ═ 1; the result shows that 6-gingerol has good linear relation between the concentration of 10.16-203.30 mug/ml, liquiritin has good linear relation between the concentration of 7.84-125.51 mug/ml, and glycyrrhizic acid has good linear relation between the concentration of 23.31-373.02 mug/ml. The results are shown in tables 11 to 13.
TABLE 116-gingerol Linear relationship examination results
TABLE 12 Liquiritin Linear relationship examination results
TABLE 13 examination of glycyrrhizic acid Linear relationship
1.9 precision test
Precisely sucking three control solutions (6-gingerol concentration is 10.16, 81.32, 203.30 μ g/ml, glycyrrhizic acid concentration is 7.84, 62.76, 156.89 μ g/ml, glycyrrhizic acid concentration is 23.31, 186.51, 466.28 μ g/ml) with different concentrations and a sample solution 10 μ l each, repeating sample injection for 6 times, measuring, and calculating sample injection precision. The results show that the precision of the instrument is good. The results are shown in tables 14 and 15.
TABLE 14 results of precision test of control solutions
TABLE 15 precision test results of test solutions
1.10 stability test
Sampling the same control solution (6-gingerol concentration: 59.34 μ g/ml; glycyrrhizic acid concentration: 101.33 μ g/ml) and sample solution to be tested at 0h, 4h, 8h, 12h, 16h, 20h, and 29h, respectively, sampling 10 μ l each time, and determining. Calculating RSD of the reference substance solution peak area to be 6-gingerol 0.13%, liquiritin 0.52%, and glycyrrhizic acid 0.92%; RSD in the peak area of the sample solution is respectively 0.09% of 6-gingerol, 0.29% of liquiritin and 0.23% of glycyrrhizic acid. The results show that the control solution and the test solution are basically stable after being placed for 29 hours at room temperature. The results are shown in tables 16 and 17.
TABLE 16 control stability test results
TABLE 17 test results of stability test of test articles
1.11 repeatability test
Taking about 0.3g of rhizoma zingiberis glabrae substance as a reference, preparing 6 parts of test solution, measuring, calculating the content, and measuring the content to obtain that the content of 6-gingerol in the sample is 3.97mg/g, the RSD is 0.70%, the content of liquiritin is 4.70mg/g, the RSD is 0.59%, the content of glycyrrhizic acid is 10.59mg/g, and the RSD is 0.64%. The results show that the method has good repeatability. The results are shown in Table 18.
TABLE 18 results of repeatability tests
1.12 recovery test
Taking a sample with known content (6-gingerol content is 3.97mg/g, glycyrrhizin content is 4.70mg/g, glycyrrhizic acid content is 10.59mg/g) about 0.15g, precisely weighing, weighing 9 parts and 3 parts in parallel, placing into a conical flask with a stopper, adding appropriate amount of 6-gingerol, glycyrrhizin and glycyrrhizic acid reference solution (6-gingerol 0.277mg/ml, glycyrrhizin 0.330mg/ml and glycyrrhizic acid 0.644mg/ml) into each group, and preparing to-be-tested sample solution. Measuring, calculating the average recovery rate of the 6-gingerol to be 98.70 percent, and the RSD to be 1.27 percent; the average recovery rate of liquiritin is 98.93%, and RSD is 0.89%; the average recovery rate of glycyrrhizic acid is 99.59%, and RSD is 1.26%. Test results show that the method has good accuracy. The results are shown in tables 19 to 21.
TABLE 196 gingerol recovery test results
TABLE 20 Glycyrrhiza glycosides recovery test results
TABLE 21 glycyrrhizic acid recovery test results
The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.
Claims (5)
1. A method for detecting content of GAN JIANG LING SHU TANG comprises subjecting a sample solution and a reference solution of GAN JIANG LING SHU TANG to HPLC detection, wherein the reference solution comprises 6-gingerol, liquiritin and ammonium glycyrrhizinate standard;
chromatographic conditions for HPLC detection include:
using a C18 chromatographic column, taking a phosphoric acid aqueous solution with the concentration of 0.1% as a mobile phase A and acetonitrile as a mobile phase B, and carrying out gradient elution procedures as follows: 0-9 min, wherein the mobile phase is 81% A; 9-30 min, wherein the mobile phase is 81-60% A; 30-45 min, wherein the mobile phase is 60-15% A;
the flow rate is 0.8-1.2 mL/min, the column temperature is 25-35 ℃, and the detection wavelength is 228-232 nm.
2. The method as claimed in claim 1, wherein the sample solution of GAN JIANG LING ZHU TANG is prepared by collecting 0.3g of base powder of GAN JIANG LING ZHU TANG, precisely adding 50% methanol, heating and refluxing for 45min, cooling, and supplementing weight.
3. The method of claim 1, wherein the chromatographic conditions comprise: the flow rate is 1.0mL/min, the column temperature is 30 ℃, and the detection wavelength is 230 nm.
4. The method of claim 1, wherein the C18 chromatography column is selected from the group consisting of an Agilent Zorbax SB-C18 chromatography column, a Phenomenex Gemini C18 chromatography column, a Thermo syncronis C18 chromatography column, an Agilent5TC-C18 chromatography column, a Kromasil 100-5-C18 chromatography column, a Welch Xtimate C18 chromatography column, a Waters symmetry C18 chromatography column, and a Phenomenex Luna C18 chromatography column.
5. The method of claim 1, wherein the method of preparing the control into a solution comprises: taking 6-gingerol, liquiritin and ammonium glycyrrhizinate as standard substances, and adding methanol to prepare single-standard or mixed-standard solution containing 20-100 mg per 1 mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010171766.1A CN111443137B (en) | 2020-03-12 | 2020-03-12 | Method for detecting content of gan Jiang Ling Zhu soup |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010171766.1A CN111443137B (en) | 2020-03-12 | 2020-03-12 | Method for detecting content of gan Jiang Ling Zhu soup |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111443137A CN111443137A (en) | 2020-07-24 |
| CN111443137B true CN111443137B (en) | 2021-06-01 |
Family
ID=71627377
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010171766.1A Active CN111443137B (en) | 2020-03-12 | 2020-03-12 | Method for detecting content of gan Jiang Ling Zhu soup |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111443137B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114441662B (en) * | 2020-11-04 | 2023-09-08 | 四川新绿色药业科技发展有限公司 | Construction method and identification method of poria, cassia, rhizoma polygonati and sweet soup and Gan Jiangling, rhizoma polygonati and Shang Tezheng atlas |
| CN113759059B (en) * | 2020-12-24 | 2023-04-18 | 北京康仁堂药业有限公司 | Process evaluation method of substance reference substance or preparation of traditional Chinese medicine compound |
| CN113759061B (en) * | 2020-12-24 | 2023-05-02 | 北京康仁堂药业有限公司 | Preparation process and quality control method of sweet Jiang Lingshu soup |
| CN112964805A (en) * | 2021-03-04 | 2021-06-15 | 青海普兰特药业有限公司 | Method for measuring contents of atractylenolide II and atractylenolide III in atractylenolide medicinal material |
| CN113156019B (en) * | 2021-04-28 | 2022-09-30 | 重庆市畜牧科学院 | Method for detecting multi-component content of liquorice heart-fire-purging decoction |
| CN113274479B (en) * | 2021-04-30 | 2022-08-19 | 上海上药杏灵科技药业股份有限公司 | Ganyang Lingzhu soup, preparation method thereof and fingerprint detection and control |
| CN115166117B (en) * | 2022-08-04 | 2024-01-26 | 成都中医药大学附属医院 | HPLC detection method for medicine for treating gastrointestinal tract complex symptom group after radiotherapy and chemotherapy |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104698096A (en) * | 2014-09-24 | 2015-06-10 | 北京康仁堂药业有限公司 | Detection method of liquorice formula granules |
| CN108107130A (en) * | 2017-12-22 | 2018-06-01 | 山东沃华医药科技股份有限公司 | The assay method of one seed ginseng branch Siberian cocklebur preparation finger |
| CN110441409A (en) * | 2019-06-21 | 2019-11-12 | 南京海昌中药集团有限公司 | A kind of quality determining method of linggui zhugan decoction |
-
2020
- 2020-03-12 CN CN202010171766.1A patent/CN111443137B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104698096A (en) * | 2014-09-24 | 2015-06-10 | 北京康仁堂药业有限公司 | Detection method of liquorice formula granules |
| CN108107130A (en) * | 2017-12-22 | 2018-06-01 | 山东沃华医药科技股份有限公司 | The assay method of one seed ginseng branch Siberian cocklebur preparation finger |
| CN110441409A (en) * | 2019-06-21 | 2019-11-12 | 南京海昌中药集团有限公司 | A kind of quality determining method of linggui zhugan decoction |
Non-Patent Citations (3)
| Title |
|---|
| Chemical profiling and quantification of Gua-Lou-Gui-Zhi decoction by high performance liquid chromatography/quadrupole-time-of-flight mass spectrometry and ultra-performance liquid chromatography/triple quadrupole mass spectrometry;Wen Xu et al.;《Journal Chromatography B》;20150208;第69-84页 * |
| 经典名方甘姜苓术汤中甘草苷含量的HPLC测定;刘鹤 等;《吉林中医药》;20200229;第263-266页 * |
| 高效液相色谱法同时测定干姜甘草配伍后6-姜辣素、甘草酸和甘草苷的含量变化;李洁 等;《中南药学》;20160930;第994-997页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111443137A (en) | 2020-07-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111443137B (en) | Method for detecting content of gan Jiang Ling Zhu soup | |
| CN111443138B (en) | Method for detecting fingerprint of ganjiang lingzhu decoction | |
| CN112098556B (en) | Detection method of angelica sinensis Liuhuang decoction | |
| CN104062374B (en) | The detection method of the Chinese medicine composition of invigorating Qi and tonifying kidney | |
| CN108802245B (en) | Method for detecting trichosanthes root or medicine prepared by taking trichosanthes root as raw material | |
| CN112763597B (en) | Multi-index content determination method for astragalus root, dendrobium stem and hawthorn granules | |
| CN112730674B (en) | Quality detection method of momordica grosvenori tea | |
| CN109521106A (en) | Method that is a kind of while detecting four kinds of component contents in five composition granule of Radix Astragali ramulus cinnamomi | |
| CN114674947B (en) | Detection method for rapidly and comprehensively controlling quality of pinellia tuber magnolia bark Shang Biaozhun decoction | |
| CN114994220B (en) | Construction method of fingerprint spectrum of Qiqingbaidu granule, determination method of component content of Qiqingbaidu granule and application of Qiqingbaidu granule | |
| CN110988238A (en) | Method for constructing HPLC (high performance liquid chromatography) characteristic map of standard decoction of lilac daphne flower bud and method for measuring component content of lilac daphne flower bud | |
| CN112858526B (en) | Centipeda minima fingerprint spectrum and construction method thereof | |
| CN114755338A (en) | Detection method of agastache rugosus and preparation thereof | |
| CN114910583A (en) | Detection method of orange-shell mixture | |
| CN111896637B (en) | Detection method of Jinqing intermediate and fingerprint spectrum construction method thereof | |
| Zhang et al. | Simultaneous determination of saponins in Radix Glycyrrhizae, Notoginseng and Ginseng by high performance liquid chromatography | |
| CN109187825B (en) | Method for measuring content of radix trichosanthis or medicine prepared by taking radix trichosanthis as raw material | |
| CN107643341B (en) | Method for determining active ingredients in cistanche deserticola total glycoside capsules | |
| CN109061002A (en) | Method that is a kind of while measuring scutelloside, rheum emodin, Physcion and Chrysophanol in seven clear Toxin-Vanquishing particles | |
| CN113759036B (en) | Method for measuring content of protodioscin in rhizoma Dioscoreae Septemlobae | |
| CN103267812B (en) | Method for detecting quality of Bazhen particles | |
| CN114563511B (en) | Detection method of bupleurum, cassia twig and dried ginger decoction | |
| CN114689710B (en) | Multi-component quality detection method for loquat lung-heat-clearing drink extract | |
| CN109856276B (en) | Content determination method of Qipi pills | |
| CN115372488A (en) | Method for detecting fingerprint of Yunv decoction |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |