CN111624285A - Quality control method for volatile components of Shenbao tablet - Google Patents
Quality control method for volatile components of Shenbao tablet Download PDFInfo
- Publication number
- CN111624285A CN111624285A CN202010734686.2A CN202010734686A CN111624285A CN 111624285 A CN111624285 A CN 111624285A CN 202010734686 A CN202010734686 A CN 202010734686A CN 111624285 A CN111624285 A CN 111624285A
- Authority
- CN
- China
- Prior art keywords
- solution
- temperature
- shenbao
- anethole
- trans
- 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.)
- Granted
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
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
Abstract
The invention relates to a quality control method of volatile components of Shenbao tablets. The quality control method comprises the following steps of measuring the content of volatile components of trans-anethole and ligustilide in the Shenbao tablet: (1) chromatographic conditions and system applicability test, (2) preparation of control solution: taking a proper amount of trans-anethole reference substance, adding an extraction solvent to prepare a reference substance solution, and (3) preparing a test substance solution: pulverizing SHENBAO tablet, adding extraction solvent, ultrasonic treating, filtering, and collecting filtrate (4) by determination: precisely sucking the reference solution and the sample solution, respectively, injecting into a liquid chromatograph, and measuring. The content determination method provided by the invention provides a reliable way for the quality control of volatile components in Shenbao tablets, and can more comprehensively control the product quality.
Description
Technical Field
The invention belongs to the field of pharmacy, relates to a quality control method of a traditional Chinese medicine preparation, and particularly relates to a content determination method of volatile components in Shenbao tablets.
Background
Shenbao tablets are a Chinese patent medicine on the market, and have the Chinese medicine standard Z20080627. The SHENBAO tablet is prepared from herba Epimedii, semen Trigonellae, fructus Rosae Laevigatae, radix rehmanniae Preparata, fructus Psoraleae, fructus Cnidii, radix Polygoni Multiflori Preparata, Cistanchis herba, fructus Lycii, semen Cuscutae, fructus Schisandrae chinensis, Rubi fructus, radix astragali, Ginseng radix Rubri, Atractylodis rhizoma, rhizoma Dioscoreae, Poria, radix Angelicae sinensis, rhizoma Ligustici Chuanxiong, fructus Foeniculi, semen plantaginis, radix Glycyrrhizae Preparata, etc. Has effects of harmonizing yin and yang, warming yang, invigorating kidney, strengthening body resistance, and consolidating constitution, and can be used for treating soreness of waist and legs, listlessness, nocturia, aversion to cold; clear and thin leucorrhea.
In the prescription process of Shenbao tablet, the Chinese angelica root, ligusticum root, fennel fruit, cnidium fruit, schisandra fruit and white atractylodes rhizome are used for independently extracting volatile oil, and in the existing standard quality control item, the control item of the volatile components of the variety only has the qualitative identification of the Chinese angelica root, the ligusticum root and the cnidium fruit, and no quantitative determination method is used, so that the quality evaluation of the product is not comprehensive.
Disclosure of Invention
The invention aims to provide a quality control method of volatile components in Shenbao tablets, which is used for quality evaluation of the Shenbao tablets, is an effective way for controlling the quality of the volatile components in the Shenbao tablets, improves the quality standard and meets the requirement of high-quality products of companies.
The quality control method comprises the step of measuring the content of volatile components of trans-anethole (derived from fennel) and ligustilide (derived from angelica and ligusticum wallichii) in the Shenbao tablets.
The method for measuring the content of trans-anethole in Shenbao tablets comprises the following steps:
(1) chromatographic conditions and the applicability test of the system,
(2) preparation of control solutions: taking appropriate amount of trans-anethole control, adding extraction solvent to obtain control solution,
(3) preparation of a test solution: pulverizing SHENBAO tablet, adding extraction solvent, ultrasonic treating, filtering, collecting filtrate,
(4) the determination method comprises the following steps: precisely sucking the reference solution and the sample solution, respectively, injecting into a liquid chromatograph, and measuring.
Wherein the chromatographic conditions are selected from Agilent ZORBAX SB-C18 (4.6X 150 mm, 5 μm) column, Kromasil100-5-C18 (4.6X 150 mm, 5 μm) column, Agilent ZORBAX Eclips XDB-C18 (4.6X 150 mm, 5 μm) column and SHISEIDO CAPCELL PAK C18 (4.6X 150 mm, 5 μm) column, preferably Agilent ZORBAX SB-C18 (4.6X 150 mm, 5 μm).
Wherein the detection wavelength is 190-400 nm,
wherein the mobile phase is selected from: a methanol-water system, a methanol-0.1% phosphoric acid water system, a methanol-0.1% formic acid water system and a methanol-0.4% glacial acetic acid water system,
wherein the proportion of the methanol to the 0.1 percent phosphoric acid water is as follows: 55-58:45,
wherein, the flow rate is: 0.8ml/min, 0.9 ml/min, 1.0ml/min, 1.1 ml/min, 1.2 ml/min,
wherein the column temperature is: 25-35 ℃.
The preparation of the test solution of the invention comprises the following steps:
the extraction solvent is selected from: absolute ethyl alcohol, normal hexane, ethyl acetate,
the kidney nourishing tablet is prepared by the following crushing modes: the pulverizer pulverizes, grinds and pulverizes,
extraction time: the time of the reaction is 20 to 45 minutes,
the extraction solvent amount is as follows: 20-50 ml.
Preferably, the method for measuring the content of trans-anethole comprises the following steps:
chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; methanol-0.1% phosphoric acid water (55-58: 45) is used as a mobile phase; the column temperature is 25-35 ℃; the flow rate is 0.8-1.0 ml/min; the detection wavelength is 254 nm; the theoretical plate number calculated according to the trans-anethole peak is not less than 5000,
preparation of control solutions: taking a proper amount of trans-anethole reference substance, precisely weighing, adding anhydrous ethanol to obtain the final product,
preparation of a test solution: pulverizing SHENBAO tablet, pulverizing with pulverizer, weighing 1-3g, placing into conical flask with plug, adding 1-40 ml absolute ethanol, sealing, weighing, ultrasonic treating, cooling, weighing again, adding absolute ethanol, shaking, filtering, collecting filtrate,
the determination method comprises the following steps: precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and measuring.
Further preferably, the method for measuring the content of trans-anethole comprises the following steps:
chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; methanol-0.1% phosphoric acid water (55: 45) is used as a mobile phase; the column temperature is 30 ℃; the flow rate is 1.0 ml/min; the detection wavelength is 254 nm; the theoretical plate number calculated according to the trans-anethole peak is not less than 5000,
preparation of control solutions: taking a proper amount of trans-anethole reference substance, precisely weighing, adding anhydrous ethanol to prepare a solution containing 14 μ g of the trans-anethole reference substance per 1ml,
preparation of a test solution: pulverizing SHENBAO tablet, weighing about 2g, precisely weighing, placing into conical flask with plug, precisely adding anhydrous ethanol 20ml, sealing, weighing, ultrasonic treating for 30 min, cooling, precisely weighing, supplementing with anhydrous ethanol, shaking, filtering, collecting filtrate,
the determination method comprises the following steps: precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and measuring.
The invention also aims to provide a method for measuring the content of another volatile component ligustilide in the Shenbao tablet.
The content determination method of ligustilide comprises the following steps:
chromatographic conditions and system applicability test: a capillary column; column temperature is programmed temperature rise, and split-flow sample injection is carried out, wherein the split-flow ratio is 1: 1-50; the flow rate is 0.7-1.5ml/min,
preparation of control solutions: taking appropriate amount of ligustilide reference substance, adding extraction solvent to obtain reference substance solution,
preparation of a test solution: taking Shenbao tablets, adding water to completely dissolve the sample, testing according to 2204 volatile oil determination method of the four general rules of the 2015 version in Chinese pharmacopoeia to obtain the Shenbao tablets,
the determination method comprises the following steps: precisely sucking 1 μ l of each of the reference solution and the sample solution, injecting into gas chromatograph, and measuring.
Wherein, the chromatographic conditions are as follows:
the gas phase capillary column is selected from: agilent HP-530 m × 0.32mm × 0.25 μm; agilent HP-INNOWax 30m × 0.32mm × 0.50 μm; agilent DB-62430 m × 0.32mm × 1.8 μm;
the temperature programming method comprises the following steps:
the preparation of the test solution comprises the following steps:
taking Shenbao tablets, adding water to completely dissolve the samples, performing 2204 volatile oil determination test according to the general rule of the four departments of the book 2015 of Chinese pharmacopoeia, adding dimethylbenzene, connecting a reflux condenser tube, heating and keeping slight boiling for 1-5 hours, cooling, after the solution is clearly layered, taking dimethylbenzene solution, placing in a 2-10ml measuring flask, washing the inner wall of a volatile oil determinator by a small amount of dimethylbenzene in times, merging the dimethylbenzene washing solution into the same measuring flask, adding absolute ethyl alcohol to scale, shaking up uniformly to obtain the Shenbao tablets,
preferably, the content determination of ligustilide according to the present invention comprises the following steps:
chromatographic conditions and system applicability test: a capillary column taking (5% -phenyl) -methyl polysiloxane as a stationary phase; column temperature is programmed temperature rise: the initial temperature is 50-70 ℃, the temperature is raised to 150 ℃ at the rate of 3-4 ℃ per minute, the temperature is maintained for 1-3 minutes, the temperature is raised to 160 ℃ at the rate of 1-3 ℃ per minute, the temperature is maintained for 6-10 minutes, the temperature is raised to 260 ℃ at the rate of 10-14 ℃ per minute, and the temperature is maintained for 1-3 minutes; the temperature of the sample inlet is 240 ℃ and 260 ℃; the temperature of the detector is 250 ℃ and 270 ℃; split-flow sample injection with a split-flow ratio of 1: 10-30; the flow rate is 0.5-1 ml/min,
preparation of control solutions: precisely weighing appropriate amount of ligustilide reference substance, adding anhydrous ethanol to obtain reference substance solution,
preparation of a test solution: taking Shenbao tablets, adding water to completely dissolve the samples, performing 2204 volatile oil determination test according to the general rule of the four departments of the book 2015 of Chinese pharmacopoeia, adding dimethylbenzene, connecting a reflux condenser tube, heating and keeping slight boiling for 1-5 hours, cooling, after the solution is clearly layered, taking dimethylbenzene solution, placing in a 2-10ml measuring flask, washing the inner wall of a volatile oil determinator by a small amount of dimethylbenzene in times, merging the dimethylbenzene washing solution into the same measuring flask, adding absolute ethyl alcohol to scale, shaking up uniformly to obtain the Shenbao tablets,
the determination method comprises the following steps: respectively sucking the reference solution and the sample solution, injecting into a gas chromatograph, and measuring.
Further preferably, the content determination of ligustilide comprises the following steps:
chromatographic conditions and system applicability test: a capillary column using (5% -phenyl) -methyl polysiloxane as a stationary phase (the column length is 30m, the inner diameter is 0.32mm, and the film thickness is 0.25 μm); column temperature is programmed temperature rise: the initial temperature is 60 ℃, the temperature is increased to 145 ℃ at the rate of 3.5 ℃ per minute, the temperature is maintained for 2 minutes, the temperature is increased to 156 ℃ at the rate of 2 ℃ per minute, the temperature is maintained for 8 minutes, and the temperature is increased to 250 ℃ at the rate of 12 ℃ per minute and the temperature is maintained for 2 minutes; the temperature of a sample inlet is 250 ℃; the temperature of the detector is 260 ℃; split-flow sample injection is carried out, and the split-flow ratio is 1: 20; the flow rate is 0.8ml/min, the number of theoretical plates is not less than 20000 calculated according to ligustilide peak,
preparation of control solutions: accurately weighing appropriate amount of ligustilide control, adding anhydrous ethanol to obtain solution containing 3.0mg per 1ml,
preparation of a test solution: taking 30 tablets of the product, placing the tablets in a round-bottom flask, adding 500ml of water, soaking until a sample is completely dissolved, testing according to a volatile oil determination method (2204 in the four Processions of the national Pharmacopeia 2015 edition), adding water from the upper end of a determinator to fill a scale part until the scale part is filled, overflowing the water into the flask, adding 1ml of dimethylbenzene, connecting a reflux condenser pipe, heating and keeping micro-boiling for 5 hours, cooling, after the solution is clearly layered, taking the dimethylbenzene liquid, placing the dimethylbenzene liquid in a 10ml measuring flask, washing the inner wall of the volatile oil determiner by a small amount of dimethylbenzene for times, merging the dimethylbenzene washing liquid into the same measuring flask, adding absolute ethyl alcohol to the scale, shaking uniformly to obtain the product,
the determination method comprises the following steps: precisely sucking 1 μ l of each of the reference solution and the sample solution, injecting into gas chromatograph, and measuring.
The invention firstly researches the content determination method of volatile components in Shenbao tablets, and establishes an HPLC (high performance liquid chromatography) content determination method of trans-anethole in the Shenbao tablets and a GC (gas chromatography) content determination method of ligustilide through a large number of experimental researches. The content determination method established by the invention has high precision, good accuracy and good stability, provides a reliable way for controlling the quality of volatile components in Shenbao tablets, can control the product quality more comprehensively, and ensures that the product is stable, uniform and controllable in quality.
Drawings
FIG. 1 shows an HPLC chromatogram of trans-anethole (a: trans-anethole, A: trans-anethole control, B: Shenbao tablet (1905288), C: fennel negative control)
FIG. 2, full wavelength scan of trans-anethole
FIG. 3, standard curve diagram of trans-anethole
FIG. 4 shows GC chromatogram of ligustilide (b: ligustilide, D: ligustilide reference, E: Shenbao tablet (1905288), F: negative control of radix Angelicae sinensis and rhizoma Ligustici Chuanxiong)
Figure 5 ligustilide standard curve chart.
Detailed Description
The invention is further illustrated by the following examples, which are not to be construed as limiting the invention.
Example 1 screening procedure for assay of Trans-Anethole in Shenbao tablets
1.1 instruments and materials
Chromatograph: agilent 1100 hplc;
a chromatographic column: agilent ZORBAX SB-C18 (4.6X 150 mm, 5 μm);
reagent: absolute ethyl alcohol, ethyl acetate, normal hexane and phosphoric acid, and analytically pure; methanol, chromatographically pure;
comparison products: trans-anethole, institute for testing and testing Chinese food and drug, purity 99.6%, batch number: 111835-201804.
Sample preparation: jiangxi Hui ren pharmaceutical industry, batch number: 1905288.
1.2 selection of the method
According to the research and research of the literature combined with the GC-MS research result of the volatile components in the Shenbao tablet, trans-anethole derived from fennel and ligustilide derived from ligusticum wallichii and angelica sinensis are selected as the index components for content determination.
By adopting GC chromatography, research results show that a trans-anethole chromatographic peak in a trans-anethole test sample solution is well separated from adjacent chromatographic peaks, but negative samples are interfered and have poor specificity. The research result of the HPLC method shows that the trans-anethole chromatographic peak has better separation and moderate retention time, so the HPLC method is adopted in the experiment to carry out content determination research on the trans-anethole in the Shenbao tablet. The chromatogram is shown in FIG. 1.
1.3 examination of preparation method of test solution
1.3.1 examination of the mode of comminution
The influence of two porphyrization methods of mortar grinding and pulverizer grinding on the content extraction of the trans-anethole is examined.
The test method comprises the following steps: taking 30 tablets of the product, grinding (grinding by a mortar or a grinder), taking about 2g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of absolute ethyl alcohol, sealing the plug, weighing, carrying out ultrasonic treatment (power 500W, frequency 40 Hz) for 30 min, cooling, precisely weighing again, supplementing the lost weight by absolute ethyl alcohol, shaking uniformly, filtering, and taking a subsequent filtrate for later use.
The results show that the Shenbao tablets are hard in texture, the Shenbao tablets crushed by the crusher are uniform in powder, and the trans-anethole content is higher; the mortar grinding time was long and laborious, and the selected method of grinding was the pulverizer grinding in consideration of the volatility of the volatile components, and the results are shown in Table 1.
Examination of extraction solvent
The influence of three solvents, namely absolute ethyl alcohol, normal hexane and ethyl acetate, on the content extraction of the trans-anethole is examined. The result shows that the test product extracted by the first method has higher trans-anethole content and simple and convenient operation, so the first method is selected as the preparation method of the test product solution. The results are shown in Table 2.
The method comprises the following steps: taking 30 tablets of the product, grinding, taking about 2g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of absolute ethyl alcohol, sealing the plug, weighing, carrying out ultrasonic treatment (power 500W and frequency 40 Hz) for 30 min, cooling, precisely weighing again, supplementing the lost weight with absolute ethyl alcohol, shaking uniformly, filtering, and taking a subsequent filtrate for later use.
The second method comprises the following steps: taking 2g of the product, grinding into fine powder, placing into a conical flask with a plug, adding 20ml of n-hexane, sealing the plug, performing ultrasonic treatment for 30 min, filtering, recovering n-hexane, transferring into a 5ml volumetric flask with absolute ethyl alcohol, and fixing the volume to the scale mark.
The third method comprises the following steps: taking 2g of the product, grinding into fine powder, placing into a conical flask with a plug, adding 20ml of ethyl acetate, sealing the plug, performing ultrasonic treatment for 30 min, filtering, recovering the ethyl acetate, transferring into a 5ml volumetric flask with absolute ethyl alcohol, and fixing the volume to the scale mark.
Investigation of extraction time
The influence of different extraction time on the extraction of the content of the trans-anethole is examined. The result shows that the ultrasonic treatment is carried out for 30 min, and the content of trans-anethole is the highest, so that 30 min is selected as the extraction time.
The test method comprises the following steps: weighing 2g of the powder, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of absolute ethyl alcohol, sealing the plug, weighing, ultrasonically treating (power 500W, frequency 40 Hz) for 30 min (20 min, 45 min), cooling, precisely weighing, supplementing the lost weight with absolute ethyl alcohol, shaking, filtering, and collecting the subsequent filtrate for use. The results are shown in Table 3.
Investigation of amount of extraction solvent
The influence of different extraction solvent amounts on the extraction of the content of the trans-anethole is examined. The result shows that the peak area and the content of the chromatographic peak of the trans-anethole extracted by 20ml of solvent are moderate, so 20ml is selected as the extracted solvent.
The test method comprises the following steps: weighing 2g of the powder, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml (30 ml and 50 ml) of absolute ethyl alcohol, sealing the plug, weighing, ultrasonically treating (power 500W and frequency 40 Hz) for 30 min, cooling, precisely weighing again, supplementing the lost weight with absolute ethyl alcohol, shaking, filtering, and collecting the subsequent filtrate for use. The results are shown in Table 4.
TABLE 4 examination of the amount of solvent extracted
1.3.5 method of preparing Final defined test solutions
Taking 30 tablets of the product, crushing and grinding, taking about 2g, precisely weighing, placing in a conical flask with a plug, precisely adding 20ml of absolute ethyl alcohol, sealing the plug, weighing, carrying out ultrasonic treatment (power 500W and frequency 40 Hz) for 30 min, cooling, precisely weighing again, supplementing the lost weight with absolute ethyl alcohol, shaking uniformly, filtering, and taking a subsequent filtrate for later use.
1.4 chromatographic conditions and System suitability test
1.4.1 investigation of the column
4 columns were investigated: agilent ZORBAX SB-C18 (4.6X 150 mm, 5 μm) column, Kromasil100-5-C18 (4.6X 150 mm, 5 μm) column, Agilent ZORBAX Eclips XDB-C18 (4.6X 150 mm, 5 μm) column and SHISEIDO CAPCELL PAK C18 (4.6X 150 mm, 5 μm) column. The result shows that the Agilent ZORBAX SB-C18 (4.6X 150 mm, 5 μm) chromatographic column has good separation effect, proper retention time and better peak shape, so the Agilent ZORBAX SB-C18 chromatographic column is selected for measuring the content of the trans-anethole.
Selection of detection wavelength
The trans-anethole control solution is subjected to full-wavelength scanning (190-400 nm) and combined with the literature, and 254nm is finally selected as the detection wavelength. The results of the spectral scan are shown in figure 2.
Investigation of mobile phase systems
Four mobile phase systems were investigated: the results of a methanol-water system, a methanol-0.1% phosphoric acid water system, a methanol-0.1% formic acid water system and a methanol-0.4% glacial acetic acid water system show that the methanol-0.1% phosphoric acid water system has the best separation effect on the trans-anethole and has proper retention time, so the methanol-0.1% phosphoric acid water system is selected as a mobile phase.
Mobile phase investigation at different ratios
The separation influence of the flowing of methanol-0.1% phosphoric acid water relative to the chromatographic peak of trans-anethole is examined, and the results show that the methanol-0.1% phosphoric acid water (55: 45) has the best separation effect on the trans-anethole and has proper retention time, so the methanol-0.1% phosphoric acid water (55: 45) is selected as the mobile phase.
Investigation of flow Rate
Five flow rates of 0.8ml/min, 0.9 ml/min, 1.0ml/min, 1.1 ml/min, and 1.2 ml/min were examined, respectively. As a result, the retention time and resolution of the chromatographic peak of trans-anethole at a flow rate of 1.0ml/min were suitable. The results of the flow rate investigation are shown in Table 5.
TABLE 5 examination of different flow rates
1.4.6 examination of column temperature
Three column temperatures of 25 deg.C, 30 deg.C and 35 deg.C were examined. The result shows that the chromatographic peak separation degree of the trans-anethole is better at the column temperature of 30 ℃, and the retention time is proper. The results of the column temperature investigation are shown in Table 6.
Preferred chromatographic conditions
Octadecylsilane chemically bonded silica is used as a filling agent; methanol-0.1% phosphoric acid water (55: 45) is used as a mobile phase; the column temperature is 30 ℃; the flow rate is 1.0 ml/min; the detection wavelength is 254 nm; the theoretical plate number is not less than 5000 calculated according to trans-anethole peak.
1.5 methodological considerations
1.5.1 Standard Curve and Linear Range
Accurately weighing 8.6 mg of trans-anethole reference substance, placing in a 50ml measuring flask, adding absolute ethyl alcohol to dissolve and dilute to scale, and shaking up to obtain reference substance mother liquor. Precisely sucking reference substance mother liquor 0.1 ml, 0.2 ml, 0.5 ml, 1ml, 2ml and 5ml, respectively placing into 10ml measuring flask, adding anhydrous ethanol to dilute to scale, shaking to obtain reference substance solutions with different concentrations, precisely sucking 10 μ l and mother liquor 10 μ l, injecting into high performance liquid chromatograph, recording peak area, drawing standard curve (see figure 3) with sample concentration (mg/ml) as abscissa and peak area as ordinate, and determining results in table 7. The regression equation is: y =78737X +9.8938, R2And = 1. The results show that the trans-anethole control shows good linear relation in the range of 0.0172 mu g to 1.72 mu g.
TABLE 7 results of Trans-anethole Standard Curve
1.5.2 precision investigation
The product (lot number: 1905288) was weighed out in an appropriate amount and precisely, a sample solution was prepared by a content measurement method, sample introduction was continued for 6 times, and peak area was recorded, and the average peak area of trans-anethole was found to be 973.7 and RSD was found to be 0.26%, and the results are shown in table 8. The results show that the precision of the instrument is good.
Stability survey
The product (lot 1905288) was weighed out in an appropriate amount and precisely weighed, and a test solution was prepared by a content measurement method, and the peak area of trans-anethole was measured at 0, 2, 4, 6, 8 and 12 hours, respectively, and the average peak area was 973.1 and the RSD was 0.58%, and the results are shown in Table 9. The results show that the test solution has good stability within 12 hours.
Repeatability test
The product (lot 1905288) was weighed out in an appropriate amount and precisely, a test solution was prepared by a content measurement method, 6 test solutions were prepared, injection was conducted, the peak area was measured, and the average content of trans-anethole was calculated to be 0.0871 mg/tablet and the RSD was calculated to be 2.27%, and the results are shown in table 10. The result shows that the repeatability of the method is better.
Accuracy test
Taking about 1.0g of the product (Shenbao tablets with known content, batch number: 1905288, trans-anethole content of 0.0871 mg/tablet), precisely weighing, precisely adding 10ml of reference solution (containing trans-anethole of 0.0115 mg/ml) according to the concentration of 1:1, adding absolute ethyl alcohol to make up to 20ml, and preparing 6 parts of test solution with the same concentration according to the preparation method of the test solution. The recovery rate was calculated by [ content determination ] method, see table 11. The result shows that the method accuracy meets the requirement of content determination.
Intermediate precision investigation
The product (batch No. 1905288) was collected, and different analysts prepared the sample solution and the reference solution according to the preparation method of the sample solution and the reference solution on different dates, and detected by Agilent and Thermo analytical instruments, and the results are shown in table 12. The results show that the trans-anethole content RSD measured by the two instruments is 2.89%.
Example 2 screening procedure for measurement of ligustilide content in Shenbao tablets
2.1 instruments and materials
Chromatograph: gas chromatograph, Agilent 7890B; the electric heating jacket with electronic temperature regulation, model DZTW, power 0.5 KW.
A chromatographic column: agilent HP-5, 30 m.times.0.32 mm.times.0.25 μm;
reagent: absolute ethyl alcohol and xylene are analytically pure, and the product is obtained by a general fine chemical company in Beijing; anhydrous sodium sulfate is analytically pure, Beijing chemical plant; water, Millipore bright and limpid-D ultrapure water integrated system preparation; nitrogen, high purity nitrogen, purity 99.999%.
Comparison products: ligustilide, lot number MUST-19041005, 99.55% pure, gendomansite biotechnology limited;
sample preparation: shenbao tablets, provided by Jiangxi Hui ren pharmaceutical industry Co., Ltd, batch number: 1905288.
2.2 selection of methods
According to the research and research of the literature combined with the GC-MS research result of the volatile components in the Shenbao tablet, trans-anethole derived from fennel and ligustilide derived from ligusticum wallichii and angelica sinensis are selected as the index components for content determination.
By adopting GC chromatography, the research result shows that the ligustilide chromatographic peak in the test solution is well separated from the adjacent chromatographic peak, and the negative sample has no interference and good specificity, so that the GC chromatography is adopted in the experiment to carry out content determination research on the ligustilide in the Shenbao tablet. The chromatogram is shown in FIG. 4.
2.3 examination of the preparation method of the test solution
2.3.1 examination of extraction methods
Through literature research, the extraction method of volatile components in the four Shenbao tablets is investigated.
The method comprises the following steps: taking 30 tablets of the product, placing the tablets in a round-bottom flask, adding 500ml of water, soaking until a sample is completely dissolved, testing according to a volatile oil determination method (2204 in the four Processions of the national pharmacopoeia 2015 edition), adding water from the upper end of a determinator until a scale part is filled, overflowing the flask, adding 1ml of xylene, connecting a reflux condenser pipe, heating and keeping micro-boiling for 5 hours, cooling, after the solution is clearly layered, taking xylene solution in parts, placing the xylene solution in a 2ml measuring flask, washing the inner wall of the volatile oil determiner by using a small amount of xylene in parts, merging xylene washing liquor into the same measuring flask, adding xylene to the scale, and shaking uniformly to obtain the product.
The second method comprises the following steps: taking 20 Shenbao tablets, grinding, placing into a separating funnel, adding 200ml of water to completely dissolve a sample, adding cyclohexane to extract for three times (30 ml. times.3), combining cyclohexane layers, volatilizing the solvent, adding cyclohexane to dissolve, and fixing the volume to a 2ml measuring flask.
The third method comprises the following steps: taking 50 Shenbao tablets, grinding, placing in a round-bottom flask, adding 200ml of ethyl acetate, heating and refluxing for extraction for 2 hours, filtering, volatilizing the solvent, adding cyclohexane for dissolution, and fixing the volume to a measuring flask of 5 ml.
The method four comprises the following steps: taking 30 tablets of the product, placing the tablets in a round-bottom flask, adding 500ml of water, soaking until a sample is completely dissolved, testing according to a volatile oil determination method (2204 in the four Processions of the Chinese pharmacopoeia 2015 edition), adding water from the upper end of a determinator until a scale part is filled, overflowing the flask, adding 1ml of ethyl acetate, connecting a reflux condenser tube, heating and keeping micro-boiling for 3 hours, cooling, after the solution is clearly layered, taking ethyl acetate liquid, placing the ethyl acetate liquid in a 2ml measuring flask, washing the inner wall of the volatile oil determinator by using a small amount of ethyl acetate for times, merging the ethyl acetate washing liquid into the same measuring flask, adding ethyl acetate to the scale, and shaking uniformly to obtain the product.
The results show that the extracting solutions of the second method and the third method are dark green, the extraction is incomplete, and the emulsification of the second method is serious; the fourth method is that the oil is light yellow, but the extraction is incomplete; the method can completely extract volatile components from SHENBAO tablet, and the oil is light yellow; therefore method one was chosen as the extraction method.
Examination of sample amount
The amount of the sample for extracting the volatile components in the Shenbao tablet is considered.
The test method comprises the following steps: taking 30 (50 and 100) tablets of the product, placing the tablets in a round-bottom flask, adding 500ml of water, soaking until a sample is completely dissolved, testing according to a volatile oil determination method (2204 in the four-part general rule of Chinese pharmacopoeia 2015 edition), adding water from the upper end of a determinator until the graduated part is filled with water and overflows into the flask, adding 1ml of xylene, connecting a reflux condenser pipe, heating and keeping micro-boiling for 5 hours, cooling, after the solution is clearly layered, taking the xylene solution in parts, placing the xylene solution in a 2ml measuring flask, washing the inner wall of the volatile oil determination device in parts by using a small amount of xylene, merging the xylene washing solution into the same measuring flask, adding the xylene to the graduation, and shaking uniformly to obtain the product.
The result shows that the sample amount of the Shenbao tablet is 30 to 50 tablets, which is more suitable, and 30 tablets are selected in the experiment. The results are shown in Table 13.
Investigation of extraction time
The influence of different extraction times on the volatile components in the Shenbao tablets is investigated.
The test method comprises the following steps: taking 30 tablets of the product, placing the tablets in a round-bottom flask, adding 500ml of water, soaking until a sample is completely dissolved, testing according to a volatile oil determination method (2204 in the four Proc. of the national pharmacopoeia 2015 edition), adding water from the upper end of a determinator until a scale part is filled, overflowing the flask, adding 1ml of dimethylbenzene, connecting a reflux condenser pipe, heating and keeping micro-boiling for 5 hours (1 hour, 2 hours, 3 hours and 4 hours), cooling, after the solution is clearly layered, taking dimethylbenzene liquid in a separate manner, placing the dimethylbenzene liquid in a 2ml measuring flask, washing the inner wall of the volatile oil determinator with a small amount of dimethylbenzene in a separate manner, merging a dimethylbenzene washing liquid into the same measuring flask, adding the dimethylbenzene to the scale, and shaking uniformly to obtain the product.
The result shows that the Shenbao tablet is extracted for 5 hours, which is suitable for 5 hours in the experiment. The results are shown in Table 14.
Investigation of constant volume
The influence of the constant volume of the test sample to different volumes on the chromatographic peak area of ligustilide is examined.
The test method comprises the following steps: taking 30 tablets of the product, placing the tablets in a round-bottom flask, adding 500ml of water, soaking until a sample is completely dissolved, testing according to a volatile oil determination method (2204 in the four Proc. of China pharmacopoeia 2015 edition), adding water from the upper end of a determinator until a scale part is filled, overflowing the flask, adding 1ml of dimethylbenzene, connecting a reflux condenser tube, heating and keeping micro-boiling for 5 hours, cooling, after the solution is clearly layered, taking the dimethylbenzene solution, placing the dimethylbenzene solution in a 2ml measuring flask (5 ml and 10 ml), washing the inner wall of the volatile oil determinator by a small amount of dimethylbenzene in times, merging the dimethylbenzene washing solution into the same measuring flask, adding absolute ethyl alcohol to the scale, and shaking up to obtain the product.
The result shows that the peak area is moderate when the volume is up to 10ml, so the volume is up to 10 ml. The results are shown in Table 15.
Method for producing a finally defined test solution
Taking 30 tablets of the product, placing the tablets in a round-bottom flask, adding 500ml of water, soaking until a sample is completely dissolved, testing according to a volatile oil determination method (2204 in the four Processions of the national pharmacopoeia 2015 edition), adding water from the upper end of a determinator to fill a scale part until the scale part is filled, overflowing the flask, adding 1ml of dimethylbenzene, connecting a reflux condenser pipe, heating and keeping micro-boiling for 5 hours, cooling, after the solution is clearly layered, taking the dimethylbenzene solution in parts, placing the dimethylbenzene solution in a 10ml measuring flask, washing the inner wall of the volatile oil determiner by using a small amount of dimethylbenzene in parts, merging the dimethylbenzene washing solution into the same measuring flask, adding absolute ethyl alcohol to the scale, and shaking uniformly to obtain the product.
2.4 chromatographic conditions and System suitability test
2.4.1 investigation of gas phase capillary columns
Three gas phase capillary columns were examined separately, including AgilentHP-530 m.times.0.32 mm.times.0.25 μm (non-polar); AgilentHP-INNOWax 30 m.times.0.32 mm.times.0.50 μm (strongly polar); AgilentDB-62430 m.times.0.32 mm.times.1.8 μm (nonpolar). The results show that under the same chromatographic condition, the difference between each chromatographic column is large, the separation effect is different, and Agilent HP-530 m multiplied by 0.32mm multiplied by 0.25 mu m chromatographic column is selected by comprehensively considering the use condition of the chromatographic column.
Investigation of split-flow sample injection
Different split ratios of 1:50, 1:20, 1:10, 1:5 and 1:1 are selected for examination, and the result shows that the split ratio is 1:20 is good, the resolution of each chromatographic peak is good, the retention time and the peak area are moderate, so the split ratio is 1: 20.
Investigation of carrier gas flow velocity
Different carrier gas flow rates of 0.7 ml/min, 0.8ml/min, 0.9 ml/min, 1.0ml/min and 1.5ml/min are selected for investigation, and as a result, the chromatographic peak separation degree of 0.7-1.0 ml/min is better, and the carrier gas flow rate of 0.8ml/min is selected in the experiment.
Investigation of column temperature
Different temperature programming modes are considered, and the result is the best temperature programming mode of the method eight. The temperature programming method is shown in table 16:
2.4.5 preferred chromatographic conditions
A capillary column using (5% -phenyl) -methyl polysiloxane as a stationary phase (the column length is 30m, the inner diameter is 0.32mm, and the film thickness is 0.25 μm); column temperature is programmed temperature rise: the initial temperature is 60 ℃, the temperature is increased to 145 ℃ at the rate of 3.5 ℃ per minute, the temperature is maintained for 2 minutes, the temperature is increased to 156 ℃ at the rate of 2 ℃ per minute, the temperature is maintained for 8 minutes, and the temperature is increased to 250 ℃ at the rate of 12 ℃ per minute and the temperature is maintained for 2 minutes; the temperature of a sample inlet is 250 ℃; the temperature of the detector is 260 ℃; split-flow sample injection is carried out, and the split-flow ratio is 1: 20; the flow rate was 0.8 ml/min. The number of theoretical plates is not less than 20000 calculated according to ligustilide peak.
2.5 methodological considerations
2.5.1 Standard Curve and Linear Range
Taking a proper amount of ligustilide reference substance as reference substance mother liquor. Precisely sucking 1ml, 0.5 ml, 0.2 ml, 0.1 ml and 0.05 ml of reference mother liquor, respectively placing the reference mother liquor in a 2ml measuring flask, adding absolute ethyl alcohol to dilute the reference mother liquor to a scale, shaking the reference mother liquor uniformly to prepare reference solutions with different concentrations, precisely sucking 1 mu l and 1 mu l of the reference mother liquor, respectively injecting the reference solutions into a gas chromatograph, recording peak areas, drawing a standard curve (see figure 5) by taking the sample injection amount (mu g) as a horizontal coordinate and the peak areas as a vertical coordinate, and obtaining the measurement results shown in a table 17. The regression equation is: y =159.59X +4.5886, R2= 0.9997. The result shows that the ligustilide reference substance has good linear relation in the range of 0.25 mu g-10.00 mu g.
Precision survey
Taking the product (batch number: 1905288), weighing, preparing sample solution by content determination method, continuously feeding sample for 6 times, recording peak area, and determining ligustilide average peak area of 563.8 and RSD of 0.21%, the results are shown in Table 18. The results show that the precision of the instrument is good.
Stability survey
The product (lot 1905288) was weighed out in an appropriate amount and precisely, and a test solution was prepared by a content measurement method, and the peak areas of ligustilide were measured at 0, 2, 4, 6, 8, and 12 hours, respectively, and the average peak area was 564.7 and the RSD was 0.34%, and the results are shown in Table 19. The results show that the test solution has good stability within 12 hours.
Repeatability test
Taking the product (batch No. 1905288), weighing, preparing test solution according to content determination method, preparing 6 test solutions, injecting sample, determining peak area, calculating ligustilide average content of 1.6329mg/g and RSD of 1.84%, and finding out the results in Table 20. The result shows that the repeatability of the method is better.
Accuracy test
Taking 15 tablets (about 10.5 g) of the product (Shenbao tablets with known content, batch No. 1905288, ligustilide content is 1.6329 mg/g), precisely adding 5ml of reference solution (containing ligustilide 3.1481 mg/ml) according to the concentration of 1:1, and preparing 6 parts of test solution with the same concentration according to the preparation method of the test solution. The recovery rate was calculated by [ content determination ] method, see table 21. The result shows that the method accuracy meets the requirement of content determination.
Example 3 method for measuring Trans-anethole content
Octadecylsilane chemically bonded silica is used as a filler in chromatographic conditions and system applicability tests; methanol-0.1% phosphoric acid water (55: 45) is used as a mobile phase; the column temperature is 30 ℃; the flow rate is 1.0 ml/min; the detection wavelength is 254 nm; the theoretical plate number calculated according to the trans-anethole peak is not less than 5000,
preparing reference substance solution by accurately weighing appropriate amount of anethole reference substance, adding anhydrous ethanol to obtain solution containing 14 μ g per 1ml,
preparing test solution by pulverizing SHENBAO tablet, pulverizing with pulverizer, weighing about 2g, precisely weighing, placing into conical flask with plug, precisely adding anhydrous ethanol 20ml, sealing, weighing, ultrasonically treating for 30 min, cooling, precisely weighing, supplementing with anhydrous ethanol, shaking, filtering, collecting filtrate,
the determination method comprises precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and determining.
Example 4 content determination of ligustilide
Chromatographic conditions and system applicability test: a capillary column using (5% -phenyl) -methyl polysiloxane as a stationary phase (the column length is 30m, the inner diameter is 0.32mm, and the film thickness is 0.25 μm); column temperature is programmed temperature rise: the initial temperature is 60 ℃, the temperature is increased to 145 ℃ at the rate of 3.5 ℃ per minute, the temperature is maintained for 2 minutes, the temperature is increased to 156 ℃ at the rate of 2 ℃ per minute, the temperature is maintained for 8 minutes, and the temperature is increased to 250 ℃ at the rate of 12 ℃ per minute and the temperature is maintained for 2 minutes; the temperature of a sample inlet is 250 ℃; the temperature of the detector is 260 ℃; split-flow sample injection is carried out, and the split-flow ratio is 1: 20; the flow rate is 0.8ml/min, the number of theoretical plates is not less than 20000 calculated according to ligustilide peak,
preparation of control solutions: accurately weighing appropriate amount of ligustilide control, adding anhydrous ethanol to obtain solution containing 3.0mg per 1ml,
preparation of a test solution: taking 30 tablets of the product, placing the tablets in a round-bottom flask, adding 500ml of water, soaking until a sample is completely dissolved, testing according to a volatile oil determination method (2204 in the four Processions of the national Pharmacopeia 2015 edition), adding water from the upper end of a determinator to fill a scale part until the scale part is filled, overflowing the water into the flask, adding 1ml of dimethylbenzene, connecting a reflux condenser pipe, heating and keeping micro-boiling for 5 hours, cooling, after the solution is clearly layered, taking the dimethylbenzene liquid, placing the dimethylbenzene liquid in a 10ml measuring flask, washing the inner wall of the volatile oil determiner by a small amount of dimethylbenzene for times, merging the dimethylbenzene washing liquid into the same measuring flask, adding absolute ethyl alcohol to the scale, shaking uniformly to obtain the product,
the determination method comprises the following steps: precisely sucking 1 μ l of each of the reference solution and the sample solution, injecting into gas chromatograph, and measuring.
Claims (9)
1. The quality control method of the volatile components of the Shenbao tablet is characterized in that the volatile components comprise trans-anethole and ligustilide, wherein the content determination method of the trans-anethole comprises the following steps:
(1) chromatographic conditions and the applicability test of the system,
(2) preparation of control solutions: adding extraction solvent into trans-anethole control to obtain control solution,
(3) preparation of a test solution: pulverizing SHENBAO tablet, adding extraction solvent, ultrasonic treating, filtering, collecting filtrate,
(4) the determination method comprises the following steps: precisely sucking the reference solution and the sample solution respectively, injecting into a liquid chromatograph, and measuring,
wherein, the chromatographic conditions are as follows:
the chromatographic column is selected from: agilent ZORBAX SB-C18 column, Kromasil100-5-C18 column, Agilent ZORBAX Eclips XDB-C18 column, SHISEIDO CAPCELL PAK C18 column,
the detection wavelength is 190-400 nm,
the mobile phase is selected from: a methanol-water system, a methanol-0.1% phosphoric acid water system, a methanol-0.1% formic acid water system and a methanol-0.4% glacial acetic acid water system,
the proportion of methanol to 0.1 percent phosphoric acid water is as follows: 55-58:45,
the flow rate is: 0.8ml/min, 0.9 ml/min, 1.0ml/min, 1.1 ml/min, 1.2 ml/min,
the column temperature was: 25-35 ℃.
2. The quality control method according to claim 1, wherein a sample solution is prepared,
the extraction solvent is selected from: absolute ethyl alcohol, normal hexane, ethyl acetate,
the kidney nourishing tablet is prepared by the following crushing modes: the pulverizer pulverizes, grinds and pulverizes,
extraction time: the time of the reaction is 20 to 45 minutes,
the extraction solvent amount is as follows: 20-50 ml.
3. The quality control method according to claim 1, wherein the method for measuring the content of trans-anethole comprises the steps of:
chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent, methanol-0.1% phosphoric acid water with the volume ratio of 55-58:45 is used as a mobile phase, the column temperature is 25-35 ℃, the flow rate is 0.8-1.0ml/min, the detection wavelength is 254nm, the theoretical plate number is not less than 5000 calculated according to a trans-anethole peak,
preparation of control solutions: taking trans-anethole reference substance, precisely weighing, adding anhydrous ethanol to obtain,
preparation of a test solution: pulverizing SHENBAO tablet, collecting 1-3g, precisely weighing, placing in conical flask with plug, precisely adding anhydrous ethanol 1-40 ml, sealing, weighing, ultrasonic treating, cooling, precisely weighing, supplementing with anhydrous ethanol, shaking, filtering, collecting filtrate,
the determination method comprises the following steps: precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and measuring.
4. The quality control method according to claim 1, wherein the method for measuring the content of trans-anethole comprises the steps of:
chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; using 55:45 methanol-0.1% phosphoric acid water as mobile phase; the column temperature is 30 ℃; the flow rate is 1.0 ml/min; the detection wavelength is 254 nm; the theoretical plate number calculated according to the trans-anethole peak is not less than 5000,
preparation of control solutions: taking a proper amount of trans-anethole reference substance, precisely weighing, adding anhydrous ethanol to prepare a solution containing 14 μ g of the trans-anethole reference substance per 1ml,
preparation of a test solution: pulverizing SHENBAO tablet, weighing 2g, precisely weighing, placing in conical flask with plug, precisely adding anhydrous ethanol 20ml, sealing, weighing, ultrasonic treating for 30 min, cooling, precisely weighing, supplementing with anhydrous ethanol, shaking, filtering, collecting filtrate,
the determination method comprises the following steps: precisely sucking 10 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and measuring.
5. The quality control method according to claim 1, wherein the content determination method of ligustilide comprises the following steps:
chromatographic conditions and system applicability test: a capillary column; column temperature is programmed temperature rise, and split-flow sample injection is carried out, wherein the split-flow ratio is 1: 1-50; the flow rate is 0.7-1.5ml/min,
preparation of control solutions: adding ligustilide as control, adding extraction solvent to obtain control solution,
preparation of a test solution: taking Shenbao tablets, adding water to completely dissolve the sample, testing according to 2204 volatile oil determination method of the four general rules of the 2015 version in Chinese pharmacopoeia to obtain the Shenbao tablets,
the determination method comprises the following steps: precisely sucking 1 μ l of each of the reference solution and the sample solution, injecting into gas chromatograph, and measuring.
6. The quality control method according to claim 5, wherein the chromatographic conditions are:
the gas phase capillary column is selected from: AgilentHP-530 m × 0.32mm × 0.25 μm; AgilentHP-INNOWax 30m × 0.32mm × 0.50 μm; AgilentDB-62430 m.times.0.32 mm.times.1.8 μm.
7. The quality control method according to claim 5,
preparation of a test solution: taking kidney essence tablets, adding water to completely dissolve the kidney essence tablets, performing 2204 volatile oil determination test according to the general rule of the four departments of the 2015 edition of Chinese pharmacopoeia, adding dimethylbenzene, connecting a reflux condenser tube, heating and keeping slight boiling for 1-5 hours, cooling, after the solution is clearly layered, taking dimethylbenzene solution, placing in a 2-10ml measuring flask, washing the inner wall of a volatile oil determinator by a small amount of dimethylbenzene in times, merging the dimethylbenzene washing solution into the same measuring flask, adding absolute ethyl alcohol to scale, and shaking up to obtain the kidney essence tablet.
8. The quality control method according to claim 5, wherein the content determination of ligustilide comprises the following steps:
chromatographic conditions and system applicability test: a capillary column taking (5% -phenyl) -methyl polysiloxane as a stationary phase; column temperature is programmed temperature rise: the initial temperature is 50-70 ℃, the temperature is raised to 150 ℃ at the rate of 3-4 ℃ per minute, the temperature is maintained for 1-3 minutes, the temperature is raised to 160 ℃ at the rate of 1-3 ℃ per minute, the temperature is maintained for 6-10 minutes, the temperature is raised to 260 ℃ at the rate of 10-14 ℃ per minute, and the temperature is maintained for 1-3 minutes; the temperature of the sample inlet is 240 ℃ and 260 ℃; the temperature of the detector is 250 ℃ and 270 ℃; split-flow sample injection with a split-flow ratio of 1: 10-30; the flow rate is 0.5-1 ml/min,
preparation of control solutions: precisely weighing ligustilide reference, adding anhydrous ethanol to obtain reference solution,
preparation of a test solution: taking Shenbao tablets, adding water to completely dissolve the samples, performing 2204 volatile oil determination test according to the general rule of the four departments of the book 2015 of Chinese pharmacopoeia, adding dimethylbenzene, connecting a reflux condenser tube, heating and keeping slight boiling for 1-5 hours, cooling, after the solution is clearly layered, taking dimethylbenzene solution, placing in a 2-10ml measuring flask, washing the inner wall of a volatile oil determinator by a small amount of dimethylbenzene in times, merging the dimethylbenzene washing solution into the same measuring flask, adding absolute ethyl alcohol to scale, shaking up uniformly to obtain the Shenbao tablets,
the determination method comprises the following steps: respectively sucking the reference solution and the sample solution, injecting into a gas chromatograph, and measuring.
9. The quality control method according to claim 5, wherein the content determination of ligustilide comprises the following steps:
chromatographic conditions and system applicability test: a capillary column taking (5% -phenyl) -methyl polysiloxane as a stationary phase, wherein the length of the capillary column is 30m, the inner diameter of the capillary column is 0.32mm, and the thickness of the membrane is 0.25 mu m; column temperature is programmed temperature rise: the initial temperature is 60 ℃, the temperature is increased to 145 ℃ at the rate of 3.5 ℃ per minute, the temperature is maintained for 2 minutes, the temperature is increased to 156 ℃ at the rate of 2 ℃ per minute, the temperature is maintained for 8 minutes, and the temperature is increased to 250 ℃ at the rate of 12 ℃ per minute and the temperature is maintained for 2 minutes; the temperature of a sample inlet is 250 ℃; the temperature of the detector is 260 ℃; split-flow sample injection is carried out, and the split-flow ratio is 1: 20; the flow rate is 0.8ml/min, the number of theoretical plates is not less than 20000 calculated according to ligustilide peak,
preparation of control solutions: accurately weighing appropriate amount of ligustilide control, adding anhydrous ethanol to obtain solution containing 3.0mg per 1ml,
preparation of a test solution: taking 30 tablets of the product, placing the tablets in a round-bottom flask, adding 500ml of water, soaking until a sample is completely dissolved, testing according to 2204 volatile oil determination method in the general rule of four parts of 2015 edition of Chinese pharmacopoeia, adding 1ml of xylene until the upper end of a determinator is filled with scale parts and overflows into the flask, connecting a reflux condenser tube, heating and keeping slightly boiling for 5 hours, cooling, after the solution is clearly layered, taking xylene solution in parts, placing the xylene solution in a 10ml measuring flask, washing the inner wall of the volatile oil determinator in parts by using a small amount of xylene, merging xylene washing liquor into the same measuring flask, adding absolute ethyl alcohol to scale, shaking uniformly to obtain the product,
the determination method comprises the following steps: precisely sucking 1 μ l of each of the reference solution and the sample solution, injecting into gas chromatograph, and measuring.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010734686.2A CN111624285B (en) | 2020-07-28 | 2020-07-28 | Quality control method for volatile components of Shenbao tablet |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010734686.2A CN111624285B (en) | 2020-07-28 | 2020-07-28 | Quality control method for volatile components of Shenbao tablet |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111624285A true CN111624285A (en) | 2020-09-04 |
| CN111624285B CN111624285B (en) | 2020-11-24 |
Family
ID=72271487
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010734686.2A Active CN111624285B (en) | 2020-07-28 | 2020-07-28 | Quality control method for volatile components of Shenbao tablet |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111624285B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112697901A (en) * | 2020-12-08 | 2021-04-23 | 安徽农业大学 | Serum metabolism marker for screening heat stress resistant mutton sheep, screening method and application |
| CN113281439A (en) * | 2021-07-25 | 2021-08-20 | 江西汇仁药业股份有限公司 | Quality control detection method of Shenbao tablets |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017207467A1 (en) * | 2016-05-30 | 2017-12-07 | Givaudan Sa | Method of resolving allergens in perfume ingredients |
| CN109580830A (en) * | 2018-12-28 | 2019-04-05 | 成都康美药业生产有限公司 | The detection method of trans-anethole content in a kind of fennel seeds |
-
2020
- 2020-07-28 CN CN202010734686.2A patent/CN111624285B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017207467A1 (en) * | 2016-05-30 | 2017-12-07 | Givaudan Sa | Method of resolving allergens in perfume ingredients |
| CN109580830A (en) * | 2018-12-28 | 2019-04-05 | 成都康美药业生产有限公司 | The detection method of trans-anethole content in a kind of fennel seeds |
Non-Patent Citations (4)
| Title |
|---|
| 南海军等: "气相色谱法测定当归挥发油中藁本内酯的含量", 《海峡药学》 * |
| 张小飞等: "当归挥发油提取工艺优化及其乳化芳香水成分分析", 《中国实验方剂学杂志》 * |
| 李越峰等: "气相色谱-质谱法测定当归片和当归超微片中藁本内酯的含量比较研究", 《中国临床药理学杂志》 * |
| 林楠: "小茴香炮制工艺及化学成分研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112697901A (en) * | 2020-12-08 | 2021-04-23 | 安徽农业大学 | Serum metabolism marker for screening heat stress resistant mutton sheep, screening method and application |
| CN113281439A (en) * | 2021-07-25 | 2021-08-20 | 江西汇仁药业股份有限公司 | Quality control detection method of Shenbao tablets |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111624285B (en) | 2020-11-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103800523B (en) | A kind of preparation method of anti virus herb composition and the assay method of finger-print | |
| CN111624285B (en) | Quality control method for volatile components of Shenbao tablet | |
| CN105842353A (en) | Establishing method of fingerprint spectrum of honeysuckle-fructus forsythiae heat-clearing tablets and fingerprint spectrum | |
| CN103245733B (en) | A kind of ageratum dripping pill authentication method | |
| CN107402265B (en) | Detection method of Kangyun granule fingerprint | |
| CN101028388B (en) | Quality inspection of Chinese-medicinal preparation for treating shortsighness and asthenopia | |
| CN102335402A (en) | Detection method of Chinese preparation mixture for invigorating the spleen and replenishing qi | |
| CN101485762A (en) | Quality control method of Chinese medicine preparation | |
| CN101513467A (en) | Method for controlling quality of dermatosis toxemia preparation | |
| CN100437112C (en) | Method for inspecting Chinese medicinal preparation quality in treatment of old man eyes dieases | |
| CN105486762B (en) | A kind of high-efficiency liquid-phase fingerprint detection method of female clever ball | |
| CN113281439B (en) | Quality control detection method of Shenbao tablets | |
| CN102441057B (en) | High performance liquid chromatography (HPLC) fingerprint detection method for blood-nourishing brain-refreshing grain | |
| CN106290599A (en) | A kind of content assaying method of Chinese medicine composition | |
| CN101816753B (en) | Method for detecting quality of compound preparation for treating cold | |
| CN108693289B (en) | Method for determining content of magnoflorine in herringbone fruit medicinal material | |
| CN103048408A (en) | Specific chromatogram determination of blood-activating and pain-relieving plaster and quality detection method thereof | |
| CN110274970A (en) | The method for building up for melting poor finger-print and its application in Yixiesheng capsule Quality Control | |
| CN114942291A (en) | Method for detecting quality of 'Zhenyang Yangyin' granule | |
| CN101008632A (en) | Standard HPLC fingerprint of schisandra fruit decoction and establishing method therefor | |
| CN109142563A (en) | A kind of construction method of guilingji capsules UPLC finger-print and its application | |
| CN111693618B (en) | Construction method and detection method of UPLC characteristic spectrum of magnolia flower medicinal material | |
| CN111060637B (en) | Quality control method of agilawood koji | |
| CN110308213B (en) | Method for constructing fingerprint spectrum of kidney-nourishing and fetus-growing pill and application of fingerprint spectrum in quality detection | |
| CN114660218A (en) | Medicine composition containing 'Qingshanjuantong decoction' and detection method |
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 |