CN113237960A - Method for detecting main drug effect components of calculus bovis factitius in pharmaceutical preparation - Google Patents
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- 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
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- 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
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- 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
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- 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
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- 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
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- 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
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Abstract
The invention provides a method for detecting main pharmacodynamic components of calculus bovis factitius in a pharmaceutical preparation, which comprises the following steps: dissolving a medicinal preparation sample in an organic solvent, carrying out ultrasonic extraction, cooling, shaking up, filtering, taking a test solution obtained from a subsequent filtrate, and detecting by adopting a high performance liquid chromatography to determine main effective components of the calculus bovis factitius in the test solution: cholic acid and hyodeoxycholic acid. The method for detecting the main medicinal components of the calculus bovis factitius in the pharmaceutical preparation provided by the invention has the advantages of good precision, reproducibility and stability, accuracy and reliability, can truly reflect the quality difference of the calculus bovis factitius in the pharmaceutical preparation, and comprehensively improves the quality control system of the pharmaceutical preparation.
Description
Technical Field
The invention belongs to the technical field of detection of traditional Chinese medicine components, relates to a detection method of main effective components of calculus bovis factitius in a pharmaceutical preparation, and particularly relates to a method for detecting the main effective components of calculus bovis factitius in the pharmaceutical preparation, which comprises the following steps: a method for detecting cholic acid and hyodeoxycholic acid.
Background
The medicinal preparation is improved from Suhe flavor pills in Song Dynasty in the 70 s, is one of the most commonly used treatment medicaments for the coronary heart disease at present by using the advantages of definite curative effect, safety and economy, has quick symptom relieving and effect taking, does not have contraindication and drug resistance of nitrates, can protect blood vessels after being taken for a long time, prevent atherosclerosis, reduce the attack frequency of angina and reduce the occurrence of serious cardiovascular events, has aromatic and warm effects, tonifies qi and strengthens heart, is clinically used for chest stuffiness caused by qi stagnation and blood stasis, and has the symptoms of precordial pain and immobility; angina pectoris and myocardial infarction caused by myocardial ischemia are the above symptoms. Because the curative effect is excellent and the safety is high, the Chinese medicine is classified as a national Chinese medicine secret variety and protected by the national secret technology, and the listed national basic medicine catalogue honors a plurality of honors such as the scientific and technical prize of the Chinese traditional and western medicine combination society.
The medicinal preparation is black brown and glossy pellet, has tan cross section after being crushed, and is bitter, pungent and cool in taste and numb and tongue feeling. The medicine preparation consists of 7 kinds of Chinese medicinal materials, including artificial musk, ginseng extract, storax, artificial bezoar, cassia, toad venom, borneol, etc. and contains both non-volatile component and volatile component. The artificial bezoar is used as a traditional Chinese medicinal material in an artificial addition proportion, the main medicinal effective component of the artificial bezoar is a bile acid compound, and the artificial bezoar is difficult to detect in a conventional ultraviolet detector because a conjugated system is lacked in a bile acid structure and the bile acid has no near ultraviolet absorption characteristic. In the early-stage calculus bovis factitius quality standard research, cholic acid and hyodeoxycholic acid are found to have pharmacological effects of relieving fever and pain, resisting inflammation, benefiting gallbladder and protecting liver, reducing blood pressure, relieving cough, strengthening heart and the like as important chemical compositions and main drug effect components in calculus bovis factitius. Therefore, cholic acid and hyodeoxycholic acid are selected as index components for determining the content of calculus bovis factitius in the pharmaceutical preparation. Therefore, it is necessary to detect the bezoar bovis, control the quality of the bezoar bovis, and improve the quality control system of the pharmaceutical preparation.
Disclosure of Invention
In view of the above-mentioned disadvantages of the prior art, the present invention aims to provide a method for detecting the major effective components of calculus bovis factitius in a pharmaceutical preparation, which is used for solving the problem that the prior art lacks the following effective components: the content determination method of cholic acid and hyodeoxycholic acid.
In order to achieve the above and other related objects, a first aspect of the present invention provides a method for detecting a major pharmaceutical effective ingredient of calculus bovis factitius in a pharmaceutical preparation, comprising: adding an organic solvent into a medicinal preparation sample for dissolving, performing ultrasonic extraction, cooling, shaking up, filtering, taking a test solution obtained from a subsequent filtrate, and detecting by adopting high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) to determine main effective components of the calculus bovis factitius in the test solution: cholic acid and hyodeoxycholic acid.
Preferably, the CAS number of the cholic acid is 81-25-4, and the CAS number of the hyodeoxycholic acid is 83-49-8.
Preferably, the pharmaceutical preparation sample is a ground pharmaceutical preparation powder.
Preferably, the ratio of the added mass of the drug preparation sample to the added volume of the organic solvent is 1:5-15 g/mL. Preferably, the ratio of the added mass of the drug preparation sample to the added volume of the organic solvent is 1:10, g/mL.
Preferably, the organic solvent is selected from methanol, ethanol, methanol: 5% formic acid (95: 5), methanol: 5% formic acid water (80: 20). Preferably, the organic solvent is methanol. The above methanol: 5% formic acid water is the volume ratio of the two. The 5% formic acid water is 5% formic acid water solution by volume percentage.
Preferably, the ultrasonic extraction time is 10-30 min. Preferably, the ultrasound extraction time is 10 min.
Preferably, the power of the ultrasonic extraction is 250-350W, and the frequency of the ultrasonic extraction is 35-45 kHz. Preferably, the power of the ultrasonic extraction is 300W, and the frequency of the ultrasonic extraction is 40 kHz.
Preferably, the pharmaceutical preparation sample is precisely weighed after the addition of the organic solvent.
Preferably, the cooling is followed by reweighing and top-up loss with organic solvent.
Preferably, the filtration means: filtering the supernatant of the shaken solution with a filter membrane, and discarding the primary filtrate to obtain the subsequent filtrate.
More preferably, the filter is a 0.45 μm filter.
Preferably, the detection is performed by using a high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) combined method, which comprises the following steps:
1) preparing a reference solution: adding organic solvent to a cholic acid and hyodeoxycholic acid reference substance for dissolving and fixing volume to prepare a reference substance solution;
2) sample detection: and (2) respectively detecting the test solution and the reference solution in the step 1) by adopting a high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD), comparing the retention time for qualitative determination, and determining the content of cholic acid and hyodeoxycholic acid in the test solution by adopting an external standard method for quantitative determination.
Preferably, in step 1), the organic solvent is selected from methanol, ethanol, methanol: 5% formic acid (95: 5), methanol: 5% formic acid (80: 20). More preferably, the organic solvent is methanol.
Preferably, in step 1), the content ranges of the components in the control solution are as follows: cholic acid is 96.90-747.68 μ g/ml, hyodeoxycholic acid is 98.92-763.30 μ g/ml.
Preferably, in step 1), the reference solution is prepared by stepwise dilution.
Preferably, in the step 2), in the combination of the high performance liquid chromatography and the evaporative light scattering detector, the detector is an Evaporative Light Scattering Detector (ELSD), the evaporation temperature is 60-90 ℃, the atomizer temperature is 40-70 ℃, and the nitrogen flow rate is 1.6-2.6 SLM. More preferably, in the combination of high performance liquid chromatography and evaporative light scattering detector, the detector is an Evaporative Light Scattering Detector (ELSD), the evaporation temperature is 90 ℃, the temperature of the atomizer is 60 ℃, and the nitrogen flow rate is 1.6 SLM.
Preferably, in the step 2), the chromatographic column in the high performance liquid chromatography-evaporative light scattering detector combination is C18A chromatographic column. More preferably, the high performance liquid chromatography-evaporative light scattering detector combined (HPLC-ELSD) chromatographic column is Kromasil C18Chromatography column (5 μm,4.6 mm. times.250 mm) with octadecylsilane bonded silica as stationary phase.
Preferably, in the step 2), the column temperature in the HPLC-evaporative light scattering detector combination is 30-40 ℃. More preferably, the column temperature in the HPLC-evaporative light scattering detector combination is 35 ℃.
Preferably, in the step 2), the flow rate in the high performance liquid chromatography-evaporative light scattering detector combination is 1.2-1.4 ml/min. More preferably, the flow rate in the HPLC-evaporative light scattering detector combination is 1.3 ml/min.
Preferably, in the step 2), the sample amount in the high performance liquid chromatography-evaporative light scattering detector combination is 10-20 μ l. More preferably, the sample size in the HPLC-evaporative light scattering detector combination is 15. mu.l.
Preferably, in the step 2), in the combination of the high performance liquid chromatography and the evaporative light scattering detector, the mobile phase is acetonitrile-0.1-0.3% formic acid aqueous solution, wherein the phase A is acetonitrile, and the phase B is 0.1-0.3% formic acid aqueous solution; the analysis time is 35 min; gradient elution.
More preferably, in the HPLC-evaporative light scattering detector combination, the mobile phase is acetonitrile-0.2% aqueous formic acid solution, wherein phase A is acetonitrile and phase B is 0.2% aqueous formic acid solution; the analysis time is 35 min; gradient elution.
The 0.1-0.3% formic acid aqueous solution is 0.1-0.3% formic acid aqueous solution by volume percentage. The 0.2% formic acid aqueous solution is 0.2% formic acid aqueous solution by volume percentage.
More preferably, as shown in table 1, the specific procedure of the gradient elution is:
0-5 min, phase A: the volume ratio of the phase B is 15: 85-41: 59;
5-25 min, phase A: the volume ratio of the phase B is 41: 59-44: 56;
25-28 min, phase A: the volume ratio of the phase B is 44: 56-100: 0;
28-35 min, phase A: the volume ratio of the phase B is 100: 0-100: 0.
TABLE 1
Preferably, in step 2), the external standard method comprises the following steps:
A) preparing a series of reference substance solutions with different concentrations according to the step 1), respectively carrying out HPLC-ELSD detection to obtain a linear relation between chromatographic peak area logarithm values of cholic acid and hyodeoxycholic acid and corresponding mass (mu g) logarithm values of the cholic acid and hyodeoxycholic acid, drawing a corresponding standard working curve, and calculating to obtain a regression equation of the standard working curve of the cholic acid and hyodeoxycholic acid;
B) and (3) carrying out HPLC-ELSD detection on the test sample solution, substituting the obtained chromatographic peak area logarithm values of the cholic acid and the hyodeoxycholic acid into the regression equation of the standard working curve of the corresponding cholic acid and hyodeoxycholic acid in the step A), and calculating to obtain the content of the cholic acid and hyodeoxycholic acid in the test sample solution.
More preferably, in the standard working curve, the logarithm of the chromatographic peak area of cholic acid and hyodeoxycholic acid is taken as the ordinate (Y axis), and the logarithm of the mass (μ g) of corresponding isocholic acid and hyodeoxycholic acid is taken as the abscissa (X axis).
The second aspect of the invention provides an application of a method for detecting main drug effect components of calculus bovis factitius in a pharmaceutical preparation in quality detection of calculus bovis factitius in the pharmaceutical preparation.
The pharmaceutical preparation is a pharmaceutical preparation containing artificial bezoar, and specifically includes but is not limited to musk heart-protecting pills and the like.
The third aspect of the invention provides a quality detection method for calculus bovis factitius in a pharmaceutical preparation, wherein the content of calculus bovis factitius in the pharmaceutical preparation is more than or equal to 0.257% calculated by cholic acid and hyodeoxycholic acid.
Preferably, the weight of each pill of the pharmaceutical preparation is 22.5mg, and the content of the calculus bovis factitius in each pill of the pharmaceutical preparation is more than or equal to 58 mu g based on the total amount of the cholic acid and the hyodeoxycholic acid.
As described above, the method for detecting the main effective components of the calculus bovis factitius in the pharmaceutical preparation provided by the invention adopts the pretreatment of optimized conditions and the instrumental detection method, and the main effective components of the calculus bovis factitius in the pharmaceutical preparation are as follows: and (3) accurately quantitatively and qualitatively detecting cholic acid and hyodeoxycholic acid. The method establishes a content determination method aiming at bile acid components without ultraviolet absorption in the pharmaceutical preparation, has good precision, reproducibility and stability, is accurate and reliable, can truly reflect the quality difference of the artificial bezoar in the pharmaceutical preparation, ensures the stability of the production process and quality among batches, is used as supplement and improvement of the existing quality control standard, and comprehensively perfects the quality control system of the pharmaceutical preparation.
Drawings
Fig. 1 shows the main effective components of the calculus bovis factitius of the present invention: high performance liquid chromatograms 1a and 1b of reference substance and test substance of cholic acid and hyodeoxycholic acid, wherein fig. 1a is the high performance liquid chromatogram of the reference substance, and fig. 1b is the high performance liquid chromatogram of the test substance; the reference mark a is cholic acid, and b is hyodeoxycholic acid.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are intended to be illustrative only and not to limit the scope of the invention.
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
The reagents and equipment used in the following examples are as follows:
1. reagent
Pharmaceutical preparations (Shanghai and Huang pharmaceuticals, Inc.); cholic acid standard substance (China pharmaceutical biologicals institute, batch number: 100078-; a hyodeoxycholic acid standard substance (China pharmaceutical biologicals institute, batch number: 100087-201411, purity 99.70%); acetonitrile (chromatographic grade, TEDIA); formic acid (chromatographic grade, CNW); ethanol (analytical grade, shanghai zhuyun chemical ltd); deionized water (Mili-Q ultrapure water preparation).
2. Instrument for measuring the position of a moving object
1260 high performance liquid chromatograph, configured with on-line vacuum degasser, autosampler, quaternary pump, DAD detector, thermostatted column oven; agilent ChemStation C01.05 chromatography workstation (Agilent Corp.); 1260Infinity ELSD evaporative light scattering detector (Agilent corporation, usa); SB-5200 ultrasonic cleaning apparatus (Ningbo Xinzhi Biotech GmbH Limited Co.); Mili-Q ultra pure water instruments (Milipore Corp.).
The content of the main effective components of the calculus bovis factitius in the pharmaceutical preparation comprises the following measuring process.
1. Preparation of test solution
Grinding the medicinal preparation sample into powder, adding organic solvent for dissolving, and precisely weighing. Wherein the ratio of the added mass of the drug preparation sample to the added volume of the organic solvent is 1:5-15 g/mL. Then, ultrasonic extraction is carried out for 10-30 min at the power of 250-350W and the frequency of 35-45 kHz. Cooled, reweighed, and made up to weight loss with organic solvent. Shaking, collecting supernatant, filtering with 0.45 μm filter membrane, and discarding the primary filtrate to obtain subsequent filtrate as sample solution. The organic solvent is selected from methanol, ethanol, methanol: 5% formic acid (95: 5), methanol: 5% formic acid (80: 20).
2. Preparation of control solutions
Adding organic solvent into cholic acid and hyodeoxycholic acid reference substances, dissolving, and diluting to constant volume to obtain reference substance solution. The content ranges of the components in the reference solution are as follows: cholic acid is 96.90-747.68 μ g/ml, hyodeoxycholic acid is 98.92-763.30 μ g/ml.
The organic solvent is selected from methanol, ethanol, methanol: 5% formic acid (95: 5), methanol: 5% formic acid (80: 20).
3. Measurement of
High performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) is adopted to respectively detect the test solution and a series of reference solutions with different concentrations, the retention time is compared for qualitative determination, and an external standard method is adopted for quantitative determination. The method comprises the steps of respectively carrying out HPLC-ELSD detection on a series of control solutions with different concentrations to obtain a linear relation between chromatographic peak area logarithm values of cholic acid and hyodeoxycholic acid and corresponding mass (mu g) logarithm values of the cholic acid and hyodeoxycholic acid, drawing corresponding standard working curves, and calculating to obtain a regression equation of the standard working curves of the cholic acid and hyodeoxycholic acid. And performing HPLC-ELSD detection on the test sample solution, substituting the obtained chromatographic peak area logarithm values of the cholic acid and the hyodeoxycholic acid into the regression equation of the standard working curve of the corresponding cholic acid and hyodeoxycholic acid, and calculating to obtain the content of the cholic acid and the hyodeoxycholic acid in the test sample solution.
The high performance liquid chromatography-evaporative light scattering detector combination comprises the following detection conditions:
the detector is an Evaporative Light Scattering Detector (ELSD), the evaporation temperature is 60-90 ℃, the temperature of the atomizer is 40-70 ℃, and the flow rate of nitrogen is 1.6-2.6 SLM; the chromatographic column is C18A chromatographic column; the column temperature is 30-40 ℃; the flow rate is 1.2-1.4 ml/min; the sample introduction amount is10-20 mul; the mobile phase is acetonitrile-0.1-0.3% formic acid water solution, wherein, the phase A is acetonitrile, and the phase B is 0.1-0.3% formic acid water solution; the analysis time is 35 min; gradient elution.
The specific procedure for gradient elution was:
0-5 min, phase A: the volume ratio of the phase B is 15: 85-41: 59;
5-25 min, phase A: the volume ratio of the phase B is 41: 59-44: 56;
25-28 min, phase A: the volume ratio of the phase B is 44: 56-100: 0;
28-35 min, phase A: the volume ratio of the phase B is 100: 0-100: 0.
example 1
1. Preparation of test solution
Grinding 120 pills of the medicinal preparation sample into powder, precisely weighing 1.0g of the powder, placing the powder into a conical flask with a plug, precisely adding 10ml of methanol for dissolution, sealing the plug, and precisely weighing. Then ultrasonic extraction is carried out for 10min under the power of 300W and the frequency of 40 kHz. Cooled, reweighed and made up to weight loss with methanol. Shaking, filtering the supernatant with 0.45 μm filter membrane, discarding the primary filtrate to obtain the subsequent filtrate 1# of the sample solution.
2. Preparation of control solutions
Adding methanol into cholic acid and hyodeoxycholic acid reference substance, dissolving, and making into reference solution No. 1. The content range of each component in the reference solution 1# is as follows: cholic acid is 96.90-747.68 μ g/ml, hyodeoxycholic acid is 98.92-763.30 μ g/ml.
3. Measurement of
High performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) is adopted to detect the sample solution 1# and a series of reference solution 1# with different concentrations respectively, retention time is compared for qualitative determination, and an external standard method is adopted for quantitative determination. The method comprises the steps of respectively carrying out HPLC-ELSD detection on a series of control solution 1# with different concentrations to obtain a linear relation between chromatographic peak area logarithm values of cholic acid and hyodeoxycholic acid and corresponding mass (mu g) logarithm values of the cholic acid and hyodeoxycholic acid, drawing a corresponding standard working curve, and calculating to obtain a regression equation of the standard working curve of the cholic acid and hyodeoxycholic acid. And performing HPLC-ELSD detection on the sample solution 1#, substituting the obtained chromatographic peak area logarithm values of the cholic acid and hyodeoxycholic acid into the regression equation of the standard working curve of the corresponding cholic acid and hyodeoxycholic acid, and calculating to obtain the content of the cholic acid and hyodeoxycholic acid in the sample solution 1 #.
Wherein, the high performance liquid chromatography comprises the following detection conditions:
the detector is an Evaporative Light Scattering Detector (ELSD), the evaporation temperature is 90 ℃, the temperature of the atomizer is 60 ℃, and the nitrogen flow rate is 1.6 SLM; the chromatographic column is Kromasil C18Chromatographic column (5 μm,4.6 mm. times.250 mm) with octadecylsilane bonded silica as stationary phase; the column temperature was 35 ℃; the flow rate is 1.3 ml/min; the sample amount is 15 mul; the mobile phase is acetonitrile-0.2% formic acid water solution, wherein, the phase A is acetonitrile, and the phase B is 0.2% formic acid water solution; the analysis time is 35 min; gradient elution.
The specific procedure for gradient elution was:
0-5 min, phase A: the volume ratio of the phase B is 15: 85-41: 59;
5-25 min, phase A: the volume ratio of the phase B is 41: 59-44: 56;
25-28 min, phase A: the volume ratio of the phase B is 44: 56-100: 0;
28-35 min, phase A: the volume ratio of the phase B is 100: 0-100: 0.
example 2
A sample of the pharmaceutical formulation of batch No. H160350 is taken and the test solution is prepared in step 1 of example 1. Meanwhile, a reference solution is prepared according to the step 2 in the example 1, wherein the cholic acid content of the reference solution is 0.3mg/ml, and the hyodeoxycholic acid content of the reference solution is 0.25 mg/ml. Precisely sucking 5 μ l and 10 μ l of control solution and 15 μ l of test solution, respectively, injecting into a liquid chromatograph, measuring according to step 2 in example 1, comparing retention time, and determining the peak appearance of cholic acid and hyodeoxycholic acid in the test solution, with specific data shown in Table 2, as shown in FIG. 1.
TABLE 2
Example 3
The detection method of 2 main effective components of the calculus bovis factitius in the pharmaceutical preparation is verified by methodology, and the performance index result is as follows.
1. Precision degree
Taking a pharmaceutical preparation (batch number: H160350), preparing a sample solution according to the step 1 in the example 1, precisely sucking 15 mu l of the same sample solution, injecting the sample solution into a gas chromatograph, continuously injecting and analyzing for 6 times according to the chromatographic conditions of the step 3 in the example 1, measuring the peak area, and calculating the content of cholic acid and hyodeoxycholic acid, wherein the specific precision result is shown in a table 3. The result shows that the chromatographic peak areas RSD of the 2 components are less than 1 percent, which indicates that the method has good precision.
TABLE 3
2. Repeatability of
6 parts of the same batch of pharmaceutical preparation (batch number: H160350) are taken, a sample solution is prepared according to the step 1 in the example 1, 15 mu l of the sample solution is precisely absorbed and injected into a gas chromatograph, sample injection analysis is respectively carried out according to the chromatographic conditions of the step 3 in the example 1, the peak area is measured, and the specific repeatability result is shown in Table 4. The result shows that the chromatographic peak areas RSD of the 2 components are all less than 2.1%, which indicates that the method has good repeatability.
TABLE 4
3. Stability of
Taking a pharmaceutical preparation (batch number: H160350), preparing a sample solution according to the step 1 in the example 1, precisely sucking 15 mu l of the same sample solution, injecting the sample solution into a gas chromatograph, injecting samples for analysis for 0, 4, 8, 12, 24 and 36H according to the chromatographic conditions of the step 3 in the example 1, measuring peak areas, and calculating the content of cholic acid and hyodeoxycholic acid, wherein specific stability results are shown in a table 5. The result shows that the chromatographic peak areas RSD of the 2 components are less than 2%, and the sample solution is basically stable within 0-36 hours, which indicates that the method has good stability.
TABLE 5
4. Linear relationship of detection method
Appropriate amount of cholic acid and hyodeoxycholic acid reference substances were precisely weighed, and added with methanol according to step 2 of example 1 to prepare a series of reference substance solutions with different concentrations. According to the chromatographic conditions of the step 3 in the example 1, 10 μ l of the reference solution is precisely sucked and injected into the gas chromatograph, the logarithmic value of the mass (μ g) of the reference solution is used as the abscissa, the logarithmic value of the peak area is used as the ordinate to perform the plotting, and the standard regression equation, the correlation coefficient and the linear range of the 2 components are obtained through the measurement and calculation, and the specific results are shown in the table 6. As is clear from Table 6, the 2 components showed good linear relationship (r) in the concentration range examined2>0.999)。
TABLE 6
5. Recovery rate
6 parts of the pharmaceutical preparation (batch number: H160350) with known concentration is precisely weighed, 0.5g of the pharmaceutical preparation is accurately added with a certain amount of 10ml of methanol solution of 2 component reference substances respectively, a sample solution is prepared according to the step 1 in the example 1, and sample injection analysis is respectively carried out according to the chromatographic conditions of the step 3 in the example 1, and the result is shown in Table 7. As can be seen from Table 7, the average sample recovery rates of cholic acid and hyodeoxycholic acid were 103.43% and 100.13%, respectively, and the RSD values were 1.42% and 1.32%, respectively, which indicates that the recovery rates of the measurement results are good, indicating that the accuracy of the method is good.
TABLE 7
6. Limit of detection (LOD) and limit of quantitation (LOQ)
The limit of detection (LOD) is the concentration of the substance to be measured at a signal-to-noise ratio (S/N) of 3; the limit of quantitation (LOQ) is the concentration of the measured substance at a signal-to-noise ratio (S/N) of 10. The results are shown in Table 8, and the LOD of cholic acid and hyodeoxycholic acid is 3.3 μ g/ml and 2.8 μ g/ml respectively; LOQ was 5.3. mu.g/ml, 6.0. mu.g/ml.
TABLE 8
Example 3
75 batches of pharmaceutical preparation samples (150104-180105) produced in 2015-2018 were collected, test solutions were prepared according to step 1 in example 1, and detection was performed according to the chromatographic conditions of step 3 in example 1. The content determination results of 75 batches of the pharmaceutical preparations are shown in table 9, and the total content of cholic acid and hyodeoxycholic acid is 0.39% -0.61%. According to the transfer rate calculation of cholic acid and hyodeoxycholic acid from artificial bezoar medicinal material to finished medicinal preparation, if the content of artificial bezoar medicinal material reaches the pharmacopeia standard, the total amount of cholic acid and hyodeoxycholic acid in the medicinal preparation is 0.367%, and on the basis, the lowest limit setting is carried out according to 70%, namely the lowest limit of cholic acid and hyodeoxycholic acid in the medicinal preparation is 0.257%. Converting into 22.5mg per pill of the medicinal preparation, wherein the content of artificial bezoar in each pill is not less than 58 μ g based on total amount of cholic acid and hyodeoxycholic acid.
TABLE 9
In conclusion, the detection method for the main active ingredients of the borneol and the artificial musk in the medicinal preparation provided by the invention has the advantages of good precision, reproducibility and stability, accuracy and reliability, can truly reflect the quality difference of the artificial musk and the borneol in the medicinal preparation, and is used for supplementing and improving the existing quality control standard. Therefore, the invention effectively overcomes various defects in the prior art and has high industrial utilization value.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which may be accomplished by those skilled in the art without departing from the spirit and scope of the present invention as set forth in the appended claims.
Claims (10)
1. A method for detecting main effective components of calculus bovis factitius in a pharmaceutical preparation comprises the following steps: adding an organic solvent into a medicinal preparation sample for dissolving, carrying out ultrasonic extraction, cooling, shaking up, filtering, taking a test solution obtained from a subsequent filtrate, and detecting by adopting a high performance liquid chromatography-evaporative light scattering detector combination to determine main drug effect components of the calculus bovis factitius in the test solution: cholic acid and hyodeoxycholic acid.
2. The method of claim 1, wherein the preparation of the test solution comprises one or more of the following conditions:
A) the ratio of the mass of the added medicinal preparation sample to the volume of the added organic solvent is 1:5-15 g/mL;
B) the organic solvent is selected from methanol, ethanol, methanol: 5% formic acid water 95: 5, methanol: 5% formic acid water 80: 20;
C) the ultrasonic extraction time is 10-30 min;
D) the power of the ultrasonic extraction is 250-350W, and the frequency of the ultrasonic extraction is 35-45 kHz.
3. The method of claim 1, wherein the detection is performed by HPLC-evaporative light scattering detector, comprising the steps of:
1) preparing a reference solution: adding organic solvent to a cholic acid and hyodeoxycholic acid reference substance for dissolving and fixing volume to prepare a reference substance solution;
2) sample detection: detecting the test solution and the reference solution in the step 1) respectively by adopting a high performance liquid chromatography-evaporative light scattering detector combination, comparing the retention time for qualitative determination, and determining the content of cholic acid and hyodeoxycholic acid in the test solution by adopting an external standard method for quantitative determination.
4. The method for detecting the main effective components of calculus bovis factitius in the pharmaceutical preparation according to claim 3, wherein in the step 1), the content ranges of the components in the reference solution are as follows: cholic acid is 96.90-747.68 μ g/ml, hyodeoxycholic acid is 98.92-763.30 μ g/ml.
5. The method for detecting the major drug effect component of calculus bovis factitius in the pharmaceutical preparation according to claim 3, wherein in the step 2), the high performance liquid chromatography-evaporative light scattering detector is used, the detector is an evaporative light scattering detector, the evaporation temperature is 60-90 ℃, the atomizer temperature is 40-70 ℃, and the nitrogen flow rate is 1.6-2.6 SLM.
6. The method according to claim 3, wherein the HPLC-evaporative light scattering detector combination used in step 2) comprises the following detection conditions: the chromatographic column is C18A chromatographic column; the column temperature is 30-40 ℃; the flow rate is 1.2-1.4 ml/min; the sample injection amount is 10-20 mu l; the mobile phase is acetonitrile-0.1-0.3% formic acid water solution, wherein, the phase A is acetonitrile, and the phase B is 0.1-0.3% formic acid water solution; the analysis time is 35 min; gradient elution.
7. The method for detecting the main effective components of calculus bovis factitius in the pharmaceutical preparation according to claim 6, wherein the specific procedure of gradient elution is as follows: 0-5 min, phase A: the volume ratio of the phase B is 15: 85-41: 59; 5-25 min, phase A: the volume ratio of the phase B is 41: 59-44: 56; 25-28 min, phase A: the volume ratio of the phase B is 44: 56-100: 0; 28-35 min, phase A: the volume ratio of the phase B is 100: 0-100: 0.
8. use of the method according to any one of claims 1-7 for detecting the major active ingredients of calculus bovis factitius in a pharmaceutical preparation for quality detection of calculus bovis factitius in a pharmaceutical preparation.
9. A quality detection method for calculus bovis factitius in a pharmaceutical preparation comprises that the content of calculus bovis factitius in cholic acid and hyodeoxycholic acid is more than or equal to 0.257%.
10. The method for detecting the quality of calculus bovis factitius in a pharmaceutical preparation according to claim 9, wherein the pharmaceutical preparation contains calculus bovis factitius in an amount of more than or equal to 58 μ g based on the total amount of cholic acid and hyodeoxycholic acid when the pharmaceutical preparation weighs 22.5mg per pill.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1598572A (en) * | 2004-08-16 | 2005-03-23 | 江苏中康药物科技有限公司 | Quality determination method for external cultivated bezoar |
| CN102944640A (en) * | 2012-11-27 | 2013-02-27 | 哈药集团中药二厂 | Quality control method of AngongNiuhuangShuan |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1598572A (en) * | 2004-08-16 | 2005-03-23 | 江苏中康药物科技有限公司 | Quality determination method for external cultivated bezoar |
| CN102944640A (en) * | 2012-11-27 | 2013-02-27 | 哈药集团中药二厂 | Quality control method of AngongNiuhuangShuan |
Non-Patent Citations (3)
| Title |
|---|
| 吴芳 等: "蒸发光散射检测器-高效液相色谱法测定小儿化毒胶囊中胆酸和猪去氧胆酸含量", 《中国药业》 * |
| 徐陆忠 等: "HPLC-ELSD法测定人工牛黄甲硝唑胶囊中胆酸和猪去氧胆酸的含量", 《医药导报》 * |
| 谭菊英 等: "HPLC-CAD法测定人工牛黄及其制剂小儿氨酚黄那敏颗粒中胆酸和猪去氧胆酸的含量", 《中国药师》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113866306A (en) * | 2021-09-28 | 2021-12-31 | 上海和黄药业有限公司 | Detection method of HPLC (high Performance liquid chromatography) characteristic spectrum of medicinal preparation |
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