Content determination method of Qianjin coptis chinensis pills

Abstract

The invention discloses a content determination method of a traditional Chinese medicine Qianjin coptis chinensis pill, which comprises the following steps: a test solution and a reference solution of the Qianjin coptis chinensis pill are taken for detection under the chromatographic conditions that a chromatographic column with octadecylsilane chemically bonded silica as a filler is adopted, a mobile phase A is selected from acetonitrile, and a mobile phase B is a buffer saline solution; and obtaining content information of the Qianjin coptis pill according to a detection result. The reference substances are catalpol, berberine hydrochloride, palmatine hydrochloride, epiberberine, coptisine, caffeic acid, ferulic acid, magnoflorine, jateorhizine and columbamine. The chemical components of the Qianjin coptis pill are comprehensively and systematically analyzed, and a technical method is provided for perfecting and improving the medicine quality standard of the Qianjin coptis pill and better controlling the medicine quality.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine analysis, and particularly relates to a method for determining the content of Qianhuanglian pills. Background Art

[0002] Type 2 diabetes mellitus (T2DM) is a globally prevalent metabolic disease characterized by insulin resistance and chronic hyperglycemia. With the changes in modern lifestyle and the aging of the population, the incidence of T2DM has increased year by year and has become one of the major diseases that seriously threaten human health. The complex pathogenesis of T2DM involves multiple factors, including genetic susceptibility, environmental influences, and lifestyle.

[0003] Qianhuangjinlian Pills, as a traditional Chinese medicine, has a long history of use and is widely used in the treatment of diabetes and its related complications. Studies have shown that Qianhuangjinlian Pills can not only effectively lower blood sugar levels, but also improve insulin sensitivity, protect pancreatic function, and have a certain preventive effect on diabetes-related complications.

[0004] Quality control of traditional Chinese medicine is a key measure to ensure the safety and effectiveness of drugs. By establishing objective, multi-dimensional and reliable quality control points through modern drug analysis technology, the quality management level of traditional Chinese medicine can be significantly improved. The current drug standards mainly focus on chemical composition, content, purity and stability to ensure the safety and effectiveness of drugs. For traditional Chinese medicine, the current standards include indicators such as chemical composition analysis, content determination, fingerprint spectrum, microbial limit test, and heavy metal and pesticide residue detection. However, there is a gap between the current standards and the actual needs of traditional Chinese medicine quality control, which is mainly reflected in the difficulty in fully reflecting the complexity of traditional Chinese medicine with multiple components and multiple functions, insufficient efficacy evaluation, and some traditional quality control methods are highly subjective, with poor accuracy and repeatability, which are difficult to meet the requirements of modern traditional Chinese medicine quality control.

[0005] Since Qianhuanglian Pills are made of raw rehmannia root and coptis root, the main active ingredients contained in the two are quite different. Therefore, the current measurement method for Qianhuanglian Pills is limited to using the HPLC detection method recorded in the pharmacopoeia to detect the main ingredients in raw rehmannia root and coptis root respectively, and there is still a lack of an overall and comprehensive method for determining the content of the main ingredients in Qianhuanglian Pills.

[0006] At present, there is no literature report on the research of the fingerprint spectrum of Qianhuang Jinlian Pills claimed for protection in the present invention. In the prior art, only the quality analysis of the single component in the Chinese medicine compound is carried out, and there is no comprehensive and systematic quality control method to reflect the quality status of the main reference sample components in the Chinese medicine compound and the finished product of the present invention, and it is impossible to effectively control its production process and product quality, and it is impossible to better guarantee its clinical efficacy. Therefore, it is necessary to adopt a fingerprint spectrum quality control method that can comprehensively control the overall quality of the Chinese medicine compound of the present invention to control its key quality. Summary of the invention

[0007] The purpose of the present invention is to provide a method for determining the content of Qianhuanglian Pills.

[0008] The object of the present invention is achieved as follows: A method for determining the content of Qianhuanglian Pills, comprising the steps of preparing a test sample, preparing a reference solution, and liquid phase detection: (1) Preparation of test solution: Grind Qianhuang Jinlian Pills into powder, add 60% methanol, and the solid-liquid ratio is 50 mL 60% methanol per 1 gram of Qianhuang Jinlian Pills; ultrasonically treat at 25°C, 35 min, and 175 W, then filter, concentrate to 50 mL, cool to room temperature, and lyophilize; add an appropriate amount of precision lyophilized powder to methanol to dissolve and prepare a Qianhuang Jinlian Pills test solution with a drug content of 0.1 g / mL.

[0009] (2) Preparation of reference solution: Accurately weigh a certain amount of catalpol, berberine hydrochloride, palmatine hydrochloride, epiberberine, coptisine, caffeic acid, ferulic acid, magnoliavine, jatrorrhizine, and fangdeline, and prepare a 0.1 mg / mL mixed reference solution in methanol.

[0010] (3) Liquid phase detection: Qianhuanglian Pills test solution and reference solution were tested. The chromatographic conditions of the test were as follows: the specifications of the chromatographic column were: column length 250 mm, inner diameter 4.0 mm, particle size 5 μm; column temperature: 25 °C; flow rate: 1 mL / min; injection volume: 10 μL; detection wavelength: 210 nm; mobile phase A was acetonitrile, mobile phase B was 0.05 mol / L potassium dihydrogen phosphate solution, and the pH of the mobile phase was 3; elution program: 0-37.5 min, 10-21.5% B; 37.5-43 min, 21.5-25% B; 43-43.5 min, 25-25.75% B; 43.5-46 min, 25.75-26.5% B; 46-48.5 min, 26.5-26.75% B; 48.5~58.5min,26.75~27.1%B; 58.5~62.5min, 27.1~27.5%B; 62.5~65.5min, 27.5~34%B; 65.5~68min,34~10%B; Based on the test results, the content information of Qianhuanglian Pills was obtained.

[0011] The standard curve of catalpol is y=2.1475x-8.4428, R 2 =0.996; the standard curve of berberine hydrochloride is y=38.6829x-152.0169, R 2 =0.999; the standard curve of caffeic acid is y=23.362x-70.052, R 2=0.997; the standard curve of magnolia alkaloids is y=58.964x-172.499, R 2 =0.998; the standard curve of ferulic acid is y=27.527x-92.427, R 2 =0.996; the standard curve of tetrandrine is y=47.796x-178.592, R 2 =0.998; the standard curve of epiberberine is y=46.312x-162.758, R 2 =0.999; the standard curve of jatrorrhizine is y=24.312x-102.446, R 2 =0.999; the standard curve of bamipine hydrochloride is y=36.045x-143.845; R 2 =0.999.

[0012] Advantages of the present invention: Compared with the prior art, the advantages of the present invention are: (1) The fingerprint of Qianhuanglian Pills was established, which overcomes the defect that the determination of the content of a single ingredient is difficult to reflect the overall content. The intrinsic quality of Qianhuanglian Pills can be controlled from a holistic and macroscopic perspective, ensuring the efficacy of the drug. By using the main means of modern drug research, the classic prescription can be passed down through fine products, so that the classic prescription can be subject to more formal quality control.

[0013] (2) The chemical composition of Qianhuanglian Pills of the present invention is complex, and it is difficult to separate its characteristic peaks. In the process of establishing the fingerprint spectrum, the present invention adopts a gradient elution method to solve the problem of difficulty in separating the fingerprint characteristic peaks and interference of impurity peaks.

[0014] (3) In the process of establishing the fingerprint spectrum of the reference sample of the Chinese medicine compound of the present invention, 9 common characteristic peaks were confirmed, and their relative retention time, relative peak area and similarity were studied, thereby ensuring the chemical composition stability and safety of the reference sample, and providing an important reference basis and quality reference for the quality control of subsequent compound preparations.

[0015] (4) The fingerprint spectra of each effective ingredient in the traditional Chinese medicine compound reference sample of the present invention are regarded as a whole, and the order and relationship of each characteristic peak are emphasized. This not only avoids the one-sidedness of judging the overall quality of the traditional Chinese medicine compound reference sample of the present invention by only measuring one or two chemical components, but also reduces the possibility of artificial processing to meet the quality standards, thus providing a new method and means for completely and accurately evaluating the quality of the traditional Chinese medicine compound reference sample of the present invention.

[0016] (5) The method of the present invention has good stability, high precision, good reproducibility, is convenient and easy to master. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is the chromatogram of Qianhuang Jinlian Pills; Figure 2 This is the superimposed spectrum of multiple batches of samples of Qianhuang Jinlian Pills; Figure 3 This is the heat map of similarity evaluation of multiple batches of Qianhuang Jinlian Pills; Figure 4 This is the chromatogram of the mixed reference solution of Shengdihuang, Huanglian and Jinhuanglian Pills; Figure 5 This is the hierarchical clustering analysis diagram of Qianhuang Jinlian Pills; Figure 6 Distribution plot for principal component analysis. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments. The specific embodiments described herein are only used to explain the present invention and are not used to limit the protection content of the present invention.

[0019] A method for determining the content of Qianhuanglian Pills, comprising the steps of preparing a test sample, preparing a reference solution, and liquid phase detection: (1) Preparation of test solution: Grind Qianhuang Jinlian Pills into powder, add 60% methanol, and the solid-liquid ratio is 50 mL 60% methanol per 1 gram of Qianhuang Jinlian Pills; ultrasonically treat at 25°C, 35 min, and 175 W, then filter, concentrate to 50 mL, cool to room temperature, and lyophilize; add an appropriate amount of precision lyophilized powder to methanol to dissolve and prepare a Qianhuang Jinlian Pills test solution with a drug content of 0.1 g / mL.

[0020] (2) Preparation of reference solution: Accurately weigh a certain amount of catalpol, berberine hydrochloride, palmatine hydrochloride, epiberberine, coptisine, caffeic acid, ferulic acid, magnoliavine, jatrorrhizine, and fangdeline, and prepare a mixed reference solution containing 0.1 mg / mL in methanol.

[0021] (3) Liquid phase detection: Qianhuanglian Pills test solution and reference solution were tested. The chromatographic conditions of the test were as follows: the specifications of the chromatographic column were: column length 250 mm, inner diameter 4.0 mm, particle size 5 μm; column temperature: 25 °C; flow rate: 1 mL / min; injection volume: 10 μL; detection wavelength: 210 nm; mobile phase A was acetonitrile, mobile phase B was 0.05 mol / L potassium dihydrogen phosphate solution, and the pH of the mobile phase was 3; elution program: 0-37.5 min, 10-21.5% B; 37.5-43 min, 21.5-25% B; 43-43.5 min, 25-25.75% B; 43.5-46 min, 25.75-26.5% B; 46-48.5 min, 26.5-26.75% B; 48.5~58.5min,26.75~27.1%B; 58.5~62.5min, 27.1~27.5%B; 62.5~65.5min, 27.5~34%B; 65.5~68min,34~10%B; Based on the test results, the content information of Qianhuanglian Pills was obtained.

[0022] The standard curve of catalpol is y=2.1475x-8.4428, R 2 =0.996; the standard curve of berberine hydrochloride is y=38.6829x-152.0169, R 2 =0.999; the standard curve of caffeic acid is y=23.362x-70.052, R 2 =0.997; the standard curve of magnolia alkaloids is y=58.964x-172.499, R 2 =0.998; the standard curve of ferulic acid is y=27.527x-92.427, R 2 =0.996; the standard curve of tetrandrine is y=47.796x-178.592, R 2 =0.998; the standard curve of epiberberine is y=46.312x-162.758, R 2 =0.999; the standard curve of jatrorrhizine is y=24.312x-102.446, R 2 =0.999; the standard curve of bamipine hydrochloride is y=36.045x-143.845; R 2 =0.999. Example

[0023] 1. Instruments and Materials 1.1 Instruments and Equipment Agilent 1200 series high performance liquid chromatograph (VWD UV detector, Agilent Technologies, USA), SK2510HP ultrasonic cleaner (Shanghai Kedao Ultrasonic Instrument Co., Ltd.), 2-4N medical centrifuge (Hunan Hengnuo Instrument Equipment Co., Ltd.), N-1200BD-OSB2100 rotary evaporator (Shanghai Quanjie Instrument Co., Ltd.), JA2003N electronic balance (actual graduation value: 1 mg, Shanghai Jinghai Instrument Co., Ltd.).

[0024] 1.2 Reagents and Materials Radix Rehmanniae (batch number: 2212021) and Rhizoma Coptidis (batch number: C230608003) were purchased from Yunnan Yunzi Pharmaceutical Co., Ltd. Berberine reference substance (batch number: 110713-202316), catalpol reference substance (batch number: 110808-202313), caffeic acid reference substance (batch number: 110725-201318), magnoliavine reference substance (batch number: 110525-201408), jatrorrhizine reference substance (batch number: 110712-201316), and epiberberine reference substance (batch number: 110713-202316) were purchased from Yunnan Yunzi Pharmaceutical Co., Ltd. : 110720-201412), ferulic acid reference substance (batch number: 110721-201311) and palmatine reference substance (batch number: 110722-201314) were purchased from China Food and Drug Inspection Institutes, tetrandrine reference substance (batch number: W14J8Z37548) was purchased from Shanghai Yuanye Biotechnology Co., Ltd., methanol (batch number: LA80X17) and acetonitrile (batch number: LA80X26) were purchased from J&K Technology Group, and other reagents were of analytical grade, and ultrapure water was used in the experiments.

[0025] 2 Experimental methods 2.1 Sample preparation The 14 batches of Coptis chinensis and Rehmannia glutinosa collected were crushed, sieved, bagged, labeled and stored according to the processing methods in the "Chinese Pharmacopoeia 2020 Edition".

[0026] Preparation of Qianhuanglian Pills: According to Table 1, accurately weigh a certain amount of 1:1 ratio of coptis chinensis and raw rehmannia powder. Reflux the raw rehmannia powder for 1 hour at a solid-liquid ratio of 1:10, and combine the filtrate after reflux three times. After concentrating the filtrate to a suitable volume, pour it into the previously prepared coptis chinensis powder and stir the powder to mix it evenly. Then place the mixture in an oven at 40°C and bake it until it is crispy, and Qianhuanghuanglian Pills are obtained.

[0027] Table 1 Qianhuangjinlian Pills Combination Table .

[0028] Preparation of test sample: Qianhuang Jinlian pills were crushed and ground into powder, and 60% methanol was added, with a solid-liquid ratio of 1:50 (g·mL-1). After ultrasonic treatment at an ultrasonic temperature of 25℃, an ultrasonic time of 35min and a power of 175w, the mixture was filtered, concentrated to 50mL, cooled to room temperature, and lyophilized to obtain the test sample.

[0029] 2.2 Solution preparation 2.2.1 Preparation of reference solution Accurately weigh a certain amount of catalpol, berberine hydrochloride, palmatine hydrochloride, epiberberine, coptisine, caffeic acid, ferulic acid, magnoliavine, jatrorrhizine, and tetrandrine, and prepare them into a 0.1 mg / mL mixed reference substance methanol solution.

[0030] 2.2.2 Preparation of test solution Accurately weigh an appropriate amount of the lyophilized powder under Item 2.1 and dissolve it in methanol to prepare a Qianhuanglian Pills test solution with a drug content of 0.1 g / mL.

[0031] 2.2.3 Preparation of negative control test solution Weigh the prescribed amount of Rehmannia root and Coptis chinensis respectively, and prepare the negative control test solution of Coptis chinensis and Rehmannia root for Qianhuangjinlian Pills by the same preparation and extraction method as Qianhuangjinlian Pills.

[0032] 2.3 Chromatographic conditions Chromatographic column: RD-C18 column (250 mm × 4.0 mm, 5 μm); column temperature: 25 ° C; flow rate: 1 mL / min; injection volume: 10 μL Detection wavelength: 210nm; elution was performed according to Table 2. Mobile phase A was acetonitrile, and mobile phase B was 0.05 mol / L potassium dihydrogen phosphate solution (pH=3) Table 2 Gradient change table .

[0033] 2.4 Methodological Review 2.4.1 Investigation of linear relationship Accurately pipette the above 9 reference substance solutions respectively, and prepare reference substance stock solutions of different concentrations after diluting them 2, 4, 6, 8, and 10 times, and record the results according to the chromatographic conditions under the above "2.3".

[0034] 2.4.2 Precision test Repeat the injection of the reference solution and the test solution 6 times according to the chromatographic conditions under "2.3" and calculate the RSD value of the characteristic peak to retention time.

[0035] 2.4.3 Repeatability test Take the same batch of raw Rehmannia root and Coptis chinensis samples, and prepare 6 groups of test sample solutions according to the same test sample preparation method. Determine according to the chromatographic conditions under "2.3" and calculate the RSD value of the characteristic peak to retention time.

[0036] 2.4.4 Stability test Take the above test solution and measure it according to the chromatographic conditions under "2.3", inject the sample at 0, 2, 4, 8, 12 and 24h respectively, and calculate the RSD value of the characteristic peak to retention time.

[0037] 2.5 System suitability test Inject samples according to the chromatographic conditions in Item 2.3. The theoretical plate number shall not be less than 9000, calculated as catalpol and berberine respectively.

[0038] 2.6 Establishment of fingerprint of Qianhuang Jinlian Pills Fifteen batches of Qianhuang Jinlian Pills samples were tested according to the method under 2.3. After the data were collected, they were imported into the Chinese medicine chromatographic fingerprint similarity evaluation system for analysis, and the common peaks were obtained and the similarity was evaluated.

[0039] 2.7 HPLC fingerprint analysis of Qianhuanglian Pills 2.7.1 Hierarchical cluster analysis The common peak information of 15 batches of Qianhuanglian Pills obtained under Item 2.6 was imported into SMICA software for hierarchical clustering analysis after processing.

[0040] 2.7.2 Principal Component Analysis The common peak information of 15 batches of Qianhuanglian Pills obtained under Item 2.6 was imported into SMICA software for principal component analysis after processing.

[0041] 3 Results The chromatogram of Qianhuanglian Pills is shown in Figure 1 .

[0042] 3.1 Precision investigation Peaks 1 to 9 are: catalpol, caffeic acid, magnolamine, ferulic acid, tertrenol, epiberberine, jatrorrhizine, palmatine hydrochloride, berberine. The relative retention time of the selected reference substances was calculated, and the results are shown in Table 3. The RDS values ​​are all less than 0.18%, indicating that the method has good precision.

[0043] Table 3 Precision of Qianhuanglian Pills .

[0044] 3.2 Repeatability study Peaks 1 to 9 are: catalpol, caffeic acid, magnolamine, ferulic acid, tertrenol, epiberberine, jatrorrhizine, palmatine hydrochloride, berberine. The relative retention time of the selected reference substances was calculated, and the results are shown in Table 4. The RDS values ​​were all less than 0.9%, indicating that the method had good repeatability.

[0045] Table 4 Repeatability of Qianjinhuanglian Pills .

[0046] 3.3 Stability investigation Peaks 1 to 9 are: catalpol, caffeic acid, magnolamine, ferulic acid, tertrenol, epiberberine, jatrorrhizine, palmatine hydrochloride, berberine. The relative retention time of the selected reference substances was calculated, and the results are shown in Table 5. The RDS values ​​are all less than 0.55%, indicating that the samples are stable and not easy to deteriorate.

[0047] Table 5 Stability of Qianjinhuanglian Pills .

[0048] 3.6 System adaptability experiment Qianhuanglian Pills of S8 were selected as the test drug for the system suitability experiment. The results showed that the theoretical plate number of catalpol was 83489, and the theoretical plate number of berberine was 289089, both greater than 9000, indicating that the chromatographic method has good system applicability.

[0049] 3.7 Linear Investigation The reference substance solutions were accurately aspirated respectively, and prepared into reference substance stock solutions of different concentrations after diluting 2, 4, 6, 8, and 10 times. The linear regression equation is shown in Table 6.

[0050] Table 6 Regression equations and linear ranges of nine reference compounds .

[0051] 3.8 Determination of the content of active ingredients in Qianhuanglian Pills Calculate the content of the main components: Calculate the peak area of ​​the corresponding chromatographic peak of the mixed reference substance in the test solution, and calculate the content of the main components by the external standard method, which is the content of the main components in Qianhuanglian Pills. The results are shown in Table 7. Table 7 Content of each ingredient in different batches .

[0052] 3.9 Establishment of fingerprint of Qianhuang Jinlian Pills Different batches of Qianhuanglian Pills were analyzed, and the superimposed fingerprints of Qianhuanglian Pills samples and the reference fingerprints were generated by the software "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)". Figure 2It can be seen that 15 batches of Qianhuang Jinlian Wan samples produced 30 common peaks.

[0053] At the same time, the similarity between 15 batches of Qianhuanglian Pills and the generated reference spectra was calculated with the help of the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (2012 Edition)" software. The similarity heat map is shown in the figure below. Figure 3 As shown. Figure 3 It can be seen that the similarity of each batch is above 0.73, indicating that the similarity of multiple batches of Qianhuang Jinlian Pills is good.

[0054] Similarly, the mixed reference solution of Radix Rehmanniae, Rhizoma Coptis and Qianhuangjinlian Pills was injected under the same chromatographic conditions as Qianhuangjinlian Pills. The results are as follows: Figure 4 As shown in the figure, 8 of the 28 common peaks of Qianhuanglian Pills belong to Shengdihuang, and the remaining 22 belong to Huanglian. The active ingredients in Shengdihuang are mostly concentrated in the first half of the chromatogram, mainly composed of highly polar substances such as organic acids and iridoid glycosides, while Huanglian is the opposite, mainly composed of alkaloids and the substances are mainly concentrated in the second half, and there is no negative interference between the two.

[0055] 3.10 HPLC fingerprint analysis of Qianhuang Jinlian Pills 3.10.1 Hierarchical Cluster Analysis Hierarchical clustering analysis was performed on the common peak data of HPLC of 15 batches of Qianhuanglian Pills samples. Figure 4 As shown. We found that sample batches S2 and S15 showed high similarity in chemical composition, while sample S6 showed obvious differences from other samples. Samples S3, S4, and S14 also showed certain similarities, while S9 clustered alone, which may mean that it was significantly different from other samples in composition. In addition, samples S10, S5, S8, and S13 clustered together with S3, S4, and S14 again to form a larger cluster, indicating that these batches have high similarity in chemical composition. Hierarchical cluster analysis uses Ward's method to merge clusters by minimizing the variance within each cluster, helping us identify batches with similar or different compositions, which is crucial for quality control, production process optimization, and ensuring the consistency of drug ingredients and the efficacy and safety of drugs.

[0056] 3.10.2 Principal Component Analysis After high performance liquid chromatography (HPLC) analysis of 15 batches of Qianhuanglian Pills samples, the common peak area data were imported into SIMCA software for principal component analysis (PCA), and we obtained a distribution diagram reflecting the differences in chemical composition between sample batches, such as Figure 5 shown. Figure 5The marks in (such as S10, S6, S1B, S5, etc.) represent samples from different batches, and the values ​​of the common peak areas indicate the relative contents of specific chemical components in these batches. The PCA results show that the first principal component explains 67.8% of the data variation, while the second principal component explains 14.4%, indicating that the main differences between samples are captured by the first principal component. In addition, through Hotelling's T2 95% confidence ellipse, we can distinguish between normal and abnormal samples, and the points outside the ellipse may represent batches with significant differences in composition. This is consistent with the results of hierarchical clustering analysis.

Claims

1. A method for determining the content of Qianhuanglian Pills, characterized in that The determination method includes the steps of test sample preparation, reference solution preparation, and liquid phase detection: (1) Preparation of test solution: Qianhuanglian pills were crushed and ground into powder, and 60% methanol was added, with the material-liquid ratio being 50 mL of 60% methanol per 1 gram of Qianhuanglian pills; ultrasonic treatment was performed at 25 °C, 35 min, and 175 W, followed by filtration, concentration to 50 mL, cooling to room temperature, and lyophilization; an appropriate amount of the lyophilized powder was dissolved in methanol to prepare a Qianhuanglian pill test solution with a drug content of 0.1 g / mL; (2) Preparation of reference solution: accurately weigh catalpol, berberine hydrochloride, palmatine hydrochloride, epiberberine, coptisine, caffeic acid, ferulic acid, magnoliavine, jatrorrhizine, and fangfang pine, and prepare a 0.1 mg / mL mixed reference solution in methanol; (3) Liquid phase detection: The Qianhuanglian Pills test solution and the reference solution were tested. The chromatographic conditions for the test were as follows: the specifications of the chromatographic column were: column length 250 mm, inner diameter 4.0 mm, particle size 5 μm; column temperature: 25 ℃; flow rate: 1 mL / min; injection volume: 10 μL; detection wavelength: 210 nm; mobile phase A was acetonitrile, mobile phase B was 0.05 mol / L potassium dihydrogen phosphate solution, and the pH of the mobile phase was 3; elution program: 0 ~ 37.5 min, 10 ~ 21.5 % B; 37.5 ~ 43 min, 21.5 ~ 25 % B; 43 ~43.5 min, 25 ~ 25.75 % B; 43.5 ~ 46 min, 25.75 ~ 26.5 % B; 46 ~ 48.5 min, 26.5 ~26.75 % B; 48.5 ~ 58.5 min,26.75 ~ 27.1% B;58.5 ~ 62.5 min,27.1 ~ 27.5% B;62.5~ 65.5 min,27.5 ~ 34 % B;65.5 ~ 68 min,34 ~ 10% B; Based on the test results, the content information of Qianhuanglian Pills was obtained.

2. The measuring method according to claim 1, characterized in that The standard curve of catalpol is y=2.1475x-8.4428, R 2 =0.996; the standard curve of berberine hydrochloride is y=38.6829x-152.0169, R 2 =0.999; the standard curve of caffeic acid is y=23.362x-70.052, R 2 =0.997; the standard curve of magnolia alkaloids is y=58.964x-172.499, R 2 =0.998; the standard curve of ferulic acid is y=27.527x-92.427, R 2 =0.996; the standard curve of tetrandrine is y=47.796x-178.592, R 2 =0.998; the standard curve of epiberberine is y=46.312x-162.758, R 2 =0.999; the standard curve of jatrorrhizine is y=24.312x-102.446, R 2 =0.999; the standard curve of bamipine hydrochloride is y=36.045x-143.845; R 2 =0.999.

Images