Quality control detection method of Yunnan red medicine

Abstract

The invention belongs to the technical field of medicine quality control, and particularly relates to a quality control detection method of Yunnan red medicine. According to the method, the UPLC fingerprint of Yunnan Hongyao is measured by adopting high performance liquid chromatography equipment through a screening extraction method and chromatographic conditions; the UPLC fingerprint spectrum is subjected to fingerprint spectrum similarity and chemical mode recognition analysis, and Yunnan Hongyao capsules of different batches or different raw material batches can be distinguished. The quality control detection method disclosed by the invention can be used for evaluating the consistency of the Yunnan red medicine, carrying out quality control on the production process and raw medicinal materials of related preparations, providing a reliable evaluation means for large-scale production of the Yunnan red medicine, and having a wide application prospect.

Description

Technical Field

[0001] The invention belongs to the technical field of drug quality control, and particularly relates to a quality control detection method for Yunnan red medicine. Background Art

[0002] As a protected variety of traditional Chinese medicine, the research and development of Yunnan Red Medicine is deeply rooted in the traditional Yi medicine theory and rich practical experience. The formula brings together many unique and precious folk Chinese herbal medicines in Yunnan. With these unique herbs, Yunnan Red Medicine has many significant effects such as hemostasis and analgesia, blood circulation and blood stasis, and wind and dampness removal. The origin, batch, preparation process and other factors of Yunnan Red Medicine have a very important influence on its efficacy. Preparing Yunnan Red Medicine into capsules, tablets and other preparations not only greatly improves the convenience of Chinese medicine, but also enhances its application value in the modern medical system.

[0003] At present, the quality control of Yunnan Hongyao mainly adopts high performance liquid chromatography, for example, quantitative determination of the content of certain active ingredients in Yunnan Hongyao capsules, such as notoginsenoside R1, ginsenoside Rg1, and ginsenoside Rb1, or limited inspection of the toxic ingredient dianconitine. Although these methods guarantee the quality of drugs to a certain extent, the existing technology can only quantitatively detect some ingredients in Yunnan Hongyao, and it is difficult to effectively distinguish the differences between different batches of products, resulting in poor accuracy in consistency evaluation.

[0004] Therefore, constructing a quality control method for Yunnan Hongyao that can accurately evaluate consistency has become a key issue that needs to be urgently addressed in this field. Summary of the invention

[0005] In view of the defects of the prior art, the present invention provides a quality control detection method for Yunnan Red Medicine, aiming at accurately performing consistency evaluation.

[0006] The present invention provides a quality control detection method for Yunnan red medicine, comprising the following steps:

[0007] Step 1, extracting the active ingredients from Yunnan red medicine to obtain a test solution;

[0008] Step 2, detecting the test solution by liquid chromatography to obtain a chromatogram;

[0009] The chromatographic conditions include:

[0010] The mobile phases were: 0.1-0.3 w / v% phosphoric acid aqueous solution as mobile phase A, acetonitrile as mobile phase B;

[0011] The chromatographic column is a C18 reverse phase column.

[0012] Preferably, in step 1, the extraction step includes: dissolving Yunnan Hongyao in a solvent, and performing reflux or ultrasonic extraction; the mass-volume ratio of Yunnan Hongyao to the solvent is 1 g: 20-100 ml; and / or, the extraction time is 30-90 min;

[0013] and / or, the Yunnan Hongyao is the powder in Yunnan Hongyao capsules.

[0014] Preferably, the mass-volume ratio of Yunnan Hongyao to the solvent is 1 g: 50 ml; and / or, the extraction is performed by ultrasonic extraction, and the extraction time is 30 min.

[0015] Preferably, the solvent is selected from methanol.

[0016] Preferably, the chromatographic conditions further include: the flow rate is 0.3-0.5 mL / min; and / or, the column temperature is 25-40 °C; and / or, the injection volume is 1-10 μL.

[0017] Preferably, the column temperature is 30 °C.

[0018] Preferably, the flow rate is 0.4 mL / min; and / or, the injection volume is 2 μL.

[0019] Preferably, the chromatographic column is selected from GIST column, 2.1×100 mm, 2 μm;

[0020] Preferably, the detection wavelength is 203-210 nm;

[0021] and / or, the elution mode of the mobile phase is gradient elution, and the elution gradient is: 0-1 min, 5%-9% mobile phase B; 1-7 min, 9% mobile phase B; 7-10 min, 9%-13% mobile phase B; 10-17 min, 13% mobile phase B; 17-18 min, 13%-14% mobile phase B; 18-32 min, 14% mobile phase B; 32-33 min, 14%-18% mobile phase B; 33-50 min, 18% mobile phase B; 50-51 min, 18%-22% mobile phase B; 51-63 min, 22% mobile phase B; 63-64 min, 22%-26% mobile phase B; 64-69 min, 26%-30% mobile phase B; 69-72 min, 30%-35% mobile phase B; 72-76 min, 35%-40% mobile phase B; 76-82 min, 40%-45% mobile phase B; 82-90 min, 45%-70% mobile phase B; 90-95 min, 70% mobile phase B;

[0022] and / or, the chromatogram is a fingerprint, containing 1-28 chromatographic peaks.

[0023] Preferably, the chromatographic peak contains at least one of (+)(−)catechin, baicalin, notoginsenoside R1, ginsenoside Rg1, wogonoside, baicalein, ginsenoside Rb1, β-asarone, wogonin, oroxylin A, paris saponin II, and paris saponin I.

[0024] Preferably, fingerprint similarity analysis and / or chemometric pattern recognition analysis is performed on the chromatogram.

[0025] Preferably, when the fingerprint similarity is greater than 0.99, it indicates good similarity between sample batches;

[0026] and / or, the chemometric pattern recognition analysis is selected from at least one of principal component analysis and partial least squares discriminant analysis.

[0027] The present invention provides a quality control detection method for Yunnan Hongyao. By screening extraction methods and chromatographic conditions, the UPLC fingerprint of Yunnan Hongyao capsules is obtained using high-performance liquid chromatography equipment; fingerprint similarity and chemometric pattern recognition analysis are performed on the UPLC fingerprint to distinguish Yunnan Hongyao capsules of different batches or different raw material batches. The quality control detection method of the present invention can be used for the consistency evaluation of Yunnan Hongyao, control the quality of the production process and raw medicinal materials of related preparations, and provides a reliable evaluation method for the large-scale production of Yunnan Hongyao, with broad application prospects.

[0028] Obviously, based on the above content of the present invention, according to common general knowledge and conventional means in the art, without departing from the above basic technical idea of the present invention, various other forms of modification, substitution, or variation can be made.

[0029] The above content of the present invention will be further described in detail below through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention belong to the scope of the present invention. Description of the Drawings

[0030] Figure 1 are the UPLC chromatograms of the reference substance (A) and the test sample (B), where the 2nd peak is (+)(−)catechin, the 3rd peak is 7-hydroxycoumarin, the 4th peak is scopoletin, the 11th peak is oroxylin A-7-O-β-D-glucuronide, the 12th peak is baicalin, the 13th peak is notoginsenoside R1, the 14th peak is ginsenoside Rg1, the 15th peak is oroxylin A-7-O-β-D-glucuronide, the 16th peak is wogonoside, the 17th peak is baicalein, the 19th peak is ginsenoside Rb1, the 21st peak is wogonin, the 23rd peak is oroxylin A, the 27th peak is paris saponin II, and the 28th peak is paris saponin I;

[0031] Figure 2 It is the UPLC fingerprint of 31 batches of samples;

[0032] Figure 3 It is the PCA score plot of 31 batches of samples;

[0033] Figure 4 It is the PLS-DA score plot of the common peaks of 31 batches of samples;

[0034] Figure 5 It is the VIP plot of the common peaks of 31 batches of samples;

[0035] Figure 6 It is the cluster analysis plot of 31 batches of samples;

[0036] Figure 7 It is the UPLC fingerprint under different extraction conditions;

[0037] Figure 8 It is the UPLC fingerprint under different extraction volumes;

[0038] Figure 9 It is the UPLC fingerprint under different flow rates;

[0039] Figure 10 It is the UPLC fingerprint under different column temperatures;

[0040] Figure 11 It is the UPLC fingerprint under different injection volumes;

[0041] Figure 12 It is the UPLC fingerprint under different mobile phases. Specific Embodiments

[0042] In the following examples and experimental examples, the reagents and materials not specifically stated are all commercially available products.

[0043] I. Instruments

[0044] Shimadzu LC-40B X3 high-performance liquid chromatograph, equipped with a CTO-40C DAD detector (Shimadzu Corporation, Japan); BSA124S analytical balance (one in a hundred thousand) (Sartorius Scientific Instruments Co., Ltd., Beijing); FA1004 analytical balance (one in ten thousand) (Shanghai Liangping Instrumentation Co., Ltd.); UPH-I ultrapure water preparation instrument (Sichuan Youpu Ultrapure Technology Co., Ltd.); SB-8200DTS dual-frequency ultrasonic instrument (Ningbo Xinyi Ultrasonic Equipment Co., Ltd.); H1650-W micro desktop high-speed centrifuge (Hunan Xiangyi Laboratory Development Co., Ltd.)

[0045] II. Reagents and Drugs

[0046] Phosphoric acid and acetonitrile were of chromatographic grade (Thermo Fisher Scientific (China) Co., Ltd.); water was ultrapure water; other reagents were all of analytical grade.

[0047] Reference standards: Purchased from Lemeitian Pharmaceutical / Deste Biotechnology Co., Ltd., Chengdu Mantest Biotechnology Co., Ltd., and Chengdu Cromar Biotechnology Co., Ltd. (+)(-) Catechin (batch number DST231108-001, purity > 98%), Baicalin (batch number DSTDH002303, purity > 98%), Notoginsenoside R1 (batch number DSTDS0050003, purity > 98%), Ginsenoside Rg1 (batch number DSTDR000902, purity > 98%), Wogonoside (batch number DSTDH002602, purity > 98%), Baicalein (batch number DSTDH002402, purity > 98%), Ginsenoside Rb1 (batch number DSTDR000603, purity > 98%), Wogonin (batch number DSTDH002501, purity > 98%), Oroxylin A (batch number DSTDQ004102, purity > 98%), Paris saponin II (batch number DSTDC002802, purity > 98%), Paris saponin I (batch number DSTDC002802, purity > 98%), Oroxylin A glucoside (batch number 57396-78-8, purity > 98%), Oroxylin A-7-O-β-D-glucuronide (batch number 36948-76-2, purity > 98%) reference standards were purchased from Lemeitian Pharmaceutical / Deste Biotechnology Co., Ltd.; 7-Hydroxycoumarin (batch number MUST-21080504, purity > 98%) reference standard was purchased from Chengdu Mantest Biotechnology Co., Ltd.; Scopolin (batch number CHB-D-091, purity > 98%) reference standard was purchased from Chengdu Cromar Biotechnology Co., Ltd.A total of 31 batches of Yunnan Hongyao Capsules were provided by Yunnan Botanic Pharmaceutical Co., Ltd., with batch numbers: 20220508 (No. S1), 20230301 (No. S2), 20230706 (No. S3), 20240303 (No. S4), 20240304 (No. S5), 20240307 (No. S6), 20240308 (No. S7), 20240309 (No. S8), 20240310 (No. S9), 20240311 (No. S10), 20240312 (No. S11), 20240801 (No. S12), 20240802 (No. S13), 20240803 (No. S14), 20240804 (No. S15), 20240805 (No. S16), 20240806 (No. S17), 20240807 (No. S18), 20240808 (No. S19), 20240809 (No. S20), 20240810 (No. S21), 20240601 (No. S22), 20240602 (No. S23), 20240603 (No. S24), 20240604 (No. S25), 20240605 (No. S26), 20240606 (No. S27), 20240607 (No. S28), 20240608 (No. S29), 20240609 (No. S30), 20240610 (No. S31).

[0048] Example 1 Quality control detection method for Yunnan Hongyao Capsules

[0049] The detection method of this example includes the following steps:

[0050] 1. Reference substance solution

[0051] Weigh an appropriate amount of the reference substance and dissolve it in methanol to obtain the solution. Among them, the concentration of (+)(-) catechin is 11.3 μg / ml, the concentration of 7-hydroxycoumarin is 13.68 μg / ml, the concentration of scopoletin is 0.1025 mg / ml, the concentration of oroxylin A glucoside is 21.75 μg / ml, the concentration of baicalin is 24.87 μg / ml, the concentration of notoginsenoside R1 is 19.87 μg / ml, the concentration of ginsenoside Rg1 is 1.93 mg / ml, the concentration of oroxylin A-7-O-β-D-glucuronide is 53.25 μg / ml, the concentration of wogonoside is 37.00 μg / ml, the concentration of baicalein is 47.5 μg / ml, the concentration of ginsenoside Rb1 is 0.1987 mg / ml, the concentration of wogonin is 3.82 μg / ml, the concentration of oroxylin A is 13.12 μg / ml, the concentration of parisposide II is 0.11 mg / ml, and the concentration of parisposide I is 11.87 μg / ml.

[0052] 2. Test solution

[0053] Take the Yunnan Hongyao Capsules to be tested, weigh about 1.0 g of the powder in the capsules, place it in a 100 mL stoppered conical flask, add 50 mL of methanol solution, weigh it, ultrasonically extract for 30 min, cool it, weigh it again, make up the lost weight with methanol solution, shake well, centrifuge (13000 r / min) for 15 min, and take the supernatant to obtain the test solution.

[0054] 3. Chromatographic conditions

[0055] Chromatographic column: Shim-pack GIST C18 (2.1X100 mm, 2 μm);

[0056] Mobile phase: 0.2% phosphoric acid (A) - acetonitrile (B),

[0057] Elution conditions: Gradient elution, and the elution gradient is as follows:

[0058] 0 - 1 min, 5% - 9% B; 1 - 7 min, 9% B; 7 - 10 min, 9% - 13% B; 10 - 17 min, 13% B; 17 - 18 min, 13% - 14% B; 18 - 32 min, 14% B; 32 - 33 min, 14% - 18% B; 33 - 50 min, 18% B; 50 - 51 min, 18% - 22% B; 51 - 63 min, 22% B; 63 - 64 min, 22% - 26% B; 64 - 69 min, 26% - 30% B; 69 - 72 min, 30% - 35% B; 72 - 76 min, 35% - 40% B; 76 - 82 min, 40% - 45% B; 82 - 90 min, 45% - 70% B; 90 - 95 min, 70% B

[0059] Flow rate: 0.4 mL / min; Column temperature 30 °C; Detection wavelength 203 nm; Injection volume 2 μL.

[0060] 4. Determination of chromatogram

[0061] Take 2 μL of the reference solution and the test solution (numbered S2) respectively, inject them into the high-performance liquid chromatograph, and obtain the UPLC fingerprint chromatogram. The results are as Figure 1 shown.

[0062] By literature review, peak 2 was identified as (+)(-) catechin, peak 3 as 7-hydroxycoumarin, peak 4 as scopolletin, peak 11 as oroxylin A-7-O-β-D-glucoside, peak 12 as baicalin, peak 13 as notoginsenoside R1, peak 14 as ginsenoside Rg1, peak 15 as oroxylin A-7-O-β-D-glucuronide, peak 16 as wogonoside, peak 17 as baicalein, peak 19 as ginsenoside Rb1, peak 21 as wogonin, peak 23 as oroxylin A, peak 27 as paris saponin II, and peak 28 as paris saponin I. Then, peak 12 (baicalin), which had good separation from adjacent chromatographic peaks and stable retention time, was selected as the reference.

[0063] 5. Similarity analysis

[0064] (1) Fingerprint similarity analysis

[0065] The chromatographic data of different batches of samples to be tested were imported into the software "Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints (2012 Edition)". The time window width was set to 0.1 min, and the multi-point calibration method was used for full-peak matching to determine the common peaks. Taking the samples to be tested numbered S1 - S31 as an example, a total of 28 common peaks were determined, as Figure 2 shown.

[0066] The chromatograms of different batches of samples to be tested were analyzed using the "Similarity Evaluation System for Traditional Chinese Medicine Fingerprints" (2012 Edition). The chromatogram of one batch of samples was set as the reference chromatogram, and the fingerprint similarities of different batches of Yunnan Hongyao Capsules were calculated. If the fingerprint similarity was greater than 0.99, it indicated good similarity among the batches of Yunnan Hongyao Capsule samples.

[0067] Taking the samples to be tested numbered S1 - S31 as an example, choosing the chromatogram of sample S2 as the reference chromatogram, the fingerprint similarities of S1 - S31 were 0.990, 0.997, 0.996, 0.997, 0.998, 0.998, 0.997, 0.999, 0.999, 0.998, 0.999, 0.994, 0.997, 0.996, 0.997, 0.996, 0.998, 0.995, 0.996, 0.996, 0.996, 0.997, 0.997, 0.997, 0.997, 0.996, 0.997, 0.996, 0.994, 0.996, 0.997, respectively, all greater than 0.99, indicating good similarity among these 31 batches of Yunnan Hongyao Capsule samples, and the similarity among batches of samples S1 - S31 was even higher.

[0068] (2) Chemical pattern recognition analysis

[0069] (2.1) Principal component analysis (PCA)

[0070] Import the common peak data into SIMICA 13.0 software for unsupervised PCA analysis. Taking the test samples numbered S1 - S31 as an example, the results are shown in Figure 3 . It can be seen therefrom that the 31 batches of samples can be divided into 3 categories with relatively obvious discrimination.

[0071] (2.2) Partial least squares discriminant analysis (PLRS-DA)

[0072] Import the common peak data into SIMICA 13.0 software, and establish a supervised PLS-DA discriminant analysis on the basis of PCA analysis to obtain the PLS-DA score plot, VIP plot, and cluster analysis plot.

[0073] Taking the test samples numbered S1 - S31 as an example, the results are respectively shown in Figures 4 - 6 .

[0074] From Figure 4 it can be seen that the 31 batches of samples can be divided into 3 regions. From Figure 5 it can be seen that the 19th peak is ginsenoside Rb1, the 23rd peak is oroxylin A, the 17th peak is baicalein, the 28th peak is paris saponin I, the 27th peak is paris saponin II, the 11th peak is oxyresveratrol, the 14th peak is ginsenoside Rg1, the 15th peak is oroxylin A-7-O-β-D-glucuronide, and the 21st peak is wogonin, and their contribution values are all greater than 1; the 18th and 22nd peaks come from Scutellaria wightiana, the 25th peak comes from processed Aconitum vilmorinianum Kom., the 26th peak is a common peak of Paris vietnamensis (Takht.) H. Li and Acorus tatarinowii Schott, the 24th peak comes from Rhynchosia volubilis Lour., and the 20th peak comes from Acorus tatarinowii Schott, which have a significant impact on classification and are the quality markers of the main components. From Figure 6 it can be seen that the 11 batches of samples can be clustered into 3 major categories, which is Figure 4 consistent. Therefore, the results of chemical pattern recognition analysis show that the above 31 batches of test samples are clearly divided into three categories, indicating that there are certain differences in the common peaks of different batches of samples.

[0075] Combined with the results of fingerprint similarity analysis and chemical pattern recognition analysis, it can be seen that the quality control method of this example can distinguish Yunnan Hongyao samples between different raw material batches.

[0076] The technical solution of the present invention will be further described below through experiments.

[0077] Experimental Example 1 Screening of Extraction Methods

[0078] 1. Screening of Extraction Conditions

[0079] Determine the active ingredients in Yunnan Hongyao Capsules (No. S1) according to the method of Example 1, with the difference that the extraction conditions are reflux for 1.5 h, ultrasonic for 1.5 h, and ultrasonic for 0.5 h respectively.

[0080] The results are as Figure 7As shown, according to the separation effect of the chromatographic peaks at 12-18min and 76-80min, and the overall peak area, the optimal extraction condition is determined to be ultrasonic 0.5h.

[0081] 2. Screening of extraction solvent volume

[0082] The effective ingredients in Yunnan Hongyao Capsules (No. S1) were determined according to the method of Example 1, except that the volumes of the extraction solvent (methanol) were 20 ml, 40 ml, 50 ml, 60 ml, 80 ml and 100 ml, respectively.

[0083] The results are as follows Figure 8 As shown, the peak area of peak 12 is used as the standard to reflect the degree of extraction. The peak area of peak 12 under different extraction solvent volumes is compared. The results show that when the methanol concentration is 50 ml, the peak area of peak 12 is the largest, and the extraction is more complete. Therefore, the optimal methanol concentration is determined to be 50 ml.

[0084] By screening the extraction conditions and the volume of the extraction solvent, the preferred extraction method of the present invention is: using 50 ml of methanol and ultrasonic extraction for 0.5 h.

[0085] Experimental Example 2 Screening of Chromatographic Conditions

[0086] 1. Flow rate screening

[0087] The effective ingredients in Yunnan Hongyao Capsules (No. S1) were determined according to the method of Example 1, except that the flow rates were set to 0.3 ml / min, 0.4 ml / min, and 0.5 ml / min, respectively.

[0088] The results are as follows Figure 9 As shown, considering the number of chromatographic peaks and response values, it was found that the flow rate had little effect on the fingerprint spectrum, and the optimal flow rate was 0.4 ml / min.

[0089] 2. Column temperature selection

[0090] The effective ingredients in Yunnan Hongyao Capsules (No. S1) were determined according to the method of Example 1, except that the column temperatures were 25°C, 30°C, and 35°C, respectively.

[0091] The results are as follows Figure 10 As shown, according to the separation of the chromatographic peaks of 12-26 min and 72-80 min, the preferred column temperature is 30°C.

[0092] 3. Screening of injection volume

[0093] The effective ingredients in Yunnan Hongyao Capsules (No. S1) were determined according to the method of Example 1, except that the injection volumes were 1 μL, 2 μL, 5 μL, and 10 μL, respectively.

[0094] The results are as follows Figure 11 As shown, when the injection volume is 1 μL, the peak area is too small. When the injection volumes are 5 μL and 10 μL, the column is overloaded. Therefore, the preferred injection volume is 2 μL.

[0095] 4. Screening of mobile phase

[0096] Determine the active ingredients in Yunnan Hongyao Capsules (No. S1) according to the method of Example 1, with the difference that the mobile phases are 0.2% phosphoric acid (A) - acetonitrile (B) and water (A) - acetonitrile (B) respectively.

[0097] The results are as follows Figure 12 As shown, when the mobile phase A is water, the separation effect of multiple chromatographic peaks is poor and the signal response value is low. Therefore, the preferred mobile phase is 0.2% phosphoric acid (A) - acetonitrile (B).

[0098] Based on the above results, the preferred chromatographic conditions are determined as follows: flow rate of 0.4 ml / min; column temperature of 30 °C; injection volume of 2 μL; mobile phase of 0.2% phosphoric acid (A) - acetonitrile (B).

[0099] Experimental Example 3 Methodology verification

[0100] I. Precision test

[0101] Intra-day precision: Take the same test solution (No. S2), inject and measure 6 times under the chromatographic conditions of Example 1. Using baicalin as the reference peak, the RSD of the relative retention time of each common chromatographic peak is 0.03% - 0.12%, and the RSD of the relative peak area is 0.54% - 2.63%, indicating good precision of the instrument.

[0102] Inter-day precision: Take the same test solution (No. S2), inject and measure 6 times on the same day under the chromatographic conditions of Example 1, continuously measure for three days and analyze the data. Using baicalin as the reference peak, the RSD of the relative retention time of each common chromatographic peak is 0.03% - 0.14%, and the RSD of the relative peak area is 0.90% - 2.49%, indicating good precision of the instrument.

[0103] II. Repeatability test

[0104] Take the content under the difference in capsule filling amount of the same batch (No. S2) of capsules (i.e., the powder in the capsules), make 6 parallel portions, prepare the test solution according to the method of the test solution in Example 1, inject and measure under the chromatographic conditions of Example 1. Using baicalin as the reference peak, the RSD of the relative retention time of each common chromatographic peak is 0.03% - 0.44%, and the RSD of the relative peak area is 1.05% - 1.55%, indicating good repeatability of the method of the present invention.

[0105] III. Stability experiment

[0106] Take the content under the difference in capsule filling amount of the same batch (batch number S2) (i.e., the powder in the capsule), prepare the test solution according to the method of the test solution in Example 1, store it at room temperature for 0, 2, 4, 8, 12, and 24 h respectively, inject and determine under the chromatographic conditions of Example 1. Using baicalin as the reference peak, the RSD of the relative retention time of each common chromatographic peak is 0.02%-0.12%, and the RSD of the relative peak area is 0.74%-2.60%, indicating that the test solution has good stability within 24 h after preparation.

[0107] The results of this experimental example show that the quality control detection method of the present invention has good precision, repeatability, and stability.

[0108] The above experimental example results show that the extraction method and chromatographic conditions of the present invention are obtained through screening. The preferred extraction method is: using 50 ml of methanol and ultrasonic extraction for 0.5 h; the preferred chromatographic conditions are a flow rate of 0.4 ml / min; a column temperature of 30 °C; an injection volume of 2 μL; 0.2% phosphoric acid (A)-acetonitrile (B). The quality control detection method of the present invention has good precision, repeatability, and stability, the method is reliable, and it helps to promote the standardization and regularization of the entire industry.

[0109] From the above examples and experimental examples, it can be seen that the present invention provides a quality control detection method for Yunnan Hongyao Capsules. The present invention screens the extraction method and chromatographic conditions, and uses a high-performance liquid chromatography device to measure the UPLC fingerprint of Yunnan Hongyao Capsules; by performing fingerprint similarity and chemical pattern recognition analysis on the UPLC fingerprint, Yunnan Hongyao Capsules of different batches or different raw material batches can be distinguished. The quality control detection method of the present invention can be used for the consistency evaluation of Yunnan Hongyao, control the quality of the production process and raw medicinal materials of related preparations, and provides a reliable evaluation means for the large-scale production of Yunnan Hongyao, with broad application prospects.

Claims

1. A quality control detection method for Yunnan Hongyao, characterized in that, It includes the following steps: Step 1: Extract the active ingredients from Yunnan Hongyao to obtain a test solution. Step 2: Detect the test solution by liquid chromatography to obtain a chromatogram. Among them, the chromatographic conditions include: The mobile phase is: 0.1 - 0.3 w / v% phosphoric acid aqueous solution as mobile phase A, and acetonitrile as mobile phase B. The chromatographic column is a C18 reversed-phase chromatographic column.

2. The quality control and detection method of Yunnan Hongyao according to claim 1, characterized in that, In Step 1, the extraction steps include: Yunnan Hongyao is dissolved in a solvent, and refluxed or ultrasonically extracted; the mass-to-volume ratio of Yunnan Hongyao to the solvent is 1 g: 20 - 100 ml; and / or, the extraction time is 30 - 90 min. And / or, the Yunnan Hongyao is the powder in Yunnan Hongyao capsules.

3. The quality control detection method of Yunnan Hongyao according to claim 2, characterized in that: The mass-to-volume ratio of Yunnan Hongyao to the solvent is 1 g: 50 ml; and / or, the extraction is by ultrasonic extraction, and the extraction time is 30 min.

4. The quality control and detection method of Yunnan Hongyao according to claim 1, characterized in that: The chromatographic conditions further include: the flow rate is 0.3 - 0.5 mL / min; and / or, the column temperature is 25 - 40 °C; and / or, the injection volume is 1 - 10 μL.

5. The quality control and detection method of Yunnan Hongyao according to claim 4, characterized in that: The column temperature is 30 °C.

6. The quality control and detection method of Yunnan Hongyao according to claim 4, characterized in that: The flow rate is 0.4 mL / min; and / or, the injection volume is 2 μL.

7. The quality control and detection method of Yunnan Hongyao according to claim 1, characterized in that: The chromatographic column is selected from GIST column, 2.1×100 mm, 2 μm.

8. The quality control and detection method of Yunnan Hongyao according to claim 1, characterized in that: The detection wavelength is 203 - 210 nm; And / or, the elution mode of the mobile phase is gradient elution, and the elution gradient is: 0 - 1 min, 5% - 9% mobile phase B; 1 - 7 min, 9% mobile phase B; 7 - 10 min, 9% - 13% mobile phase B; 10 - 17 min, 13% mobile phase B; 17 - 18 min, 13% - 14% mobile phase B; 18 - 32 min, 14% mobile phase B; 32 - 33 min, 14% - 18% mobile phase B; 33 - 50 min, 18% mobile phase B; 50 - 51 min, 18% - 22% mobile phase B; 51 - 63 min, 22% mobile phase B; 63 - 64 min, 22% - 26% mobile phase B; 64 - 69 min, 26% - 30% mobile phase B; 69 - 72 min, 30% - 35% mobile phase B; 72 - 76 min, 35% - 40% mobile phase B; 76 - 82 min, 40% - 45% mobile phase B; 82 - 90 min, 45% - 70% mobile phase B; 90 - 95 min, 70% mobile phase B; And / or, the chromatogram is a fingerprint, including 1 - 28 chromatographic peaks.

9. The quality control and detection method of Yunnan Hongyao according to claim 1, characterized in that: The chromatogram is subjected to fingerprint similarity analysis, and / or chemical pattern recognition analysis.

10. The quality control detection method of Yunnan Hongyao according to claim 9, characterized in that: When the fingerprint similarity is greater than 0.99, it indicates that the similarity between sample batches is good; And / or, the chemical pattern recognition analysis is selected from at least one of principal component analysis and partial least squares - discriminant analysis.

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