Quality detection method of bulbus fritillariae cirrhosae lung-heat-clearing syrup

Abstract

The invention provides a quality detection method of bulbus fritillariae cirrhosae lung-heat-clearing syrup. According to the quality detection method, the contents of amygdalin, sipeimine, liquiritin and platycodin in the bulbus fritillariae cirrhosae lung-heat-clearing syrup are detected by adopting an HPLC-ELSD (High Performance Liquid Chromatography-Evaporative Light Scattering Detector) chromatography. According to the method disclosed by the invention, the contents of four components, namely amygdalin, sipeimine, liquiritin and platycodin D, in the bulbus fritillariae cirrhosae lung-heat-clearing syrup are determined by adopting an HPLC-ELSD method, so that the quality of the bulbus fritillariae cirrhosae lung-heat-clearing syrup can be relatively rapidly evaluated, and reference is expected to be provided for quality control of a preparation. According to the method, interference of strong ultraviolet absorption substances is effectively avoided, the research cost is reduced, the operation process is simplified, an economic and efficient analysis means is provided for determining the content of saponin compounds, and meanwhile, a reference is provided for formulating a quality control standard of the bulbus fritillariae cirrhosae lung-heat-clearing syrup.

Description

Technical Field

[0001] The invention provides a quality detection method for Chuanbei Qingfei syrup, belonging to the field of drug detection. Background Art

[0002] Chuanbei Qingfei Syrup (also known as Chuanbei Qingfei Dew) is composed of eight Chinese medicinal herbs: Fritillaria cirrhosa, loquat leaves, bitter almonds, ophiopogon japonicus, platycodon grandiflorum, licorice, mint, and rehmannia root. It has the effects of clearing the lungs and moistening dryness, relieving cough and reducing phlegm. It is used to treat dry cough, dry throat, sore throat and bronchitis caused by phlegm-heat in the lungs, and has shown good anti-inflammatory effects in clinical practice. Its quality standards are included in the second volume of the Ministry of Health's Drug Standards (Chinese Medicine Formula Preparations) WS3-B-0184-90, but the quality standards are relatively simple, only including properties, relative density and routine items under syrup. At present, its research mainly focuses on clinical application and pharmacological activity, and its indicator active ingredients are less studied, such as: Sun Hongde, Yu Xiaofang, Huang Yan, et al. Effect of Chuanbei Qingfei Syrup on Serum IFN-γ and IL-2 in Children with Mycoplasma Pneumonia [J]. China Practical Medicine, 2012, 7(16): 177-178. Xue Yijia. Determination of glycyrrhizic acid in Chuanbei Qingfei syrup by HPLC[J]. Strait Pharmacy, 2014, 26(12):86-88. Gong Xueyuan, et al., Study on the detection of adulterated Ophiopogon japonicus in Chuanbei Qingfei syrup based on HPLC-Q-TOF-MS technology, Strait Pharmacy, Vol. 34, No. 5, 2022, established a detection method for adulterated Ophiopogon japonicus in Chuanbei Qingfei syrup using high performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry (HPLC-Q-TOF-MS) technology. Chen Jianqin, Determination of amygdalin, liquiritin and glycyrrhizic acid in Chuanbei Qingfei syrup by dual wavelength UPLC method, Chinese Journal of Pharmaceutical Industry, 2015, 46(8), established an ultra-high performance liquid chromatography method for simultaneous determination of amygdalin, liquiritin and glycyrrhizic acid in Chuanbei Qingfei syrup. Application number 202011430478.X, invention name: HPLC method for simultaneous detection of 6 active ingredients in Chuanbei Qingfei Syrup, provides an HPLC method for simultaneous detection of 6 active ingredients in Chuanbei Qingfei Syrup, the 6 active ingredients include ursolic acid, oleanolic acid, amygdalin, catalpol, glycyrrhizic acid and liquiritin; the steps include: measuring the Chuanbei Qingfei Syrup sample to be tested, adding an alcohol aqueous solution and then extracting to obtain a test solution; injecting it into a high performance liquid chromatograph, and obtaining a chromatogram after testing by HPLC, wherein mobile phase A and mobile phase B are acetonitrile and 0.08-0.12% phosphoric acid aqueous solution, respectively; calculating the contents of the 6 active ingredients in Chuanbei Qingfei Syrup by external standard method. Application number 202111623401.9, invention name: A detection method for Chuanbei Qingfei Syrup, provides a detection method for Chuanbei Qingfei Syrup, specifically relates to a detection method for Chinese medicine preparations, the detection method identifies loquat leaves by thin layer chromatography, and determines the content of ammonium glycyrrhizinate in licorice. Through methodological investigation, the detection method has the effects of strong specificity, good durability, good recovery rate, etc., and can effectively detect the quality of Chuanbei Qingfei Syrup.

[0003] Amygdalin is a common cyanogenic glycoside compound with multiple active effects such as immunomodulation, immunosuppression, antitussive, antiasthmatic, and anti-inflammatory. Zhong Changhui, Qian Jin, Yang Yuanzheng, Xian Lina, Xiao Xing, Hu Zhihua. Effect of amygdalin on the Kelch-like epichlorohydrin-related protein-1 / nuclear factor E2-related factor 2 signaling pathway in rats with bronchial asthma [J]. Chinese Journal of Clinical Pharmacology, 2022, 38(18): 2157-2161. Studies have shown that amygdalin can effectively inhibit airway remodeling and prevent the progression of asthma. Its effect may be related to the activation of the Keap-1 / Nrf2 antioxidant signaling pathway; chaetoclax glycoside belongs to the alkaloid glycoside compound, Zhang Hongli. Evaluation of the anti-inflammatory efficacy of chaetoclax Vb and chaetoclax glycoside in vitro [D]. Henan University, 2020. The in vitro anti-inflammatory effect of licorice was studied and it was found that xibei glycosides regulated inflammatory gene expression by blocking TLR4-mediated MyD88-dependent and TRIF-dependent signaling pathways; liquiritigenin is one of the main flavonoid components in licorice. Zhu Hongjiang, Pang Lijian, Wang Tianjiao, Zang Ningzi, Yang Li, Wang Mei, Qiu Yihe, Si Qi, Lv Xiaodong. Research progress on the treatment of lung diseases with Adenophora Root and Radix Ophiopogonis Decoction and its quality marker (Q-marker) prediction analysis [J]. Chinese Journal of Traditional Chinese Medicine, 1-18. It is believed that liquiritigenin can be used as one of the quality markers of Adenophora Root and Radix Ophiopogonis Decoction, which can treat chronic bronchitis, pneumonia, chronic obstructive pulmonary disease (COPD) and many other lung diseases. Licorice glycosides exert anti-inflammatory effects by inhibiting the extracellular signal-related kinase (ERK) / mitogen-activated protein kinase (MAPK) pathway, reducing the expression of inflammatory factors such as TNF-α and IL-1β mRNA, and inhibiting the gene and protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) (Cui Yuanyuan, Liu Jiaxin, Xing Boyu, et al. Research progress on the basis of anti-inflammatory active substances and their mechanism of action of licorice [J]. Chinese Journal of Traditional Chinese Medicine, 2024, 42(06): 99-103+276.); Platycodon saponin D is a pentacyclic triterpenoid saponin isolated from Platycodon grandiflorum, which can prevent and treat lung diseases by reducing lung inflammation, alleviating lung oxidative stress, inhibiting lung tissue fibrosis, and preventing lung tissue cell apoptosis. ( Chai Yanqun, Mao Di, Chai Yanping. Research progress on the pharmacological effects of platycoside D in preventing and treating lung diseases [J]. Modern Drugs and Clinics, 2024, 39(11): 2996-3000.). Since there are many glycoside components in the four ingredients and the ultraviolet absorption differences are large, there is currently no literature reporting the detection of the four ingredients under the same conditions. Summary of the invention

[0004] In order to solve the above technical problems, the present invention adopts HPLC-ELSD method to determine the contents of four components, amygdalin, fritillary glycosides, liquiritin and platycodon saponin in Chuanbei Qingfei syrup, so as to provide a reference for comprehensively evaluating the quality of Chuanbei Qingfei syrup.

[0005] A quality detection method for Chuanbei Qingfei syrup, characterized in that: it uses HPLC-ELSD chromatography to detect the contents of amygdalin, fritillary glycosides, liquiritin and platycodon saponin in Chuanbei Qingfei syrup, and the chromatographic conditions are:

[0006] The chromatographic column was Waters d C18 (4.6 mm × 250 mm, 5 μm); the mobile phase was acetonitrile (A)-1% acetic acid aqueous solution (B); gradient elution: 0-22 min, 10% → 40% A; 22-30 min, 40% → 60% A; the flow rate was 1.0 mL / min; the column temperature was 30-40°C;

[0007] ELSD detection: drift tube temperature 55-110°C, gas flow rate 2.0-3.0 L / min, injection volume 10 μL.

[0008] Further preferably, the column temperature is 40°C; and the ELSD detection conditions are: drift tube temperature 55°C, gas flow rate 2.5 L / min.

[0009] Specifically, it includes the following steps:

[0010] a. Preparation of reference solution: accurately weigh the reference substances of amygdalin, xibei glycosides, liquiritin and platycoside D, and add methanol to prepare reference substance stock solutions with concentrations of 0.95, 0.52, 0.58 and 0.61 mg / mL, respectively; then accurately measure 1 mL of the stock solutions, place them in the same 5 mL volumetric flask, and add methanol to prepare mixed reference substance solutions with concentrations of 190, 104, 116 and 122 μg / mL for amygdalin, xibei glycosides, liquiritin and platycoside D, respectively;

[0011] b. Preparation of test solution:

[0012] Take Chuanbei Qingfei syrup, shake and extract with n-butanol saturated with water, combine the n-butanol solution, evaporate to dryness, add water to the residue to dissolve, let it cool and pass it through a D101 macroporous adsorption resin column, elute with water, discard the water eluate, and then elute with 70% ethanol until the eluate is colorless, collect the eluate, evaporate to dryness, add 3 mL of methanol to dissolve the residue, filter it with a microporous filter membrane, and take the filtrate as the test solution;

[0013] c. Detection using the HPLC-ELSD chromatographic conditions described in claim 1 or 2.

[0014] Among them, the contents of amygdalin, fritillary glycoside, liquiritin and platycodon saponin in the Chuanbei Qingfei syrup are: amygdalin 19.20±1.20μg / mL, fritillary glycoside 7.17±0.50μg / mL, liquiritin 91.45±5.32μg / mL and platycodon saponin 49.3±1.28μg / mL respectively; the relative retention times of amygdalin, fritillary glycoside, liquiritin and platycodon saponin are: 9.272min, 12.678min, 14.257min, 21.193min respectively, and the relative retention time fluctuates up and down by 0.05min.

[0015] The present invention adopts HPLC-ELSD method to determine the content of four components of amygdalin, xibei glycoside, liquiritin and platycodon saponin D in Chuanbei Qingfei syrup, which can quickly evaluate the quality of Chuanbei Qingfei syrup, in order to provide a reference for the quality control of the preparation. The method effectively avoids the interference of strong ultraviolet absorbing substances, reduces the research cost, simplifies the operation process, provides an economical and efficient analytical means for the determination of saponin compound content, and also provides a reference for the formulation of quality control standards for Chuanbei Qingfei syrup. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 HPLC-ELSD chromatogram (wherein, A-Chuanbei Qingfei syrup; B-mixed reference; C-negative sample lacking bitter almond; D-negative sample lacking Chuanbei; E-negative sample with licorice; F-negative sample lacking platycodon; 1-amygdalin; 2-citracin glycoside; 3-liquiritigenin; 4-platycodonoside D);

[0017] Figure 2 The column temperature was 30°C for the chromatogram;

[0018] Figure 3 Chromatograms under different gradient elution;

[0019] Figure 4 Chromatograms with changing mobile phases;

[0020] Figure 5 Chromatogram after changing the drift tube temperature;

[0021] Figure 6 Chromatogram after changing the drift tube temperature;

[0022] Figure 7 Chromatogram using HPD-100 macroporous resin;

[0023] Figure 8 Chromatogram using DM13 macroporous resin. DETAILED DESCRIPTION

[0024] Example 1 Detection method of the present invention

[0025] Instruments and Materials

[0026] 1.1 Instruments: Agilent 1260Ⅱ high performance liquid chromatograph (Agilent, USA); Alltech CHROMELSD3300 evaporative light detector (Alltech, USA); XS105DU 1 / 100000 electronic balance (Mettler Toledo).

[0027] 1.2 Materials Reference Medicinal materials: loquat leaf (batch number: 231201), purchased from Beijing Bencao Fangyuan Pharmaceutical Technology Co., Ltd.; bitter almond (batch number: 21091501), purchased from Anguo Yifang Pharmaceutical Co., Ltd.; licorice (batch number: 230501), platycodon (batch number: 220401-01), both purchased from Hongya County Wawushan Pharmaceutical Co., Ltd.; mint (batch number: 23070401), purchased from Bozhou Qiaojuntang Chinese Medicine Beverage Co., Ltd.; ophiopogon (batch number: 231000081), Fritillaria cirrhosa (batch number: 240201531), and rehmannia glutinosa (batch number: 180706281), all purchased from Kangmei Pharmaceutical Co., Ltd. Reference substances: liquiritin (batch number: 111610-202209), purchased from China Food and Drug Inspection Institute; amygdalin (batch number: Y08O10K96808), xibei glycoside (batch number:

[0028] J26IB218361), Platycodon saponin D (batch number: IB236178), were purchased from Shanghai Yuanye Biotechnology Co., Ltd. There were 3 batches of Chuanbei Qingfei syrup samples, and batch numbers 1 to 3 were 230301, 230302, and 230303, respectively, all provided by Fujian Yannian Pharmaceutical Co., Ltd. D101 macroporous adsorption resin (batch number: 14240513002) was purchased from Beijing Solaibao Technology Co., Ltd. Methanol and acetonitrile were chromatographically pure, glacial acetic acid and n-butanol were analytically pure, and water was ultrapure water.

[0029] 2 Methods and Results

[0030] 2.1 Chromatographic conditions

[0031] The chromatographic column was Waters d C18 (4.6 mm × 250 mm, 5 μm); the mobile phase was acetonitrile (A)-1% acetic acid aqueous solution (B); gradient elution: 0-22 min, 10% → 40% A; 22-30 min, 40% → 60% A; flow rate was 1.0 mL / min; column temperature was 40°C; ELSD detection: drift tube temperature was 55°C, gas flow rate was 2.5 L / min, and injection volume was 10 μL. The chromatograms of the mixed reference solution, Chuanbei Qingfei syrup sample solution, and negative sample solution are shown in Figure 1 .

[0032] 2.2 Solution preparation

[0033] 2.2.1 Reference solution:

[0034] Accurately weigh appropriate amounts of amygdalin, xibei glycosides, liquiritin and platycoside D, and add methanol to prepare reference stock solutions with concentrations of 0.95, 0.52, 0.58 and 0.61 mg / mL, respectively. Then accurately measure 1 mL of the stock solution, place it in the same 5 mL volumetric flask, and add methanol to prepare mixed reference solutions with concentrations of 190, 104, 116 and 122 μg / mL for amygdalin, xibei glycosides, liquiritin and platycoside D, respectively.

[0035] 2.2.2 Preparation of test solution

[0036] Take 50mL of Chuanbei Qingfei syrup, shake and extract it 3 times with water-saturated n-butanol, 30mL each time, combine the n-butanol liquid, evaporate to dryness, add 2mL of water to the residue to dissolve it, cool it and pass it through a D101 macroporous adsorption resin column (inner diameter 1.5cm, column height 8cm), elute with 30mL of water, discard the water eluate, and then elute with 70% ethanol until the eluate is colorless, collect the eluate, evaporate to dryness, add 3mL of methanol to the residue to dissolve it, filter it with a microporous filter membrane (0.45μm), and take the filtrate as the test solution.

[0037] 2.2.3 Negative control solution:

[0038] According to the prescription ratio of Chuanbei Qingfei Syrup, prepare negative samples lacking bitter almond, Fritillaria cirrhosa, liquorice and Platycodon grandiflorum respectively, and prepare negative control solution according to the method under "2.2.2".

[0039] Example 2 Methodological Study of the Detection Method of the Present Invention

[0040] 2.3.1 Specificity test:

[0041] Take the mixed reference solution, test solution and negative control solution respectively, and measure according to the chromatographic conditions under "2.1", and record the chromatogram. The results show that the chromatographic peaks of amygdalin, xibeimu glycosides, liquiritin and platycodon saponin D in the test solution are consistent with the retention time in the reference solution, and there is no interference peak at the same retention time of amygdalin, xibeimu glycosides, liquiritin and platycodon saponin D reference in the negative control solution, indicating that other medicinal flavors in the prescription have no effect on the determination results, and the method has good specificity.

[0042] 2.3.2 Linear relationship investigation:

[0043] Accurately pipette appropriate amount of amygdalin, xibeimu glycoside, liquiritin and platycodon saponin D reference substance stock solutions under "2.2.1", and dilute them with methanol to make 6 concentrations of mixed reference substance solutions (the mass concentrations of amygdalin are 7.60, 19.00, 38.00, 95.00, 190.00 and 380.00 μg / mL, the mass concentrations of xibeimu glycoside are 4.16, 10.40, 20.80, 52.00, 104.00 and 208.00 μg / mL, and the mass concentrations of liquiritin are 4.16, 10.40, 20.80, 52.00, 104.00 and 208.00 μg / mL, respectively). The mass concentrations of the tested components (X, μg / mL) were 4.64, 11.60, 23.20, 58.00, 116.00, and 232.00 μg / mL, respectively, and the mass concentrations of platycodon saponin D were 4.88, 12.20, 24.40, 61.00, 122.00, and 244.00 μg / mL, respectively) and mixed. The samples were injected and analyzed according to the chromatographic conditions under "2.1". The peak areas were recorded. A standard curve was drawn with the peak area (Y) as the ordinate and the mass concentration of the tested component (X, μg / mL) as the abscissa. The regression equations of the components are shown in Table 1.

[0044] Table 1 Linear regression equations of the four chemical components in Chuanbei Qingfei syrup

[0045]

[0046] 2.3.3 Precision test:

[0047] Take the mixed reference solution and repeat the injection and determination 6 times according to the chromatographic conditions under "2.1". The RSDs (n=6) of the peak areas of amygdalin, xibei glycoside, liquiritin and platycodon saponin D are calculated to be 0.60%, 0.92%, 0.77% and 0.62%, respectively, indicating that the instrument has good precision.

[0048] 2.3.4 Stability test The test solution of Chuanbei Qingfei syrup was sampled and measured at 0, 4, 8, 12, 16 and 20 h, and the RSDs (n=6) of the peak areas of amygdalin, fritillary glycosides, liquiritigenin and platycodon saponin D were calculated to be 0.62%, 0.83%, 0.67% and 0.72%, respectively, indicating that the test solution had good stability within 20 h.

[0049] 2.3.5 Repeatability test:

[0050] Take 6 portions of the same batch of Chuanbei Qingfei syrup for testing, prepare the test solution according to the method under "2.2.2", inject and measure, and record the peak area. The RSD (n=6) of the peak areas of amygdalin, xibei glycoside, liquiritin, and platycodon saponin D were 0.86%, 0.75%, 0.78%, and 1.12%, respectively, indicating that the method has good repeatability.

[0051] 2.4 Sample determination

[0052] Weigh appropriate amounts of Chuanbei Qingfei syrup from three different batches, prepare the test solution according to the method under "2.2.2", and determine according to the chromatographic conditions under "2.1". Calculate the contents of amygdalin, xibei glycoside, liquiritigenin and platycodon saponin D in the three batches of samples by external standard method. The results are shown in Table 2.

[0053] Table 2 Average content of four components in three batches of samples (n = 3)

[0054]

[0055] Example 3 Condition screening test of the detection method of the present invention

[0056] In this study, four components, namely, amygdalin, xibei glycoside, liquiritigenin and platycoside D, were determined in Chuanbei Qingfei syrup, most of which were glycosides. Glycosides are usually extracted with alcohol. In this study, water-saturated n-butanol was selected as the extraction solvent, and the number of extractions (1, 2, 3 times), the volume of the extraction solvent (30 ml, 40 ml, 50 ml) and the type of macroporous resin (D101, HPD-100, DM130) were investigated. The results showed that the best extraction effect was achieved by shaking and extracting with water-saturated n-butanol for 3 times, 30 mL each time, and using a D101 macroporous adsorption resin column.

[0057] According to relevant literature, since there are many glycosides in the four components to be tested, the ultraviolet absorption is quite different, the ultraviolet absorption of amygdalin and liquiritin is strong, and the ultraviolet absorption of xibeimu glycoside and platycodon saponin D is weak or no ultraviolet absorption, ELSD detection is adopted. The experiment compares two different mobile phases, acetonitrile-0.1% acetic acid and acetonitrile-1% acetic acid solution. The results show that the separation effect is better when acetonitrile-1% acetic acid solution is used as the mobile phase. The experiment adopts gradient elution, and the retention time of the four components to be tested is appropriate by optimizing the time-proportion curve of the mobile phase.

[0058] This study also compared three different chromatographic columns: Diamonsil C18 (150×4.6mm, 5μm), Diamonsil C18 (250×4.6mm, 5μm) and Waters d C18 (4.6mm×250mm, 5μm). The Waters d C18 (4.6mm×250mm, 5μm) chromatographic column had a higher resolution and no interference with the target components. At the same time, the effect of column temperature (30℃, 35℃ and 40℃) on the resolution of chromatographic peaks was investigated. The results showed that the resolution was best at 40℃, so 40℃ was selected as the operating column temperature.

[0059] Wherein, when the column temperature is 30°C, other conditions are the same as those in Example 1, such as Figure 2 As shown, the separation between peaks 1 and 2 is poor and the baseline is unstable.

[0060] The evaporative light scattering detector plays an important role in the determination of gas flow and drift tube temperature. This study investigated the methods with gas flow rates of 2.0, 2.5, and 3.0 L / min and drift tube temperatures of 55°C, 60°C, and 110°C. The results showed that when the drift tube temperature was 55°C and the gas flow rate was 2.5 L / min, the baseline was stable and the overall noise was small. The chromatograms of amygdalin, xibei glycoside, liquiritin, and platycodon saponin D had good chromatographic peak responses.

[0061] The gradient elution conditions were changed, and the other conditions were the same as in Example 1. Figure 3 As shown, 0-10min, 10-20% A; 10-15min, 20-30% A; 15-22min, 30-40% A; 22-30min, 40-60% A; Results: The separation degree of peak 1, peak 2 and peak 4 was poor.

[0062] The mobile phase (acetonitrile (A)-0.1% acetic acid water) was changed, and the other conditions were the same as in Example 1. Figure 4 As shown, the separation between peaks 1 and 4 is poor, and peak 2 is not detected.

[0063] The drift tube temperature was changed to 110°C, and other conditions were the same as in Example 1. Figure 5 As shown, peaks 2 and 4 were poorly separated, and peak 1 was not detected.

[0064] The temperature of the drift tube was changed to 60°C, and other conditions were the same as in Example 1. Figure 6 As shown, the separation of peaks 1, 2, and 4 is poor.

[0065] HPD-100 macroporous resin was used for pretreatment of the sample, and other conditions were the same as those in Example 1. Figure 7 As shown, the result: glycoside components were not eluted.

[0066] DM13 macroporous resin was used for pretreatment of the sample, and other conditions were the same as those in Example 1. Figure 8 As shown, the result: glycoside components were not eluted.

[0067] In summary, there are fewer inspection items in the current standards of Chuanbei Qingfei Syrup, and manufacturers cannot effectively control product quality, and regulatory authorities cannot evaluate its quality. The present invention uses HPLC combined with ELSD (HPLC-ELSD) method to successfully determine the contents of 4 ingredients of amygdalin, xibei glycosides, liquiritin and platycodon saponin D in Chuanbei Qingfei Syrup. This method effectively avoids the interference of strong ultraviolet absorbing substances, reduces research costs, simplifies the operating procedures, and provides an economical and efficient analytical means for the determination of saponin compound content, and also provides a reference for the formulation of quality control standards for Chuanbei Qingfei Syrup.

Claims

1. A quality detection method for Chuanbei Qingfei syrup, characterized in that: It uses HPLC-ELSD chromatography to detect the contents of amygdalin, xibei glycoside, liquiritin and platycodon saponin in Chuanbei Qingfei syrup, and the chromatographic conditions are: The chromatographic column was Waters d C18 (4.6 mm × 250 mm, 5 μm); the mobile phase was acetonitrile (A)-1% acetic acid aqueous solution (B); gradient elution: 0-22 min, 10% → 40% A; 22-30 min, 40% → 60% A; the flow rate was 1.0 mL / min; the column temperature was 30-40°C; ELSD detection: drift tube temperature 55-110°C, gas flow rate 2.0-3.0 L / min, injection volume 10 μL.

2. The quality detection method of Chuanbei Qingfei syrup according to claim 1, characterized in that: The column temperature is 40°C; the ELSD detection conditions are: drift tube temperature 55°C, gas flow rate 2.5L / min.

3. The quality detection method of Chuanbei Qingfei syrup according to claim 1 or 2, characterized in that: It includes the following steps: a. Preparation of reference solution: accurately weigh the reference substances of amygdalin, xibei glycosides, liquiritin and platycoside D, and add methanol to prepare reference substance stock solutions with concentrations of 0.95, 0.52, 0.58 and 0.61 mg / mL, respectively; then accurately measure 1 mL of the stock solutions, place them in the same 5 mL volumetric flask, and add methanol to prepare mixed reference substance solutions with concentrations of 190, 104, 116 and 122 μg / mL for amygdalin, xibei glycosides, liquiritin and platycoside D, respectively; b. Preparation of test solution: Take Chuanbei Qingfei syrup, shake and extract with n-butanol saturated with water, combine the n-butanol solution, evaporate to dryness, add water to the residue to dissolve, let it cool and pass it through a D101 macroporous adsorption resin column, elute with water, discard the water eluate, and then elute with 70% ethanol until the eluate is colorless, collect the eluate, evaporate to dryness, add 3 mL of methanol to dissolve the residue, filter it with a microporous filter membrane, and take the filtrate as the test solution; c. Detection using the HPLC-ELSD chromatographic conditions described in claim 1 or 2.

4. The quality detection method of Chuanbei Qingfei syrup according to claim 3 is characterized in that: The contents of amygdalin, xibei glycoside, liquiritin and platycodon saponin in the Chuanbei Qingfei syrup are: amygdalin 19.20±1.20μg / mL, xibei glycoside 7.17±0.50μg / mL, liquiritin 91.45±5.32μg / mL and platycodon saponin 49.3±1.28μg / mL, respectively; the relative retention times of amygdalin, xibei glycoside, liquiritin and platycodon saponin are: 9.272min, 12.678min, 14.257min, 21.193min, respectively, and the relative retention time fluctuates up and down by 0.05min.

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