| 摘要: |
| 目的：建立泻白散基准样品特征图谱方法，以LC-MS/MS指认各特征峰并对其进行归属，建立基准样品各组成药味质控指标的含量测定方法综合评价其质量。方法：以色谱柱Hypersil ODS2(250 mm×4.6 mm，5μm)、乙腈-0.1%磷酸/甲酸梯度洗脱、检测波长280nm、流速1.0 ml/min、柱温25℃、进样量10μl进行指纹图谱研究，以同一色谱条件使用LC-MS/MS联用进行峰归属并进行化学成分分析，以指认的各药味专属性峰建立基准样品质控指标含量测定方法。结果：泻白散基准样品特征图谱重复性良好，供试品溶液48 h内稳定，多批次检测显示泻白散指纹图谱共有峰14个，相似度均＞0.9;峰归属确认其中5个归属于炒桑白皮，5个归属于炙甘草，3个归属于地骨皮;并通过LC-MS/MS指认出其中11个色谱峰;选取桑皮苷A、甘草酸、枸杞素B分别建立含量测定方法，结果显示方法均具有专属性，重复性良好，加样回收率分别为94.9%、93.5%、95.2%。结论：该指纹图谱方法经验证可行，可有效鉴别泻白散三味饮片;结合LC-MS/MS成分分析可有效选取基准样品各药味的质控成分;建立的检测桑皮苷A、甘草酸、枸杞素B含量测定方法经方法学考察确认可行。 |
| 关键词: 泻白散 基准样品 LC-MS/MS 成分分析 高效液相色谱 |
| DOI： |
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| Liquid chromatography-tandem mass spectrometry combined with high-performance liquid chromatography for evaluating the quality of Xiebai powder reference sample |
| DENG Yun,ZHAO Sinan,PENG Yunzhu |
| (Hangzhou Vintage Medication Co.，Ltd.，Hangzhou 311202，Zhejiang，China) |
| Abstract: |
| Objective: To establish a method for determining the specific chromatogram of Xiebai powder reference sample using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the identification and attribution of each characteristic peak，and to evaluate the quality of the reference sample by establishing the methods for content determination of quality control indicators for each traditional Chinese medicine (TCM) drug in the reference sample.Methods: The fingerprint study was conducted on a Hypersil ODS22 column (250 mm × 4.6 mm，5 μm) with the mobile phase of acetonitrile-0.1% phosphoric acid/formic acid for gradient elution at a detection wavelength of 280 nm，a flow rate of 1.0 ml/min，a column temperature of 25°C，and a sample size of 10 μl，and LC-MS/MS with the same chromatograph conditions was performed for peak attribution and chemical component analysis.The specific peak identified for each TCM drug was used to establish the methods for content determination of quality control indicators for the reference sample.Results: The specific chromatogram of Xiebai powder reference sample had a good reproducibility，and the solution of the test article remained stable within 48 hours.Detection of multiple batches obtained 14 common peaks on the fingerprint of Xiebai powder，with a similarity of >0.9，and peak attribution showed that 5 were attributed to fried Cortex Mori，5 were attributed to Radix Glycyrrhizae Preparata，and 3 were attributed to Cortex Lycii.A total of 11 chromatographic peaks were identified by LC-MS/MS.The method for content determination were established for mulberroside A，glycyrrhizic acid，and lyciumin B，and the results showed that the method had good specificity and reproducibility，with a recovery rate of 94.9%，93.5%，and 95.2%，respectively.Conclusion: The method for determining the fingerprint is feasible after validation and can effectively identify Xiebai powder decoction pieces with three TCM drugs，and its combination with LC-MS/MS component analysis can effectively determine the quality control components for each drug.Methodological investigation shows that the method for determining the content of mulberroside A，glycyrrhizic acid，and lyciumin B is feasible. |
| Key words: Xiebai powder reference sample liquid chromatography-tandem mass spectrometry component analysis high-performance liquid chromatography |
