清肺合剂薄层色谱鉴别、HPLC指纹图谱及多指标成分质量研究

Quality control of Qingfei Mixture：thin-layer chromatography identification，high-performance liquid chromatography fingerprint development，and multi-component quantification

关键词:  清肺合剂  薄层色谱  高效液相色谱  指纹图谱  有效成分  含量测定

Key words:  Qingfei Mixture  TLC  HPLC  fingerprint  effective ingredients  content determination

摘要:

[目的] 建立薄层色谱（TLC）鉴别、高效液相色谱（HPLC）指纹图谱及柚皮苷、新橙皮苷、黄芩苷及盐酸麻黄碱含量测定方法以控制清肺合剂质量。[方法] 采用薄层色谱法对鱼腥草、青黛及甘草进行定性鉴别。采用Agilent Eclipse XDB-C₁₈（4.6 mm×250 mm，5 μm）色谱柱；以乙腈-0.1%磷酸水溶液进行梯度洗脱，柱温为30 ℃；进样量为10 μL；流速1.0 mL/min；检测波长280 nm。建立了15批清肺合剂的HPLC指纹图谱，采用中药色谱指纹图谱相似度评价系统（A版）进行相似度分析和评价，同时测定其中4种有效成分的含量。[结果] 鱼腥草、青黛及甘草的薄层色谱鉴别方法专属性强，无其他成分干扰。清肺合剂HPLC指纹图谱及4种有效成分测定方法的方法学考察结果均符合相关规定的要求。不同批次的清肺合剂指纹图谱相似度均超过0.999。共标定了18个共有峰，对其中的7个成分进行了定性，包括盐酸麻黄碱、苦杏仁苷、甘草苷、柚皮苷、新橙皮苷、黄芩苷及白花前胡甲素。其中柚皮苷、新橙皮苷、黄芩苷和盐酸麻黄碱的平均含量分别为：3.517、2.137、6.417、0.204 mg/mL。[结论] 本文建立的清肺合剂薄层鉴别方法、HPLC指纹图谱及4种成分含量测定方法专属性强、稳定可靠、可操作性强，可以用于清肺合剂的整体质量控制。

Abstract:

[Objective] To establish quality control methods for Qingfei mixture, including thin-layer chromatography(TLC) identification, high-performance liquid chromatography(HPLC) fingerprint analysis, and content determination of naringin, neohesperidin, baicalin, and ephedrine hydrochloride.[Methods] TLC was used for qualitative identification of Houttuynia cordata, Indigo naturalis, and Glycyrrhiza uralensis. HPLC analysis was performed on an Agilent Eclipse XDB-C18 column(4.6 mm×250 mm, 5 μm) with a gradient elution of acetonitrile and 0.1% aqueous phosphoric acid solution. The column temperature was 30℃, with an injection volume of 10 μL, a flow rate of 1.0 mL/min, and a detection wavelength of 280 nm. HPLC fingerprints of 15 batches of Qingfei mixture were established and evaluated using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version A). The contents of four active components were simultaneously determined.[Results] The TLC methods for Houttuynia cordata, Indigo naturalis, and Glycyrrhiza uralensis showed strong specificity without interference. Methodological validation of the HPLC fingerprint and content determination met regulatory requirements. The similarity of fingerprints from different batches reached 0.999. Eighteen common peaks were labeled, and seven components were identified:ephedrine hydrochloride, amygdalin, liquiritin, naringin, neohesperidin, baicalin, and peucedanum praeruptorum A. The average contents of naringin, neohesperidin, baicalin, and ephedrine hydrochloride were 3.517, 2.137, 6.417, and 0.204 mg/mL, respectively.[Conclusion] The established TLC identification, HPLC fingerprint, and content determination methods are specific, stable, reliable, and practical for comprehensive quality control of Qingfei mixture.