[关键词]
[摘要]
目的 建立芪参颗粒的HPLC指纹图谱分析方法,并对其中的主要成分进行同时定量分析,为芪参颗粒的质量控制提供依据。方法 采用HPLC法,用Agilent Eclipse plus C18(250 mm×4.6 mm,5 μm)色谱柱,0.1%甲酸乙腈溶液-0.1%甲酸水溶液为流动相,梯度洗脱,体积流量1.0 mL/min,检测波长254 nm,柱温30℃。建立10批芪参颗粒样品的HPLC指纹图谱,并进行相似度评价;通过与各组方药味的比对,对共有峰进行归属,通过LC-Q TOF-MS对所有共有峰进行鉴定,并对通过对照品比对确认的9个共有峰进行定量测定。结果 10批样品指纹图谱与对照指纹图谱的相似度均大于0.991;对标定的18个共有峰进行归属和鉴定,发现1、2、8、14、18号峰来自黄芪,3~7、9~11号峰来自金银花,12、13、15、16号峰来自丹参,17、18号峰来自甘草;通过对照品的指认,发现4号峰为绿原酸、8号峰为毛蕊异黄酮葡萄糖苷、9号峰为异绿原酸B、10号峰为异绿原酸A、14号峰为芒柄花苷、15号峰为丹酚酸B、16号峰为丹酚酸A、17号峰为甘草酸及18号峰为芒柄花素,并对这9种成分进行了定量测定,测定结果分别为6.676~10.213、0.628~0.963、1.018~1.886、1.082~1.972、0.477~0.790、11.327~17.788、0.519~0.908、2.000~3.638、0.010~0.016 mg/g。结论 建立的芪参颗粒HPLC指纹图谱和9个指标成分的同时定量测定方法专属性强,简便、可靠,重复性好,可为芪参颗粒质量控制和评价提供参考。
[Key word]
[Abstract]
Objective To establish the HPLC fingerprint and determine main components of Qishen Granules (QG), so as to provide a scientific basis for its quality control. Methods HPLC analysis was performed on an Agilent Eclipse plus C18 column (250 mm×4.6 mm, 5 μm). The gradient elution was performed by the mobile phase consisting of acetonitrile-0.1% formic acid and 0.1% formic acid aqueous with the flow rate of 1.0 mL/min, the detection wavelength was set at 254 nm, and the column temperature was 30℃. Fingerprints of ten batches of QG were determined, and the similarities among fingerprints were evaluated. Attributing analysis of the common peaks was achieved by comparing the retention times with the chromatograms of single constituent drugs, and identifications of common peaks were performed on LC-Q TOF-MS and nine components were further confirmed by the reference substances, the content of the nine compounds was subsequently analyzed by HPLC. Results The similarities of 10 batches of QG were all greater than 0.991. There were 18 common peaks marked in total, peaks 1, 2, 8, 14 and 18 from Astragalus membranaceus var. mongholicus, peaks 3, 4, 5, 6, 7, 9, 10 and 11 from Lonicera japonica, peaks 12, 13, 15 and 16 from Salvia miltiorrhiza, and peaks 17 and 18 from Glycyrrhiza uralensis. Based on the identification of the common peaks, nine components such as chlorogenic acid (peaks 4), calycosin-7-glucoside (peaks 8), isochlorogenic acid B (peaks 9), isochlorogenic acid A (peaks 10), ononin (peaks 14), salvianolic acid B (peaks 15), salvianolic acid A (peaks 16), glycyrrhizic acid (peaks 17), and formononetin (peaks 18) were identified and quantified. The content of the nine components was determined as 6.676-10.213, 0.628-0.963, 1.018-1.886, 1.082-1.972, 0.477-0.790, 11.327-17.788, 0.519-0.908, 2.000-3.638, and 0.010-0.016 mg/g, respectively. Conclusion The method established in this study shows good characteristics, specificity, and repeatability, which can provide scientific basis for the quality control of QG.
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[基金项目]
国家"重大新药创制"科技重大专项资助项目(2018ZX09201008-001);国家"重大新药创制"科技重大专项资助项目(2018ZX09101002-002-004);国家自然科学基金项目(81573839);国家自然科学基金项目(81774155)