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目的建立高效液相色谱法同时测定25批蜘蛛香中3-O-咖啡酰基奎宁酸、橙皮苷和4,5-O-二咖啡酰基奎宁酸含量的方法。方法以Phenomenex Gemini C18(4.6 mm×250 mm,5μm)为色谱柱,乙腈(A)-0.1%磷酸水溶液(B)为流动相系统梯度洗脱,流速为1.0 mL·min-1,检测波长340 nm,柱温25℃,进样量10μL。运用该方法对25批蜘蛛香药材进行测定,并通过聚类分析对结果进行分析。结果蜘蛛香中3-O-咖啡酰基奎宁酸、橙皮苷和4,5-O-二咖啡酰基奎宁酸的线性范围分别为0.33~1.97μg(r=0.999 9),1.14~6.84μg(r=0.999 9),0.11~0.66μg(r=0.999 9),平均加样回收率(n=6)分别为97.02%(RSD 0.6%),100.60%(RSD 1.7%),96.79%(RSD 1.0%)。采用聚类分析能将不同产地的25批蜘蛛香药材分成3类。结论该方法简便、快速、准确,可用于蜘蛛香药材的质量评价和控制。
OBJECTIVE To establish a method for the simultaneous determination of 3-O-caffeoylquinic acid, hesperidin and 4,5-O-dicofecaloylquinic acid in 25 batches of spider incense by high performance liquid chromatography. Methods The mobile phase consisted of a gradient of Phenomenex Gemini C18 (4.6 mm × 250 mm, 5 μm) and acetonitrile (A) -0.1% phosphoric acid solution (B) with a flow rate of 1.0 mL · min -1. The detection wavelength was 340 nm, column temperature 25 ℃, injection volume 10μL. Using this method, 25 batches of spider incense herbs were determined, and the results were analyzed by cluster analysis. Results The linear ranges of 3-O-caffeoylquinic acid, hesperidin and 4,5-O-dis Caffeoylquinic acid were 0.33-1.97μg (r = 0.999 9), 1.14-6.84μg (r = 0.999 9) and 0.11-0.66 μg (r = 0.999 9) respectively. The average recoveries were 97.02% (RSD 0.6%), 100.60% (RSD 1.7%) and 96.79% 1.0%). Cluster analysis of 25 batches of spider incense from different origins can be divided into three categories. Conclusion The method is simple, rapid and accurate and can be used for the quality evaluation and control of spider incense.