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目的:建立测定川东獐牙菜中雏菊叶龙胆苷、苦龙胆酯苷、去甲基雏菊叶龙胆酮和雏菊叶龙胆酮含量的高效液相色谱方法。方法:HYPERSIL BDS C_(18)(4.6mm×200mm,5μm)色谱柱,流动相为乙腈-磷酸二氢钠(0.03mol·L~(-1),pH=3.0),梯度洗脱(CH_3CN%:0 min 10%,15min 20%,30min 35%,50min 55%),流速为1.0mL·min~(-1),检测波长为250nm,柱温为25℃。结果:雏菊叶龙胆苷,苦龙胆酯苷,去甲基雏菊叶龙胆酮和雏菊叶龙胆酮分别在0.0403-0.3024μg,0.0604-0.4527μg,0.1320-0.9900μg,0.1610-1.2072μg范围内线性良好,相关系数r分别为0.9999,0.9999,0.9999,0.9999,其平均回收率均在98%-103%范围内,RSD均小于1%(n=5)。结论:本方法操作简便,分离效果好,灵敏度高,重现性好,为全面控制川东獐牙菜质量提供一个可靠的方法。
OBJECTIVE: To establish a high performance liquid chromatographic method for the determination of glycosides, gentiopicrin, desmethyl daisy, gentianone, and daisy gentisin in Swertia chuanxiong. Method: HYPERSIL BDS C_(18) (4.6mm×200mm, 5μm) column with mobile phase of acetonitrile-sodium dihydrogen phosphate (0.03mol·L-1, pH=3.0), gradient elution (CH_3CN%) : 0 min 10%, 15 min 20%, 30 min 35%, 50 min 55%), flow rate 1.0 mL·min -1, detection wavelength 250 nm, and column temperature 25°C. RESULTS: Daisy gentioside, gentioglycoside, demethyl-daisy gentisone and daisy geranone were in the range of 0.0403-0.3024 μg, 0.0604-0.4527 μg, 0.1320-0.9900 μg, and 0.1610-1.2072 μg, respectively. The internal linearity was good, and the correlation coefficients r were 0.9999, 0.9999, 0.9999, and 0.9999, respectively. The average recoveries were within the range of 98%-103%, and the RSDs were all less than 1% (n=5). Conclusion: The method is easy to operate, with good separation effect, high sensitivity and good reproducibility. It provides a reliable method for comprehensive control of the quality of Swertia arborescens.