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目的:建立RP-HPLC法同时测定龙胆中龙胆苦苷、獐芽菜苦苷和獐芽菜苷的含量。方法:采用Diamasil C18(2)柱(250mm×4.6 mm,5μm);流动相为乙腈(A)-0.1%磷酸水溶液(B)梯度洗脱(0～22 min,10%～16%A;22～25 min,16%～10%A);流速为1.0 mL.min-1;检测波长为240 nm;柱温25℃。结果:龙胆苦苷、獐芽菜苦苷和獐芽菜苷进样量分别在1.0～20μg(r=0.9997)、1.0～20μg(r=0.9995)、0.12～2.4μg(r=0.9995)范围内与峰面积呈良好的线性关系;平均回收率(n=9)分别为99.1%,98.8%和98.6%。结论:该方法准确、简便、具有良好的重现性和稳定性,适合于龙胆的质量控制研究。
OBJECTIVE: To establish a RP-HPLC method for the simultaneous determination of gentiopicroside, swertia glycoside and romanin in gentian. METHODS: Diamasil C18 (2) column (250 mm × 4.6 mm, 5 μm) was used as the mobile phase. The mobile phase consisted of gradient elution from acetonitrile to A 0.1% phosphoric acid solution (0-22 min, 10-16% A; ~ 25 min, 16% ~ 10% A). The flow rate was 1.0 mL.min-1. The detection wavelength was 240 nm. The column temperature was 25 ℃. Results: The injection rates of gentiopicroside, swertia glycosides and romaine were 1.0 ~ 20μg (r = 0.9997), 1.0 ~ 20μg (r = 0.9995), 0.12 ~ 2.4μg There was a good linear relationship between the internal area and the peak area. The average recoveries (n = 9) were 99.1%, 98.8% and 98.6% respectively. Conclusion: The method is accurate, simple and has good reproducibility and stability. It is suitable for the quality control of gentian.