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目的:建立反相高效液相色谱法同时测定喉毛花植物中绿原酸、獐牙菜苦苷、龙胆苦苷、獐牙菜苷、槲皮素的含量。方法:采用ZORBAX SB-C18(250mm×4.6mm,5μm)色谱柱,以流动相甲醇和水(含0.04%磷酸)的比例在0-45min内由10∶90至60∶40线性梯度洗脱,45-50min内60∶40等度洗脱,流速1m L/min,检测波长254nm,柱温30℃。结果:5种成分均达到基线分离,绿原酸、獐牙菜苦苷、龙胆苦苷、獐牙菜苷、槲皮素的线性范围分别为0.0495-0.99μg(r=0.9998),0.0515-1.03μg(r=0.9999),0.06-1.2μg(r=0.9998),0.0495-0.99μg(r=0.9998),0.0495-0.99μg(r=0.9999);回收率为100.8%(RSD=2.51%),100.7%(RSD=0.91%),100.2%(RSD=2.89%),99.6%(RSD=0.85%),99.8%(RSD=1.36%)。结论:该方法测定快速,结果准确、可靠。
Objective: To establish a reversed-phase high performance liquid chromatography (HPLC) method for the simultaneous determination of chlorogenic acid, swertiamazide, gentiopicroside, swertiagenin and quercetin in laryngx plants. Methods: A linear gradient of 10:90 to 60:40 was performed on a ZORBAX SB-C18 (250 mm × 4.6 mm, 5 μm) column with a mobile phase of methanol and water (0.04% phosphoric acid) over a period of 0-45 min. 45-50min 60:40 isocratic elution, flow rate 1m L / min, detection wavelength 254nm, column temperature 30 ℃. Results: The five components all achieved baseline separation. The linear ranges of chlorogenic acid, swertiarin, gentiopicroside, swertiagenin and quercetin were 0.0495-0.99μg (r = 0.9998), 0.0515- (R = 0.9998), 0.0495-0.99 μg (r = 0.9998), 0.0495-0.99 μg (r = 0.9999); recovery of 100.8% (RSD = 2.51%), 100.7% (RSD = 0.91%), 100.2% (RSD = 2.89%), 99.6% (RSD = 0.85%), 99.8% (RSD = 1.36%). Conclusion: The method is rapid, accurate and reliable.