离子液体超声辅助萃取HPLC法测定天山雪莲中的4个有效成分

Sensitive determination of 4 active ingredients in Herba Saussureae Involucratae by UAE-HPLC method

目的：建立以离子液体1-丁基-3-甲基咪唑六氟磷酸盐（[BMIm]PF6）为萃取剂超声辅助萃取,高效液相色谱法分离测定天山雪莲中紫丁香苷、芦丁、绿原酸和咖啡酸4个有效成分的方法。方法：采用Phenomenex C18色谱柱（250 mm×4.6 mm,5μm）,流动相为甲醇-0.1%醋酸水溶液,梯度洗脱（0～5 min,30%B→40%B;5～10 min,40%B→60%B;10～15 min,60%B→70%B）,流速0.8 mL·min-1,紫外检测波长270 nm,柱温35℃。结果：该方法对紫丁香苷、芦丁、绿原酸和咖啡酸浓度分别在0.0011～108.0μg·mL-1（r=0.9999）、0.0011～106.0μg·mL-1（r=0.9999）、0.0021～208.0μg·mL-1（r=0.9999）、0.0015～154.0μg·mL-1（r=0.9999）范围内线性关系良好,定性检测限（S/N=3）依次为0.0374、0.0094、0.1018、0.0549 ng·mL-1;样品回收率为96.12%～101.1%。结论：本法灵敏度高、定量准确、操作简便、成本低,且具有环保优点,可作为天山雪莲中紫丁香苷、芦丁、绿原酸和咖啡酸的分离测定的有效方法。

Objective： To develop a simple method based on high performance liquid chromatography（ HPLC） and ultrasonic assisted extraction（ UAE） using 1-butyl-3-methylimidazolium hexafluorophosphate as extractant for the rapid determination of syringin,rutin,chlorogenic acid,and caffeic acid in Herba Saussureae Involucratae. Methods： Four compounds were isolated using a Phenomenex C18column with a gradient elution（ 0-5 min,30% B→40% B; 5-10 min,40% B→60% B; 10-15 min,60% B→70% B）; the mobile phase was methanol-0. 1% acetic acid solution with the UV detection wavelength at 270 nm; the flow rate of 0. 80 mL·min-1and column temperature at 35 ℃. Results： The linearity range for syringin,rutin,chlorogenic acid,and caffeic acid was found to be 0. 0011-108. 0 μg·mL-1（ r =0. 9999）,0. 0011-106. 0 μg·mL-1（ r =0. 9999）,0. 0021-208. 0 μg·mL-1（ r = 0. 9999）,and 0. 0015-154. 0 μg·mL-1（ r =0. 9999）,respectively. It was indicative of good linear dependence of peak area on concentration. The limits of detection（ LOD） were 0. 0374,0. 0094,0. 1018,0. 0549 ng·mL-1,respectively. The recovery ranged from 96. 12% to 101. 1%. Conclusion： The proposed method was found to be sensitive,simple,accurate,and environmental friendly,which can be an effective method for the determination of syringin,rutin,chlorogenic acid,and caffeic acid in Herba Saussureae Involucratae.