高效液相色谱-串联四极杆质谱法测定人参组培不定根中11种皂苷成分

Determination of 11 saponins in adventitious roots of ginseng tissue cultures by high performance liquid chromatography-tandem mass spectrometry

建立高效液相色谱-串联四极杆质谱(HPLC-MS/MS)法定量分析人参组培不定根中11种皂苷成分。样品经粉碎研磨,以70%甲醇水溶液为溶剂进行超声提取,离心过滤后测定。采用C_(18)色谱柱(100 mm×2.1 mm,1.8μm),用水和乙腈进行梯度洗脱,流量为0.3 mL/min,柱温为35℃。质谱采用电喷雾电离源,负离子扫描,多反应监测模式进行检测。11种皂苷的质量浓度在0.1~10μg/mL(Rb1为0.2~10μg/mL)范围内和响应强度线性相关,相关系数均大于0.995,各皂苷的定量限为0.001~0.010 g/kg,加标回收率为90.43%~97.82%,相对标准偏差为1.93%~6.33%(n=6)。该方法操作简单,分析时间短,灵敏度高,准确可靠,适用于人参组培不定根中多种皂苷类成分的同时测定。

A high performance liquid chromatography-tandem mass spectrometry method was established for the determination of 11 saponins in tissue culture adventitious root of ginseng.The samples were crushed and ground,and were subjected to ultrasonic extraction with 70%methanol as the solvent,and then were determined after centrifugal filtration.Using a C_(18) column(100 mm×2.1 mm,1.8μm),gradient elution was performed by using mobile phase water and acetonitrile,the flow rate was 0.3 mL/min,the column temperature was 35℃.The analytes were detected by electrospray ionization source,negative ion scanning and multi reaction monitoring mode.The mass concentration and response intensity of 11 saponins had a good linearity in the range of 0.1-10μg/mL(Rb1 was 0.2-10μg/mL),linear correlation coefficients were greater than 0.995,the quantitation limits of saponins ranged from 0.001-0.010 g/kg,the average recoveries of saponins were from 90.43%to 97.82%,and the relative standard deviations were from 1.93%to 6.33%(n=6).The method has the advantages of simple operation,short analysis time,high sensitivity,accuracy and reliability,and provides an effective way for the determination of 11 saponins in tissue culture roots of ginseng.