LC-MS/MS法测定酸枣仁中酸枣仁皂苷A、B和斯皮诺素的含量

Determination of Jujuboside A,Jujuboside B and Spinosin in Semen Ziziphi Spinosae by LC-MS/MS

目的建立简单、快速、灵敏同时测定酸枣仁中酸枣仁皂苷A、B和斯皮诺素的LC-MS/MS法。方法采用Phenomenex Gemini C1(850 mm×2.0 mm,5μm)色谱柱;流动相:甲醇-(5 mmol.L-1甲酸铵-0.1%甲酸缓冲液)梯度洗脱;流速为0.25 mL.min-1;柱温为40℃;质谱条件:采用负离子的模式,以MRM配对离子方式进行定量,酸枣仁皂苷A、B和斯皮诺素的选择检测离子质荷比分别为m/z 1251.9→1251.9,1089.9→1089.9和607.3→427.1。结果酸枣仁皂苷A、B和斯皮诺素分别在50～1000 ng.mL-1、25～500 ng.mL-1和12.5～500 ng.mL-1的浓度范围内,线性关系良好;酸枣仁皂苷A、B和斯皮诺素的最低检测限(LOQ)分别为5,2.5和0.5 ng.mL-1;平均回收率为99.35%～101.10%,RSD<5.01%(n=9)。三批酸枣仁中酸枣仁皂苷A、B和斯皮诺素的含量分别为1.28～1.38 mg.g-1、1.02～1.11 mg.g-1和1.05～1.24 mg.g-1。结论该法专属性强、快速灵敏,可用于酸枣仁中酸枣仁皂苷A、B和斯皮诺素含量的测定。

Objective To develop a LC-MS/MS method for the determination of jujuboside A,jujuboside B and spinosin in Semen Zizyphi Spinosae.Methods The column was Phenomenex Gemini C18（50 mm × 2.0 mm,5 μm）,mobile phase consisted of methanol： 5 mmol/L ammonium formate-0.1% formic acid buffer at a flow rate of 0.25 mL/min.The temperature of column was 40 ℃.The LC-MS/MS system was operated by using an electrospray ionization probe in the negative ion mode.Scan mode was in multiple reaction ion monitoring（MRM）mode.The mass charge ration of ion（m/z） was 1251.9→1251.9 for jujuboside A,1089.9→1089.9 for jujuboside B and 607.3→427.1 for spinosin,respectively.Results The linear ranges of jujuboside A,jujuboside B and spinosin were 50 ～ 1000 ng·mL-1,25 ～ 500 ng·mL-1 and 12.5 ～ 500 ng·mL-1,respectively.The lowest limit of quantification（LOQ） of jujuboside A,jujuboside B and spinosin was 5 ng·mL-1,2.5 ng·mL-1and 0.5 ng·mL-1（S/N3）,respectively.The average recovery was 99.35 % ～ 101.10%,and RSD 5.01 %（n=9）.The average content of jujuboside A,jujuboside B and spinosin in three batches of Semen Zizyphi Jujubae was in the range of 1.28 ～ 1.38 mg·g-1,1.02 ～ 1.11 mg·g-1 and 1.05 ～ 1.24 mg·g-1,respectively.Conclusion A sensitive,accuracy and suitable LC-MS/MS method has been developed,and the method can be applied for the detemination of jujuboside A,jujuboside B and spinosin in Semen Zizyphi Spinosae.