UPLC-DAD-ELSD切换波长法同时测定复方酸枣仁颗粒中8种成分的含量

Simultaneous determination of eight active components in compound Semen Ziziphi Spinoase granules by UPLC-DAD-ELSD wavelength switching

目的:建立UPLC-DAD-ELSD切换波长法测定酸枣仁颗粒中芒果苷、阿魏酸、甘草苷、斯皮诺素、知母皂苷BⅡ、酸枣仁皂苷A、白桦脂酸、白桦脂醇的含量。方法:采用Agilent Zorbax Eclipse plus C18色谱柱(2.1 mm×50 mm,1.8μm),流动相为乙腈-0.1%醋酸溶液,梯度洗脱,流速0.4mL·min^(^(-1)),检测波长250 nm(0~5.8 min,芒果苷,阿魏酸)、280 nm(5.8~25 min,甘草苷,斯皮诺素),柱温40℃;蒸发光散射检测器(知母皂苷-BⅡ、酸枣仁皂苷A、白桦脂酸、白桦脂醇),雾化器温度为40℃,漂移管温度为100℃,气体流速1.60 L·min^(^(-1))。结果:各待测组分分离度良好;芒果苷、阿魏酸、甘草苷、斯皮诺素、知母皂苷BⅡ、酸枣仁皂苷A、白桦脂酸、白桦脂醇的进样量分别在0.021~0.412μg(r=0.999 9)、0.004~0.083μg(r=0.999 0)、0.020~0.402μg(r=0.999 9)、0.019~0.348μg(r=0.999 9)、0.094~1.890μg(r=0.999 0)、0.020~0.410μg(r=0.999 2)、0.051~1.025μg(r=0.999 1)和0.013~0.270μg(r=0.999 5)范围内与各自峰面积呈良好线性关系;芒果苷含量为-0.534~0.911 mg·g^(-1),阿魏酸含量为0.114~0.213 mg·g^(-1),甘草苷含量为0.534~0.552 mg·g1,斯皮诺素含量为0.641~0.733 mg·g^(-1),知母皂苷BⅡ含量为1.066~1.208 mg·g^(-1),酸枣仁皂苷A含量为0.644~0.845 mg·g^(-1),白桦脂酸含量为2.181~2.211 mg·g^(-1),白桦脂醇含量为0.307~0.309 mg·g^(-1)。结论:经方法学验证,本法可用于复方酸枣仁颗粒的质量控制。

Objective：To establish a method for simultaneous determination of mangiferin,ferulic acid,liquiritin,spinosin,timosaponin B Ⅱ,jujuboside A,betulinic acid,betulin in compound Semen Ziziphi Spinoase granules by UPLC-DAD-ELSD wavelength switching. Methods：Agilent Zorbax Eclipse plus chromatographic column（2.1 mm×50 mm,1.8 μm） was adopted. The mobile phase was acetonitrile-0.1% acetic acid with gradient elution at a flow rate of 0.4 m L·min-1. The detection wavelength was 250 nm（0-5.8 min,for mangiferin and ferulic acid）,280 nm（5.8-25 min,for liquiritin,spinosin）. The column temperature was 40 ℃. The nebulizer temperature of ELSD for timosaponin B Ⅱ,jujuboside A,betulinic acid and betulin was maintained at 40 ℃,the drift tube temperature was 100 ℃ and the gas flow rate was maintained at 1.60 L·min-1. Results：The complete separation of 8 compounds was obtained. The method had a good linearity in the ranges of 0.021-0.412 μg（r=0.999 9） for mangiferin,0.004-0.083 μg（r=0.999 0） for ferulic acid,0.020-0.402 μg（r=0.999 9） for liquiritin and 0.019-0.348 μg（r=0.999 9） for spinosin,0.094-1.890 μg（r=0.999 0） for timosaponin BII,0.020-0.410 μg（r=0.999 2） for jujuboside A,0.051-1.025 μg（r=0.999 1） for betulinic acid and 0.013-0.270 μg（r=0.999 5） for betulin. The average recovery were 103.0%,101.2%,102.6%,100.6%,100.6%,100.7%,98.0%,101.2%（n=6）,respectively. Th e contents of three batches of samples were 0.534-0.911 mg·g-1 for mangiferin,0.114-0.213 mg·g-1 for ferulic acid,0.534-0.552 mg·g-1 for liquiritin,0.641-0.733 mg·g-1 for spinosin,1.066-1.208 mg·g-1 for timosaponin B Ⅱ,0.644-0.845 mg·g-1 for jujuboside A,2.181-2.211 mg·g-1 for betulinic acid,and 0.307-0.309 mg·g-1 for betulin. Conclusion：This method is proved to be valid for the quality control of compound Semen Ziziphi Spinoase granules.