基于HPLC-DAD-ELSD串联法同时测定麝香保心丸中8种成分的含量

Simultaneous Determination of Eight Components in Shexiang Baoxin Pill(麝香保心丸)by HPLC-DAD-ELSD Series Method

目的:建立麝香保心丸中苯甲酸苄酯、胆酸、蟾毒他灵、蟾毒灵、人参皂苷Rb1、人参皂苷Rb2、华蟾酥毒基和酯蟾毒配基8种成分的含量测定方法,为麝香保心丸的质量评价提供实验参考。方法:采用HPLC-DADELSD串联法同时测定麝香保心丸中苯甲酸苄酯、胆酸、蟾毒他灵、蟾毒灵、人参皂苷Rb1、人参皂苷Rb2、华蟾酥毒基和酯蟾毒配基8种化学成分的含量,色谱柱为Waters XBridgeTMC18柱(250 mm×4.6 mm,5μm);流动相为乙腈-0.5%甲酸水溶液,梯度洗脱;检测波长为280 nm,柱温为30℃;蒸发光散射检测器(ELSD)漂移管温度为40℃;载气流量1.5 L/min。结果:苯甲酸苄酯、胆酸、蟾毒他灵、蟾毒灵、人参皂苷Rb1、人参皂苷Rb2、华蟾酥毒基、酯蟾毒配基分离度良好,分别在2.769~55.370μg/mL(r=0.9998)、2.285~45.700μg/mL(r=0.9999)、2.251~45.021μg/mL(r=0.9997)、2.445~48.902μg/mL(r=0.9998)、1.912~38.230μg/mL(r=0.9999)、2.350~46.990μg/mL(r=0.9998)、2.275~45.500μg/mL(r=0.9999)、2.335~46.700μg/mL(r=0.9998)的范围内线性关系良好;平均回收率分别为99.98%(RSD=0.86%)、100.09%(RSD=0.65%)、99.98%(RSD=0.66%)、101.95%(RSD=0.92%)、99.50%(RSD=0.40%)、100.09%(RSD=0.69%)、99.89%(RSD=0.66%)、99.99%(RSD=0.62%)。结论:该法操作方便、准确、重复性好,可客观、全面地对麝香保心丸的内在质量进行有效评价。

Objective:To establish a method for the determination of benzyl benzoate,cholic acid,bufalin,bufalin,ginsenoside Rb1,ginsenoside Rb2,cinobufagin and ester bufalin ligand in Shexiang Baoxin Pill,so as to provide experimental reference for the quality evaluation of Shexiang Baoxin Pill.Methods:The contents of benzyl benzoate,cholic acid,bufalin,bufalin,ginsenoside Rb1,ginsenoside Rb2,cinobufagin and resibufogenin in Shexiang Baoxin Pill were determined by HPLC-DAD-ELSD series method.The chromatographic column was Waters XBridgeTMC18 column(250 mm)x 4.6 mm,5μm).The mobile phase was acetonitrile-0.5%formic acid aqueous solution in a gradient elution manner.The detection wavelength was 280 nm and the column temperature was 30℃.The drift tube temperature of evaporative light scattering detector(ELSD)was 40℃,and the carrier gas flow was 1.5 L/min.Results:Benzyl benzoate,cholic acid,bufotaline,bufalin,ginsenoside Rb1,ginsenoside Rb2,cinobufagin,and resibufogenin displayed good linear relationships within the range of 2.769~55.370μg/mL(r=0.9998),2.285~45.700μg/mL(r=0.9999),2.251~45.021μg/mL(r=0.9997),2.445~48.902μg/mL(r=0.9998),1.912~38.230μg/mL(r=0.9999),2.350~46.990μg/mL(r=0.9998),2.275~45.500μg/mL(r=0.9999),2.335~46.700μg/mL(r=0.9998).And the average recoveries were 99.98%(RSD=0.86%),100.09%(RSD=0.65%),99.98%(RSD=0.66%),101.95%(RSD=0.92%),99.50%(RSD=0.40%),100.09%(RSD=0.69%),99.89%(RSD=0.66%),99.99%(RSD=0.62%),respectively.Conclusion:The method is convenient,accurate and reproducible.It can objectively and comprehensively evaluate the internal quality of Shexiang Baoxin Pill.