HPLC-ELSD法测定黑种草总皂苷提取物中黑种草皂苷A的含量

Content Determination of Nigeglanoside A in Total Saponin Extract of Nigella glandulifera by HPLC-ELSD

目的:建立测定黑种草总皂苷提取物中黑种草皂苷A含量的方法。方法:采用高效液相色谱-蒸发光散射检测器(HPLC-ELSD)法。色谱柱为Cosmosil C18(250mm×4.6mm,5μm),流动相为乙腈-水(梯度洗脱),流速为1.0ml/min,柱温为30℃,进样量为10μl,ELSD的漂移管温度为65℃,气体流速为2.0L/min。结果:黑种草皂苷A的质量浓度在0.183～2.562mg/ml范围内与其峰面积积分值呈良好的线性关系(r=0.9990);精密度、稳定性、重复性试验的RSD均<2%;平均加样回收率为99.93%,RSD=1.60%(n=6)。结论:该方法操作简便、重复性好、准确性高,可用于黑种草总皂苷提取物中黑种草皂苷A的含量测定。

OBJECTIVE： To establish a method for the determination of nigeglanoside A in total saponin extract of Nigella glan-dulifera. METHODS： HPLC-ELSD method was adopted. The separation was performed on Cosmosil C18（250mm×4.6mm,5μm） column with mobile phase consisted of acetonitrile-water （gradient elution） at the flow rate of 1.0 ml/min. The column temperature was 30 ℃ and injection volume was 10 μl. The drift tube temperature of ELSD was set at 65 ℃ with the flow rate of 2.0 L/rain. RESULTS： The linear range of nigeglanoside A were 0.183-2.562 mg/ml （r = 0.999 0） with an average recovery of 99.93 % （RSD=1.60%, n=6）. RSDs of precision, stability and reproducibility tests were all lower than 2%. CONCLUSIONS： The method is sim-ple, reproducible and accurate, and it can be used for the content determination of nigeglanoside A in total saponin extract of N. glandulifera.