UPLC-MS/MS法同时测定裸花紫珠干浸膏中5种有效成分的含量

Simultaneous Determination of 5 Effective Components in Luohua Zizhu Dry Extract by UPLC-MS/MS

目的:建立同时测定裸花紫珠干浸膏中5种有效成分含量的方法。方法:采用超高效液相色谱-串联质谱法。色谱条件:色谱柱为Zorbax Eclipse Plus C_(18),流动相为乙腈-水(梯度洗脱),流速为0.3 mL/min,柱温为40℃,进样量为2μL;质谱条件:多反应监测离子扫描模式,干燥气/雾化气为氮气,干燥气温度为325℃,干燥气流量为6 L/min,雾化气压力为45 psi,鞘流气温度为350℃,鞘流气流量为12 L/min,毛细管电压为4 000 V(+)、3 500 V(-),喷嘴电压为500 V。结果:木犀草苷、毛蕊花糖苷、槲皮素、木犀草素、芦丁检测质量浓度线性范围分别为0.504 8~252.4 ng/mL(r=0.999 9)、0.712 4~356.2 ng/mL(r=0.999 0)、0.509 4~254.7 ng/mL(r=0.996 2)、0.303 0~151.5 ng/mL(r=0.999 8)、0.602 2~301.1 ng/mL(r=0.999 6);精密度、稳定性、重复性试验的RSD<3.0%;定量限分别为0.42、0.87、0.33、0.12、0.76 ng/mL;加样回收率分别为97.99%~101.20%(RSD=1.3%,n=6)、96.50%~101.20%(RSD=1.7%,n=6)、94.81%~99.34%(RSD=1.7%,n=6)、97.54%~100.51%(RSD=1.2%,n=6)、93.37%~98.70%(RSD=1.9%,n=6)。结论:该方法操作简便,精密度、稳定性、重复性好,可用于裸花紫珠干浸膏中5种有效成分含量的同时测定。

OBJECTIVE： To establish a method for simultaneous determination of 5 effective components in Luohua zizhu dry extract. METHODS： UPLC-MS/MS was conducted. The separation was performed on an Zorbax Eclipse Plus C18 column with mobile phase of acetonitrile-water （gradient elution） at the flow rate of 0.3 mL/min. The column temperature was set at 40 ℃ and sample size was 2 μL. The analytes were detected in the multiple reaction monitoring （MRM） mode. Nitrogen was used as drying gas and atomized gas. The temperature and flow rate of drying gas were 325 ℃ and 6 L/min. The pressure of atomized gas was 45 psi. The temperature and flow rate of sheath gas were 350 ℃ and 12 L/min. The voltage of capillary were 4 000 V（＋） and 3 500 V（-）. The voltage of nozzle was 500 V. RESULTS：The linear ranges of luteoloside,acteoside,quercetin,luteolin and rutin were 0.504 8- 252.4 ng/mL（r=0.999 9）,0.712 4-356.2 ng/mL（r=0.999 0）,0.509 4-254.7 ng/mL（r=0.996 2）,0.303 0-151.5 ng/mL（r=0.999 8） and 0.602 2-301.1 ng/mL （r=0.999 6）, respectively. RSDs of precision, stability and reproducibility tests were all less than 3.0%. The limit of quantitation were 0.42,0.87,0.33,0.12,0.76 ng/mL. The recoveries were 97.99%-101.20% （RSD=1.3%,n=6）, 96.50%-101.20% （RSD=1.7%,n=6）, 94.81%-99.34% （RSD=1.7%,n=6）, 97.54%-100.51% （RSD=1.2%,n=6）,93.37%-98.70% （RSD=1.9%,n=6）, respectively. CONCLUSIONS： The method is simple, precise, stable and reproducible, and can be used for simultaneous determination of 5 effective components in Luohua zizhu dry extract.