HPLC-MS/MS法同时测定健胃消食片中8种化学成分的含量

目的: 建立HPLC-MS/MS法同时测定健胃消食片中山柰酚、槲皮素、柚皮素、柚皮苷、芦丁、芹菜素、橙皮苷和绿原酸的含量。方法: 采用Sapphire C₁₈色谱柱(150 mm×4.6 mm,5 μm),以甲醇-水(0.03%的甲酸)为流动相进行梯度洗脱,流速0.8 mL·min^-1,进样量10 μL,柱温30℃;采用电喷雾离子源进行负离子模式监测,多反应监测模式(MRM)用于定量分析,源喷射电压-4.5 kV,离子源温度650℃;雾化气为(GS1,N₂)压力413.7 kPa,辅助气(GS2,N₂)压力448.2 kPa,气帘气(N₂)压力206.8 kPa,接口加热,全程通入氮气,MRM模式定量,扫描范围为m/z 100~1 000。结果: 健胃消食片中8种成分的峰面积和浓度呈良好的线性关系,r>0.999 1;平均加样回收率(n=9)为97.25%~100.8%。5批样品中山柰酚、槲皮素、柚皮素、柚皮苷、芦丁、芹菜素、橙皮苷、绿原酸的含量依次为2.135~4.616、0.244 3~0.319 4、0.257 2~0.361 7、20.01~34.65、7.561~11.82、0.020 58~0.025 58、259.2~300.2、40.52~46.62 μg·g^-1。结论: 本法简便、准确,重现性好,专属性高,可用于健胃消食片的质量控制。

Objective: To develop an HPLC-MS/MS method for the determination of campherol,quercetin,naringenin,naringin,rutin,apigenin,hesperidin and chlorogenic acid in Jianwei Xiaoshi tablets. Methods: Analysis was performed on a Sapphire C₁₈ column(150 mm×4.6 mm,5 μm)with a gradient elution of methanol and water containing 0.03% formic acid at a flow rate of 0.8 mL·min^-1.The injection volume was 10 μL,and the column temperature was controlled at 30℃.The multiple-reaction monitoring scanning was employed for the quantification with switching electrospray ion source polarity in negative mode.The ion spray voltage was set at-4.5 kV and the turbo spray temperature was maintained at 650℃;nebuliser gas(gas 1)and heater gas(gas 2)were set at 413.7 and 448.2 kPa,respectively.The curtain gas was kept at 206.8 kPa and the interface heater was on.Nitrogen was used in all cases.The full-scan mass covered the range from m/z 100 to 1 000.MRM mode was employed for quantification. Results: There was significant correlation between the peak area and concentration of each compound within the test range(r>0.999 1).The average recoveries(n=9)of the compounds ranged from 97.25% to 100.8%.The contents of campherol,quercetin,naringenin,naringin,rutin,apigenin,hesperidin and chlorogenic acid in 5 batches of Jianwei Xiaoshi tablets were 2.135-4.616,0.244 3-0.319 4,0.257 2-0.361 7,20.01-34.65,7.561-11.82,0.020 58-0.025 58,259.2-300.2,and 40.52-46.62 μg·g^-1,respectively. Conclusion: The established method is simple,accurate and highly reproducible,which can be used for quality control of Jianwei Xiaoshi tablets.