LC-MS/MS法同时测定芪雪浸膏剂中9种成分的含量

Determination of 9 components in Qixue concrete by LC-MS/MS

目的建立同时测定芪雪浸膏剂中大黄素、毛蕊异黄酮、大黄酸、槲皮素、杨梅素、虎杖苷、金丝桃苷、大黄素甲醚、黄芪甲苷成分的LC-MS/MS方法。方法采用液质联用方法测定;Thermo Hypersil GOLD PFP C_(18)色谱柱(100 mm×2.1 mm,1.9μm);流动相为0.1%甲酸水溶液(A)-乙腈(B),梯度洗脱:0~1 min,25%B;1~5.5 min,25%B^90%B;5.5~9 min,90%B;9~9.5 min,90%B^25%B;9.5~13 min,25%B;流速:0.2 mL·min^(-1);进样量20μL;柱温:30℃;以电喷雾离子源(ESI)正负离子模式将样品离子化,多反应监测(MRM)模式进行测定;喷雾电压(+)4000/(-)3000 V;源内温度200℃;毛细管温度270℃;鞘气(N2)压力20 Arb,辅助气(N2)压力为5 Arb。结果大黄素、毛蕊异黄酮、大黄酸、槲皮素、杨梅素、虎杖苷、金丝桃苷、大黄素甲醚、黄芪甲苷浓度分别在19.88~1988 ng·mL^(-1)(r=0.9990)、20.92~2092 ng·mL^(-1)(r=0.9995)、19.99~1999.4ng·mL^(-1)(r=0.9987)、40.56~4056 ng·mL^(-1)(r=0.9983)、39.68~3968 ng·mL^(-1)(r=0.9974)、20.72~2072 ng·mL^(-1)(r=0.9999)、19.64~1964 ng·mL^(-1)(r=0.9993)、20.04~2004 ng·mL^(-1)(r=0.9998)、20.80~2080 ng·mL^(-1)(r=0.9988)内与峰面积呈良好的线性关系,精密度、稳定性、重复性和加样回收结果均符合方法学测定要求。结论利用建立的方法可同时测定芪雪浸膏剂中9种成分的含量,操作简单、重复性好、准确可靠,能为芪雪浸膏剂的质量控制提供依据。

Objective To determine the content of emodin, calycosin, rhein, quercetin, myricetin, polydatin, hyperin, physcion and astragaloside Ⅳ in Qixue concrete by LC-MS/MS. Methods The target sample was determined on a Thermo Hypersil GOLD PFP C（18） column（100 mm×2.1 mm, 1.9 μm） with gradient elution（0.1% formic acid aqueous as solution A and acetonitrile as solution B）： 0-1 min, 25%B; 1-5.5 min, 25%B-90%B; 5.5-9 min, 90%B; 9-9.5 min, 90%B-25%B; 9.5-13 min, 25%B. The flow rate was 0.2 mL·min^（-1）, the sample quantity was 20 μL, and the column temperature was 30 ℃. The eletrospray ionization（ESI） with positive source of the mass spectrometer and multiple reaction monitor（MRM） mode were used to detect the 9 components in Qixue concrete; Spray voltage：（ ＋） 4000/（-） 3000 V. Temperature within the source at 200 ℃; The temperature of the capillary was 270 ℃; the sheath gas pressure was 20 Arb; and the auxiliary air pressure was 5 Arb. Results Emodin, calycosin, rhein, quercetin, myricetin, polydatin, hyperin, physcion and astragalo-side Ⅳ showed good linearity at 19.88-1988 ng·mL^（-1）（r = 0.9990）, 20.92-2092 ng·mL^（-1）（r = 0.9995）, 19.99-1999.4 ng·mL^（-1）（r = 0.9987）, 40.56-4056 ng·mL^（-1）（r = 0.9983）, 39.68-3968 ng·mL^（-1）（r = 0.9974）, 20.72-2072 ng·mL^（-1）（r = 0.9999）, 19.64-1964 ng·mL^（-1）（r = 0.9993）, 20.04-2004 ng·mL^（-1）（r = 0.9998）, 20.80-2080 ng·mL^（-1）（r = 0.9988）, respectively. The method was precision, stability, repeatability and the recovery test results conformed to the requirements of the methodology. Conclusion The method is sensitive, accurate and suitable, and can be applied for the determination of 9 components in Qixue concrete.