LC-MS/MS同时测定痛安注射液中13个成分的含量

Simultaneous determination of thirteen components in Tong'an injection by LC-MS/MS

利用超高效液相色谱-质谱联用仪(UPLC-MS/MS)建立了同时测定痛安注射液(TAI)中丁香脂素、N-反式阿魏酰酪胺、白屈菜红碱、青藤碱、黄连碱、血根碱、白屈菜碱、木兰花碱、别隐品碱、原阿片碱、杜鹃素、次罂粟碱、二氢血根碱共13个有效成分的定性定量检测方法,用于痛安注射液的质量控制。采用Agilent Zorbax SB-Aq C_(18)柱(2.1 mm×150 mm,3.5μm)色谱柱,流动相A为0.1%甲酸水溶液,B为乙腈,梯度洗脱(0.01~2 min,5%B;2~8 min,5%~30%B;8~11 min,30%~95%B;11~13 min,95%B;13~13.1 min,95%~5%B;13.1~14 min,5%B),流速0.5 m L·min^(-1),柱温25℃;电喷雾离子源(ESI)正离子条件下检测,多反应离子监测(MRM)进行定量。结果所测上述13个待测成分在测定的质量浓度范围内均具有良好的线性关系(r>0.999 0);加样回收率在95.70%~104.8%,RSD均小于1.9%,10个批次样品中上述13种成分的质量浓度依次为0.021 2~0.029 0,0.001 7~0.002 3,0.000 9~0.001 3,5.952~6.205 2,0.195 4~0.240 5,0.002 0~0.002 9,0.693~0.798 2,0.069 3~0.078 2,0.089 29~0.102 9,0.386 5~0.420 1,0.014 3~0.015 9,0.755 3~0.842 1,0.008 2~0.011 2 g·L^(-1)。经方法学验证,该方法简便、快速、准确、重复性好,可以为综合评价痛安注射液的质量提供参考。通过对10个批次的痛安注射液进行分析测定,并根据含量测定结果表明不同批次痛安注射液的质量并无明显差异,从而为痛安注射液的质量控制提供依据。

The aim of this study was to develop a simple, sensitive ultra performance liquid chromatography mass spectrometry （UPLC-MS/MS） method for the determination of syringaresinol, N-trans-feruloyltyramine, chelerythrine chloride, sinomenine, coptisine chloride, sanguinarine, chelidonine, magnolflorine, allocryptopine, protopine, farrerol, stylopine and dihydrosanguin-arine in Tong＇an injection （TAI）, which could be used for the quality control of TAI. The UPLC analysis was performed on Agilent Zorbax SB-Aq column （2.1 mm×150 mm,3.5 μm）, with 0.1% formic acid solution （A） -acetonitrile （B） as the mobile phase for gradient elution （0.01-2 min, 5%B; 2-8 min, 5%-30%B; 8-11 min, 30%-95%B; 11-13 min, 95%B; 13-13.1 min, 95%-5%B; 13.1-14 min, 5%B）. The flow rate was 0.5 mL·min-1, and the column temperature was 25℃; multiple reaction monitoring （MRM） was performed in electrospray ion source positive ion mode for quantitative determination. The calibration curves for the above thirteen compounds showed good linear relationship in corresponding mass concentration range （r〉0.999 0）. The average recovery rate of the compounds ranged from 95.70% to 104.8%, with RSD of less than 1.9%. The contents of thirteen active components in 10 batches of TAI were 0.021 2-0.029 0, 0.001 7-0.002 3, 0.000 9-0.001 3, 5.952-6.205 2, 0.195 4-0.240 5, 0.002 0-0.002 9, 0.693-0.798 2, 0.069 3-0.078 2, 0.089 29-0.102 9, 0.386 5-0.420 1, 0.014 3-0.015 9, 0.755 3-0.842 1, and 0.008 2-0.011 2 g·L^-1 respectively. Methodology validation proved that this method was simple, rapid, sensitive and accurate, which can be used to provide reference for the comprehensive evaluation of TAI quality. The determination results of 10 batches of TAI showed the content of each batch had no significant difference. The results may provide a basis for the quality control of TAI.