UHPLC-MS/MS法同时测定六合氨基酸注射液中6个有效成分的含量

目的：建立UHPLC-MS/MS法同时检测六合氨基酸注射液中6个有效成分（L-精氨酸、L-谷氨酸、L-门冬氨酸、L-亮氨酸、L-异亮氨酸和L-缬氨酸）的含量。方法：采用Agilent ZORBAX SB-C₁₈（3.0mm×150 mm，5 μm）色谱柱分离，以含0.2%甲酸和0.02%七氟丁酸的甲醇溶液-含0.2%甲酸和0.02%七氟丁酸的水溶液为流动相，梯度洗脱，流速0.4 mL·min^-1，柱温50℃；以电喷雾离子源（ESI），正离子扫描方式，多反应监测扫描模式进行定量分析。结果：所测6个氨基酸浓度在一定范围内呈良好的线性关系，r>0.999 3；实验精密度、稳定性、重复性良好；平均加样回收率为95.8%~103.2%，RSD<4.2%。10批样品中L-精氨酸、L-谷氨酸、L-门冬氨酸、L-亮氨酸、L-异亮氨酸和L-缬氨酸的含量检测结果分别为21.64~22.24、18.12~18.64、3.84~4.24、16.40~16.88、10.76~11.48和11.96~12.48 mg·mL^-1。结论：经方法学验证，所建立的分析方法简便、快速、准确，灵敏度高，可作为六合氨基酸注射液的质量控制和评价方法。  

Objective: To establish a method for simultaneous determination of six effective components (L-arginine, L-glutamic acid, L-aspartic acid, L-leucine, L-isoleucine and L-valine) in the compound amino acid injection by UHPLC-MS/MS. Methods: Separation was performed on an Agilent Zorbax SB-C₁₈ column (3.0 mm×150 mm, 5 μm) using a linear gradient solvent system of methanol(0.2% formic acid and 0.02% heptafluorobutyric acid) and water(0.2% formic acid and 0.02% heptafluorobutyric acid) as mobile phase, at a flow rate of 0.4 mL·min^-1 and the column temperature was set at 50℃. Detection was executed by MS/MS in the multiple reaction monitoring(MRM) mode using a positive electrospray ionization interface. Results: This method exhibited excellent linearity for all of the analytes with regression coefficients r>0.999 3. The precision,stability, and repeatability of the method were good for all the six amino acids. The average recoveries varied between 95.8% and 103.2%,with the RSD were less than 4.2%. The components of L-arginine,L-glutamic acid,L-aspartic acid, L-leucine, L-isoleucine and L-valine in 10 samples were 21.64~22.24,18.12~18.64, 3.84~4.24, 16.40~16.88, 10.76~11.48 and 11.96~12.48 mg·mL^-1, respectively. Conclusion: The method was proved to be convenient, accurate, sensitive and good in reproducibility, which could be applied to the quality control and assessment of compound amino acid injection. 

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