固相萃取LC-MS/MS法测定疏血通注射液中次黄嘌呤浓度

SPE-LC-MS/MS method for determination of hypoxanthine concentration in Shuxuetong injection

关键词:  液质联用  固相萃取  疏血通注射液  次黄嘌呤  定量分析

Key words:  LC-MS/MS  solid phase extraction  Shuxuetong injection  hypoxanthine  quantitative determination

摘要:

[目的]建立固相萃取液相色谱串联质谱法（LC-MS/MS）测定疏血通注射液中次黄嘌呤浓度。[方法]疏血通注射液经固相萃取柱处理后，采用LC-MS/MS进行含量测定。色谱柱为Acquity BEH C₁₈分析柱（100 mm×2.1 mm，1.7 μm），以甲醇-水（含0.1%甲酸）为流动相进行梯度洗脱，流速0.3 mL/min。于ESI负离子模式下，以多反应离子监测（MRM）扫描方式进行监测，用于定量分析的离子为m/z 134.9→92.0（次黄嘌呤）和m/z 151.0→92.1（内标6-巯基嘌呤）。[结果]次黄嘌呤在质量浓度0.1~0.5 μg/mL范围内与待测物和内标的峰面积比值呈良好线性关系（r=0.999 8），平均加样回收率为98.7%，RSD为0.31%。[结论]该方法简便、准确、灵敏，可用于疏血通注射液中次黄嘌呤的浓度测定。

Abstract:

[Objective] Solid phase extraction with liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) was applied to determine the concentration of hypoxanthine in Shuxuetong injection.[Methods] After extracted with SPE column,Shuxuetong injection was detected by LC-MS/MS. This injection was separated on an Acquity BEH C₁₈ column (100 mm×2.1 mm,1.7 μm),and methanol-water (containing 0.1% formic acid) was used as a mobile phase for gradient elution at a flow rate of 0.3 mL/min. Mass spectrometric detection was performed in negative ion and multiple reaction monitoring mode with an electrospray ionization (ESI) source. The ion pairs used for quantitative analysis were m/z 134.9→92.0 for hypoxanthine and m/z 151.0→92.1 for 6-mercaptopurine (internal standard,IS).[Results] Hypoxanthine had a good linear relationship with the peak area ratio of the analyte and the IS in the range of 0.1~0.5 μg/mL (r=0.999 8). The average recovery rate was 98.7%,and the RSD was 0.31%.[Conclusion] This method described here is a simple,accurate and sensitive method that can be used for the determination of hypoxanthine concentrationin Shuxuetong injection.