HPLC-MS/MS法测定人血浆中瓜氨酸浓度及多发伤患者胃肠道损伤的相关性研究

Correlation between citrulline concentration and gastrointestinal injury in patients with multiple injuries by HPLC-MS/MS

目的建立高效液相色谱-串联质谱(HPLC-MS/MS)法测定瓜氨酸含量,评价瓜氨酸与多发伤患者胃肠道损伤的关系。方法色谱柱:Shim-pack XR-ODS(2.0 mm×3 mm,3.5 μm);流动相:水(含0.1%甲酸)(A)-乙腈溶液(B)(90∶10);流速:0.2 mL·min-1;柱温:40℃;进样量:10 μL;色谱分析时间:5 min。质谱采用电喷雾离子源(ESI)正离子模式下,以多反应监测模式进行正离子扫描。结果瓜氨酸浓度在0.236~59 μg·mL-1与峰面积线性关系良好,3个不同浓度质控样本日内和日间精密度RSD值均在±15%内,方法学回收率为95.83%~103.75%。结论本方法操作简便、快速、准确、专属性强,为评价瓜氨酸的活性提供方法学基础,可用于患者血浆中瓜氨酸活性的常规监测。

Objective To establish a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method to determine citrulline,and to evaluate the relationship between citrulline and gastrointestinal injury in patients with multiple injuries.Methods The chromatographic column was Shim-pack XR-ODS (2.0 mm×3 mm,3.5 μm).The mobile phase was water (containing 0.1% formic acid)-acetonitrile solution (90∶10) with elution and volume flow rate of 0.2 mL·min-1.The column temperature was 40℃ and the chromatographic analysis time was 5 min.Mass spectrometry was used for positive ion scanning with electrospray ionization (ESI) and selected ion reaction monitoring (SIM).Results The calibration curves showed good linearity at 0.236-59 μg·mL-1.The intra-day and inter-day precision and accuracy values of citrulline at 3?levels of QC samples were less than 15%.The recovery rate of citrulline was 95.83%-103.75%.Conclusion An accurate,simple and sensitive method to determine the plasma concentration of citrulline by HPLC-MS/MS is established.The proposed method helps identify and quantify citrulline in clinical studies.