基于GC-MS的心肌缺血大鼠血浆中内源性代谢物测定方法的建立

目的：建立GC-MS测定心肌缺血大鼠血浆中内源性代谢物的方法。方法：皮下注射异丙肾上腺素复制大鼠心肌缺血模型，采用DB-5MS Ultra Inert（30m×250μm，0.25μm）色谱柱，GC-MS进行程序升温，氦气（纯度>99.999%）为载气；无分流进样；流速为1mL·min-1；进样口温度设为250℃；进样量为2μL。质谱参数：离子源温度设为230℃；电子能量为70eV；电子倍增器电压1847V；电离方式EI。质谱采用全扫描方式（扫描范围30～550m/z）进行数据采集。自建对照品物质库，并参考气质工作站的数据库对内源性生物标志物进行鉴定指认，然后进行方法学考察，包括精密度、重现性、稳定性。结果：仪器与方法精密度、重复性和稳定性试验的相对峰面积RSD值均小于20%。结论：该方法快速、灵敏、准确、重复性好，可用于心肌缺血大鼠血浆中内源性代谢物的测定，该方法可应用于心肌缺血模型的代谢组学研究。

Determination of endogenous metabolites in plasma of myocardial ischemia rat model by GC-MS

Objective: To establish a GS-MS method for determination of endogenous metabolites in plasma of myocardial ischemia rats. Methods:subcutaneous injection of isoproterenol replication model of myocardial ischemia rats, using DB-5MS Ultra Inert(30m×250m, 0.25μm) column chromatography, GC-MS by temperature programmed, carrier gas was helium (purity >99.999%), the flow rate was 1mL·min-1; splitless injection; into the injector temperature of 250℃; sample injection volume was 2μL. MS parameters: ion source temperature was 230℃; EI ionization; electron energy of 70, electron multiplier voltage of 1847 V eV. MS scan mode wtih full data acquisition, and the scanning range of 30~550m/z . Using the database of temperament work station, combined with the self-built reference material library to identify endogenous biomarkers, select the endogenous biomarkers with high matching degree as the identification results. Results:Instrument and method precision, repeatability and stability test of relative peak area of RSD values were less than 20% .Conclusion:The method is rapid, sensitive, accurate, reproducible, and can be used for the determination of endogenous metabolites in plasma of rats with myocardial ischemia. The method can be applied to myocardial ischemia model of blood metabolic group study.