LC-ESI-MS测定大鼠血浆中同型半胱氨酸的含量

LC-ESI-MS determination of homocysteine in rat plasma

目的：建立LC-ESI-MS法测定大鼠体内的同型半胱氨酸（HCY）。方法：色谱柱为Zorbax SB-C18（2.1mm×100mm,3.5μm）,乙腈-水（40∶60）为流动相,流速为0.3mL·min-1,柱温为30℃。ESI（电喷雾离子源）,正离子检测,雾化气压力设为172.375kPa;干燥气（N2）流速设为7L·min-1,干燥气温度设为350℃,[M＋H]＋,m/z136。结果：HCY在2.5~100μmol.L-1范围内线性关系良好,线性方程C=0.0079X-8.1011,r=0.9981,最低检测限为1μmol.L-1。HCY日内及日间精密度RSD均低于6%。12周龄的Wistar-Kyoto大鼠,自发性高血压大鼠体内HCY的浓度分别为（9.57±3.49）μmol.L-1和（11.59±3.13）μmol.L-1。结论：该法操作简便、快速,灵敏度高,可用于大鼠体内HCY的检测。

Objective：To establish an LC-ESI-MS method for determination of homocysteine in rat plasma.Methods：A Zorbax SB-C18（2.1 mm×100 mm,3.5 μm）column was used,with mobile phase of methanol-water（40∶60）,flow rate 0.3 mL·min-1,column temperature 30 ℃.Electrospray ionization（ESI）source was applied and operated in positive ion mode.The pressure of atomizing gas set up 172.375 kPa;the flow rate of drying gas（N2）was 7 L·min-1 and the temperature was 350 ℃,[M＋H]＋,m/z 136.Results：The calibration curve of homocysteine in plasma was linear in the concentration range of 2.5-100 μmol·L-1.C= 0.0079X-8.1011,r=0.9981.The LLOQ was 1 μmol·L-1.The precision of intra-day and inter-day was both below 6%.The concentration of HCY in 12 weeks Wistar-Kyoto and SHR（Spontaneously hypertensive rats）were（9.57±3.49）μmol·L-1 and（11.59±3.13）μmol·L-1,respectively.Conclusion：The analytical method for homocysteine is accurate,sensitive and suitable for analysis of rat plasma concentration.