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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 a LC-ESI-MS method for the determination of homocysteine (HCY) in rats. Methods: The chromatographic column was Zorbax SB-C18 (2.1 mm × 100 mm, 3.5 μm). Acetonitrile-water (40:60) was used as the mobile phase at a flow rate of 0.3 mL · min-1 and the column temperature was 30 ℃. ESI, electrospray ionization, positive ion detection, atomizing gas pressure 172.375 kPa; drying gas N2 flow rate 7 L · min -1, drying temperature 350 ° C. [M + H] +, m / z 136. Results: The linearity of HCY in the range of 2.5 ~ 100μmol.L-1 was good, the linear equation C = 0.0079X-8.1011, r = 0.9981, the lowest detection limit was 1μmol.L-1. The intra-day and inter-day HCI RSDs were less than 6%. The concentration of HCY in Wistar-Kyoto rats of 12 weeks old was (9.57 ± 3.49) μmol.L-1 and (11.59 ± 3.13) μmol.L-1 respectively in spontaneously hypertensive rats. Conclusion: The method is simple, rapid, sensitive and can be used for the detection of HCY in rats.