HPLC法测定肾移植患者血清中内源性氢化可的松浓度

Determination of the Serum Concentration of Endogenous Cortisol in Patients Underwent Renal Transplantation by HPLC

目的:建立测定肾移植患者血清中内源性氢化可的松浓度的方法。方法:采用高效液相色谱法。色谱柱为Diamonsil C18,流动相为乙腈-水-甲醇(51∶136∶77),流速为1.2 mL·min-1,紫外检测波长为245 nm,柱温为30 ℃,进样量为30 μL。结果:氢化可的松血药浓度在50～300 μg·L-1范围内线性关系良好,最低检测限为8 μg·L-1;日内、日间RSD分别为4.04%～6.81%、4.09%～9.16%,低、中、高3种浓度的方法回收率分别为(103.12±4.53)%、(100.77±4.07)%、(101.61±6.92)%。肾移植患者内源性氢化可的松术后1～3、3～6、6～12、>12个月的浓度分别为(31.89±21.42)、(45.26±29.26)、(49.69±22.00)、(53.56±29.86)μg·L-1。结论:本法操作简便、灵敏度高、快速可靠,适用于肾移植患者血清中内源性氢化可的松浓度的测定。

OBJECTIVE：To establish the method for the determination of serum concentration of endogenous cortisol in patients underwent renal transplantation. METHODS： HPLC method was adopted. The separation was performed on C18（250 mm×4.6 mm, 5 μm） column with mobile phase consisted of acetonitrile-water-methanol （51 ∶ 136 ∶ 77） at flow rate of 1.2 mL·min-1. UV detection wavelength was set at 245 nm and column temperature was 30 ℃. The injection volume was 30 μL. RESULTS： The linear range of cortisol was 50～300 μg·L-1 with lowest detection limit of 8 μg·L-1. The RSDs of intra-day and inter-day were 4.04%～6.81% and 4.09%～9.16%. Average recoveries of cortisol at low, medium and high concentrations were （103.12±4.53）%,（100.77±4.07）% and（101.61±6.92）%. The concentrations of cortisol 1～3 months, 3～6 months, 6～12 months, more than 12 months after operation were （31.89±21.42）μg·L-1, （45.26±29.26）μg·L-1,（49.69±22.00）μg·L-1,（53.56±29.86）μg·L-1. CONCLUSION： The method is simple, sensitive, rapid, reliable and suitable for the determination of serum concentration of endogenous cortisol in patients underwent renal transplantation.