反相HPLC法测定血浆中的氢化可的松和甲基强的松龙

Determination of Hydrocortisone and Methylprednisolone in Plasma by Reversed--Phase HPLC

作者用反相高效液相层析法测定血浆中甲基强的松龙和氢化可的松的浓度。样品用2.5ml乙酸乙酯萃取,分出有机相后,通空气挥干,残渣用甲醇溶解进样。采用ODS分析柱,以甲醇:醋酸盐缓冲液(pH3.0,0.01mol/L)=64:36(V/V)为流动相,地塞米松为内标,在254nm波长下监测洗提液。标准曲线用牛血浆代替人血浆,不仅使成本降低,而且避免了人血浆中内源性氢化可的松的干扰,本法具有操作简便、快速、灵敏、准确等特点。

A sensitive and rapidmethod for the simultaneous measurementof both hydrocortisone (HC) and methyl-prednisolone (MP) using dexamethasone(DX) as the internal standard was pre-sented. High-performance liquid chroma-tograph Model 344(Beckman, USA) with a254nm wavelength UV detector and re-versed-phase Ultrasphere ODS (5μm)column was used .Sample wasextracted withethyl acetate and the organic layer wasevaporated to dryness under an air streamin a 37 C water bath. The residue wasdissolved in 30μl mobile phase and 20μlinjected. The mobile phase of methanoland 0.01mol/L acetate buffer (pH 3.0)(64:36) was pumped at 1 .0ml/min throughthe column. The detector was operated at0.005 aufs. The retention time for HC,MP and DX was 5.43min, 7.7 min and11.5min, respectively. Standard curve was linear in the con-centration range of 0.02 to 1.28mg/L forHC and 0.05 to 12.8mg/L for MP. Sensi-tivity was 1ng for HC and 1.5ng for MP;extraction recovery 76-83%;within dayCV less than 4%, and inter-day CV lessthan 6% for both steroids.