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目的建立儿童经过体外循环后血浆异丙酚的浓度测定方法方法采用反相高效液相色谱—荧光检测技术,血浆样品经过加入内标(麝香草酚)的甲醇溶液沉淀蛋白质后,取上清液20μl进行HPLC分析,流动相为乙腈:水=56:44(V/V),流速为1ml/min,荧光检测条件为λex276nm,λem=312nm,机器采用Agilent1100系列高效色谱仪,分离色谱柱采用DikmaDiamonsilC18(150mm×4.6nm,5.0um),保护柱为PhenomenexSecurity(4mm×3mm)。结果在0.02~8.0mg/L的范围内线性关系好(r=0.9993,n=9),最低检测浓度是0.005ug/ml,(S/N=2),日内,日间RSD分别为0.810%~5.73%和0.932%~7.337%,平均回收率为96.99%。结论本法灵敏度高,方法简单,实验快速,线性关系好,专一性强,可用于儿童经过体外循环后血浆异丙酚的浓度测定。
Objective To establish a method for the determination of plasma propofol concentrations in children after cardiopulmonary bypass. Methods The plasma samples were precipitated by adding methanol (thymol) methanol solution with reversed-phase high performance liquid chromatography-fluorescence detection. The supernatant 20μl for HPLC analysis, the mobile phase of acetonitrile: water = 56:44 (V / V), flow rate of 1ml / min, fluorescence detection conditions λex276nm, λem = 312nm machine using Agilent1100 series of high performance chromatographic separation column using DikmaDiamonsilC18 (150 mm × 4.6 nm, 5.0 μm) and the guard column was Phenomenex Security (4 mm × 3 mm). The results showed good linearity (r = 0.9993, n = 9) in the range of 0.02-8.0 mg / L and the lowest detection limit was 0.005ug / ml (S / N = 2). The intra- and interday RSD were 0.810% ~ 5.73% and 0.932% ~ 7.337%, the average recovery was 96.99%. Conclusion This method has high sensitivity, simple method, rapid experiment, good linearity and specificity. It can be used for the determination of plasma propofol in children after cardiopulmonary bypass.