摘要
采用固相萃取纯化血清样品,荧光胺柱前衍生,建立了反相高效液相色谱/荧光检测血清中胰岛素的方法。色谱条件:LunaC5柱(100 mm×4.6 mm,5μm)分离,流动相为乙腈-Tris缓冲溶液(10 mmol/L,pH 8.5)梯度洗脱,流速:1 mL/min。激发波长(λex)273 nm,发射波长(λem)476 nm,进样量20μL。结果表明:胰岛素在10~800μg/L范围内峰面积和质量浓度呈良好的线性关系,相关系数r为0.9971;检出限为5μg/L。日内、日间精密度分别为1.5%、1.0%,回收率为92%。
A method for the determination of insulin in plasma by reversed-phase high performance liquid chromatography with fluorescence detection was developed. The method involved solid-phase extraction of insulin from plasma and pre-cotumn derivatization of insulin with fluorescamine. The derivatives were separated on a LunaC5 column using a acetonitrile -Tris buffer solution（ 10 mmol/L, pH 8.5 ） with gradient elution and detected at λex ： 273 nm, λem ： 476 nm. The linearity range of the assay for insulin was 10 - 800 i^g/L with correlation coefficient of 0. 997 1, the detection limit of 5 μg/L, the intra- and inter- day precision of 1.5% and 1.0% , and the average recovery of 92% .
出处
《分析测试学报》
CAS
CSCD
北大核心
2008年第8期898-900,共3页
Journal of Instrumental Analysis
基金
重庆市科技计划资助项目(2005BB5226)
关键词
固柑萃取
反相高效液相色谱
胰岛素
荧光胺
solid phase extraction
reversed-phase high performance liquid chromatography
insulin
fluorescamine
