高效液相色谱评价血清胆固醇试剂中酶的性能  

Evaluation of enzyme properties in cholesterol reagent kits by HPLC

作　　者：陈文祥[1] 董军[1] 唐蔚青[1] 李培瑛[1] 李健斋[1] 

机构地区：[1]卫生部北京老年医学研究所

出　　处：《中华医学检验杂志》1995年第4期208-210,共3页

摘　　要：为了评价国内市场上血清胆固醇试剂中胆固醇酯酶（ＣＥＨ）和胆固醇氧化酶（ＣＯＤ）的性能，设计了测定反应液（血清加试剂）中未被水解的胆固醇酯（ＵＨＣＥ）和未被氧化的游离胆固醇（ＵＯＦＣ）的方法。该法用正己烷提取未经皂化的和皂化的反应液中的游离胆固醇，衍生为苯氨基甲酸酯，高效液相色谱测定其含量。未经皂化的反应液中的游离胆固醇是ＵＯＦＣ，皂化后的反应液中的游离胆固醇是ＵＯＦＣ与ＵＨＣＥ之和，从中减去ＵＯＦＣ即为ＵＨＣＥ。用此法检查了１２种国产和进口试剂盒，多数试剂的ＣＯＤ和ＣＥＨ性能良好，少数较差，表现为反应液中ＵＨＣＥ较高，其中有２种试剂为１％～２％，２种为２％～４％，１种高达５．４％，ＣＥＨ是影响试剂性能的主要因素。该法可以有效地鉴定胆固醇试剂中两种酶的性能是否能达到要求。n order to evaluate the activities of cholesterol ester hydrolase and cholesterol oxidase in cholesterolreagent kits,we prepared a HPLC method for measuring unhydrolysed cholesterol ester(UHCE )andunoxidated free cholesterol(UOFC )levels in reaction mixture(serum-reagent ).Free chOlesterol con-tents in non-saponified and saponified reaction mixtures were determined by HPLC after extration withhexane and derivatization into phenylurethane. Free cholesterol in the non-saponified reaction mixture isUOFC;free cholesterol in the saponified reaction mixture is the sum of UOFC and UHCE, from whichUHCE was calculated by deducting UOFC. The efficiencing of COD and CEH varied in twelve commer-cial cholesterol reagent kits,most of them are good and CEH is the main factor determining the qualityof cholesterol reagents.This method can appraise effectively whether the two enzymes in cholesterolreagent kits fulfill the quality requirements.

关 键 词：胆固醇氧化酶 胆固醇酯酶 高效液相色谱法 

分 类 号：R446.112[医药卫生—诊断学]