高效液相色谱检测全血贮存对血清总胆固醇和高密度脂蛋白胆固醇的影响

Stability of serum total and high density lipoprotein cholesterol during whole blood incubation examined by high performance liquid chromatography

目的检查全血贮存对血清总胆固醇(TC)和高密度脂蛋白胆固醇(HDLC)的影响,并探讨其可能机制。方法建立高效液相色谱(HPLC)方法,测定不同个体全血样品经不同温育后血清TC、总游离胆固醇(TFC)、HDLC和高密度脂蛋白游离胆固醇(HDLFC),计算有关参数或指标并做统计分析。结果建立的HPLC方法平均批内变异系数0.22%～0.51%;全血温育造成血清TC和HDLC变化(平均0.6%～2.1%和2.0%～4.1%),有些个体变化明显(TC>±3%,HDLC>±5%)。结论建立血清TC、TFC、HDLC和HDLFC的精密测定方法;血清TC和HDLC在不同条件下的全血贮存中,受血清体积变化、血细胞-脂蛋白间胆固醇交换或转移等许多复杂因素的影响而发生变化;血脂测定标本应尽量减少不必要的贮存。

Objective To examine the changes of serum total and high density lipoprotein cholesterol during whole blood incubation and to investigate the possible mechanisms responsible for the instability.Methods A method for the measurement of serum total cholesterol (TC), total free cholesterol (TFC), high density lipoprotein cholesterol (HDLC) and high density lipoprotein free cholesterol (HDLFC) by high performance liquid chromatography (HPLC) was developed. Whole blood specimens were incubated at 25℃ and 4℃ for various period of time and serum TC, TFC, HDLC and HDLFC were measured.Results The new HPLC method showed a mean within-run CV of 0.22%~0.51%. The averaged changes during the incubations ranged 0.6%~2.0% for TC and 2.0%~4.1% for HDLC.Conclusion An HPLC method has been established that is highly precise and can be used for detecting subtle cholesterol changes in biological samples. Different cholesterol exchanges or transfers between blood cells and lipoproteins exist during various incubations. Prolonged whole blood storage that causes serum TC and HDLC changes should be avoided in clinical lipid measurements.