高效液相色谱法测定血清葡萄糖  

Determination of glucose in human serum by high performance liquid chromatography

作　　者：陈忠余[1] 张天娇[2] 张传宝[2] 张江涛[2] 周伟燕[2] 闫颖[2] 陈文祥[2] 

机构地区：[1]重庆市第一人民医院检验科,重庆400011 [2]卫生部北京医院卫生部临床检验中心,北京100730

出　　处：《临床检验杂志》2011年第9期660-662,共3页Chinese Journal of Clinical Laboratory Science

基　　金：国家科技支撑计划资助项目(2007BAI05B09);国家高技术研究发展计划(863)资助(2006AA020909)

摘　　要：目的建立一种测定血清葡萄糖(glucose,Glu)的高效液相色谱法(HPLC),探讨其能否作为血清Glu测定的参考方法。方法以D-半乳糖(D-galactose)为内标物,用无水乙醇沉淀、去除血清中的蛋白质,在pH 9.1条件下与1-苯基-3-甲基-5-吡唑啉酮(PMP)反应,用HPLC测定血清Glu衍生产物、D-半乳糖衍生产物,用标准曲线法定量;对建立的方法进行方法学评价。结果本法测定血清Glu的批内变异系数(CV)为0.33%～0.67%,平均0.51%;批间CV为0.02%～0.85%,平均0.38%;总CV为0.47%～1.03%,平均0.68%。回收率为98.22%～101.96%。分析Glu的参考物质SRM 965a,测定结果与认定值的偏差为-0.928%～0.347%,平均偏差为-0.072%。结论初步建立了测定血清Glu的HPLC法;该法准确、精密,有望作为血清Glu测定的候选参考方法。Objective To establish a candidate reference method for the determination of glucose in human serum based on high performance liquid chromatography（HPLC）.Method D-galactose as the internal standard was added to the serum samples,and glucose standard was also added to equilibrate.The samples were treated with anhydrous ethanol to remove proteins by precipitation.Glucose and galactose reacted with 1-phenyl-3-methyl-5-pyrazolone in serum under weakly alkaline condition and the derivatives were analyzed by high performance liquid chromatography（HPLC）.The concentration of glucose in serum was quantified by a calibration curve.Results The methodology features of the established HPLC method for determination of serum glucose were evaluated and the results were as follows.The within-run,between-run and total coefficients of variation ranged from 0.33% to 0.67%,0.02 % to 0.85% and 0.47% to 1.03%,and the averages were 0.51%,0.38% and 0.68%,respectively.The analytical recovery rate ranged from 98.22% to 101.96%.The average bias of certified reference material SRM 965 was-0.072%,ranged from-0.928% to 0.347%.Conclusion The established HPLC method for serum glucose determination in this study is accurate,precise and practicable,and may be used as candidate reference method for determination of glucose in serum.

关 键 词：葡萄糖 高效液相色谱 1-苯基-3-甲基-5-吡唑啉酮 

分 类 号：R286.0[医药卫生—中药学]