高效毛细管电泳法快速测定血清假尿嘧啶核苷  

Rapid Analysis of Pseudouridine in Human Serum by High Performance Capillary Electrophoresis

作　　者：程明刚[1] 郝艳华[1] 邹亚红 梁统[2] 周克元[2] 凌光鑫[2] 

机构地区：[1]深圳市宝安人民医院,广东深圳518101 [2]广东医学院生化教研室,广东湛江524023

出　　处：《现代检验医学杂志》2004年第2期11-13,共3页Journal of Modern Laboratory Medicine

基　　金：广东省深圳市宝安区科学技术局;广东省深圳市科学技术局资助(课题编号:199904002)项目

摘　　要：目的 建立血清中假尿嘧啶核苷(?)的高效毛细管电泳快速测定方法。方法 用Bio-rad BioFo-cusTM 3000型高效毛细管电泳仪,仪器配有二极管阵列检测器,24 cm×25μm(i.d.)涂层石荧毛细管柱(Bio-rad公司:148-3011)及硼酸盐缓冲液(200mmol/L,pH8.2),应用电压为-16 kV,采用68.95 kPa·s压力进样。用磺基水杨酸作内标,血清标本经超滤后,超滤液直接进样分析。结果 血清中(?)可在5 min内与其它杂峰基线分离。在(?)浓度为0～100μmol/L的范围内,用(?)与内标的峰面积比值(X)对相应的(?)浓度(Y)得标准曲线为:Y=42.68X+1.23(r=0.999 2),血清中(?)最低检测限为3μmol/L。采用此法,尚可分析血清中尿酸、马尿酸等。结论 采用高效毛细管电泳法检测人血清中的(?),血清标本处理简单、精密度高、试剂消耗低、全自动化,可用于假尿嘧啶核苷等的临床应用价值探讨。Objective To establish a rapid method to analysis pseudouridine in human serum by high performance capillary electrophoresis (HPCE). Methods A Bio-rad BioFocus?3000 high performance capillary electrophoresis system with a diode-array UV detector.a 24 cm×25 μm(i. d. )coated capillary,a borate buffer solution(200 mmol/L,pH8. 2) were used. Sulfosalicylic acid was added as an internal standard to serum and the mixture was ultrafiltered by centrifugation using a centrifree micropartition system(Mrcut-off 30kDa),the ultrafiltrate was directly used for HPCE analysis. Results In this experiment,pseudouridine could be baseline resolved with uridine and anti-tumor drugs and other endogenous substance in serum within 5 minutes. Linearity between the concentrations of pseudouridine and the corresponding peak area ratios of pseudouridine and the internal standard,was demonstrated in the 0-100 μmol/L (r=0. 999 2). The detection limit of concentration for pseudouridine was 3μmol/L. Conclusion The method,which had simple sample preperation.good assay precision, low cost and full-automation,is shown to have promising clinical utility for quantitative determination of a few important metabolites (such as pseudouridine, uric acid).

关 键 词：假尿嘧啶核苷 尿酸 血清 超滤法 高效毛细管电泳法 

分 类 号：R446.112[医药卫生—诊断学]