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开发和验证毛细管区带电泳法简单测定尿液代谢产物(草酸盐、柠檬酸盐、尿酸和肌酐)。

Development and validation of a simple determination of urine metabolites (oxalate, citrate, uric acid and creatinine) by capillary zone electrophoresis.

机构信息

Centre Grup de Tècniques de Separació en Química (GTS), Química Analítica, Departament de Química, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain.

出版信息

Talanta. 2010 Apr 15;81(1-2):392-7. doi: 10.1016/j.talanta.2009.12.014. Epub 2009 Dec 21.

Abstract

Oxalate, citrate, uric acid and creatinine are important urine markers for the evaluation and treatment of urolithiasic patients. They have been traditionally analysed by enzymatic and chromatographic techniques which present practical drawbacks, mainly in the sample pre-treatment step. The purpose of this study was to evaluate those markers in urine samples, by an easy multi-analyte assay using capillary zone electrophoresis. The four urine metabolites were determined, at 25 degrees C, by using a 50 cm x 75 microm capillary in 50 mmol l(-1) phosphate buffer (pH 6.5), at constant voltage of -30 kV and UV detection at 195 nm (for oxalate and citrate) or 30 kV and 234 nm (for creatinine and uric acid). The sample pre-treatment was minimum, 5- and 20-fold dilution of the urine sample and acidification to pH 3-4. Validation parameters (linear range, sensitivity, accuracy, precision and detection limits) were statistically comparable to those obtained with the official methods normally used in the clinical practice. The effect of freezing as a conservation method of urine samples is also discussed in terms of recoveries of the analytes. The analytical method developed is highly useful as a diagnostic tool for detecting metabolic renal disorders due to its simplicity, time consuming, easy automation, cost efficiency and analytical effectiveness, accomplishing with the clinical requirements.

摘要

草酸盐、柠檬酸盐、尿酸和肌酸是评估和治疗尿路结石患者的重要尿液标志物。它们传统上通过酶和色谱技术进行分析,这些技术在样品预处理步骤中存在实际的缺点。本研究的目的是通过使用毛细管区带电泳的简单多分析物测定法来评估尿液样本中的这些标志物。在 25°C 下,使用 50cm×75μm 的毛细管,在 50mmol/L 磷酸盐缓冲液(pH6.5)中,在-30kV 的恒电压和 195nm(用于草酸盐和柠檬酸盐)或 30kV 和 234nm(用于肌酸和尿酸)处进行检测,测定四种尿液代谢物。样品预处理最少,尿液样品稀释 5-20 倍并酸化至 pH3-4。验证参数(线性范围、灵敏度、准确性、精密度和检测限)与临床实践中通常使用的官方方法获得的参数在统计学上具有可比性。还讨论了冷冻作为尿液样品保存方法的效果,以及对分析物回收率的影响。由于其简单、耗时、易于自动化、成本效益和分析效果,所开发的分析方法可作为检测代谢性肾脏疾病的诊断工具,符合临床要求。

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