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[使用西格玛试剂盒对尿草酸估计方法的一种改进]

[A modification of the estimation of urinary oxalate using a Sigma kit].

作者信息

Ebisuno S, Ohkawa T

机构信息

Department of Urology, Wakayama Medical College.

出版信息

Hinyokika Kiyo. 1988 Feb;34(2):225-30.

PMID:3376811
Abstract

The Sigma kit for estimating urinary oxalate is an enzymatic procedure. However, some errors were encountered using the standard assay system of the kit. Firstly, an overestimate of oxalate may arise from the oxidation of ascorbate during the alkaline wash stage of the extraction of oxalate from urine. Secondly, an underestimate of oxalate may occur because of incomplete extraction of oxalate. A modified assay system for measurement of urinary oxalate using the kit is reported. The following points were modified: urine was diluted two-fold with 0.2 M EDTA and and 0.2 M citrate buffer (pH 3.0), oxalate from urine was extracted with 0.06 N sodium hydroxide to prevent the overestimation by the oxidation of ascorbate, and a plate mixer and addition of a small magnet to the vial were used in the steps of both absorption and extraction of oxalate to keep the accuracy of the estimation. The linearity of standard curve, reproducibility and recovery rates of the modified method were studied, and good results were obtained (linearity; r = 0.999, CV of reproducibility; 5.3%, recovery rate; 101% (300 microM) and 103% (600 microM). A good correlation was seen between the modified Sigma method and high performance liquid chromatography (r = 0.991).

摘要

用于估算尿草酸的西格玛试剂盒是一种酶促方法。然而,使用该试剂盒的标准检测系统时遇到了一些误差。首先,在从尿液中提取草酸盐的碱性洗涤阶段,抗坏血酸的氧化可能导致草酸盐被高估。其次,由于草酸盐提取不完全,可能会出现草酸盐被低估的情况。本文报道了一种使用该试剂盒测量尿草酸的改良检测系统。以下几点进行了改进:用0.2M EDTA和0.2M柠檬酸盐缓冲液(pH 3.0)将尿液稀释两倍,用0.06N氢氧化钠提取尿液中的草酸盐,以防止因抗坏血酸氧化而导致的高估,并且在草酸盐的吸收和提取步骤中使用平板混合器并在小瓶中添加小磁铁以保持估算的准确性。研究了改良方法的标准曲线线性、重现性和回收率,结果良好(线性;r = 0.999,重现性CV;5.3%,回收率;101%(300 microM)和1%(600 microM)。改良的西格玛方法与高效液相色谱法之间具有良好的相关性(r = 0.991)。

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