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毛细管电泳法测定鸡胚血浆和尿囊液中的尿酸

Determination of uric acid in plasma and allantoic fluid of chicken embryos by capillary electrophoresis.

作者信息

Matejcková Jana, Tůma Petr, Samcová Eva, Zemanová Zdenka

机构信息

Department of Analytical Chemistry, Charles University, Ruská 87, 100 00 Prague 10, Czech Republic.

出版信息

J Sep Sci. 2007 Aug;30(12):1947-52. doi: 10.1002/jssc.200700046.

Abstract

Capillary electrophoresis with diode array detection (DAD) was used to determine uric acid (UA) in chicken plasma and the allantoic fluid of chicken embryos. Complete separation of uric and ascorbic acids was attained in less than 10 min in the optimized BGE containing 60 mM MES + 30 mM Tris + 0.001% (w/v) polybrene (pH 6.1). The limit of UA detection (0.2 mg/L) was found to be low enough for sensitive analysis of native plasma and allantoic fluid samples. Range of linearity (1-200 mg/L), repeatability for peak area (CV <4.1%) and migration time (CV < 2.5%), as well as recovery of UA from biological samples (97-100%), were found to be satisfactory. The method was applied to detect the elevated UA concentrations (hyperuricemia) in chicken embryos with induced unilateral renal agenesis. CE/DAD analysis of the chicken plasma can be carried out with a relatively small volume of samples (1 microL).

摘要

采用二极管阵列检测(DAD)的毛细管电泳法测定鸡血浆和鸡胚尿囊液中的尿酸(UA)。在含有60 mM MES + 30 mM Tris + 0.001%(w/v)聚乙烯亚胺(pH 6.1)的优化背景电解质(BGE)中,10分钟内即可实现尿酸和抗坏血酸的完全分离。发现尿酸检测限(0.2 mg/L)足够低,可用于天然血浆和尿囊液样品的灵敏分析。线性范围(1 - 200 mg/L)、峰面积重复性(CV < 4.1%)和迁移时间重复性(CV < 2.5%)以及生物样品中尿酸的回收率(97 - 100%)均令人满意。该方法用于检测单侧肾发育不全诱导的鸡胚中升高的尿酸浓度(高尿酸血症)。鸡血浆的CE/DAD分析可使用相对少量的样品(1微升)进行。

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