[Liquid chromatographic determination of free thiamine and its esters in whole blood].

Dakar medical Pub Date : 2006-01-01

M Ake, Y N Soko, K A Malan

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Abstract

Introduction: To appreciate vitamin B1 status in normal and pathophysiological states in local ivorian populations, a high-performance liquid chromatographic method for the simultaneous determination of thiamine and its phosphate esters: thiamine monophosphate and thiamine diphsophate or cocarboxylase in whole blood has been developed.

Methods: The method involves extraction with diethylether, followed by pre-column alkaline derivatization by bromide cyanogen. The extract was analysed by isocratic reversed-phase isocratic high performance liquid chromatography with fluorescent detection. System suitability tests were applied to assess the method's continuing suitability for use. The main tests of method validation: linearity, precision, accuracy and sensibility were applied to the analytical procedure.

Results: Thiamine and its esters were eluted within 10 minutes with a good resolution. System suitability tests showed that the chromatographic system was suitable for continuing use. Results of validation tests show the reliability of the method : linearity domain, satisfying precision and accuracy, detection limits between 0.067 pg/l et 0.92 pg/l (0.160 fmol/l et 1.822 fmol/l).

Conclusion: The proposed method is suitable for determination of thiamine and its phosphate esters in whole blood.

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全血中游离硫胺素及其酯的液相色谱测定。

为了了解科特迪瓦当地人群在正常和病理生理状态下维生素B1的状态，建立了一种高效液相色谱法同时测定全血中硫胺素及其磷酸酯:单磷酸硫胺素和二磷酸硫胺素或辅酶羧化酶。方法:二乙醚萃取，溴化氰柱前碱衍生。采用荧光检测等容反相等容高效液相色谱法对提取物进行分析。系统适用性测试用于评估该方法的持续适用性。方法验证的主要试验:线性、精密度、准确度和灵敏度。结果:硫胺素及其酯类在10分钟内洗脱，洗脱效果良好。系统适用性试验表明，该色谱系统适合继续使用。验证结果表明，该方法在线性范围内，精密度和准确度均较好，检出限为0.067 pg/l ~ 0.92 pg/l (0.160 fmol/l ~ 1.822 fmol/l)。结论:该方法适用于全血中硫胺素及其磷酸酯的测定。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Dakar medical

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