采用内标法 HPLC 分析黄素腺嘌呤二核苷酸的自动化工作流程方法。

An automated workflow approach for the analysis of flavin adenine dinucleotide by HPLC with internal standard.

机构信息

NSW Health Pathology, Department of Chemical Pathology, Royal Prince Alfred Hospital, Camperdown, NSW, 2050, Australia; School of Medicine, University of Sydney, Camperdown, NSW, 2006, Australia.

NSW Health Pathology, Department of Chemical Pathology, Royal Prince Alfred Hospital, Camperdown, NSW, 2050, Australia.

出版信息

Anal Biochem. 2020 Apr 1;594:113616. doi: 10.1016/j.ab.2020.113616. Epub 2020 Feb 7.

DOI:10.1016/j.ab.2020.113616

PMID:32035844

Abstract

Flavin adenine dinucleotide (FAD) is an active coenzyme of vitamin B2 involved in oxidation and reduction reactions. In this study we have developed a fully automated method for the analysis of FAD by fluorescence detection. FAD is extracted from whole blood samples by trichloroacetic acid, with diethyl ribityl isoalloxazine used as an internal standard. Linearity for FAD was above 0.99 (r) up to a concentration range of 1000 nmol/L. Precision of the method (intra-day and inter-day) compared against commercial quality control material was below 8% (coefficient of variation) with recovery of FAD exceeding 90%. Accuracy of the protocol was compared against a previous cycle from an external quality assurance program (RCPAQAP) (n = 12) with satisfactory agreement. Overall, this method has increased laboratory workflow and reduced manual labour required in performing FAD analysis.

摘要

黄素腺嘌呤二核苷酸（FAD）是维生素 B2 的一种活性辅酶，参与氧化还原反应。在本研究中，我们开发了一种通过荧光检测分析 FAD 的全自动方法。FAD 从全血样本中用三氯乙酸提取，二乙基核糖异咯嗪用作内标。FAD 的线性范围超过 1000 nmol/L，线性度大于 0.99（r）。与商业质量控制材料相比，该方法的精密度（日内和日间）低于 8%（变异系数），FAD 的回收率超过 90%。与外部质量保证计划（RCPAQAP）的先前周期（n = 12）相比，该方案的准确性具有令人满意的一致性。总的来说，该方法提高了实验室的工作流程，并减少了进行 FAD 分析所需的人工劳动。