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辅酶Q分析的技术层面:一种新型高效液相色谱 - 电化学检测法的验证

Technical Aspects of Coenzyme Q Analysis: Validation of a New HPLC-ED Method.

作者信息

Paredes-Fuentes Abraham J, Oliva Clara, Montero Raquel, Alcaide Patricia, Ruijter George J G, García-Villoria Judit, Ruiz-Sala Pedro, Artuch Rafael

机构信息

Division of Inborn Errors of Metabolism-IBC, Biochemistry and Molecular Genetics Department, Hospital Clínic de Barcelona, 08028 Barcelona, Spain.

Clinical Biochemistry Department, Institut de Recerca Sant Joan de Déu, Hospital Sant Joan de Déu, Esplugues de Llobregat, 08950 Barcelona, Spain.

出版信息

Antioxidants (Basel). 2022 Mar 10;11(3):528. doi: 10.3390/antiox11030528.

Abstract

The biochemical measurement of the CoQ status in different tissues can be performed using HPLC with electrochemical detection (ED). Because the production of the electrochemical cells used with the Coulochem series detectors was discontinued, we aimed to standardize a new HPLC-ED method with new equipment. We report all technical aspects, troubleshooting and its performance in different biological samples, including plasma, skeletal muscle homogenates, urine and cultured skin fibroblasts. Analytical variables (intra- and inter-assay precision, linearity, analytical measurement range, limit of quantification, limit of detection and accuracy) were validated in calibrators and plasma samples and displayed adequate results. The comparison of the results of a new ERNDIM external quality control (EQC) scheme for the plasma CoQ determination between HPLC-ED (Lab 1) and LC-MS/MS (Lab 2) methods shows that the results of the latter were slightly higher in most cases, although a good consistency was generally observed. In conclusion, the new method reported here showed a good analytical performance. The global quality of the EQC scheme results among different participants can be improved with the contribution of more laboratories.

摘要

不同组织中辅酶Q状态的生化测定可采用高效液相色谱法(HPLC)结合电化学检测(ED)进行。由于与库仑化学系列检测器配套使用的电化学池已停止生产,我们旨在使用新设备规范一种新的HPLC-ED方法。我们报告了该方法的所有技术细节、故障排除情况及其在不同生物样本(包括血浆、骨骼肌匀浆、尿液和培养的皮肤成纤维细胞)中的性能。在校准品和血浆样本中对分析变量(批内和批间精密度、线性、分析测量范围、定量限、检测限和准确度)进行了验证,结果显示良好。对血浆辅酶Q测定的新的欧洲神经肌肉疾病研究和诊断国际联盟(ERNDIM)外部质量控制(EQC)方案中,HPLC-ED方法(实验室1)和液相色谱-串联质谱法(LC-MS/MS,实验室2)的结果比较表明,尽管总体上观察到良好的一致性,但在大多数情况下,后者的结果略高。总之,本文报道的新方法具有良好的分析性能。随着更多实验室的参与,不同参与者之间EQC方案结果的整体质量有望得到提高。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/02ea/8944485/c7adc9cad6bf/antioxidants-11-00528-g001.jpg

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