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一种快速且经过验证的 GC-MS/MS 方法,用于同时定量血清肌醇和 D-手性肌醇异构体。

A rapid and validated GC-MS/MS method for simultaneous quantification of serum Myo- and D-chiro-inositol isomers.

机构信息

Department of Laboratory Medicine, the Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, China; Department of Clinical Laboratory, Taizhou Central Hospital (Taizhou University Hospital), Taizhou, Zhejiang Province, 318000, China.

Physical and chemical laboratory of Hunan Provincial Center for Disease Control and Prevention, Changsha, Hunan 410005, China.

出版信息

J Chromatogr A. 2024 Sep 13;1732:465246. doi: 10.1016/j.chroma.2024.465246. Epub 2024 Aug 8.

Abstract

BACKGROUND

Myo-inositol (MI) and D-chiro-inositol (DCI) are two paramount isomers of inositol, both vital in glucose and steroid metabolism. Deficits in MI, DCI or MI/DCI ratio are expressly concerned with several pathological process, whereas MI and DCI lack practical measurement for human specimen.

METHODS

To quantify MI and DCI in serum samples simultaneously, a gas chromatography tandem mass spectrometry (GC-MS/MS) method was established. The process flow was optimized in ion source, derivative agent volume and reaction time. The performance characteristics were verified by commercial standards and clinical serums.

RESULTS

This method was confirmed to be sensitive (LOD ≤ 30 ng/mL of MI, ≤3 ng/mL of DCI) and reproducible (RSD < 6 % for repeated analyses). Quantitative determinations performed good linearity within the measurement range of 0.500-10.00 and 0.005-0.500 μg/mL for MI and DCI respectively (R > 0.999). The recoveries of MI and DCI were 97.11-99.35 % and 107.82-113.09 %, respectively. This method was successfully applied to 114 clinical specimens. No significant matrix effect was observed in serum samples under current conditions.

摘要

背景

肌醇(MI)和 D-手性肌醇(DCI)是肌醇的两种主要异构体,在葡萄糖和类固醇代谢中都至关重要。MI、DCI 或 MI/DCI 比值的不足与几种病理过程明显相关,而 MI 和 DCI 缺乏对人体标本的实际测量方法。

方法

为了同时定量血清样本中的 MI 和 DCI,建立了一种气相色谱串联质谱(GC-MS/MS)方法。在离子源、衍生剂体积和反应时间方面优化了工艺流程。通过商业标准品和临床血清验证了性能特征。

结果

该方法被证实具有灵敏度(MI 的 LOD≤30ng/mL,DCI 的 LOD≤3ng/mL)和重现性(重复分析的 RSD<6%)。MI 和 DCI 的定量测定在各自的测量范围内均具有良好的线性关系(MI:0.500-10.00μg/mL,R>0.999;DCI:0.005-0.500μg/mL,R>0.999)。MI 和 DCI 的回收率分别为 97.11%-99.35%和 107.82%-113.09%。该方法成功应用于 114 份临床标本。在当前条件下,血清样本中未观察到明显的基质效应。

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