Abstract The coupling of high performance liquid chromatography (HPLC) with post column derivatisation (HPLC-PCD) will increase the sensitivity of an instrument to the target molecule, improve specificity by selective derivatisation to remove potential interferences from the matrix, and be cost-effective; all these factors may well prove that PCD is an alternative to existing techniques for routine analysis. Calcidiol (25(OH)D) concentrations are used in the determination of vitamin D deficiency in humans, and there is growing interest and demand in its analysis with some controversy surrounding the sensitivity and specificity of its detection. 25(OH)D analysis is currently performed using either competitive binding assays or HPLC analysis, coupled with detection by a variety of means, such as, UV–Vis absorption, electrochemical or mass spectroscopy (MS) detection. HPLC coupled with MS (HPLC-MS) detection is the gold standard for 25(OH)D analysis due to its sensitivity and selectivity compared to all other techniques, however, its utility is limited by its high cost. This article reviews the current status of HPLC based techniques in the detection of 25(OH)D (and its metabolites), and further explores the use of HPLC-PCD.

中文翻译：

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液相色谱分离的未来应包括柱后衍生：基于维生素 D 分析视角的讨论观点

摘要 高效液相色谱 (HPLC) 与柱后衍生化 (HPLC-PCD) 的结合将提高仪器对目标分子的灵敏度，通过选择性衍生化去除基质中的潜在干扰来提高特异性，并且具有成本效益; 所有这些因素都可以很好地证明 PCD 是现有常规分析技术的替代品。Calcidiol (25(OH)D) 浓度用于确定人类维生素 D 缺乏症，并且对其分析的兴趣和需求越来越大，但围绕其检测的灵敏度和特异性存在一些争议。25(OH)D 分析目前使用竞争性结合分析或 HPLC 分析进行，并结合多种方法进行检测，例如 UV-Vis 吸收、电化学或质谱 (MS) 检测。HPLC 联用 MS (HPLC-MS) 检测是 25(OH)D 分析的黄金标准，因为与所有其他技术相比，它具有灵敏度和选择性，但其实用性受到其高成本的限制。本文回顾了基于 HPLC 技术检测 25(OH)D（及其代谢物）的现状，并进一步探讨了 HPLC-PCD 的使用。