商用25-基维生素D套件的问题和开发简单,稳固和更快的HPLC方法
Syed Muhammad Saad1, Abdul Rafiq Khan1,2, Khalid Mohammed Khan3
1Department of Chemistry, University of Karachi, University Road, Karachi-75270, Pakistan.
Journal of chromatographic science
|June 24, 2024
概括
一种新的,快速的高性能液态染色学 (HPLC) 方法简化了维生素D测试. 这种强大的测定准确地测量了血清中的25-基维生素D2和D3,克服了当前商业套件的局限性.
科学领域:
- 临床化学 临床化学
- 分析化学 分析化学
- 内分泌学 在内分泌学.
背景情况:
- 维生素D对骨健康和矿物质代谢至关重要.
- 现有的维生素D试验 (HPLC,免疫试验) 存在诸如复杂性,大量样本和干扰等挑战.
- 限制包括试剂的使用,长的提取时间,蒸发步骤和抗体交叉反应.
研究的目的:
- 开发一种简化,快速和强大的高性能液体染色学 (HPLC) 方法来确定25-基维生素D2 (25-OHD2) 和25-基维生素D3 (25-OHD3).
- 解决与当前商业维生素D测试套件和已发表的测试方法相关的常见问题.
主要方法:
- 作为内部标准,将血清样本 (300μl) 与含有劳拉的乙二结合.
- 使用C18弹进行提取,然后用70%的甲醇清洗,然后用乙醇/异醇混合物化.
- 通过HPLC-UV系统进行分析,10分钟运行时间.
主要成果:
- 该方法证明了25-OHD2和25-OHD3在10-750nmol/L范围内的线性.
- 对于维生素D的两种形式,测试内部和测试间的精度都低于10%.
- 与LC-MS/MS.相比,观察到较高的相关系数 (R2 > 0.94),与LC-MS/MS.相比.
结论:
- 开发的HPLC方法是简单的,快速的和强大的,可以同时确定25-OHD2和25-OHD3.
- 它有效地克服了现有的商业套件和测试的常见限制.
- 这种方法适用于大量实验室进行常规维生素D分析.
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