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High throughput UHPLC-MS/MS method for the simultaneous quantification of six vitamin D metabolites: application for vitamin D determination in patients after liver or kidney transplantation
Analytical Methods ( IF 2.7 ) Pub Date : 2020-09-28 , DOI: 10.1039/d0ay01088j Ting Hu 1, 2, 3, 4 , Han Li 1, 2, 3, 4 , Hongchuan Liu 1, 2, 3, 4 , Ling Cong 1, 2, 3, 4 , Lihong Liu 1, 2, 3, 4 , Zhuoling An 1, 2, 3, 4
Analytical Methods ( IF 2.7 ) Pub Date : 2020-09-28 , DOI: 10.1039/d0ay01088j Ting Hu 1, 2, 3, 4 , Han Li 1, 2, 3, 4 , Hongchuan Liu 1, 2, 3, 4 , Ling Cong 1, 2, 3, 4 , Lihong Liu 1, 2, 3, 4 , Zhuoling An 1, 2, 3, 4
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Vitamin D metabolites are fat-soluble vitamins that regulate broad spectrum of physiological and pathological processes. Accurate and high-throughput methods for the detection of vitamin D metabolites are essential to elucidate body functions. In this study, a sensitive and high throughput ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) method was proposed for the accurate quantification of six vitamin D metabolites, including vitamin D2, vitamin D3, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, 1,25-dihydroxyvitamin D2, and 1,25-dihydroxyvitamin D3. Through the optimization of chromatographic and mass spectrometric conditions, only 20 μL serum or plasma could satisfy the quantification of six vitamin D metabolites. The limit of detection (LOD) was determined to be 0.02–0.05 pg mL−1. The validation method was carried out following the International Council for Harmonization (ICH) guidelines. All quantification performances, including linearity, accuracy, precision, extraction recovery and matrix effect, were investigated and were satisfactory for the accurate detection of vitamin D metabolites. A disease of the liver or kidney, the main organs for vitamin D metabolism, could lead to abnormal levels of vitamin D. Here, the established UHPLC-MS/MS method was further used for determination of vitamin D levels in plasma samples of patients after liver or kidney transplantation. Thirty-three liver transplant recipients (LTRs) and 63 kidney transplant recipients (KTRs) were included in this study. Vitamin D deficiency or insufficiency is common in KTRs and LTRs, with a prevalence of more than 99%.
中文翻译：
高通量UHPLC-MS / MS方法同时定量六种维生素D代谢物：在肝或肾移植后患者中用于维生素D测定的应用
维生素D代谢产物是脂溶性维生素，可调节广泛的生理和病理过程。准确，高通量的维生素D代谢物检测方法对于阐明人体功能至关重要。在这项研究中，敏感和高通量与串联质谱法提出了六个维生素d代谢物的精确定量（UHPLC-MS / MS）的方法，包括维生素d超高效液相色谱2，维生素d 3，25-羟基d 2，25-羟基维生素d 3，1,25-二羟基维生素d 2，和1,25-二羟基维生素d 3。通过优化色谱和质谱条件，只有20μL的血清或血浆可以满足6种维生素D代谢产物的定量要求。检测限（LOD）确定为0.02–0.05 pg mL -1。验证方法是按照国际协调理事会（ICH）指南进行的。研究了所有定量性能，包括线性，准确性，精密度，提取回收率和基质效应，这些结果对于准确检测维生素D代谢物是令人满意的。肝脏或肾脏疾病是维生素D代谢的主要器官，可能导致维生素D含量异常。在此，已建立的UHPLC-MS / MS方法进一步用于测定患者血浆后维生素D的含量。肝或肾移植。这项研究包括33位肝移植受者（LTR）和63位肾移植受者（KTR）。维生素D缺乏或不足在KTR和LTR中很常见，患病率超过99％。
更新日期：2020-11-12
中文翻译：
高通量UHPLC-MS / MS方法同时定量六种维生素D代谢物：在肝或肾移植后患者中用于维生素D测定的应用
维生素D代谢产物是脂溶性维生素，可调节广泛的生理和病理过程。准确，高通量的维生素D代谢物检测方法对于阐明人体功能至关重要。在这项研究中，敏感和高通量与串联质谱法提出了六个维生素d代谢物的精确定量（UHPLC-MS / MS）的方法，包括维生素d超高效液相色谱2，维生素d 3，25-羟基d 2，25-羟基维生素d 3，1,25-二羟基维生素d 2，和1,25-二羟基维生素d 3。通过优化色谱和质谱条件，只有20μL的血清或血浆可以满足6种维生素D代谢产物的定量要求。检测限（LOD）确定为0.02–0.05 pg mL -1。验证方法是按照国际协调理事会（ICH）指南进行的。研究了所有定量性能，包括线性，准确性，精密度，提取回收率和基质效应，这些结果对于准确检测维生素D代谢物是令人满意的。肝脏或肾脏疾病是维生素D代谢的主要器官，可能导致维生素D含量异常。在此，已建立的UHPLC-MS / MS方法进一步用于测定患者血浆后维生素D的含量。肝或肾移植。这项研究包括33位肝移植受者（LTR）和63位肾移植受者（KTR）。维生素D缺乏或不足在KTR和LTR中很常见，患病率超过99％。
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