Rapid liquid chromatography-tandem mass spectrometry method for determination of 24,25(OH)(2)D and 25OHD with efficient separation of 3-epi analogs
Rapid liquid chromatography-tandem mass spectrometry method for determination of 24,25(OH)(2)D and 25OHD with efficient separation of 3-epi analogs
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快速液相色谱-串联质谱法测定 24,25(OH)(2)D 和 25OHD 并有效分离 3-epi 类似物
DOI：
10.1016/j.jsbmb.2018.11.012
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发表时间：
2019
影响因子：
4.1
通讯作者：
Qiu Ling
中科院分区：
文献类型：
--
作者：
Yu Songlin;Zhou Weiyan;Wang Danchen;Yin Yicong;Cheng Qian;Xie Shaowei;Sun D;an;Li Honglei;Cheng Xinqi;Qiu Ling
本研究建立并验证了一种基于液相色谱-串联质谱（LC-MS/MS）的快速方法，无需衍生化步骤，可同时测量24，25（OH）2D 2、24，25（OH）2D 3、25 OHD 2和25 OHD 3，同时有效分离3-epi类似物。通过沉淀和液-液萃取制备样品。根据临床和实验室标准协会指南，对该方法的线性、精密度、准确度、回收率和基质效应进行了全面评价。此外，还对38名健康志愿者血清中的四种维生素D代谢物进行了评价。总分析时间为8.0 min，3-epi 24，25（OH）2D 3和3-epi 25 OHD 3有效分离，不受异构体如23，25（OH）2D 3或1，25（OH）2D 2，1，25（OH）2D 3的干扰。25 OHD 3、25 OHD 2、24，25（OH）2D 3和24，25（OH）2D 2的批内变异系数（CV）分别为4.07%-6.55%、4.26%-7.84%、2.46%-7.21%和4.90%-6.87%，总CV为4.29%-6.64%。6.14%~ 7.84%、4.33%~ 7.21%、5.82%~ 9.90%。25 OHD 3和25 OHD 2的定量限为0.625 ng/mL，24，25（OH）2D 3和24，25（OH）2D 2的定量限为0.5 ng/mL。LC-MS/MS方法与SRM 972 a认证结果相比，25 OHD 3、25 OHD 2和24，25（OH）2D 3的相对偏倚分别为-2.21%至1.01%、3.38%至6.73%和-7.72%至3.9%。志愿者中25 OHD、24，25（OH）2D和25 OHD/24，25（OH）2D的平均值±SD分别为13.5±4.4 ng/mL（范围：7.6-27.5 ng/mL）、0.84±0.42 ng/mL（范围：0.26-2.1 ng/mL）和18±7（范围：8-37）。因此，开发了一种简单，精确的LC-MS/MS方法，用于适当保留和分离维生素D代谢物及其表观类似物。
This study establishes and validates a rapid method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) without derivatization steps to simultaneously measure of 24,25(OH)2D2, 24,25(OH)2D3, 25OHD2, and 25OHD3, while efficiently separating the 3-epi analogs. Samples were prepared by precipitation and liquid-liquid extraction. The linearity, precision, accuracy, recovery and matrix effect of the method were thoroughly evaluated according to the Clinical & Laboratory Standards Institute guidelines. Additionally, the four vitamin D metabolites in the serum of 38 apparently healthy Chinese volunteers were evaluated. The total analysis time was 8.0 min, with efficient separation of 3-epi 24,25(OH)2D3and 3-epi 25OHD3, without interference from isomers such as 23,25(OH)2D3or 1,25(OH)2D2, 1,25(OH)2D3. Good reproducibility was obtained for all four metabolites with within-run coefficient variations (CVs) of 4.07%–6.55%, 4.26%–7.84%, 2.46%–7.21%, and 4.90%–6.87% for 25OHD3, 25OHD2, 24,25(OH)2D3, and 24,25(OH)2D2, respectively, and the total CVs were 4.29%–6.64%, 6.14%–7.84%, 4.33%–7.21%, 5.82%–9.90%, respectively. The limit of quantification was 0.625 ng/mL for 25OHD3and 25OHD2, and 0.5 ng/mL for 24,25(OH)2D3and 24,25(OH)2D2. The relative bias of the LC-MS/MS method compared to the certified results of SRM 972a for 25OHD3, 25OHD2and 24,25(OH)2D3was -2.21% to 1.01%, 3.38% to 6.73%, and -7.72% to -3.9%, respectively. The mean±SD values for 25OHD, 24,25(OH)2D and 25OHD/24,25(OH)2D in the volunteers were 13.5±4.4 ng/mL(range:7.6–27.5 ng/mL), 0.84±0.42 ng/mL (range:0.26–2.1 ng/mL), and 18±7(range:8–37), respectively. Thus, a simple, precise LC-MS/MS method for appropriate retention and separation of vitamin D metabolites and their epi analogs was developed.