Determination of 1,25-dihydroxyvitamin D2 and 1,25-dihydroxyvitamin D3 in human serum using liquid chromatography with tandem mass spectrometry
Determination of 1,25-dihydroxyvitamin D2 and 1,25-dihydroxyvitamin D3 in human serum using liquid chromatography with tandem mass spectrometry
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DOI：
10.1016/j.jchromb.2016.04.034
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发表时间：
2016-08-01
影响因子：
3
通讯作者：
Qiu, Ling
中科院分区：
文献类型：
--
作者：
Fang, Huiling;Yu, Songlin;Qiu, Ling
Vitamin D plays important roles in skeletal metabolism and many other diseases, including chronic renal failure, hypoparathyroidism, sarcoidosis and rickets. 1 alpha,25-dihydroxy vitamin D (1 alpha,25(OH)(2)D), the active form of vitamin D, exhibits an extremely low serum concentration, which makes its quantification very challenging. High performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) is considered to be the "gold standard" for the determination of these chemicals, which are found in low concentrations in the serum, but conventionally, it needs tedious sample pretreatment procedures, such as solid phase extraction and derivatization. Herein, we describe a simple and rapid HPLC-MS/MS method for the simultaneous quantification of 1 alpha,25-dihydroxy vitamin D-3 (1 alpha,25(OH)(2)D-3) and 1 alpha,25-dihydroxy vitamin D-2 (1 alpha,25(OH)(2)D-2). The analytes were extracted from the matrix by liquid-liquid extraction, centrifuged to dryness and reconstituted with 75% methanol. Lithium acetate was employed to improve the ionization efficiency of 1 alpha,25(OH)(2)D. The assay was sensitive with a low limit of quantitation of 10.0 pg/mL for both 1 alpha,25(OH)(2)D-3 and 1 alpha,25(OH)(2)D-2 using a 0.5 mL sample aliquot. Linearity was obtained over the range of 10.0 pg/mL to 500 pg/mL. Both the inter-assay and intra-assay precisions were
