Identification of conjugation positions of urinary glucuronidated vitamin D 3 metabolites by LC/ESI‐MS/MS after conversion to MS/MS‐fragmentable derivatives
Identification of conjugation positions of urinary glucuronidated vitamin D 3 metabolites by LC/ESI‐MS/MS after conversion to MS/MS‐fragmentable derivatives
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转化为 MS/MS 可裂解衍生物后，通过 LC/ESI-MS/MS 鉴定尿葡萄糖醛酸化维生素 D 3 代谢物的结合位置
DOI：
10.1002/bmc.4538
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发表时间：
2019
影响因子：
1.8
通讯作者：
Higashi Tatsuya
中科院分区：
文献类型：
--
作者：
Yoshimura Yusuke;Hibi Ryuichiro;Nakata Akiho;Togashi Moeka;Ogawa Shoujiro;Ishige Takayuki;Satoh Mamoru;Nomura Fumio;Higashi Tatsuya
开发了一种基于液相色谱/电喷雾电离-串联质谱的方法，用于鉴定人尿液中25-羟基维生素D3[25（OH）D3]和24，25-二羟基维生素D3[24，25（OH）2D 3]的单葡糖苷酸结合位置。该方法采用4-（4-二甲氨基苯基）-1，2，4-三唑啉-3，5-二酮衍生化，将葡萄糖醛酸苷转化为可裂解的衍生物，这为MS/MS过程中识别结合位置提供了有用的产物离子。衍生化还提高了尿液样本分析的检测灵敏度和特异性。在优化的LC条件下，位置异构体单葡糖苷酸、25（OH）D3 - 3-和-25-葡糖苷酸或24，25（OH）2D 3 - 3-、-24-和-25-葡糖苷酸彼此完全分离。使用该方法，成功确定了葡萄糖醛酸化25（OH）D3和24，25（OH）2D 3的结合位置分别为C3和C24位置。与25（OH）D3不同，24，25（OH）2D 3不存在3-葡糖苷酸，因此我们发现24，25（OH）2D 3的C24-羟基发生选择性葡糖苷酸化。
A liquid chromatography/electrospray ionization–tandem mass spectrometry‐based method was developed for the identification of the conjugation positions of the monoglucuronides of 25‐hydroxyvitamin D3[25(OH)D3] and 24,25‐dihydroxyvitamin D3[24,25(OH)2D3] in human urine. The method employed derivatization with 4‐(4‐dimethylaminophenyl)‐1,2,4‐triazoline‐3,5‐dione to convert the glucuronides into fragmentable derivatives, which provided useful product ions for identifying the conjugation positions during the MS/MS. The derivatization also enhanced the assay sensitivity and specificity for urine sample analysis. The positional isomeric monoglucuronides, 25(OH)D3‐3‐ and ‐25‐glucuronides, or 24,25(OH)2D3‐3‐, ‐24‐ and ‐25‐glucuronides, were completely separated from each other under the optimized LC conditions. Using this method, the conjugation positions were successfully determined to be the C3 and C24 positions for the glucuronidated 25(OH)D3and 24,25(OH)2D3, respectively. The 3‐glucuronide was not present for 24,25(OH)2D3, unlike 25(OH)D3, thus we found that selective glucuronidation occurs at the C24‐hydroxy group for 24,25(OH)2D3.