A specific LC/ESI-MS/MS method for determination of 25-hydroxyvitamin D3 in neonatal dried blood spots containing a potential interfering metabolite, 3-epi-25-hydroxyvitamin D3
✍ Scribed by Tatsuya Higashi; Masahiro Suzuki; Junji Hanai; Shinsuke Inagaki; Jun Zhe Min; Kazutake Shimada; Toshimasa Toyo'oka
- Publisher
- John Wiley and Sons
- Year
- 2011
- Tongue
- English
- Weight
- 260 KB
- Volume
- 34
- Category
- Article
- ISSN
- 1615-9306
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✦ Synopsis
A specific LC/ESI-MS/MS method for determination of 25-hydroxyvitamin D 3 in neonatal dried blood spots containing a potential interfering metabolite, 3-epi-25hydroxyvitamin D 3 Vitamin D deficiency in an infant is associated with a wide range of adverse health outcomes in later life. A method for the quantification of 25-hydroxyvitamin D 3 [25(OH)D 3 , the best-established indicator of vitamin D status] in neonatal dried blood spots (DBSs) using LC/ESI-MS/MS has been developed and validated. The method employed two steps of derivatization, a Diels-Alder reaction with 4-phenyl-1,2,4-triazoline-3,5-dione followed by acetylation, to enhance the detectability of 25(OH)D 3 in ESI-MS/MS and to separate 25(OH)D 3 from 3-epi-25-hydroxyvitamin D 3 [3-epi-25(OH)D 3 ], a potent interfering metabolite. 25(OH)D 3 was extracted from two DBS punches (3 mm in diameter, equivalent to 5.3 mL of whole blood), purified using an Oasis HLB s cartridge, and subjected to derivatization prior to analysis with LC/ESI-MS/MS. 25-Hydroxyvitamin D 4 was used as the internal standard. This method was reproducible (intra-and interassay RSDs, o6.9%) and accurate (analytical recovery, 95.2-102.7%), and the LOQ was 3.0 ng/mL. The developed method enabled specific quantification of 25(OH)D 3 in neonatal DBSs and detection of vitamin D deficiency without interference from 3-epi-25(OH)D 3 .