LC-MS/MS analysis of vitamin D3 metabolites in human serum using a salting-out based liquid-liquid extraction and DAPTAD derivatization
| Autor: | Doug A. Brooks, Amanda Bergamin, Shane M. Hickey, Deepti K Sharma, Michael D. Wiese, Peter D. O’Loughlin, Paul H. Anderson, Ali K. Alshabrawy |
|---|---|
| Přispěvatelé: | Alshabrawy, Ali K, Bergamin, Amanda, Sharma, Deepti K, Hickey, Shane M, Brooks, Doug A, O'Loughlin, Peter, Wiese, Michael D, Anderson, Paul H |
| Rok vydání: | 2021 |
| Předmět: |
Vitamin
liquid-liquid extraction Formic acid Clinical Biochemistry 030226 pharmacology & pharmacy 01 natural sciences Biochemistry High-performance liquid chromatography Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Liquid–liquid extraction Protein precipitation LC-MS/MS Acetonitrile Derivatization Chromatography 010401 analytical chemistry DAPTAD derivatization Cell Biology General Medicine vitamin D3 metabolites 0104 chemical sciences chemistry salting-out Salting out |
| Zdroj: | Journal of Chromatography B. 1173:122654 |
| ISSN: | 1570-0232 |
| DOI: | 10.1016/j.jchromb.2021.122654 |
| Popis: | LC-MS/MS has recently emerged as the best-practice for simultaneous analysis of vitamin D metabolites. We have developed and validated an LC-MS/MS method for simultaneous quantification of 25(OH)D3, 24,25(OH)2D3, and 3-epi-25(OH)D3 in human serum. These three metabolites were extracted from 50 μL of serum by acetonitrile protein precipitation followed by salting-out of acetonitrile. DAPTAD (4-(4′-dimethylaminophenyl)-1,2,4-triazoline-3,5-dione) was used to derivatize the extracted metabolites and their deuterated isotope internal standards. Chromatographic separation was achieved on a UPLC C18 column (Waters® ACQUITY 100 × 2.1 mm, 1.7 µm) utilizing 0.1% formic acid and acetonitrile as mobile phases. Limits of quantification were 1 ng/mL for 25(OH)D3 and 0.1 ng/mL for 24,25(OH)D3 and 3-epi-25(OH)D3. In-house and external Vitamin D External Quality Assessment Scheme (DEQAS) quality control sample analysis revealed satisfactory method accuracy. Within-analytical batch and between analytical batches precision were |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect