LC/MS/MS Quantitative Analysis Of Vitamin D: Hidden Analytical Errors Overlooked By Validation | 10239
ISSN: 2155-9872

Journal of Analytical & Bioanalytical Techniques
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LC/MS/MS quantitative analysis of vitamin D: Hidden analytical errors overlooked by validation

4th International Conference and Exhibition on Analytical & Bioanalytical Techniques

Eduard Rogatsky

Accepted Abstracts: J Anal Bioanal Tech

DOI: 10.4172/2155-9872.S1.015

Vitamin D deficiency is a widespread clinical problem and has been associated with many adverse health outcomes. Analysis of Vitamin D2 (ergocalciferol) and D3 (cholecalciferol) and their major metabolites 25(OH)D2 and 25(OH)D3 has become a high priority topic in clinical analysis. Since water loss is a favored process, in-source self dehydration of vitamin D can decrease S/N and such an undesirable process was tightly controlled. In-source water loss was found to be insignificant (~8%) within the assay linear range, using a methanol/water mobile phase under our experimental conditions. In contrast, if acetonitrile/ water was used, severe in-source water loss was observed and ions m/z 395.2 and m/z 383.2 resulting from analyte in-source dehydration became predominant. When acetonitrile was used as mobile phase, and if only the transitions m/z 395.2?→?377.2 for 25(OH)D2 and m/z 383.2?→?377.2 for 25(OH)D3 were used for quantitation, the assay standard curve was still acceptably linear; however the area ratio between analyte and its in-source water loss product was non-linear at different analyte concentrations. The concentration-dependent water loss varied by 20%, resulting in a significant quantitation error in this provisionally valid assay. For an acetonitrile-based mobile phase, both transitions (for analyte and for its in-source dehydrated product) should be summed to maintain true full range assay linearity.
Eduard Rogatsky completed his M.Sc in physical chemistry from Belarus State University, Ph.D. in Bioanalytical chemistry from Bar-Ilan University (Israel) in 1999, and postdoctoral studies at Albert Einstein College of Medicine, NY. He joined the faculty there in 2001, and is currently a Senior Associate Scientist and Director of Mass Spectrometry in the Biomarker Analytical Resource Core Laboratory, Einstein-Montefiore Institute for Clinical and Translational Research, Bronx, NY, USA