Quantitative Analysis of 25-OH Vitamin D Using Supported Liquid Extraction and Liquid Chromatography - Mass Spectrometry
Eduard Rogatsky*, Shaynah Browne, Min Cai, Harsha Jayatillake and Daniel Stein
Biomarker Analytical Resource Core, Einstein-Montefiore Institute for Clinical and Translational Research. Albert Einstein College of Medicine, USA
- *Corresponding Author:
- Eduard Rogatsky
Biomarker Analytical Resource Core
Einstein-Montefiore Institute for Clinical and Translational Research
Albert Einstein College of Medicine, USA
Received date: March 27, 2014; Accepted date: May 20, 2014; Published date: May 25, 2014
Citation: Rogatsky E, Browne S, Cai M, Jayatillake H, Stein D (2014) Quantitative Analysis of 25-OH Vitamin D Using Supported Liquid Extraction and Liquid Chromatography - Mass Spectrometry. J Chromatograph Separat Techniq 5:224. doi:10.4172/2157-7064.1000224
Copyright: © 2014 Rogatsky E, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
We report a low sample volume LC/MS method for 25OH Vitamin D analysis. The method requires only 40 μl serum, is fully automatable, fast and sensitive. The method was successfully implemented for clinical analysis of infants for vitamin D deficiency. The 6 min method has a sensitivity range from 2 to 100 ng/ml. A 3 min method was validated from 5 to 100 ng/ml range. Fast analysis, low sample volume and high sensitivity were achieved by combination of supported liquid extraction sample preparation, UHPLC chromatography and fused-core chromatographic column for separation, and highly sensitive mass spectrometry for detection. We emphasize the importance of sample preparation quality for rugged LC/MS analysis. Using SLE-based sample preparation we successfully used only 40 μl of serum while achieving a LLOQ of 2 ng/ml. We found from assayed samples, that 3-12 month old infants were not Vitamin D deficient, compared to adults. The average level of 25(OH)D3 in infants was 57.3 compared to 38.0 ng/ml for adults in a Bronx, NY patient population.