I'm going to extract vitamin D from mammalian cells and quantify using HPLC. Did anybody do such thing? I don't want to use ultrasounds because of variaty of compounds in cells which I don't need. I'm thinking about DMSO or propanol extraction.
HI,
I would suggest you read the followings
Overview of extraction methods for analysis of vitamin D and its metabolites in biological samples., Musteata ML1, Musteata FM., Bioanalysis. 2011 Sep;3(17):1987-2002. doi: 10.4155/bio.11.195.
Recent trends in the determination of vitamin D. Gomes FP1, Shaw PN, Whitfield K, Koorts P, Hewavitharana AK., Bioanalysis. 2013 Dec;5(24):3063-78. doi: 10.4155/bio.13.283.
The last method I optimized for NP-HPLC detection of Vit D in tablets was a simple adaptation of the European Pharmacopoeia (5th edition)
http://lib.njutcm.edu.cn/yaodian/ep/EP5.0/16_monographs/monographs_a-c/Cholecalciferol.pdf
The most common way to extract Vit D from biological samples is based on Chloroform/methanol 1st extraction and chloroform/water for phase separation.
THX a lot! I'll read it and try to do my experiment :)
I found another useful text: Metabolic profiling of major vitamin D metabolites using
Diels–Alder derivatization and ultra-performance liquid
chromatography–tandem mass spectrometry (Pavel A. Aronov & Laura M. Hall & Katja Dettmer & Charles B. Stephensen & Bruce D. Hammock)
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