67804-03-9Relevant academic research and scientific papers
Regioselective hydroxylation in the production of 25-hydroxyvitamin D by Coprinopsis cinerea peroxygenase
Babot, Esteban D.,Del R??o, Jos?? C.,Kalum, Lisbeth,Mart??nez, Angel T.,Guti??rrez, Ana
, p. 283 - 290 (2015)
Monohydroxylated metabolites of vitamin D3 (cholecalciferol) and vitamin D2 (ergocalciferol), generically known as 25-hydroxycalciferol, are better for several diseases, and other applications, than vitamin D (calciferol). This work describes a novel biotechnological approach for the preparation of 25-hydroxycalciferols, starting from readily available cholecalciferol and ergocalciferol. This approach enables the regioselective (100%) hydroxylation of these compounds (at the C-25 position) under mild and environmentally friendly conditions by using a peroxidase from the fungus Coprinopsis cinerea (gene model CC1G-08427T0 from the sequenced genome), which catalyzes monooxygenation with H2O2 as the only co-substrate (peroxygenase). Hydroxylation of cholecalciferol and ergocalciferol is a true peroxygenation, as demonstrated by incorporation of 18O from H218O2 into the products. The peroxygenase has additional advantages related to its recombinant nature, enabling enzyme engineering and low-cost overexpression in an industrial host. Therefore, the peroxygenase is a promising biocatalyst for the production of vitamin D active metabolites.
Mass fragmentographic assay for 25-hydroxyvitamin D in plasma without derivatizatfon: Enhanced sensitivity for metabolites of vitamins D2 and D3 after pre-column dehydration
Coldwell,Trafford,Makin
, p. 348 - 356 (2007/10/02)
A mass fragmentographic method for the measurement in plasma of underivatized, dehydrated 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 was developed. Quantitative dehydration of vitamin D metabolites was achieved prior to gas chromatography using a new high-temperature injection system and a small precolumn packed with aluminium powder in the injection port, followed by mass fragmentography on an inexpensive bench-top mass spectrometer. Plasma (2 ml) was incubated with hexadeuteriated 25-hydroxyvitamin D3 prior to acetonitrile extraction and purification using cartridges pre-packed with microparticulate silica using reversed- and normal-phase solvent systems. After purification, capillary gas chromatography mass spectrometry was carried out following dehydration of the secosteroids on the aluminium powder pre-column at 400°C. High-intensity dehydrated molecular ions were produced which were used for selected ion monitoring. The assay sensitivity for 25-hydroxyvitamin D was approximately 1 ng ml-1. The intra-assay variation was less than 7% and the recovery of added standard was quantitative. It is suggested that this method may be used to provide target values for much needed quality control schemes to monitor assays routinely used in the estimation of 25-hydroxyvitamin D. The behaviour of a number of other hydroxylated vitamin D metabolites,when injected without derivatization on to a gas chromatographic column, was also investigated.
