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New approach for the clinical monitoring of 25‐hydroxyvitamin D 3 and 25‐hydroxyvitamin D 2 by ultra high performance liquid chromatography with MS/MS based on the standard reference material 972
Author(s) -
Plíšek Jiří,
Krčmová Lenka Kujovská,
Aufartová Jana,
Morales Tanausú V.,
Esponda Sarah M.,
Oros Roman,
Kasalová Eva,
SantanaRodriguez Jose J.,
Sobotka Luboš,
Solich Petr,
Solichová Dagmar
Publication year - 2013
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201300553
Subject(s) - vitamin d and neurology , high performance liquid chromatography , osteoporosis , chromatography , vitamin d deficiency , chemistry , medicine
Biomarkers, 25‐hydroxyvitamin D 3 and 25‐hydroxyvitamin D 2 , are important indicators of the vitamin D general status and are monitored in several pathophysiological disorders, such as osteoporosis, diabetes, heart disease, etc. A novel ultra‐HPLC with MS/MS methodology for the analysis of 25‐hydroxyvitamin D derivatives coupled with a very simple and highly rapid sample preparation step was developed. Analytical parameters obtained showed linearity ( R 2 ) above 0.999 for both vitamins with accuracies between 95.8 and 102%. The LODs were as low as 0.22 and 0.67 nmol/L for 25‐hydroxyvitamin D 3 and 25‐hydroxyvitamin D 2 , respectively. Intra‐assay precision (%RSD) was lower than 4.5%, and inter‐assay precision (%RSD) was lower than 6.5%. The feasibility of the developed methodology to be applied in clinical routine analysis has been proved by its application in blood samples from non‐agenarian patients, patients with familial hypercholesterolemia and patients suffering from age‐related macular degeneration.