New approach for the clinical monitoring of 25-hydroxyvitamin D sub(3) and 25-hydroxyvitamin D sub(2) by ultra high performance liquid chromatography with MS/MS based on the standard reference material 972

• Published in: Journal of separation science Vol. 36; no. 23; pp. 3702 - 3708
• Main Authors: Plisek, Jiri, Krcmova, Lenka Kujovska, Aufartova, Jana, Morales, Tanausu V, Esponda, Sarah M, Oros, Roman, Kasalova, Eva, Santana-Rodriguez, Jose J, Sobotka, Lubos, Solich, Petr, Solichova, Dagmar
• Format: Journal Article
• Language: English
• Published: 01.12.2013
• Subjects: Derivatives; Disorders; Heart diseases; High performance liquid chromatography; Linearity; Methodology; Patients; Vitamins
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.201300553

Biomarkers, 25-hydroxyvitamin D sub(3) and 25-hydroxyvitamin D sub(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 super(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 sub(3) and 25-hydroxyvitamin D sub(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.