BACKGROUND: Rapid, easy and reliable measurement of the major vitamin D metabolites is required in order to fulfill the needs of a clinical routine laboratory. To overcome these challenges, we have developed and validated a LC-MS/MS method for the quantification of 25-hydroxyvitamin D2 and D3, epi-25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3. METHODS: Sample preparation was based on precipitation and centrifugation of 100μL of patient serum, followed by injection into the LC-MS/MS system. Samples from Vitamin D Standardization Program (n=80) and patient samples (n=281) have been compared with a reference LC-MS/MS method. For the analytical validation NIST and Labquality quality control materials were used. RESULTS: Mean intra-assay and inter-assay imprecision were <6.0 and 6.4% and mean recoveries were within 95-104%. LOQ's were 0.5μg/L for 24,25(OH)2D3, 1.1μg/L for 25(OH)D3 and epi-25(OH)D3 and 1.7μg/L for 25(OH)D2. A 3% bias obtained between the proposed and the reference method satisfies Vitamin D Standardization Program recommendations. CONCLUSIONS: We present a rapid, easy, reliable and cost-effective method completely adequate for routine testing, which permits the measurement of the ratio of 24,25-dihydroxyvitamin D to 25-hydroxyvitamin D, Vitamin D Metabolite Ratio (VMR), in serum samples.
BACKGROUND: Rapid, easy and reliable measurement of the major vitamin D metabolites is required in order to fulfill the needs of a clinical routine laboratory. To overcome these challenges, we have developed and validated a LC-MS/MS method for the quantification of 25-hydroxyvitamin D2 and D3, epi-25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3. METHODS: Sample preparation was based on precipitation and centrifugation of 100μL of patient serum, followed by injection into the LC-MS/MS system. Samples from Vitamin D Standardization Program (n=80) and patient samples (n=281) have been compared with a reference LC-MS/MS method. For the analytical validation NIST and Labquality quality control materials were used. RESULTS: Mean intra-assay and inter-assay imprecision were <6.0 and 6.4% and mean recoveries were within 95-104%. LOQ's were 0.5μg/L for 24,25(OH)2D3, 1.1μg/L for 25(OH)D3 and epi-25(OH)D3 and 1.7μg/L for 25(OH)D2. A 3% bias obtained between the proposed and the reference method satisfies Vitamin D Standardization Program recommendations. CONCLUSIONS: We present a rapid, easy, reliable and cost-effective method completely adequate for routine testing, which permits the measurement of the ratio of 24,25-dihydroxyvitamin D to 25-hydroxyvitamin D, Vitamin D Metabolite Ratio (VMR), in serum samples.
Authors: Christopher T Sempos; Annemieke C Heijboer; Daniel D Bikle; Jens Bollerslev; Roger Bouillon; Patsy M Brannon; Hector F DeLuca; Glenville Jones; Craig F Munns; John P Bilezikian; Andrea Giustina; Neil Binkley Journal: Br J Clin Pharmacol Date: 2018-07-17 Impact factor: 4.335
Authors: Toni E Ziegler; Amita Kapoor; Neil C Binkley; Karen S Rice; Jeffrey Rogers; Clifford J Jolly; Jane E Phillips-Conroy Journal: Am J Primatol Date: 2018-12 Impact factor: 2.371
Authors: Stephen A Wise; Johanna E Camara; Christopher T Sempos; Pierre Lukas; Caroline Le Goff; Stephanie Peeters; Carolyn Q Burdette; Federica Nalin; Grace Hahm; Ramón A Durazo-Arvizu; Adam J Kuszak; Joyce Merkel; Étienne Cavalier Journal: J Steroid Biochem Mol Biol Date: 2021-05-16 Impact factor: 5.011
Authors: Konstantinos Makris; Harjit P Bhattoa; Etienne Cavalier; Karen Phinney; Christopher T Sempos; Candice Z Ulmer; Samuel D Vasikaran; Hubert Vesper; Annemieke C Heijboer Journal: Clin Chim Acta Date: 2021-03-10 Impact factor: 6.314
Authors: Niek F Dirks; Mariëtte T Ackermans; Paul Lips; Renate T de Jongh; Marc G Vervloet; Robert de Jonge; Annemieke C Heijboer Journal: Nutrients Date: 2018-04-13 Impact factor: 5.717