Quantitation of 25-hydroxyvitamin D in dried blood spots by 2D LC-MS/MS without derivatization and correlation with serum in adult and pediatric studies

被引：27
作者：
Jensen, Bent P. [1 ]
Saraf, Rajneeta [2 ]
Ma, Jing [1 ]
Berry, Sarah [2 ]
Grant, Cameron C. [2 ,3 ]
Camargo, Carlos A., Jr. [4 ,5 ]
Sies, Christiaan W. [1 ]
机构：
[1] Canterbury Hlth Labs, Specialist Biochem, POB 151, Christchurch, New Zealand
[2] Univ Auckland, Ctr Longitudinal Res He Ara Ki Mua & Growing New, Auckland, New Zealand
[3] Univ Auckland, Dept Paediat Child & Youth Hlth, Auckland, New Zealand
[4] Massachusetts Gen Hosp, Boston, MA 02114 USA
[5] Harvard Med Sch, Boston, MA USA
来源：
CLINICA CHIMICA ACTA | 2018年 / 481卷
关键词：
Vitamin D; Dried blood spots; 2D LC-MS/MS; 25-hydroxyvitamin D-3; 25-hydroxyvitamin D-2; VITAMIN-D; ACCURATE QUANTIFICATION; D METABOLITES;
D O I：
10.1016/j.cca.2018.02.024
中图分类号：
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号：
1001 ;
摘要：
Background: Demand for measurement of 25-hydroxyvitamin D (25OHD) is growing and dried blood spot (DBS) sampling is attractive as samples are easier to collect, transport and store. Methods: A 2D LC-MS/MS assay without derivatization was developed. DBS punches (3.2 mm) were ultrasonicated with d(6)-250HD(3) in 70% methanol followed by hexane extraction, dry-down and reconstitution. The assay was validated and applied to two studies comparing whole blood adult DBS with serum samples (n = 40) and neonatal whole blood DBS with cord serum samples (n = 80). Results: The assay was validated in whole blood DBS over the range 13-106 nmol/L 25OHD(3) and 11-91 nmol/L 25OHD(2) with a limit of detection of 3 nmol/L. Intra-and inter-day imprecision was < 13% CV and bias < 12%. The assay had high recovery and minimal matrix effects. Triplicate DBS study samples had a mean CV of 13% for 25OHD(3). No 25OHD(2) was detected. DBS calculated serum 25OHD(3) concentrations correlated strongly with serum concentrations in the adult DBS/serum study (r = 0.94) and moderately in the neonatal DBS/cord serum study (r = 0.69). Conclusions: Direct quantitation of 25OHD in DBS by 2D LC-MS/MS without derivatization was found to be an alternative to serum quantitation applicable to clinical research studies on adult DBS samples.
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页码：61 / 68
页数：8
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