Measurement of serum 3-epi-25-hydroxyvitamin D3,25-hydroxyvitamin D3and 25-hydroxyvitamin D2in infant, paediatric and adolescent populations of Korea using ultra-performance liquid chromatography-tandem mass spectrometry.
Background We evaluated the performance of ultra-performance liquid chromatography-tandem mass spectrometry to measure serum 3-epi-25-hydroxyvitamin D3, 25-hydroxyvitamin D3and 25-hydroxyvitamin D2concentrations in 519 infant, paediatric and adolescent serum samples in Korea. Methods We used a Kinetex XB-C18 column and isocratic methanol/water (77.5/22.5, v/v) with 0.025% (v/v) high-performance liquid chromatography solvent additive flowing at 0.25 mL/min, yielding an 11 min/sample run time. A TQD triple quadrupole mass spectrometer in electrospray ionization positive ion mode with multiple reaction monitoring transition via an MSMS vitamin D kit was used to evaluate precision, carryover, ion suppression and linearity. Samples were prepared using the 4-phenyl-1,2,4-triazoline-3,5-dione derivatization method. Results Intra- and inter-run precisions were 1.23-13.28% and 1.02-10.08%, respectively. Group carryovers were -0.27% and 0.10%, respectively. There was no ion suppression. The calibration curve showed good linearity from calibrator Level 1 (11.75 nmol/L) to 6 (375 nmol/L) with R2> 0.9999. The 3-epi-25-hydroxyvitamin D3and 25-hydroxyvitamin D3peaks were clearly separated in the extracted ion chromatogram. Infant serum samples 3-epi-25-hydroxyvitamin D3concentrations were significantly higher than paediatric and adolescent concentrations. Conclusions The ultra-performance liquid chromatography-tandem mass spectrometry assay performed acceptably, clearly separating 3-epi-25-hydroxyvitamin D3from 25-hydroxyvitamin D3. High 3-epi-25-hydroxyvitamin D3concentrations were observed in infant but not in paediatric and adolescent serum samples.
1 LabGenomics Clinical Laboratories, Seongnam, Korea.
Sung E
SE
Cho
2 Department of Laboratory Medicine, Ilsan Paik Hospital, Inje University College of Medicine, Goyang, Korea.
Sollip
S
Kim
3 Department of Pediatrics, Ilsan Paik Hospital, Inje University College of Medicine, Goyang, Korea.
Young D
YD
Kim
1 LabGenomics Clinical Laboratories, Seongnam, Korea.
Hyojung
H
Lee
1 LabGenomics Clinical Laboratories, Seongnam, Korea.
Dong H
DH
Seo
4 Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea.|5 Department of Laboratory Medicine, Seoul National University Bundang Hospital, Seongnam, Korea.
Junghan
J
Song
2 Department of Laboratory Medicine, Ilsan Paik Hospital, Inje University College of Medicine, Goyang, Korea.
Tae H
TH
Um
2 Department of Laboratory Medicine, Ilsan Paik Hospital, Inje University College of Medicine, Goyang, Korea.
Chong R
CR
Cho
3 Department of Pediatrics, Ilsan Paik Hospital, Inje University College of Medicine, Goyang, Korea.
Nam H
NH
Kim
3 Department of Pediatrics, Ilsan Paik Hospital, Inje University College of Medicine, Goyang, Korea.
Jong H
JH
Hwang
D015652:25-Hydroxyvitamin D 2 / Q000097:blood*; D000293:Adolescent; D001774:Blood Chemical Analysis / Q000379:methods*; D002112:Calcifediol / Q000097:blood*; D002648:Child; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007223:Infant; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D015203:Reproducibility of Results; D056910:Republic of Korea; D053719:Tandem Mass Spectrometry; D055815:Young Adult
D015652:25-Hydroxyvitamin D 2; D002112:Calcifediol
25-hydroxyvitamin D2; 25-hydroxyvitamin D3; 3-epi-25-hydroxyvitamin D3; Korean paediatric population; ultra-performance liquid chromatography-tandem mass spectrometry