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Isotope dilution ultra performance liquid chromatography-tandem mass spectrometry method for simultaneous measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in human serum.
Clin Chim Acta. 2011 Aug 17; 412(17-18):1594-9.CC

Abstract

BACKGROUND

An ultra performance liquid chromatography-tandem mass spectrometry method with calibration traceable to NIST SRM was developed and validated to measure concentrations of 25-hydroxyvitamin D(2) (25OHD(2)), 25-hydroxyvitamin D(3) (25OHD(3)) and the C-3 epimer of 25OHD(3) (epi-25OHD(3)) in human serum.

METHODS

Tri- and hexa-deuterated internal standards were added to serum (100 μl) to monitor recovery. Liquid-liquid extraction was used to extract the hexane-soluble materials. Calibration solutions [8-100 nmol/L 25OHD(2,) 12-150 nmol/L 25OHD(3), and 4-50 nmol/L epi-25OHD(3)] prepared in phosphate-buffered saline containing 4% albumin were similarly processed. Using a pentafluorophenyl column (2.1×100 mm) and isocratic methanol/water (72/28, v/v) flowing at 0.4 ml/min, run time was 14 min per sample; 25OHD(3) and epi-25OHD(3) were baseline separated. Atmospheric pressure chemical ionization in the positive ion mode with selected reaction monitoring captured the following transitions: 25OHD(2), m/z 395.3>377.3 (209.1 qualifier); (epi-)25OHD(3), m/z 383.3>365.3 (105.1 qualifier); d(3)-25OHD(2), m/z 398.3>380.3; and d(6)-25OHD(3), m/z 389.3>371.3.

RESULTS

Recovery averaged ≥98%. Total imprecision was ≤10% when concentrations were ≥20 nmol/l. Bias averaged <5%. Detection limits were <5 nmol/l. Median (nmol/l) 25OHD(2), 25OHD(3) and epi-25OHD(3) were quantitated in 98 blood donors (<LOD, 56.0, <LOD) and 35 pregnant women (<LOD, 87.6, 3.70).

CONCLUSIONS

This method is highly accurate, precise and specific.

Authors+Show Affiliations

Centers for Disease Control & Prevention, NCEH/DLS/NBB, Atlanta, GA 30341, United States. zwa5@cdc.govNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Validation Study

Language

eng

PubMed ID

21601563

Citation

Schleicher, Rosemary L., et al. "Isotope Dilution Ultra Performance Liquid Chromatography-tandem Mass Spectrometry Method for Simultaneous Measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in Human Serum." Clinica Chimica Acta; International Journal of Clinical Chemistry, vol. 412, no. 17-18, 2011, pp. 1594-9.
Schleicher RL, Encisco SE, Chaudhary-Webb M, et al. Isotope dilution ultra performance liquid chromatography-tandem mass spectrometry method for simultaneous measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in human serum. Clin Chim Acta. 2011;412(17-18):1594-9.
Schleicher, R. L., Encisco, S. E., Chaudhary-Webb, M., Paliakov, E., McCoy, L. F., & Pfeiffer, C. M. (2011). Isotope dilution ultra performance liquid chromatography-tandem mass spectrometry method for simultaneous measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in human serum. Clinica Chimica Acta; International Journal of Clinical Chemistry, 412(17-18), 1594-9. https://doi.org/10.1016/j.cca.2011.05.010
Schleicher RL, et al. Isotope Dilution Ultra Performance Liquid Chromatography-tandem Mass Spectrometry Method for Simultaneous Measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in Human Serum. Clin Chim Acta. 2011 Aug 17;412(17-18):1594-9. PubMed PMID: 21601563.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isotope dilution ultra performance liquid chromatography-tandem mass spectrometry method for simultaneous measurement of 25-hydroxyvitamin D2, 25-hydroxyvitamin D3 and 3-epi-25-hydroxyvitamin D3 in human serum. AU - Schleicher,Rosemary L, AU - Encisco,Sara E, AU - Chaudhary-Webb,Madhulika, AU - Paliakov,Ekaterina, AU - McCoy,Leslie F, AU - Pfeiffer,Christine M, Y1 - 2011/05/13/ PY - 2011/03/17/received PY - 2011/05/04/revised PY - 2011/05/05/accepted PY - 2011/5/24/entrez PY - 2011/5/24/pubmed PY - 2011/10/19/medline SP - 1594 EP - 9 JF - Clinica chimica acta; international journal of clinical chemistry JO - Clin Chim Acta VL - 412 IS - 17-18 N2 - BACKGROUND: An ultra performance liquid chromatography-tandem mass spectrometry method with calibration traceable to NIST SRM was developed and validated to measure concentrations of 25-hydroxyvitamin D(2) (25OHD(2)), 25-hydroxyvitamin D(3) (25OHD(3)) and the C-3 epimer of 25OHD(3) (epi-25OHD(3)) in human serum. METHODS: Tri- and hexa-deuterated internal standards were added to serum (100 μl) to monitor recovery. Liquid-liquid extraction was used to extract the hexane-soluble materials. Calibration solutions [8-100 nmol/L 25OHD(2,) 12-150 nmol/L 25OHD(3), and 4-50 nmol/L epi-25OHD(3)] prepared in phosphate-buffered saline containing 4% albumin were similarly processed. Using a pentafluorophenyl column (2.1×100 mm) and isocratic methanol/water (72/28, v/v) flowing at 0.4 ml/min, run time was 14 min per sample; 25OHD(3) and epi-25OHD(3) were baseline separated. Atmospheric pressure chemical ionization in the positive ion mode with selected reaction monitoring captured the following transitions: 25OHD(2), m/z 395.3>377.3 (209.1 qualifier); (epi-)25OHD(3), m/z 383.3>365.3 (105.1 qualifier); d(3)-25OHD(2), m/z 398.3>380.3; and d(6)-25OHD(3), m/z 389.3>371.3. RESULTS: Recovery averaged ≥98%. Total imprecision was ≤10% when concentrations were ≥20 nmol/l. Bias averaged <5%. Detection limits were <5 nmol/l. Median (nmol/l) 25OHD(2), 25OHD(3) and epi-25OHD(3) were quantitated in 98 blood donors (<LOD, 56.0, <LOD) and 35 pregnant women (<LOD, 87.6, 3.70). CONCLUSIONS: This method is highly accurate, precise and specific. SN - 1873-3492 UR - https://www.unboundmedicine.com/medline/citation/21601563/Isotope_dilution_ultra_performance_liquid_chromatography_tandem_mass_spectrometry_method_for_simultaneous_measurement_of_25_hydroxyvitamin_D2_25_hydroxyvitamin_D3_and_3_epi_25_hydroxyvitamin_D3_in_human_serum_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0009-8981(11)00266-X DB - PRIME DP - Unbound Medicine ER -