Abstract
We developed and evaluated a candidate serum reference material to help improve clinical routine measurement, and to provide traceability of the measurement results. D8-Homocystine, dithiothreitol, and acetonitrile were used as an internal standard, the reducing agent, and the protein precipitating agent, respectively. A triple quadrupole mass spectrometer with an electrospray ionization source was used for monitoring the transitions (m/z 140.0 → 94.0, 136.0 → 90.0) in multiple-reaction-monitoring mode. We used a calibration model relying on bracketing and gravimetric measurements to give SI-traceability and higher accuracy to serum value assignments. The method was evaluated for accuracy using NIST Standard Reference Material SRM1955. The results of the three concentrations (1, 2, and 3) of total homocysteine in human serum samples were determined by an isotope-dilution liquid chromatography-tandem mass spectrometry method; tHcy 1 is 28.8 ± 1.1 μmol/L, tHcy 2 is 17.93 ± 0.57 μmol/L, and tHcy 3 is 14.38 ± 0.46 μmol/L.
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Acknowledgements
This work is financially supported by the National Natural Science Foundation of China (No. 21275134) and National Key Technology Research and Development Program of the Ministry of Science and Technology of China (2013BAK10B01, 2011FY130100, 2011BAI02B05, 2011AA02A111).
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All procedures performed in studies involving human participants were in accordance with the ethical standards of the Ethics Committee of Beijing Aerospace General Hospital and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
This article does not contain any studies with animals performed by any of the authors.
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Liu, Y., Song, D., Xu, B. et al. Development of a matrix-based candidate reference material of total homocysteine in human serum. Anal Bioanal Chem 409, 3329–3335 (2017). https://doi.org/10.1007/s00216-017-0272-3
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DOI: https://doi.org/10.1007/s00216-017-0272-3