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Simple and accurate candidate reference measurement procedure for total testosterone in human serum by one-step liquid-liquid extraction coupled with isotope dilution mass spectrometry.

Abstract

Testosterone in human serum is commonly tested in clinical laboratories using immunoassay methods as well as liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. To standardize and ensure the accuracy of the measurement results, reference procedures with higher metrological order are required. A simple measurement procedure based on one-step liquid-liquid extraction (LLE) and liquid chromatography-isotope dilution tandem mass spectrometry (LC-IDMS/MS) was developed for total testosterone in human serum. The procedure involved serum spiked with 13C3-testosterone, equilibration for 2 h, and extraction with an organic solvent. Testosterone certified reference material (CRM) was used as the calibration standard to ensure the traceability to the International System of Units (SI). Testosterone in serum CRMs from the National Institute for Standards and Technology (NIST) and LGC were used to validate the accuracy of the newly developed method. The deviations of the obtained values from the NIST and LGC certified values ranged from -0.55% to 0.45%. Similarly, the coefficient of variations (CVs) of the replicate measurements were in the range of 0.55% and 0.78%, respectively. The relative expanded uncertainties were comparable with those of the certified materials. The newly developed LC-IDMS/MS procedure demonstrated adequate trueness and precision, and was simple to perform. The method can be used for value assignment of testosterone in external quality assessment (EQA) materials as well as certification of CRMs in the future. Graphical abstract.

Authors+Show Affiliations

Chemical Metrology Division, Applied Sciences Group, Health Sciences Authority, 1 Science Park Road #01-05/06, The Capricorn Singapore Science Park II, Singapore, 117528, Singapore.Chemical Metrology Division, Applied Sciences Group, Health Sciences Authority, 1 Science Park Road #01-05/06, The Capricorn Singapore Science Park II, Singapore, 117528, Singapore.Chemical Metrology Division, Applied Sciences Group, Health Sciences Authority, 1 Science Park Road #01-05/06, The Capricorn Singapore Science Park II, Singapore, 117528, Singapore.Department of Laboratory Medicine, National University Hospital, 5 Lower Kent Ridge Road, Singapore, 119074, Singapore.Chemical Metrology Division, Applied Sciences Group, Health Sciences Authority, 1 Science Park Road #01-05/06, The Capricorn Singapore Science Park II, Singapore, 117528, Singapore. liu_qinde@hsa.gov.sg.Chemical Metrology Division, Applied Sciences Group, Health Sciences Authority, 1 Science Park Road #01-05/06, The Capricorn Singapore Science Park II, Singapore, 117528, Singapore.Chemical Metrology Division, Applied Sciences Group, Health Sciences Authority, 1 Science Park Road #01-05/06, The Capricorn Singapore Science Park II, Singapore, 117528, Singapore.Department of Laboratory Medicine, National University Hospital, 5 Lower Kent Ridge Road, Singapore, 119074, Singapore.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31630222

Citation

Chen, Yizhao, et al. "Simple and Accurate Candidate Reference Measurement Procedure for Total Testosterone in Human Serum By One-step Liquid-liquid Extraction Coupled With Isotope Dilution Mass Spectrometry." Analytical and Bioanalytical Chemistry, 2019.
Chen Y, Teo HL, Liu H, et al. Simple and accurate candidate reference measurement procedure for total testosterone in human serum by one-step liquid-liquid extraction coupled with isotope dilution mass spectrometry. Anal Bioanal Chem. 2019.
Chen, Y., Teo, H. L., Liu, H., Loh, T. P., Liu, Q., Teo, T. L., ... Sethi, S. K. (2019). Simple and accurate candidate reference measurement procedure for total testosterone in human serum by one-step liquid-liquid extraction coupled with isotope dilution mass spectrometry. Analytical and Bioanalytical Chemistry, doi:10.1007/s00216-019-02152-y.
Chen Y, et al. Simple and Accurate Candidate Reference Measurement Procedure for Total Testosterone in Human Serum By One-step Liquid-liquid Extraction Coupled With Isotope Dilution Mass Spectrometry. Anal Bioanal Chem. 2019 Oct 19; PubMed PMID: 31630222.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Simple and accurate candidate reference measurement procedure for total testosterone in human serum by one-step liquid-liquid extraction coupled with isotope dilution mass spectrometry. AU - Chen,Yizhao, AU - Teo,Hui Ling, AU - Liu,Hong, AU - Loh,Tze Ping, AU - Liu,Qinde, AU - Teo,Tang Lin, AU - Lee,Tong Kooi, AU - Sethi,Sunil Kumar, Y1 - 2019/10/19/ PY - 2019/04/16/received PY - 2019/09/12/accepted PY - 2019/08/27/revised PY - 2019/10/21/entrez PY - 2019/10/21/pubmed PY - 2019/10/21/medline KW - Human serum KW - Isotope dilution mass spectrometry KW - Liquid chromatography-tandem mass spectrometry KW - Liquid-liquid extraction KW - Reference measurement procedure KW - Testosterone JF - Analytical and bioanalytical chemistry JO - Anal Bioanal Chem N2 - Testosterone in human serum is commonly tested in clinical laboratories using immunoassay methods as well as liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. To standardize and ensure the accuracy of the measurement results, reference procedures with higher metrological order are required. A simple measurement procedure based on one-step liquid-liquid extraction (LLE) and liquid chromatography-isotope dilution tandem mass spectrometry (LC-IDMS/MS) was developed for total testosterone in human serum. The procedure involved serum spiked with 13C3-testosterone, equilibration for 2 h, and extraction with an organic solvent. Testosterone certified reference material (CRM) was used as the calibration standard to ensure the traceability to the International System of Units (SI). Testosterone in serum CRMs from the National Institute for Standards and Technology (NIST) and LGC were used to validate the accuracy of the newly developed method. The deviations of the obtained values from the NIST and LGC certified values ranged from -0.55% to 0.45%. Similarly, the coefficient of variations (CVs) of the replicate measurements were in the range of 0.55% and 0.78%, respectively. The relative expanded uncertainties were comparable with those of the certified materials. The newly developed LC-IDMS/MS procedure demonstrated adequate trueness and precision, and was simple to perform. The method can be used for value assignment of testosterone in external quality assessment (EQA) materials as well as certification of CRMs in the future. Graphical abstract. SN - 1618-2650 UR - https://www.unboundmedicine.com/medline/citation/31630222/Simple_and_accurate_candidate_reference_measurement_procedure_for_total_testosterone_in_human_serum_by_one-step_liquid-liquid_extraction_coupled_with_isotope_dilution_mass_spectrometry L2 - https://dx.doi.org/10.1007/s00216-019-02152-y DB - PRIME DP - Unbound Medicine ER -