Evaluation of uncertainty sources in the determination of testosterone in urine by calibration-based and isotope dilution quantification using ultra high performance liquid chromatography tandem mass spectrometry
Pitarch-Motellón, Jorge; Roig-Navarro, Antoni F.; Sancho Llopis, Juan V.et al.
[en] Three quantification methodologies, namely calibration with internal standard (Cal-IS, non-weighted), weighted calibration with internal standard (wCal-IS) and isotope pattern deconvolution (IPD) have been used for the determination of testosterone in urine by LC-MS/MS. Uncertainty has been calculated and compared for the three methodologies through intra- and inter-laboratory reproducibility assays. IPD showed the best performance for the intra-laboratory reproducibility, with RSD and combined uncertainty values below 4% and 9% respectively. wCal-IS showed similar performance, while Cal-IS where not constant and clearly worse at the lowest concentration assayed (2 ng/mL) reaching RSD values up to 16%. The inter-laboratory assay indicated similar results although wCal-IS RSD (20%) was higher than IPD (10%) and Cal-IS get worse with RSD higher than 40% for the lowest concentration level. Uncertainty budgets calculated for the three procedures revealed that intercept and slope were the most important factors contributing to uncertainty for Cal-IS. The main factors for wCal-IS and IPD were the volumes of sample and/or standard measured.
Disciplines :
Chemistry
Author, co-author :
Pitarch-Motellón, Jorge
Roig-Navarro, Antoni F.
Sancho Llopis, Juan V.
Ibañez, Maria
Fabregat Cabello, Neus ; Université de Liège - ULiège > Département de pharmacie > Chimie médicale
Evaluation of uncertainty sources in the determination of testosterone in urine by calibration-based and isotope dilution quantification using ultra high performance liquid chromatography tandem mass spectrometry
Gosetti, F., Mazzucco, E., Gennaro, M.C., Marengo, E., Ultra high performance liquid chromatography tandem mass spectrometry determination and profiling of prohibited steroids in human biological matrices. A review. J. Chromatogr. B. 927 (2013), 22–36, 10.1016/j.jchromb.2012.12.003.
Botrè, F., New and old challenges of sports drug testing. J. Mass Spectrom. 43 (2008), 903–907, 10.1002/jms.1455.
Jarek, A., Kowalczyk, K., Chołbiński, P., Chajewska, K., Turek-Lepa, E., Pokrywka, A., Bulska, E., Kwiatkowska, D., Analytical procedure for steroid profiling valid for Athlete Biological Passport. Chem. Pap. 69 (2015), 254–261, 10.1515/chempap-2015-0030.
Saudan, C., Testosterone and doping control. Br. J. Sports Med. 40 (2006), i21–i24, 10.1136/bjsm.2006.027482.
Badoud, F., Guillarme, D., Boccard, J., Grata, E., Saugy, M., Rudaz, S., Veuthey, J.-L., Analytical aspects in doping control: challenges and perspectives, Forensic Sci. Int 213 (2011), 49–61, 10.1016/j.forsciint.2011.07.024.
Nicoli, R., Guillarme, D., Leuenberger, N., Baume, N., Robinson, N., Saugy, M., Veuthey, J.-L., Analytical strategies for doping control purposes: needs, challenges, and perspectives. Anal. Chem. 88 (2016), 508–523, 10.1021/acs.analchem.5b03994.
Trout, G.J., Kazlauskas, R., Sports drug testing − an analyst's perspective. Chem. Soc. Rev., 33, 2004, 1, 10.1039/b201476a.
World Anti-Doping Agency, WADA Technical Document TD2016EAAS, (2016) 1–16.
Thevis, M., Thomas, A., Pop, V., Schänzer, W., Ultrahigh pressure liquid chromatography–(tandem) mass spectrometry in human sports drug testing: possibilities and limitations. J. Chromatogr. A 1292 (2013), 38–50, 10.1016/j.chroma.2012.12.048.
Taylor, P.J., Matrix effects: the Achilles heel of quantitative high-performance liquid chromatography–electrospray–tandem mass spectrometry. Clin. Biochem. 38 (2005), 328–334, 10.1016/j.clinbiochem.2004.11.007.
Furey, A., Moriarty, M., Bane, V., Kinsella, B., Lehane, M., Ion suppression; A critical review on causes, evaluation, prevention and applications. Talanta 115 (2013), 104–122, 10.1016/j.talanta.2013.03.048.
Trufelli, H., Palma, P., Famiglini, G., Cappiello, A., An overview of matrix effects in liquid chromatography-mass spectrometry. Mass Spectrom. Rev. 30 (2011), 491–509, 10.1002/mas.20298.
González-Antuña, A., Rodríguez-González, P., Centineo, G., García Alonso, J.I., Evaluation of minimal 13C-labelling for stable isotope dilution in organic analysis. Analyst, 135, 2010, 953, 10.1039/b924432 h.
Fabregat-Cabello, N., Pitarch-Motellón, J., Sancho, J.V., Ibáñez, M., Roig-Navarro, A.F., Method development and validation for the determination of selected endocrine disrupting compounds by liquid chromatography mass spectrometry and isotope pattern deconvolution in water samples. Comparison of two extraction techniques. Anal. Methods 8 (2016), 2895–2903, 10.1039/C6AY00221H.
Fabregat-Cabello, N., Zomer, P., Sancho, J.V., Roig-Navarro, A.F., Mol, H.G.J., Comparison of approaches to deal with matrix effects in LC-MS/MS based determinations of mycotoxins in food and feed. World Mycotoxin J. 9 (2016), 149–161, 10.3920/WMJ2014.1872.
Fabregat-Cabello, N., Sancho, J.V., Vidal, A., González, F.V., Roig-Navarro, A.F., Development and validation of a liquid chromatography isotope dilution mass spectrometry method for the reliable quantification of alkylphenols in environmental water samples by isotope pattern deconvolution. J. Chromatogr. A. 1328 (2014), 43–51, 10.1016/j.chroma.2013.12.077.
Spirito, E., Botre, F., The role of measurement uncertainty in doping analysis. Int. J. Risk Assess. Manag., 5, 2005, 374, 10.1504/IJRAM.2005.007178.
Barroso, O., Miller, J., Squirrell, A., Westwood, S., Measurement uncertainty in anti-doping quantitative analysis for prohibited threshold substances. Bioanalysis 4 (2012), 1653–1665, 10.4155/bio.12.127.
García-Alonso, J.I., Rodríguez-González, P., Uncertainty evaluation in IDMS. Isot. Dilution Mass Spectrom, 2013, RSC Publishing, 379–433.
Ramaley, L., Herrera, L.C., Software for the calculation of isotope patterns in tandem mass spectrometry, Rapid Commun. Mass Spectrom. 22 (2008), 2707–2714, 10.1002/rcm.3668.
García-Alonso, J.I., Rodríguez-González, P., IDMS procedures and calculation methods. Isot. Dilution Mass Spectrom, 2013, RSC Publishing, 41–76.
Rodríguez-González, P., García Alonso, J.I., Recent advances in isotope dilution analysis for elemental speciation. J. Anal. At. Spectrom., 25, 2010, 239, 10.1039/b924261a.
Castillo, Á., Gracia-Lor, E., Roig-Navarro, Á.F., Sancho, J.V., Rodríguez-González, P., Alonso, J.I.G., Isotope pattern deconvolution-tandem mass spectrometry for the determination and confirmation of diclofenac in wastewaters. Anal. Chim. Acta. 765 (2013), 77–85, 10.1016/j.aca.2012.11.033.
World Anti-Doping Agency, Technical Document-TD2014DL Decision Limits for the Quantification of Threshold Substances, (n.d.).
Nordtest, Handbook for Calculation of Measurement Uncertainty in Environmental Laboratories, (2012).
Kragten, J., Tutorial review. Calculating standard deviations and confidence intervals with a universally applicable spreadsheet technique. Analyst, 119, 1994, 2161, 10.1039/an9941902161.
