Determination of antipsychotic drugs in oral fluid using dried saliva spots by gas chromatography-tandem mass spectrometry
The present work describes the optimization and validation of an analytical method for the determination of six antipsychotic drugs (chlorpromazine, levomepromazine, cyamemazine, clozapine, haloperidol, and quetiapine) in oral fluid samples after solvent extraction from dried saliva spots, by gas chromatography coupled to tandem mass spectrometry. The method was fully validated, and the included parameters were selectivity, linearity, limits of quantification, precision and accuracy, stability, and recovery. The method was linear for all compounds from 10 to 400 ng/mL, except for haloperidol (5–100 ng/mL), presenting coefficients of determination higher than 0.99. Inter- and intra-day precision and accuracy were in conformity with the criteria usually seen in bioanalytical method validation; i.e., coefficients of variation were lower than 15% and an accuracy of 15% or better for all studied drugs. The recoveries obtained with this miniaturized technique ranged from 63 to 97%. The herein described method is the first to be reported using the dried saliva spots approach for the analysis of these antypshychotic drugs, proving great sensitivity apart from its simple and fast procedure. The method was considered a good alternative to the conventional techniques to be applied in clinical and toxicological analyses, even more taking into account the extremely low sample volume used (50 μL).
KeywordsDried saliva spots Oral fluid GC-MS/MS Antipsychotic drugs
The authors acknowledge all the nursing team of the Departamento de Psiquiatria e Saúde Mental from the Centro Hospitalar Cova da Beira.
This work is supported by FEDER funds through the POCI-COMPETE 2020 - Operational Programme Competitiveness and Internationalization in Axis I – Strengthening Research, Technological Development and Innovation (Project POCI-01- 0145-FEDER-007491) and National Funds by FCT – Foundation for Science and Technology (UID / Multi / 00709/2019).
Compliance with ethical standards
The present study was approved by the ethics committee from Centro Hospitalar Cova da Beira and has been conducted according to ethical standards. The analyzed authentic samples belonged to individuals who provided an informed consent for their use, and all analyses were carried out according to the ethical standards of the institution.
Conflict of interest
The authors declare that they have no conflicts of interest.
- 2.Mohammadi M, Akhondzadeh S. Schizophrenia: etiology and pharmacotherapy. IDrugs. 2001;4:1167–72.Google Scholar
- 3.Da Fonseca BM, Moreno IED, Barroso M, Costa S, Queiroz JA, Gallardo E. Determination of seven selected antipsychotic drugs in human plasma using microextraction in packed sorbent and gas chromatography – tandem mass spectrometry. Anal Bioanal Chem. 2013;405:3953–63. https://doi.org/10.1007/s00216-012-6695-y.CrossRefGoogle Scholar
- 11.Patteet L, Maudens KE, Morrens M, Sabbe B, Dom G, Neels H. Determination of common antipsychotics in quantisal-collected oral fluid by UHPLC-MS/MS: method validation and applicability for therapeutic drug monitoring. Ther Drug Monit. 2016;38:87–97. https://doi.org/10.1097/FTD.0000000000000242.CrossRefGoogle Scholar
- 12.Di Corcia D, Lisi S, Pirro V, Gerace E, Salomone A, Vincenti M. Determination of pharmaceutical and illicit drugs in oral fluid by ultra-high performance liquid chromatography-tandem mass spectrometry. J Chromatogr B. 2013;927:133–41. https://doi.org/10.1016/j.jchromb.2013.01.025.CrossRefGoogle Scholar
- 18.Carvalho J, Rosado T, Barroso M, Gallardo E, Investigação C De, Interior B, D AI, Fármaco-toxicologia L De, Beira U. Determination of antiepileptic drugs using dried saliva spots. J Anal Toxicol. 2018:1–11. https://doi.org/10.1093/jat/bky064.
- 19.Ribeiro A, Prata M, Vaz C, Rosado T, Restolho J, Barroso M, et al. Determination of methadone and EDDP in oral fluid using the dried saliva spots sampling approach and gas chromatography-tandem mass spectrometry. Anal Bioanal Chem. 2019:1–11. https://doi.org/10.1007/s00216-019-01654-z.
- 21.Chen X, Zheng S, Le J, Qian Z, Zhang R, Hong Z, et al. Ultrasound-assisted low-density solvent dispersive liquid–liquid microextraction for the simultaneous determination of 12 new antidepressants and 2 antipsychotics in whole blood by gas chromatography–mass spectrometry. J Pharm Biomed Anal. 2017;142:19–27. https://doi.org/10.1016/j.jpba.2017.04.032.CrossRefGoogle Scholar
- 22.Rosado T, Oppolzer D, Cruz B, Barroso M, Varela S, Oliveira V, et al. Development and validation of a gas chromatography/tandem mass spectrometry method for simultaneous quantitation of several antipsychotics in human plasma and oral fluid. Rapid Commun Mass Spectrom. 2018;32:2081–95. https://doi.org/10.1002/rcm.8087.CrossRefGoogle Scholar
- 23.Koller D, Zubiaur P, Saiz-Rodríguez M, Abad-Santos F, Wojnicz A. Simultaneous determination of six antipsychotics, two of their metabolites and caffeine in human plasma by LC-MS/MS using a phospholipid-removal microelution-solid phase extraction method for sample preparation. Talanta. 2019;198:159–68. https://doi.org/10.1016/j.talanta.2019.01.112.CrossRefGoogle Scholar
- 25.Khelfi A, Azzouz M, Abtroun R, Reggabi M, Alamir B. Determination of chlorpromazine, haloperidol, levomepromazine, olanzapine, risperidone, and sulpiride in human plasma by liquid chromatography/tandem mass spectrometry (LC-MS/MS). Int J Anal Chem. 2018;2018:1–13. https://doi.org/10.1155/2018/5807218.CrossRefGoogle Scholar
- 27.Food and Drug Administration, Bioanalytical Method Validation Guidance for Industry, (2018)http://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/default.htm (accessed 4 Feb 2019).
- 28.International Conference on Harmonisation (1996) Validation of analytical procedures: text and methodology Q2 (R1). Int Conf Harmon 11–12 . doi: http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf accessed 4 Feb 2019).
- 33.World Anti-doping Agency, International standard for laboratories: identification criteria for qualitative assays incorporating column chromatography and mass spectrometry, (2010). https://www.wada-ama.org/sites/default/files/resources/files/WADA_TD2010IDCRv1.0_Identification%20Criteria%20for%20Qualitative%20Assays_May%2008%202010_EN.doc.pdf (accessed 6 Jan 2019).
- 34.Pujadas M, Pichini S, Civit E, Santamariña E, Perez K, de la Torre R. A simple and reliable procedure for the determination of psychoactive drugs in oral fluid by gas chromatography-mass spectrometry. J Pharm Biomed Anal. 2007;44:594–601. https://doi.org/10.1016/j.jpba.2007.02.022.CrossRefGoogle Scholar
- 35.De Bièvre P, Günzler H. Validation in chemical measurement. 1st edition. Berlin: Springer-Verlag Berlin Heidelberg; 2005.Google Scholar