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Development of analytical method for the determination of methylphenidate, the analog ethylphenidate and their metabolite ritalinic acid in oral fluid samples by micro-QuEChERS and liquid chromatography-tandem mass spectrometry

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Author(s):
Chinaglia, Kaue de Oliveira ; Arantes, Ana Carolina Furiozo ; da Cunha, Kelly Francisco ; de Campos, Eduardo Geraldo ; Kahl, Julia Martinelli Magalhaes ; Rodrigues, Leonardo Costalonga ; Costa, Jose Luiz
Total Authors: 7
Document type: Journal article
Source: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES; v. 1205, p. 7-pg., 2022-06-15.
Abstract

The aim of this study was to develop a quantitative method for the analysis of methylphenidate, the analog ethylphenidate and their metabolite ritalinic acid in oral fluid, using micro-QuEChERS extraction and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Oral fluid samples were collected with QuantisalTM device, extracted by micro-QuEChERS technique and analyzed by LC-MS/MS. The developed method met the validation criteria of Academy Standards Board (ASB) Standard Practices for Method Validation in Forensic Toxicology (Standard 036, 2019) with limits of detection and quantification of 0.5 ng/mL and calibration curve from 0.5 to 50 ng/mL. Within-run imprecision was greater than 18.7% while between-run imprecision was greater than 17.0 % for all analytes. Bias did not vary more than 7.7 %. No evidence of carryover was found. Stability studies presented satisfactory results for 24 h on autosampler (10 ?), after 3 cycles of freeze/thaw, 7 days on freezer (-20 C) and until 7 days on refrigerator (4 C) for methylphenidate. The validated method was further successfully applied to the analysis of 5 authentic oral fluid samples collected from volunteers at parties and music festivals from different cities in Brazil. Four samples had positive results for methylphenidate and ritalinic acid, and only one sample was positive for methylphenidate. Ethylphenidate was not detected in the samples. The method showed acceptable analytical performance and is environmentally friendly, requiring reduced use of solvents and reagents, with potential to be applied to clinical and forensic analyses. (AU)

FAPESP's process: 19/02866-1 - Development of an analytical method for the determination of methylphenidate, ethylphenidate and ritalinic acid in oral fluid samples using liquid chromatography-tandem mass spectrometry
Grantee:Kauê de Oliveira Chinaglia
Support Opportunities: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 20/10809-5 - Microextraction techniques in forensic toxicology
Grantee:José Luiz da Costa
Support Opportunities: Regular Research Grants