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Development and Validation of a Method Using Dried Oral Fluid Spot to Determine Drugs of Abuse
Ana Laura Bemvenuti Jacques1, Maíra Kerpel Dos Santos1, Renata Pereira Limberger1
1Graduate Program of Pharmaceutical Sciences, Faculty of Pharmacy, Federal University of Rio Grande do Sul, 2752 Ipiranga Ave, Porto Alegre, RS, 90610-000, Brazil.
Abstract:
A liquid chromatography-mass spectrometry method using dried oral fluid spots was developed and validated for the simultaneous quantification of cocaine, benzoylecgonine, cocaethylene, amphetamine, and 3,4-methylenedioxymethamphetamine. The oral fluid was applied to a Whatman 903 grade paper and submitted to a drying time of 2.5 h. The extraction procedure was optimized by chemometric approach using simplex centroid design. Spots were extracted with a mixture of acetonitrile, buffer, and methanol. Calibration curves covered a linear concentration range of 40-500 ng/mL. Validation parameters of linearity, precision, accuracy, selectivity, carryover, matrix effects, and stability were evaluated and showed satisfactory results. Spot homogeneity was also satisfactory, with less than 15% of deviation from nominal concentration. Spot volume did not influence accuracy when less than 100 μL of the sample was applied to the spot. The validation of the proposed method suggests a potential application in different scenarios in toxicology.
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