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Development and Validation of a Method Using Dried Oral Fluid Spot to Determine Drugs of Abuse
Authors:Ana Laura Bemvenuti Jacques B.S.  Maíra Kerpel dos Santos Ph.D.  Renata Pereira Limberger Ph.D.
Affiliation:1. Graduate Program of Pharmaceutical Sciences, Faculty of Pharmacy, Federal University of Rio Grande do Sul, 2752 Ipiranga Ave, Porto Alegre, RS, 90610‐000 BrazilCorresponding Author: Ana Laura Bemvenuti Jacques, B.S. E‐mail:;2. Graduate 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.
Keywords:forensic science  forensic toxicology  investigative techniques  substance abuse detection  analytic sample preparation methods  analytical chemistry techniques  dried oral fluid spot
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