Cargando…

A Human Oral Fluid Assay for D- and L- Isomer Detection of Amphetamine and Methamphetamine Using Liquid-Liquid Extraction

Medical providers are increasingly confronted with clinical decision-making that involves (meth)amphetamines. And clinical laboratories need a sensitive, efficient assay for routine assessment of D- and L-isomers to determine the probable source of these potentially illicit analytes. This paper pres...

Descripción completa

Detalles Bibliográficos
Autores principales: Robbins, Brian, Carpenter, Rob E., Long, Mary, Perry, Jacob
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9734005/
https://www.ncbi.nlm.nih.gov/pubmed/36507104
http://dx.doi.org/10.1155/2022/4819599
_version_ 1784846497109508096
author Robbins, Brian
Carpenter, Rob E.
Long, Mary
Perry, Jacob
author_facet Robbins, Brian
Carpenter, Rob E.
Long, Mary
Perry, Jacob
author_sort Robbins, Brian
collection PubMed
description Medical providers are increasingly confronted with clinical decision-making that involves (meth)amphetamines. And clinical laboratories need a sensitive, efficient assay for routine assessment of D- and L-isomers to determine the probable source of these potentially illicit analytes. This paper presents a validated method of D- and L-isomer detection in human oral fluid from an extract used for determination of a large oral fluid assay (63 analytes) on an older AB SCIEX 4000 instrument. Taken from the positive extract, D- and L-analytes were added. The method for extraction included addition of internal standard and a 2-step liquid-liquid extraction and dry-down step to concentrate and clean the samples. The samples were suspended in 50% MeOH in water, diluted with mobile phase, with separation and detection accomplished using LC-MS/MS to determine analyte concentration. Once samples were confirmed positive for (meth)amphetamine from the large oral fluid assay, they were further examined for the enantiomeric forms with 50 μl aliquots of the standards and samples of interest combined with 450 μl of D- and L-assay mobile phase, then analyzed using chiral column separation, and LC-MS/MS detection with standard curve spanning the range from 2.5 to 1000 ng/mL. The result is a sensitive and accurate detection of D- and L-isomers of amphetamine and methamphetamine in human oral fluid performed on an older model mass spectrometer (AB SCIEX 4000). The novelty of this assay is twofold (a) the 2-step liquid-liquid extraction and dry-down step to concentrate and clean the samples, and (b) its adoption characteristics as a reflex test from a large ODT panel without the need to invest in newer or expensive LC-MS/MS instruments. Finally, this assay also has potential to add a valuable option to high-throughput laboratories seeking a D- and L-testing alternative to urine drug testing methods.
format Online
Article
Text
id pubmed-9734005
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Hindawi
record_format MEDLINE/PubMed
spelling pubmed-97340052022-12-10 A Human Oral Fluid Assay for D- and L- Isomer Detection of Amphetamine and Methamphetamine Using Liquid-Liquid Extraction Robbins, Brian Carpenter, Rob E. Long, Mary Perry, Jacob J Anal Methods Chem Research Article Medical providers are increasingly confronted with clinical decision-making that involves (meth)amphetamines. And clinical laboratories need a sensitive, efficient assay for routine assessment of D- and L-isomers to determine the probable source of these potentially illicit analytes. This paper presents a validated method of D- and L-isomer detection in human oral fluid from an extract used for determination of a large oral fluid assay (63 analytes) on an older AB SCIEX 4000 instrument. Taken from the positive extract, D- and L-analytes were added. The method for extraction included addition of internal standard and a 2-step liquid-liquid extraction and dry-down step to concentrate and clean the samples. The samples were suspended in 50% MeOH in water, diluted with mobile phase, with separation and detection accomplished using LC-MS/MS to determine analyte concentration. Once samples were confirmed positive for (meth)amphetamine from the large oral fluid assay, they were further examined for the enantiomeric forms with 50 μl aliquots of the standards and samples of interest combined with 450 μl of D- and L-assay mobile phase, then analyzed using chiral column separation, and LC-MS/MS detection with standard curve spanning the range from 2.5 to 1000 ng/mL. The result is a sensitive and accurate detection of D- and L-isomers of amphetamine and methamphetamine in human oral fluid performed on an older model mass spectrometer (AB SCIEX 4000). The novelty of this assay is twofold (a) the 2-step liquid-liquid extraction and dry-down step to concentrate and clean the samples, and (b) its adoption characteristics as a reflex test from a large ODT panel without the need to invest in newer or expensive LC-MS/MS instruments. Finally, this assay also has potential to add a valuable option to high-throughput laboratories seeking a D- and L-testing alternative to urine drug testing methods. Hindawi 2022-12-02 /pmc/articles/PMC9734005/ /pubmed/36507104 http://dx.doi.org/10.1155/2022/4819599 Text en Copyright © 2022 Brian Robbins et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Robbins, Brian
Carpenter, Rob E.
Long, Mary
Perry, Jacob
A Human Oral Fluid Assay for D- and L- Isomer Detection of Amphetamine and Methamphetamine Using Liquid-Liquid Extraction
title A Human Oral Fluid Assay for D- and L- Isomer Detection of Amphetamine and Methamphetamine Using Liquid-Liquid Extraction
title_full A Human Oral Fluid Assay for D- and L- Isomer Detection of Amphetamine and Methamphetamine Using Liquid-Liquid Extraction
title_fullStr A Human Oral Fluid Assay for D- and L- Isomer Detection of Amphetamine and Methamphetamine Using Liquid-Liquid Extraction
title_full_unstemmed A Human Oral Fluid Assay for D- and L- Isomer Detection of Amphetamine and Methamphetamine Using Liquid-Liquid Extraction
title_short A Human Oral Fluid Assay for D- and L- Isomer Detection of Amphetamine and Methamphetamine Using Liquid-Liquid Extraction
title_sort human oral fluid assay for d- and l- isomer detection of amphetamine and methamphetamine using liquid-liquid extraction
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9734005/
https://www.ncbi.nlm.nih.gov/pubmed/36507104
http://dx.doi.org/10.1155/2022/4819599
work_keys_str_mv AT robbinsbrian ahumanoralfluidassayfordandlisomerdetectionofamphetamineandmethamphetamineusingliquidliquidextraction
AT carpenterrobe ahumanoralfluidassayfordandlisomerdetectionofamphetamineandmethamphetamineusingliquidliquidextraction
AT longmary ahumanoralfluidassayfordandlisomerdetectionofamphetamineandmethamphetamineusingliquidliquidextraction
AT perryjacob ahumanoralfluidassayfordandlisomerdetectionofamphetamineandmethamphetamineusingliquidliquidextraction
AT robbinsbrian humanoralfluidassayfordandlisomerdetectionofamphetamineandmethamphetamineusingliquidliquidextraction
AT carpenterrobe humanoralfluidassayfordandlisomerdetectionofamphetamineandmethamphetamineusingliquidliquidextraction
AT longmary humanoralfluidassayfordandlisomerdetectionofamphetamineandmethamphetamineusingliquidliquidextraction
AT perryjacob humanoralfluidassayfordandlisomerdetectionofamphetamineandmethamphetamineusingliquidliquidextraction