Thomas Berg1, Elin Eliassen1, Benedicte Jørgenrud1, Saranda Kabashi1, Alexey Petukhov2,3, Stig Tore Bogstrand1,4. 1. Section of Drug Abuse Research, Department of Forensic Sciences, Division of Laboratory Medicine, Oslo University Hospital, Oslo, Norway. 2. Moscow Scientific Practical Center of Drug Addiction, Moscow Municipal Department of Healthcare, Moscow, Russia. 3. I.M. Sechenov First Moscow State Medical University, Moscow, Russia. 4. Lovisenberg Diaconal University College, Oslo, Norway.
Abstract
BACKGROUND: Phosphatidylethanols (PEths) are specific, direct alcohol biomarkers that can be determined in human blood to distinguish between heavy and social drinking. PEth 16:0/18:1 is among the most predominant PEth homologues in human blood. The aim of the study was to develop a high throughput and sensitive UHPLC-MS/MS method for the determination of PEth 16:0/18:1 in whole blood. METHODS: Whole blood samples were prepared by 96-well supported liquid extraction (SLE). Extracted samples were analyzed for PEth 16:0/18:1 by reversed phase UHPLC-MS/MS. RESULTS: The developed UHPLC-MS/MS method was fully validated in whole blood with PEth 16:0/18:1-D5 as internal standard. Intermediate precision and intermediate accuracy were within ≤± 12% and ≤± 17%, respectively, at PEth 16:0/18:1 concentrations of 1.4-2112 ng/mL (2.0-3004 nmol/L). Limit of quantification (LOQ) was 1.7 ng/mL (2.4 nmol/L). CONCLUSION: For the first time, 96-well SLE was used for preparation of a PEth homologue in biological samples. A mixture of tert-butyl methyl ether and 2-propanol (5:1, v:v) was chosen as organic eluent based on an evaluation of extraction recovery, purity of extracts, and evaporation time. The developed UHPLC-MS/MS method can be used for high throughput analyses and sensitive determinations of PEth 16:0/18:1 in whole blood.
BACKGROUND:Phosphatidylethanols (PEths) are specific, direct alcohol biomarkers that can be determined in human blood to distinguish between heavy and social drinking. PEth 16:0/18:1 is among the most predominant PEth homologues in human blood. The aim of the study was to develop a high throughput and sensitive UHPLC-MS/MS method for the determination of PEth 16:0/18:1 in whole blood. METHODS: Whole blood samples were prepared by 96-well supported liquid extraction (SLE). Extracted samples were analyzed for PEth 16:0/18:1 by reversed phase UHPLC-MS/MS. RESULTS: The developed UHPLC-MS/MS method was fully validated in whole blood with PEth 16:0/18:1-D5 as internal standard. Intermediate precision and intermediate accuracy were within ≤± 12% and ≤± 17%, respectively, at PEth 16:0/18:1 concentrations of 1.4-2112 ng/mL (2.0-3004 nmol/L). Limit of quantification (LOQ) was 1.7 ng/mL (2.4 nmol/L). CONCLUSION: For the first time, 96-well SLE was used for preparation of a PEth homologue in biological samples. A mixture of tert-butyl methyl ether and 2-propanol (5:1, v:v) was chosen as organic eluent based on an evaluation of extraction recovery, purity of extracts, and evaporation time. The developed UHPLC-MS/MS method can be used for high throughput analyses and sensitive determinations of PEth 16:0/18:1 in whole blood.
Authors: Mark Boterman; Mira Doig; Massimo Breda; Steve Lowes; Jim Jersey; Ronald Shoup; Fabio Garofolo; Isabelle Dumont; Suzanne Martinez; Shane Needham; Maria Cruz Caturla; Philippe Couerbe; Joelle Guittard; John Maltas; Tim Lansing; Masood Bhatti; Christine Schiebl; Petra Struwe; Curtis Sheldon; Roger Hayes; Timothy Sangster; Colin Pattison; Johanne Bouchard; Lee Goodwin; Rafiq Islam; Rudi Segers; Zhongping John Lin; Jim Hillier; Wei Garofolo; Dieter Zimmer; Lois Folguera; Thomas Zimmermann; Maria Pawula; Marc Moussallie; Leonardo de Souza Teixeira; Thais Rocha; Daniel Tang; Paula Jardieu; James Truog; Jenny Lin; Richard Lundberg; Chris Cox; Alan Breau; Chiara Bigogno; Dick Schoutsen; Carmen Dilger; Mohammed Bouhajib; Ann Levesque; Sofi Gagnon-Carignan; Robert Nicholson; Rand Jenkins; Ming Hung Lin; Shane Karnik; Theo De Boer; Richard Houghton; Rachel Green; William DeMaio; Romuald Sable; Kirk Smith; Christoph Siethoff; Laura Cojocaru; Mike Allen; Tammy Harter; Saadya Fatmi; Farhad Sayyarpour; Michele Malone; Stuart Best; Xinping Fang Journal: Bioanalysis Date: 2012-03 Impact factor: 2.681
Authors: E N Sauve; M Langødegård; D Ekeberg; A M L Øiestad Journal: J Chromatogr B Analyt Technol Biomed Life Sci Date: 2011-11-03 Impact factor: 3.205