Chiral Analysis of Methadone and its Main Metabolite, EDDP, in Postmorten Brain and Blood by Automated SPE and Liquid Chromatography-Mass Spectrometry

Karen Marie Dollerup Holm; Kristian Linnet

Chiral Analysis of Methadone and its Main Metabolite, EDDP, in Postmorten Brain and Blood by Automated SPE and Liquid Chromatography-Mass Spectrometry

Bidragets oversatte titel: Chiral Analyse af methadone og dennes hovedmetabolit, EDDP, i Postmortem Hjerne og Blod ved Automatisk SPE og Væskekromatografi-Massespektrometri

Karen Marie Dollerup Holm, Kristian Linnet

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Abstract

Vi udviklede en metode baseret på væskekromatografi
med tandem-massespektrometri til kvantificering af de individuelle enantiomer
af metadon og dets primære metabolit, R/S-2-ethyl-1,5-dimethyl-
3,3-diphenylpyrrolinium (EDDP), i postmortem blod og hjernevæv. Prøvebehandlingen blev udført på et Tecan Evo robotsystem. Udfældning blev efterfulgt af fastfase-ekstraktion, inddampning og rekonstituering i den mobile fase. Enantiomerne var fuldt
adskilt med væskekromatografi på en chiral a-(1)-syreglycoprotein kolonne. En Quattro mikro massespektrometer blev anvendt til detektion i positiv ion-mode med en elektrospray-kilde. Den nedre grænse for kvantificering i hjernevæv var 0,005 mg/kg for metadon og 0,001 mg/kg for EDDP enantiomerne; den maksimale præcision var 17% for begge stoffer, korrektheden varierede fra 94 til 101%. I blod, var kvantificeringsgrænsen 0,001 mg/kg for alle stoffer, den samlede relative standardafvigelse var <15%, og korrekthed varierer fra 95 til 109%. Hjerne- (n=11) og blod-(n=15) prøver blev analyseret med intermediær korrekthed varierende fra 7,5 til 15% ved 0,005 mg/kg og 6,8 til 11,3% ved 0,25 mg/kg for alle stoffer. Metode blev udvikling med fokus på fremstilling af en ren ekstrakt, især fra hjerneprøver. Metoden blev testet på autentisk hjerne og femorale blodprøver

Bidragets oversatte titel	Chiral Analyse af methadone og dennes hovedmetabolit, EDDP, i Postmortem Hjerne og Blod ved Automatisk SPE og Væskekromatografi-Massespektrometri
Originalsprog	Engelsk
Tidsskrift	Journal of Analytical Toxicology
Vol/bind	36
Udgave nummer	21
Sider (fra-til)	487-496
Antal sider	10
ISSN	0146-4760
Status	Udgivet - 1 sep. 2012

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Det Sundhedsvidenskabelige Fakultet
metadon
chiral analyse
hjerne
blod
toksikologi

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title = "Chiral Analysis of Methadone and its Main Metabolite, EDDP, in Postmorten Brain and Blood by Automated SPE and Liquid Chromatography-Mass Spectrometry",

abstract = "We developed a method based on liquid chromatography coupled with tandem mass spectrometry to quantify individual enantiomers of methadone and its primary metabolite, R/S-2-ethyl-1,5-dimethyl- 3,3-diphenylpyrrolinium (EDDP), in postmortem blood and brain tissue. Samples were prepared with a Tecan Evo robotic system. Precipitation was followed by solid-phase extraction, evaporation and reconstitution in the mobile phase. Enantiomers were fully separated with liquid chromatography on a chiral a(1)-acid glycoprotein column. A Quattro micro mass spectrometer was used for detection in the positive ion mode with an electrospray source. The lower limit of quantification in brain tissue was 0.005 mg/kg for methadone and 0.001 mg/kg for EDDP enantiomers; the maximum precision was 17% for both compounds; accuracy ranged from 94 to 101%. In blood, the limit of quantification was 0.001 mg/kg for all compounds, the total relative standard deviation was <15%, and the accuracy varied from 95 to 109%. Brain (n 5 11) and blood (n 5 15) samples were analyzed with intermediate precision that varied from 7.5 to 15% at 0.005 mg/kg and from 6.8 to 11.3% at 0.25 mg/kg for all compounds. Method development focused on producing a clean extract, particularly from brain samples. The method was tested on authentic brain and femoral blood samples.",

keywords = "Faculty of Health and Medical Sciences, metadon, chiral analyse, hjerne, blod, toksikologi",

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T1 - Chiral Analysis of Methadone and its Main Metabolite, EDDP, in Postmorten Brain and Blood by Automated SPE and Liquid Chromatography-Mass Spectrometry

AU - Holm, Karen Marie Dollerup

AU - Linnet, Kristian

PY - 2012/9/1

Y1 - 2012/9/1

N2 - We developed a method based on liquid chromatography coupled with tandem mass spectrometry to quantify individual enantiomers of methadone and its primary metabolite, R/S-2-ethyl-1,5-dimethyl- 3,3-diphenylpyrrolinium (EDDP), in postmortem blood and brain tissue. Samples were prepared with a Tecan Evo robotic system. Precipitation was followed by solid-phase extraction, evaporation and reconstitution in the mobile phase. Enantiomers were fully separated with liquid chromatography on a chiral a(1)-acid glycoprotein column. A Quattro micro mass spectrometer was used for detection in the positive ion mode with an electrospray source. The lower limit of quantification in brain tissue was 0.005 mg/kg for methadone and 0.001 mg/kg for EDDP enantiomers; the maximum precision was 17% for both compounds; accuracy ranged from 94 to 101%. In blood, the limit of quantification was 0.001 mg/kg for all compounds, the total relative standard deviation was <15%, and the accuracy varied from 95 to 109%. Brain (n 5 11) and blood (n 5 15) samples were analyzed with intermediate precision that varied from 7.5 to 15% at 0.005 mg/kg and from 6.8 to 11.3% at 0.25 mg/kg for all compounds. Method development focused on producing a clean extract, particularly from brain samples. The method was tested on authentic brain and femoral blood samples.

AB - We developed a method based on liquid chromatography coupled with tandem mass spectrometry to quantify individual enantiomers of methadone and its primary metabolite, R/S-2-ethyl-1,5-dimethyl- 3,3-diphenylpyrrolinium (EDDP), in postmortem blood and brain tissue. Samples were prepared with a Tecan Evo robotic system. Precipitation was followed by solid-phase extraction, evaporation and reconstitution in the mobile phase. Enantiomers were fully separated with liquid chromatography on a chiral a(1)-acid glycoprotein column. A Quattro micro mass spectrometer was used for detection in the positive ion mode with an electrospray source. The lower limit of quantification in brain tissue was 0.005 mg/kg for methadone and 0.001 mg/kg for EDDP enantiomers; the maximum precision was 17% for both compounds; accuracy ranged from 94 to 101%. In blood, the limit of quantification was 0.001 mg/kg for all compounds, the total relative standard deviation was <15%, and the accuracy varied from 95 to 109%. Brain (n 5 11) and blood (n 5 15) samples were analyzed with intermediate precision that varied from 7.5 to 15% at 0.005 mg/kg and from 6.8 to 11.3% at 0.25 mg/kg for all compounds. Method development focused on producing a clean extract, particularly from brain samples. The method was tested on authentic brain and femoral blood samples.

KW - Faculty of Health and Medical Sciences

KW - metadon

KW - chiral analyse

KW - hjerne

KW - blod

KW - toksikologi

M3 - Journal article

SN - 0146-4760

VL - 36

SP - 487

EP - 496

JO - Journal of Analytical Toxicology

JF - Journal of Analytical Toxicology

IS - 21

ER -

Chiral Analysis of Methadone and its Main Metabolite, EDDP, in Postmorten Brain and Blood by Automated SPE and Liquid Chromatography-Mass Spectrometry

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