Simultaneous UPLC–MS/MS quantification of the endocannabinoids 2-arachidonoyl glycerol (2AG), 1-arachidonoyl glycerol (1AG), and anandamide in human plasma: Minimization of matrix-effects, 2AG/1AG isomerization and degradation by toluene solvent extraction

Zoerner, Alexander A.; Bátkai, Sándor; Suchy, Maria-Theresia; Gutzki, Frank‐Mathias; Engeli, Stefan; Jordan, Jens; Tsikas, Dimitrios

doi:10.1016/j.jchromb.2011.06.025

Cited by 102 publications

(104 citation statements)

References 36 publications

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“…Among them, LLE based on toluene was defi nitively chosen by us because of several key advantages: poor or absent contamination and minimal isomerization, best recovery and selectivity. A similar recovery was reported by Zoerner et al ( 18 ) for 2AG when extracted with toluene, but we achieved a higher recovery for AEA. The further on line purifi cation helped in avoiding any phenomena of matrix interference, guaranteeing the vigorousness of the system across large batches.…”

Section: Discussionsupporting

confidence: 91%

“…Several preanalytical and analytical pitfalls, potentially explaining part of the discrepancy, were described in the last decade (13)(14)(15)(16)(17)(18); in addition, the defi nition of "normality", and thus, the selection of control subjects, may have represented a major source of variability among the reported values.…”

Section: Specimensmentioning

confidence: 99%

“…Different extraction approaches were tested to achieve the best recovery and stability, minimizing contamination derived from common solvents and disposables. First, we assessed the stability of 2AG in terms of isomerization to 1AG and of total loss of 1/2AG during evaporation in different solvents, observing that both effects were promoted by the protic ambient typical of MeOH ( 18 ). The huge loss observed in MeOH and MeOH-containing mixtures could be explained only in part by the adhesion to the tube surface, but mostly by degradation, probably by hydrolysis to arachidonic acid and glycerol.…”

Section: Downloaded Frommentioning

confidence: 99%

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Estimation of reference intervals of five endocannabinoids and endocannabinoid related compounds in human plasma by two dimensional-LC/MS/MS

Fanelli

Lallo

Belluomo

et al. 2012

Journal of Lipid Research

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Supplementary key words endocannabinoids • validation • reference intervalsThe endocannabinoids (ECs) are bioactive lipid mediators derived from membrane phospholipids. Since the discovery of the fi rst lipid mediator of the endocannabinoid system (ECS), arachidonoyl-ethanolamide (AEA), also called anandamide ( 1 ), several molecules belonging to this family were identifi ed, the most important being 2-arachidonoyl-glycerol (2AG) and its isomer 1AG among MAGs ( 2, 3 ), palmitoyl-ethanolamide (PEA) and oleoylethanolamide (OEA) among the NAEs, to which AEA belongs ( 4 ). Both 2AG and AEA act on cannabinoid receptor type 1 (CB1) and type 2 (CB2), whereas PEA and OEA act by infl uencing AEA metabolism and binding the peroxisome proliferator-activated receptor ␣ ( 5, 6 ), thus defi ned endocannabinoid related compounds (ERC ).Abstract The elucidation of the role of endocannabinoids in physiological and pathological conditions and the transferability of the importance of these mediators from basic evidence into clinical practice is still hampered by the indefi niteness of their circulating reference intervals. In this work, we developed and validated a two-dimensional LC/ MS/MS method for the simultaneous measurement of plasma endocannabinoids and related compounds such as arachidonoyl-ethanolamide, palmitoyl-ethanolamide, and oleoylethanolamide, belonging to the N-acyl-ethanolamide (NAE) family, and 2-arachidonoyl-glycerol and its inactive isomer 1-arachidonoyl-glycerol from the monoacyl-glycerol (MAG) family. We found that several pitfalls in the endocannabinoid measurement may occur, from blood withdrawal to plasma processing. Plasma extraction with toluene followed by online purifi cation was chosen, allowing high-throughput and reliability. We estimated gender-specifi c reference intervals on 121 healthy normal weight subjects fulfi lling rigorous anthropometric and hematic criteria. We observed no gender differences for NAEs, whereas signifi cantly higher MAG levels were found in males compared with females. MAGs also signifi cantly correlated with triglycerides. NAEs increased with age in females, and arachidonoyl-ethanolamide correlated with adiposity and metabolic parameters in females. This work paves the way to the establishment of defi nitive reference intervals for circulating endocannabinoids to help physicians move from the speculative research fi eld into the clinical fi eld. -Fanelli,F

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Section: Discussionsupporting

confidence: 91%

Section: Specimensmentioning

confidence: 99%

Section: Downloaded Frommentioning

confidence: 99%

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Estimation of reference intervals of five endocannabinoids and endocannabinoid related compounds in human plasma by two dimensional-LC/MS/MS

Fanelli

Lallo

Belluomo

et al. 2012

Journal of Lipid Research

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“…2-AG was prepared in acetonitrile which compared to ethanol is known to reduce the isomerization of 2-AG to 1-AG [30]. All stock solutions were stored at -70°C.…”

Section: Preparation Of Standardsmentioning

confidence: 99%

Identification of Lipid Mediators in Peripheral Human Tissues Using an Integrative In Vivo Microdialysis Approach

2016

J Anal Bioanal Tech

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Endocannabinoids and related N-acylethanolamines (NAEs) are lipid mediators involved in a number of physiological and pathological mechanisms in peripheral tissues. Microdialysis (MD) technique allows continues sampling of endogenous substances in the interstitial fluids of the tissues. The main limitation of MD sampling of lipophilic compounds is low recovery due to adsorption to the MD system and particularly to the catheter membranes. In this in vivo study microdialysate samples were collected from human trapezius muscle and forearm skin. The levels of arachidonoylethanolamide (AEA), 2-arachidonoylglycerol (2-AG), oleoylethanolamide (OEA), palmitoylethanolamide (PEA), and stearoylethanolamide (SEA) were analyzed in both microdialysate and in catheter membrane samples using liquid chromatography tandem mass spectrometry. OEA, PEA and SEA were identified in all microdialysate and catheter membrane samples from trapezius and skin. 2-AG was found in all catheter membrane samples from both tissues but not in the actual microdialysate. In conclusion sampling of OEA, PEA and SEA was achievable from trapezius and skin with the presented MD set-up. 2-AG is present in both trapezius muscle and skin tissue but adsorbs to the membranes in a higher extent than the NAEs. Furthermore, consideration of data conserved in the membrane during an MD experiment could be a relevant and more broadly applicable extension of MD sampling methodology which could fill an "information gap" and enhance an adequate interpretation of microdialysate data outcomes

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“…The advantages of LC over GC include less time-consuming sample preparation, mainly due to the fact that pre-derivatization of non-volatile analytes (such as ECs and NAEs) is usually not needed in LC, and shorter retention times, which makes it possible to analyse many samples in a relatively short time [157]. Modified LCs or ultra-pressure LCs (UPLC) have also been used for EC analysis [173] as these instruments can operate at very high back pressure with the advantage of high chromatographic resolution, high sensitivity, and shorter analysis times. In addition, the ionization techniques ESI and APCI have frequently been used for ECs and NAEs [157].…”

Section: Lc-ms/msmentioning

confidence: 99%

Endocannabinoids and Related Lipids in Chronic Pain: Analytical and Clinical Aspects

Stensson¹

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Simultaneous UPLC–MS/MS quantification of the endocannabinoids 2-arachidonoyl glycerol (2AG), 1-arachidonoyl glycerol (1AG), and anandamide in human plasma: Minimization of matrix-effects, 2AG/1AG isomerization and degradation by toluene solvent extraction

Cited by 102 publications

References 36 publications

Estimation of reference intervals of five endocannabinoids and endocannabinoid related compounds in human plasma by two dimensional-LC/MS/MS

Estimation of reference intervals of five endocannabinoids and endocannabinoid related compounds in human plasma by two dimensional-LC/MS/MS

Identification of Lipid Mediators in Peripheral Human Tissues Using an Integrative In Vivo Microdialysis Approach

Endocannabinoids and Related Lipids in Chronic Pain: Analytical and Clinical Aspects

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