Simultaneous ultra-high performance liquid chromathograpy-electrospray ionization-quadrupole-time of flight mass spectrometry quantification of endogenous anandamide and related N-acylethanolamides in bio-matrices.
We describe and validate a sensitive UHPLC-ESI-QTOF-MS method for the simultaneous quantification of seven endocannabinoids and non-endocannabinoids related N-acylethanolamides: N-arachidonoylethanolamide, N-palmitoylethanolamide, N-stearoylethanolamide, N-oleoylethanolamide, N-linoleoylethanolamide, N-α-linolenoylethanolamide and N-eicosapentaenoylethanolamide in several bio-matrices for the purpose of research and clinical application. We examined effects of different liquid-liquid and solid phase extraction on the recovery of endocannabinoids and N-acylethanolamides. Protein precipitation with cooled acetone and extraction with acetonitrile (1% v/v formic acid) using OASIS HLB cartridge gave better results. Separation was performed on a Waters Acquity UPLC HSST3 column using a 9min elution gradient coupled with high resolution mass spectrometry (QTOF/MS). The high sensitivity of the developed method allow its application on sample with low volumes or low levels of endocannabinoids and N-acylethanolamides and make the method suitable for routine measurement in human bio-matrices, such as plasma, serum (500μL), urine (1mL) and tissues (10-30mg). Its application in clinical research could contribute to unravel pathophysiological roles of these family of lipid mediators and disclose novel diagnostic and prognostic markers.
Dipartimento di Scienze Biomediche e Cliniche L. Sacco, Università degli Studi di Milano, Via G.B. Grassi, 74-20157 Milano, Italy. Electronic address: roberta.ottria@unimi.it.
Roberta
R
Ottria
Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, Sezione di Tossicologia Forense, Università degli Studi di Milano, Via Mangiagalli, 37-20133 Milano, Italy.
Alessandro
A
Ravelli
Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, Sezione di Tossicologia Forense, Università degli Studi di Milano, Via Mangiagalli, 37-20133 Milano, Italy.
Fausto
F
Gigli
Dipartimento di Scienze Biomediche e Cliniche L. Sacco, Università degli Studi di Milano, Via G.B. Grassi, 74-20157 Milano, Italy. Electronic address: pierangela.ciuffreda@unimi.it.
Pierangela
P
Ciuffreda
D000577:Amides; D000818:Animals; D001095:Arachidonic Acids / Q000032:analysis / Q000097:blood* / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D063388:Endocannabinoids / Q000032:analysis / Q000097:blood* / Q000652:urine*; D004983:Ethanolamines / Q000032:analysis / Q000097:blood / Q000652:urine; D006801:Humans; D057230:Limit of Detection; D008041:Linoleic Acids / Q000032:analysis / Q000097:blood / Q000652:urine; D008297:Male; D010169:Palmitic Acids / Q000032:analysis / Q000097:blood / Q000652:urine; D053284:Polyunsaturated Alkamides / Q000032:analysis / Q000097:blood* / Q000652:urine*; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013229:Stearic Acids / Q000032:analysis / Q000097:blood / Q000652:urine; D053719:Tandem Mass Spectrometry / Q000379:methods
D000577:Amides; D001095:Arachidonic Acids; D063388:Endocannabinoids; D004983:Ethanolamines; D008041:Linoleic Acids; D010169:Palmitic Acids; D053284:Polyunsaturated Alkamides; D013229:Stearic Acids; C466089:stearoylethanolamide; C109347:linoleoyl ethanolamide; C005958:palmidrol; C078814:anandamide
Anandamide, Bio-matrices; Endocannabinoids; LC–MS/MS; N-acylethanolamides; Quantitative analysis