Anandamide levels in human female reproductive tissues: Solid-phase extraction and measurement by ultraperformance liquid chromatography tandem mass spectrometry

• Published in: Analytical biochemistry Vol. 400; no. 2; pp. 155 - 162
• Main Authors: Marczylo, Timothy H., Lam, Patricia M.W., Amoako, Akwasi A., Konje, Justin C.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.05.2010
• Subjects: Amniotic Fluid - chemistry; Anandamide; Arachidonic Acids - analysis; Arachidonic Acids - blood; Arachidonic Acids - isolation & purification; Chromatography, High Pressure Liquid - methods; Endocannabinoid; Endocannabinoids; Extraembryonic Membranes - chemistry; Female; Humans; Placenta - chemistry; Polyunsaturated Alkamides - analysis; Polyunsaturated Alkamides - blood; Polyunsaturated Alkamides - isolation & purification; Pregnancy; Solid Phase Extraction; Tandem Mass Spectrometry - methods; Umbilical Arteries - chemistry; Umbilical Veins - chemistry; Endocannabinoid; Solid-phase extraction; Anandamide
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2009.12.025

Anandamide ( N-arachidonoylethanolamide), a bioactive lipid, is reported to play a role in pregnancy maintenance and parturition. Our aims were to (1) evaluate AEA levels at the human maternal:fetal interface and (2) validate the use of solid-phase extraction of AEA from tissues. AEA was analyzed in cord and maternal blood, amniotic fluid, placenta, and fetal membranes collected during Caesarean section ( n = 14). Extraction efficiencies were 42 and 36% for the placenta and the fetal membranes, respectively. Tissue AEA was quantified using an isotope-dilution method and UPLC-ESI-MS/MS giving intra- and inter-day variability for tissues spiked with 0.2, 1, and 5 pmol/g AEA of less than 12%. Accuracy for these spiked samples was between 95% and 103% for fetal membranes and between 99% and 114% for placenta. Mean AEA concentrations were 2.72 ± 1.04 pmol/g for placenta and 1.19 ± 0.68 pmol/g for fetal membranes, and 0.93 ± 0.28, 0.88 ± 0.33, 0.77 ± 0.30, and 0.06 ± 0.04 nM for maternal, umbilical vein, and umbilical artery plasma and amniotic fluid. Higher AEA concentrations were found in placenta compared to fetal membranes ( P < 0.0001), in umbilical vein compared with umbilical artery ( P = 0.0015), and in plasma from maternal circulation compared with umbilical artery ( P = 0.0152). The relevance of these changes in AEA concentrations at the maternal:fetal interface requires further investigation.