Abstract
In this study, a novel approach was developed to quantify endocannabinoids (eCBs), and was based on the liquid biosensor BIONOTE. This device is composed of a probe that can be immersed in a solution, and an electronic interface that can record a current related to the oxy-reductive reactions occurring in the sample. The two most representative members of eCBs have been analysed in vitro by BIONOTE: anandamide (N-arachidonoylethanolamine, AEA) and 2-arachidonoylglycerol (2-AG). Bovine serum albumin was used to functionalize the probe and improve the sensibility of the whole analytical system. We show that BIONOTE is able to detect both AEA and 2-AG at concentrations in the low nanomolar range, and to discriminate between these eCBs and their moieties arachidonic acid, ethanolamine and glycerol. Notably, BIONOTE distinguished these five different molecules, and it was also able to quantify AEA in human plasma. Although this is just a proof-of-concept study, we suggest BIONOTE as a cheap and user-friendly prototype sensor for high throughput quantitation of eCB content in biological matrices, with an apparent diagnostic potential for tomorrow’s medicine.
Highlights
The development of more and more sophisticated technologies and analytical techniques for the identification and quantitation of small molecules contributed to the study of lipid species, and to the understanding of their biological roles under health and disease conditions
When the Screen-Printed Electrode probe (SPE) is immersed in a solution, an input signal consisting of a triangular waveform from +1 V to −1 V is applied and oxy-reduction phenomena involving the analytes dissolved in the aqueous media are induced
The preliminary phase of this study was aimed at evaluating the feasibility of the BIONOTE device coupled with unmodified SPE to analyze eCBs
Summary
The development of more and more sophisticated technologies and analytical techniques for the identification and quantitation of small molecules contributed to the study of lipid species, and to the understanding of their biological roles under health and disease conditions. Anandamide (N-arachidonoylethanolamine, AEA) and 2-arachidonylglycerol (2-AG) [1,2,3], two derivatives of the polyunsaturated fatty acid arachidonic acid (AA), were identified as endogenous ligands of the type-1 (CB1 ) and type-2 (CB2 ) cannabinoid receptors [4,5], and named endocannabinoids (eCBs) [6]. This fundamental breakthrough led to the identification of additional eCBs and eCB-binding receptors, as well as to Nacylphosphatidylethanolamine (NAPE)-specific phospholipase D [7] and diacylglycerol lipases α and β [8] as the main biosynthetic enzymes for AEA and 2-AG formation, respectively. Fatty acid amide hydrolase [9] and monoacylglycerol lipase [10] were recognized as the main responsible for AEA and 2-AG hydrolysis to AA and ethanolamine or glycerol, respectively
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