Enhanced chemiluminescent assays for acetylcholine

Abstract

Acetylcholine and choline chemiluminescent assays have limitations when these compounds are detected in small areas of mammalian nervous tissue. Use of 7-dimethyl-aminonaphthalene-1,2-dicarbonic acid hydrazide (7-DMAN), instead of luminol, gives a threefold increase in emitted light in the chemiluminescent assay for acetylcholine based on the coupled choline oxidase-peroxidase reaction. Addition of light enhancers, such as para-iodophenol or D-luciferin, to luminol or 7-DMAN further increased the light emission. Under these conditions the detection limit for acetylcholine was 650 femtomoles. This enhanced chemiluminescent assay should be convenient for the detection of in vivo and in vitro acetylcholine release from mammalian neurons.

Influence of acetylcholinesterase on embryonic spinal rat motoneurones growth in culture: a quantitative morphometric study

“…The last traces of ether were evaporated and the pH of the extract was approximately 4. The chemiluminescent determination of ACh, as described in detail by Israel & Lesbats 1981) and slightly modified by Ternaux & Chamoin 1994), is based on the following principle: the enzyme choline oxidase from Arthrobacter globiformis , catalyses the oxidation of choline, generating betaïne and H 2 O 2 with consumption of oxygen. The hydrogen peroxide formed is continuously monitored by measuring the light produced by the oxidation of luminol in the presence of peroxidase.…”

Analysis of acetylcholine from extracellular fluid in brain by in vivo microdialysis and LC–ESI-MS/MS with the stable isotope-labeled internal standard

“…In vivo microdialysis has been used extensively since the late 1960s and early 1970s. Reported methods for the measurement of ACh were based on the use of chemiluminescence [7], electrochemical (ECD) [8], fluorescence [9], MS [10,11] and UV [12] detections. A review by Tsai [13] surveyed various analytical techniques that have been adopted for the measurement of ACh.…”

Development of a liquid chromatography/tandem mass spectrometry method for the quantitation of acetylcholine and related neurotransmitters in brain microdialysis samples

“…Analytical methods for monitoring the levels of ACh were initially based upon the use of chemiluminescence [22] and/or electrochemical, fluorescence and ultraviolet detection systems [23][24][25][26]. Tsai reviewed the various analytical techniques employed for the measurement of ACh [27].…”