Determination of Adenylate Nucleotides in Amphipod Gammarus fossarum by Ion-Pair Reverse Phase Liquid Chromatography: Possibilities of Positive Pressure Micro-Solid Phase Extraction

Adenine nucleotides—adenosine monophosphate, diphosphate, and triphosphate—are of utmost importance to all living organisms, where they play a critical role in the energy metabolism are tied allosteric regulation various regulatory enzymes. Adenylate charge represents precise relationship between concentrations adenosine triphosphate indicates amount metabolic available an organism. The experimental conditions adenylate extraction freshwater amphipod Gammarus fossarum reported here for first time crucial qualitative quantitative determination nucleotides using efficient sensitive ion-pair reverse phase LC. It was shown that calcified exoskeleton impeded neutralization homogenate. highest yield obtained by homogenization perchloric acid subsequent addition potassium hydroxide phosphate buffer achieve pH around 11. This method enables separation accurate detection adenylates. Our study provides new insight into complexity quantification is application as confident physiological measure environmental stress health index G. fossarum.