Determination of Adenylate Nucleotides in Amphipod Gammarus fossarum by Ion-Pair Reverse Phase Liquid Chromatography: Possibilities of Positive Pressure Micro-Solid Phase Extraction
Autor: | Marijana Erk, Ema Svetličić, Amela Hozić, Mario Cindrić, Lucija Dončević, Zuzana Redžović, Sanja Gottstein |
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Rok vydání: | 2021 |
Předmět: |
0106 biological sciences
Adenosine monophosphate Adenylate kinase Filtration and Separation ecosystem monitoring 01 natural sciences Analytical Chemistry lcsh:Chemistry chemistry.chemical_compound ecological friendly solutions heterocyclic compounds Nucleotide Solid phase extraction Energy charge Biology chemistry.chemical_classification Chromatography Chemistry 010401 analytical chemistry Extraction (chemistry) Reversed-phase chromatography lcsh:QC1-999 0104 chemical sciences freshwater amphipods 010601 ecology lcsh:QD1-999 adenylate extraction adenylate energy charge lcsh:Physics Homogenization (biology) |
Zdroj: | Separations Volume 8 Issue 2 Separations, Vol 8, Iss 20, p 20 (2021) |
ISSN: | 2297-8739 |
DOI: | 10.3390/separations8020020 |
Popis: | 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 and are tied to allosteric regulation in various regulatory enzymes. Adenylate energy charge represents the precise relationship between the concentrations of adenosine monophosphate, diphosphate, and triphosphate and indicates the amount of metabolic energy available to an organism. The experimental conditions of adenylate extraction in freshwater amphipod Gammarus fossarum are reported here for the first time and are crucial for the qualitative and quantitative determination of adenylate nucleotides using efficient and sensitive ion-pair reverse phase LC. It was shown that amphipod calcified exoskeleton impeded the neutralization of homogenate. The highest adenylate yield was obtained by homogenization in perchloric acid and subsequent addition of potassium hydroxide and phosphate buffer to achieve a pH around 11. This method enables separation and accurate detection of adenylates. Our study provides new insight into the complexity of adenylate extraction and quantification that is crucial for the application of adenylate energy charge as a confident physiological measure of environmental stress and as a health index of G. fossarum. |
Databáze: | OpenAIRE |
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