Next Article in Journal
ESI–LC–MS/MS for Therapeutic Drug Monitoring of Binary Mixture of Pregabalin and Tramadol: Human Plasma and Urine Applications
Previous Article in Journal
Bio-Guided Fractionation of Oil Palm (Elaeis guineensis) Fruit and Interactions of Compounds with First-Line Antituberculosis Drugs against Mycobacterium tuberculosis H37Ra
Article

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

1
Ruđer Bošković Institute, 10000 Zagreb, Croatia
2
Faculty of Food Technology and Biotechnology, University of Zagreb, 10000 Zagreb, Croatia
3
Faculty of Science, University of Zagreb, 10000 Zagreb, Croatia
*
Author to whom correspondence should be addressed.
Academic Editor: Fabio Gosetti
Separations 2021, 8(2), 20; https://doi.org/10.3390/separations8020020
Received: 31 December 2020 / Revised: 6 February 2021 / Accepted: 7 February 2021 / Published: 13 February 2021
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. View Full-Text
Keywords: adenylate energy charge; freshwater amphipods; ecosystem monitoring; adenylate extraction; ecological friendly solutions adenylate energy charge; freshwater amphipods; ecosystem monitoring; adenylate extraction; ecological friendly solutions
Show Figures

Graphical abstract

MDPI and ACS Style

Redžović, Z.; Erk, M.; Svetličić, E.; Dončević, L.; Gottstein, S.; Hozić, A.; Cindrić, M. Determination of Adenylate Nucleotides in Amphipod Gammarus fossarum by Ion-Pair Reverse Phase Liquid Chromatography: Possibilities of Positive Pressure Micro-Solid Phase Extraction. Separations 2021, 8, 20. https://doi.org/10.3390/separations8020020

AMA Style

Redžović Z, Erk M, Svetličić E, Dončević L, Gottstein S, Hozić A, Cindrić M. Determination of Adenylate Nucleotides in Amphipod Gammarus fossarum by Ion-Pair Reverse Phase Liquid Chromatography: Possibilities of Positive Pressure Micro-Solid Phase Extraction. Separations. 2021; 8(2):20. https://doi.org/10.3390/separations8020020

Chicago/Turabian Style

Redžović, Zuzana, Marijana Erk, Ema Svetličić, Lucija Dončević, Sanja Gottstein, Amela Hozić, and Mario Cindrić. 2021. "Determination of Adenylate Nucleotides in Amphipod Gammarus fossarum by Ion-Pair Reverse Phase Liquid Chromatography: Possibilities of Positive Pressure Micro-Solid Phase Extraction" Separations 8, no. 2: 20. https://doi.org/10.3390/separations8020020

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop