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

Redžović, Zuzana; Erk, Marijana; Svetličić, Ema; Dončević, Lucija; Gottstein, Sanja; Hozić, Amela; Cindrić, Mario

doi:10.3390/separations8020020

Cited by 2 publications

(2 citation statements)

References 38 publications

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“…Composite samples of S. ambulans for quantification of adenylates were prepared as described by Redžović et al [37], who applied a modified method used by Díaz Enrich et al [38]. Amphipods were pooled due to their small size and mass (10-50 specimens per sample, mass of a sample: 0.012-0.034 g; Table S4).…”

Section: Preparation Of Synuella Ambulans Samplesmentioning

confidence: 99%

“…The AMP, ADP and ATP concentrations determined in the S. ambulans samples were calculated from the peak area of the respective nucleotide and expressed as µmol g −1 wet weight (w.w.). The final result was obtained by multiplying the adenylate concentration by the extraction recovery of internal standard ADN [37].…”

Section: Separation and Quantification Of Adenylates And Aec Calculationmentioning

confidence: 99%

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Energy Status of Stygophilous Amphipod Synurella ambulans as a Promising Biomarker of Environmental Stress in the Hyporheic Zone

Redžović,

Erk,

Gottstein

et al. 2023

Water

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The hyporheic zone (HZ) is a sensitive ecotone in river ecosystems because of its biodiversity and susceptibility to human activities. Hyporheic fauna are exposed to multiple stressors that affect the physiology and metabolism of organisms and ultimately ecosystem functioning and biodiversity. This study aimed to quantify adenine nucleotides in the stygophilous freshwater amphipod Synurella ambulans and to evaluate the potential of adenylate energy charge (AEC) as a physiological biomarker of general stress in the HZ of the Sava River at sites with different intensities of environmental and anthropogenic stress. Field studies were conducted seasonally (in December 2018 and April, July, and October 2019) at two sampling sites, one upstream (1-UP) and the other downstream (2-DOWN) of the discharge from the wastewater treatment plant using the standard piston pump. The amphipod population from site 1-UP had significantly higher AEC values than the population from site 2-DOWN in all seasons except summer. Coordinated changes in AEC values with the ATP/ADP ratio indicated differences in energy status between the two populations. However, no changes in the apparent equilibrium constant of adenylate kinase were observed, except in spring at site 1-UP. Multiple linear regression models showed the strongest associations of AEC with Fe and Zn accumulated in amphipods, followed by environmental factors (conductivity, dissolved O2, and concentrations of nitrites and phosphates in the interstitial water). AEC was shown to be a useful index of environmental stress in S. ambulans because it can directly measure the change in available energy and thus the metabolic stress to which the organism is exposed. Finally, seasonal and spatial variations in AEC values reflected ecological status in the HZ.

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Section: Preparation Of Synuella Ambulans Samplesmentioning

confidence: 99%

Section: Separation and Quantification Of Adenylates And Aec Calculationmentioning

confidence: 99%

Energy Status of Stygophilous Amphipod Synurella ambulans as a Promising Biomarker of Environmental Stress in the Hyporheic Zone

Redžović,

Erk,

Gottstein

et al. 2023

Water

Self Cite

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Advancements in Adenine Nucleotides Extraction and Quantification from a Single Drop of Human Blood

Popović,

Dončević,

Biba

et al. 2024

Molecules

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Adenine nucleotides (ANs)—adenosine 5′-triphosphate (ATP), adenosine 5′-diphosphate (ADP), and adenosine 5′-monophosphate (AMP)—are essential for energy transfer and the supply of countless processes within cellular metabolism. Their concentrations can be expressed as adenylate energy charge (AEC), a measure of cellular metabolic energy that directly correlates with the homeostasis of the organism. AEC index has broad diagnostic potential, as reduced ATP levels are associated to various conditions, such as inflammatory diseases, metabolic disorders, and cancer. We introduce a novel methodology for rapid isolation, purification, and quantification of ANs from a single drop of capillary blood. Of all the stationary phases tested, activated carbon proved to be the most efficient for the purification of adenine nucleotides, using an automated micro-solid phase extraction (µ-SPE) platform. An optimized µ-SPE method, coupled with RP-HPLC and a run time of 30 min, provides a reliable analytical framework for adenine nucleotide analysis of diverse biological samples. AN concentrations measured in capillary blood samples were 1393.1 µM, 254.8 µM, and 76.9 µM for ATP, ADP, and AMP molecules aligning with values reported in the literature. Overall, this study presents a streamlined and precise approach for analyzing ANs from microliters of blood, offering promising applications in clinical diagnostics.

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

Cited by 2 publications

References 38 publications

Energy Status of Stygophilous Amphipod Synurella ambulans as a Promising Biomarker of Environmental Stress in the Hyporheic Zone

Energy Status of Stygophilous Amphipod Synurella ambulans as a Promising Biomarker of Environmental Stress in the Hyporheic Zone

Advancements in Adenine Nucleotides Extraction and Quantification from a Single Drop of Human Blood

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