The TVB-N content was assessed as described previously [27 (link)]. For this, a fish muscle fraction (10 g) was extracted with 6% perchloric acid and brought up to 50 mL. TVB-N value was determined by titration of the distillate with 10 mM HCl after steam-distillation of the acid extracts rendered alkaline to pH 13 with 20% NaOH. The resulting values were expressed as mg TVB-N·kg−1 fish muscle.
Nucleotide extracts were obtained following the method proposed by Ryder [28 (link)]. The analysis was carried out by HPLC following the procedure proposed by Aubourg et al. [29 (link)]. The K value (%) was calculated on the basis of the following molar concentration ratio, in which the concentrations of the different molecules involved in the adenosine-triphosphate degradation pathway are taken into account: K value (%) = 100 × [hypoxanthine + inosine]/[adenosine-triphosphate + adenosine-diphosphate + adenosine-monophosphate + inosine-monophosphate + inosine + hypoxanthine].
Nucleotide extracts were obtained following the method proposed by Ryder [28 (link)]. The analysis was carried out by HPLC following the procedure proposed by Aubourg et al. [29 (link)]. The K value (%) was calculated on the basis of the following molar concentration ratio, in which the concentrations of the different molecules involved in the adenosine-triphosphate degradation pathway are taken into account: K value (%) = 100 × [hypoxanthine + inosine]/[adenosine-triphosphate + adenosine-diphosphate + adenosine-monophosphate + inosine-monophosphate + inosine + hypoxanthine].
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