: Changes in the phosphorus-containing metabolites were monitored by 31P nuclear magnetic resonance in the developing embryos of Clarias batrachus. Phosphomonoester, yolk phosphoprotein, phosphocreatine, ATP, and inorganic phosphate (Pi) were consistently observed in all the developmental stages of C. batrachus. None of these phosphometabolites exhibited any significant change in their concentration up to the blastula stage, whereas distinct decrease in all except inorganic phosphate was observed in the fry stage. Concomitantly an increase in the concentration of inorganic phosphate was observed. Further, from the resonance positions of alpha, beta, and gamma phosphate groups of ATP, it was evident that the ATP molecules in vivo were liganded either to Ca2+ or Mg2+. This study also revealed that the intracellular pH of the developing embryos was approximately 7.05 up to the gastrula stage, after which it decreased in the fry stage to 6.98 units.