Resumen

MIYARES-ESTRADA, Maelys et al. Application of Neesler reagent in the ammonium quantification used in the fermentations of biotechnology product. Vaccimonitor [online]. 2015, vol.24, n.1, pp.33-44. ISSN 1025-028X.

The ammonium salts are used in fermentations to supplement the deficient amounts of nitrogen and stabilize the pH of the culture medium. The excess ammonium ion exerts a detrimental effect on the fermentation process inhibiting microbial growth. An analytical method based on Neesler reagent was developed for monitoring and controlling the concentration of ammonium during the fermentation process. The test was standardized, by means of the selection of measuring equipment, and the reaction time as well as comparing standards of ammonium salts. The method was characterized with the evaluation of the next parameters: Specificity, Linearity and Range, Quantification Limit, Accuracy and Precision. The method proved to be specific. Two linear curves were defined in the ranges of concentrations of ammonium chloride salt (2-20 µg/ml) and ammonium sulfate salt (5-30 µg/ml). The limits of quantification were the lowest points of each one. The method proved to be accurate and precise. This assay was applied to samples of the yeast culture and bacteria of the genus Saccharomyces and E. coli respectively . A novel method in micro plate for quantification and analytical control of ammonia was developed. T his method is used to control this fundamental chemical component in the fermentations , to optimize the culture medium. Thus, an appropriate expression of recombinant proteins and proper vaccine candidates for clinical use are achieved. The ammonium salts are used in fermentations to supplement the deficient amounts of nitrogen and stabilize the pH of the culture medium. The excess ammonium ion exerts a detrimental effect on the fermentation process inhibiting microbial growth. An analytical method based on Neesler reagent was developed for monitoring and controlling the concentration of ammonium during the fermentation process. The test was standardized, by means of the selection of measuring equipment, and the reaction time as well as comparing standards of ammonium salts. The method was characterized with the evaluation of the next parameters: Specificity, Linearity and Range, Quantification Limit, Accuracy and Precision. The method proved to be specific. Two linear curves were defined in the ranges of concentrations of ammonium chloride salt (2-20 µg/ml) and ammonium sulfate salt (5-30 µg/ml). The limits of quantification were the lowest points of each one. The method proved to be accurate and precise. This assay was applied to samples of the yeast culture and bacteria of the genus <em>Saccharomyces </em>and <em>E. coli </em>respectively . A novel method in micro plate for quantification and analytical control of ammonia was developed. T his method is used to control this fundamental chemical component in the fermentations , to optimize the culture medium. Thus, an appropriate expression of recombinant proteins and proper vaccine candidates for clinical use are achieved. <p></p>

Palabras clave : ammonium; Neesler; analytic control; fermentation.

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