Formulations for Protecting the Probiotic Saccharomyces boulardii from Degradation in Acidic Condition

Abstract

Saccharomyces boulardii is a nonpathogenic yeast with proven health benefits, some of them depending on its viability. However, the living yeast is sensitive to environmental conditions and its viability is less than 1% in the faeces after oral administration. Therefore, we assessed the survival conditions of S. boulardii in aqueous suspension and in its freeze-dried form and we formulated microspheres with the former and tablets with the latter in order to preserve the viability of the probiotic. While the viability of the yeast in aqueous suspension could be maintained for one year at ؊20°C and ؉5°C, increasing the temperature led to almost total mortality within 14 d at ؉40°C and 4 d at ؉60°C. The viability of the freeze-dried yeast was preserved for one year at ؉25°C without moisture. With 75% relative humidity, the mortality was significant at 28 d at ؉25°C and almost total within 1 d at ؉60°C. In vitro, whereas less than 1% of non-encapsulated or non-tabletted S. boulardii survived after 120 min at pH 1.1, both formulations in microspheres and direct compression enabled to protect the yeast from degradation in HCl and to release it viable at pH 6.8. However, despite a similar release profile from both dosage forms, the compression led to a significant decrease in the viability of the freeze-dried yeast. In conclusion, although both formulations are efficient in protecting S. boulardii in acidic condition, microspheres provide a higher entrapment efficiency and a faster release of the viable probiotic in intestinal condition than matrix tablets.Key words microsphere; tablet; Saccharomyces boulardii; viability conditions; probiotic Biol. Pharm. Bull. 31(2) 266-272 (2008) © 2008 Pharmaceutical Society of Japan * To whom correspondence should be addressed. e-mail: sandrine.graff@univ-paris5.ft Assessment of the Survival of S. boulardii under Different Handling/Storage Conditions Samples of freeze-dried yeast (batch 4) were stored at ϩ25 or ϩ60°C either under aerobic conditions with 75% humidity or with no relative humidity, respectively (using saturated NaCl solution to maintain a constant humidity within the enclosure, or silicate gel to maintain the enclosure without humidity) or anaerobic conditions (nitrogen) for one year. A control sample was stored in ambient conditions for one year (temperature between ϩ15 and ϩ35°C and relative humidity from 35 to 90%). At 0,1,4,7,14,28, 90, 180 d and one year, the freezedried yeast was sampled to evaluate its viability.Samples of the aqueous suspension (batch 1) were stored at Ϫ20, ϩ5, ϩ25, ϩ40 and ϩ60°C under aerobic or anaerobic conditions for one year. At the times cited above, the aqueous suspension was sampled to assess the yeast viability. All the experiments were carried out in triplicate.Determination of Viable Yeasts The viability of the yeast has been defined as the proportion of the cells that are capable of multiplication after incubation in standard microbiological conditions. It was evaluated after serial 10-fold dilutions by a plate culture method u...

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