The aim of this study was to investigate whether L-Ascorbic acid would facilitate the anti-obesity effects of chitosan and psyllium husk in vivo. The study was carried out with male Hartley guinea pigs for 5 weeks. The results show that chitosan itself did not influence body weight gain and food efficiency ratio (FER). However, the addition of L-Ascorbic acid to chitosan decreased these parameters; the body weight gain and FER in the chitosan-2 group (high-fat diet group with 5 % chitosan containing 0.5 % L-Ascorbic acid) was significantly (p < 0.05) lower than that in F-controls (high-fat diet group), and was similar to that in controls (normal diet group). L-Ascorbic acid enhanced significantly (p < 0.05) the increases of total fecal weight and fecal fat excretion by chitosan. The addition of L-Ascorbic acid to psyllium husk did not differ from psyllium husk alone in terms of changes in weight gain, plasma lipid levels, and fat pad weight. We found that the addition of L-Ascorbic acid to chitosan influenced the reduction in body weight gain and FER, and the increase in total fecal weight and fecal fat excretion in guinea pigs fed a high-fat diet.