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The artificial sweetener isomalt is widely used due to its low caloric, non-diabetogenic and non-cariogenic properties. Although the sweetening potency of isomalt has been reported to be lower than that of sucrose, no data on the sensitivity of humans for this polyol are available. Using an up-down, two-alternative forced choice staircase procedure we therefore determined taste detection thresholds for isomalt in human subjects (n = 10; five females and five males) and compared them to taste preference thresholds, determined using a two-bottle preference test of short duration, in a highly frugivorous nonhuman primate, the spider monkey (n = 4; one female, three males). We found that both species detected concentrations of isomalt as low as 20 mM. Both humans and spider monkeys are less sensitive to isomalt than to sucrose, which is consistent with the notion of the former being a low-potency sweetener. The spider monkeys clearly preferred all suprathreshold concentrations tested over water, suggesting that, similar to humans, they perceive isomalt as having a purely sweet taste that is indistinguishable from that of sucrose. As isomalt, like most sweet-tasting polyols, may elicit gastric distress when consumed in large quantities, the present findings may contribute to the choice of appropriate amounts and concentrations of this sweetener when it is employed as a sugar substitute or food additive for human consumption. Similarly, the taste preference threshold values of spider monkeys for isomalt reported here may be useful for determining how much of it should be used when it is employed as a low-caloric sweetener for frugivorous primates kept on a vegetable-based diet, or when medication needs to be administered orally.