FULL COMMUNICATION Evaluation of the Antihyperglycemic Effect of Minor Steviol Glycosides in Normoglycemic and Induced-Diabetic Wistar Rats Irma Aranda-Gonza ´lez, 1,2 Yolanda Moguel-Ordo ´n ˜ez, 3 Luis Chel-Guerrero, 2 Maira Segura-Campos, 2 and David Betancur-Ancona 2 1 Faculty of Medicine, Autonomous University of Yucatan, Merida, Mexico. 2 Faculty of Chemical Engineering, Autonomous University of Yucatan, Merida, Mexico. 3 National Institute of Forestry, Agricultural and Livestock Research, Mococha Experimental Campus, Mococha, Mexico. ABSTRACT Steviol glycosides are a family of compounds found in Stevia rebaudiana Bertoni that are responsible for sweetness capacity. The antihyperglycemic effect of the two major steviol glycosides, Rebaudioside A and Stevioside, has been studied and it has been found that despite having the same common structure, only Stevioside exerts an antihyperglycemic effect. Although other steviol derivatives are found in smaller amounts (minor steviol glycosides) in S. rebaudiana, whether or not they possess antihyperglycemic activity has not been evaluated. The aim of this study was to evaluate the antihyperglycemic effect of minor steviol glycosides in normoglycemic and diabetic (streptozotocin/nicotinamide) Wistar rats. Rats were subjected to an intraperitoneal glucose tolerance test (IPGTT) both before and after chronic treatment (28 days). After 6 h of fasting, IPGTT was conducted in pentobarbital-anesthetized rats using 1 g/kg of glucose plus 20 mg/kg of the minor glycoside (Dulcoside A, Rebaudioside B, C, D, or Steviolbioside) or control treatment (distilled water, glibenclamide, or metformin); the blood of the tip of the tail was collected at time 0, 15, 30, 60, and 120min.; and blood glucose was measured, and its net area under the curve (AUCnet) was calculated. After 28-day chronic oral administration, IPGTT was again performed. Differences were considered significant at P < .05 by one-way ANOVA. Acute intraperitoneal or chronic oral administration of 20 mg/kg of minor steviol glycosides had no antihyperglycemic effect in normoglycemic or induced-diabetic Wistar rats. Considering the dose tested, it is unlikely that these glycosides have an effect on glucose in diabetic or normoglycemic humans. KEYWORDS:  antihyperglycemic  dulcoside  rebaudioside  Stevia rebaudiana  steviol glycosides  steviolbioside INTRODUCTION S tevia rebaudiana Bertoni is a plant belonging to the genus Stevia, which are native to tropical and subtropical regions of South and Central America and possess a high sweetening capacity. 1 A family of compounds derived from steviol, named steviol glycosides, is responsible for this sweetener property. Steviol is glycosylated at C-19 and C-13 and replaces the carboxyl hydrogen atom of the hydroxyl group with combi- nations of glucose, xylose, and rhamnose, 2 leading to dif- ferent steviol glycosides. To date, nine steviol glycosides have been fully identified by the Joint FAO/WHO Expert Committee for Food Additives, which are: Stevioside, Re- baudioside A, Rebaudioside B, Rebaudioside C, Rebaudio- side D, Rebaudioside E, Rebaudioside F, Dulcoside A, and Steviolbioside. 3 Among the steviol glycosides, Stevioside and Rebaudioside A are found in greater quantities, whereas the remaining glycosides can be found in small quantities in the leaves of S. rebaudiana. 1,4,5 In 2008, the food and drug administration in the United States granted the status to Rebaudioside A of ‘‘generally recognized as safe’’ for use as a sweetener and not only as a supplement. 6 In the same year, the JECFA and The Food Standards Australia and New Zealand (FSANZ) established the acceptable daily intake (ADI) at 0–4 mg/kg bw per day (based on steviol equivalents). 6 Several studies have shown that Stevioside has an anti- hyperglycemic effect in Goto-Kakizaki rats, with increased insulin and decreased glucagon production, 7–9 or even a hypoglycemic effect when administered subcutaneously. 10 Moreover, Stevioside has also shown an antihyperglycemic effect in people with diabetes 11 ; notably, Stevioside ex- erts its effect on glucose at doses of 5 mg/kg, just above the ADI. 12 Interestingly, although both Rebaudioside A and Ste- vioside are members of the steviol glycoside family, Rebaudioside A does not have the same effect on GK rats, 13 or humans, 14 when tested at the same dose as Stevioside. Manuscript received 1 February 2016. Revision accepted 5 July 2016. Address correspondence to: Dr. David Betancur-Ancona, PhD, Facultad de Ingenierı´a Quı´mica, Universidad Auto´noma de Yucata´n, Perife´rico Norte Km. 33.5, Tablaje Cat- astral 13615, Col. Chuburna´ de Hidalgo Inn, Me´rida, Yucata´n, C.P. 97203, Mexico, E-mail: bancona@correo.uady.mx JOURNAL OF MEDICINAL FOOD J Med Food 00 (0) 2016, 1–9 # Mary Ann Liebert, Inc., and Korean Society of Food Science and Nutrition DOI: 10.1089/jmf.2016.0014 1