Green coffee fruit is rich in phenolic acids; however, the compounds’ effectiveness is dependent of their preservation. Besides preservation, microencapsulation technique can improve benefic effects and security of phytonutrients. Oral acute toxicity and lethal dose 50% (LD50) hydroethanolic extract from whole coffee fruit microencapsulated by spray dryer (MCE) was conducted under OECD Principles of Good Laboratory Practices (OECD, OCDE) 423, Paris, 2001). Male and female mice (n= 6/sex/dose), were treated with single dose by gavage of vehicle (water) or 321.47, 1637.35, 3214.7, 8036.75 and 16073.5 mg/kg bw of MCE equivalent to 11.753, 58.77, 117.54, 293.84 and 587.68 mg of 5-O-caffeoylquinic acid (5-CQA) and 8.74, 43.7, 87.4, 218.5, 437 mg of caffeine, respectively. The animals were monitored individually for clinical signs of toxicity or mortality by using the Hippocratic screen according to Malone & Robichaud (1962) in the open field 5, 10, 15, 30, 60, 120, 240 min and up to 48 hours after administration of ME, and daily for 14 days. Feed (PMI LabDiet Purina Certified Rodent Meal) and filtered tap water were provided ad libitum. Body weights, feed consumption and daily mean water consume were recorded regularly throughout the study and calculated. After 14 days, animals were euthanized by carbon dioxide asphyxiation and subjected to gross necropsy. Liver, heart, lungs, kidneys, stomach, and spleen were weight and relative weight of the organs was determined [(organ weight / body weight) x 100]. In Hippocratic screening, doses of MCE up to 16073.5 mg/kg bw did not cause any relevant behavioral changes or deaths thus making it impossible to establish the LD50. Mean of feed intake showed significant difference (p<0.05) only among male groups. Relative weight of liver of female treated with MCE ranging from 8036.75 to 16073.5 mg/kg bw showed a significant difference (p<0.05), but had no corresponding histopathological alteration.