Natural products are gaining clinical significance in modern day health care systems to prevent diseases. Bitter melon, a health promoting vegetable, is traditionally used for medical nutrition therapy to cure diabetes but to reap maximum health claims, vigilant control of its substances in diet is crucial as part of curative action for effective diabetes management. In the present research, first phase focused on detection of key bioactive components, i.e., charantin and vicine in different parts of its fruit. In the second phase, normal and hyperglycemic Sprague Dawley rats were fed on skin, flesh and whole fruit of bitter melon at 150 and 300 mg/kg body weight and assessed for diabetes prophylaxis and treatment. The highest amount of charantin (0.16 ± 0.02 mg/g) was recorded in flesh while vicine was present in abundance in whole fruit (0.21 ± 0.01 μg/100 g). In normal rats, bitter melon supplementation was helpful in managing the onset of diabetes. Hyperglycemic rats showed diabetic complications including polydipsia, polyuria, glycosuria, renal hypertrophy and increased glomerular filtration rate. However, bitter melon consumption showed significant improvements in these parameters. The most potent dose was 300 mg/kg whole fruit that resulted in 31.64% lowering of blood glucose level and 27.35% increase in insulin level in hyperglycemic rats.