Melatonin is a neurohormone that regulates circadian rhythms in the human body. It can also be taken to alleviate insomnia and sleep disorders. Pasteurized milk is a good source of nutrients and some bioactive compounds. Recently, the growing trend of healthy foods has resulted in higher competition with regard to milk products. Functional milk has been developed with higher bioactive compounds to respond to consumer demand. High melatonin pasteurized milk was developed using selected edible grains and mulberry leaves to fortify melatonin in pasteurized milk. Melatonin and free tryptophan of fourteen edible grains and mulberry leaves were determined using HPLC-FD. Highest melatonin concentrations were observed in white sesame, sunflower and soybean (75.24, 67.45 and 56.49 ng/g dry weight (dw), with highest concentrations of free tryptophan in soybean, red bean and mung bean (2617.83, 1527.23 and 845.27 ng/g dw, respectively), while melatonin and free tryptophan contents in fresh mulberry leaves were 51.57 ng/g and 210.53 ng/g dw, respectively. Soymilk powder and mulberry leaf tea were supplemented to prepare high melatonin pasteurized milk. Results showed that chemical compositions, melatonin and free tryptophan contents significantly increased (P < 0.05) with increasing amounts of soymilk powder and mulberry leaf tea. Sensory evaluation gave highest overall liking score for the treatment consisting of mulberry leaf tea (4.00%), soymilk powder (4.00%) and milk (89.80%). Findings indicated that mulberry leaves and soybean are both good sources of melatonin and free tryptophan and can be applied to prepare high melatonin pasteurized milk.