Ayurveda scholars have given comprehensive descriptions about characteristics of pure and vitiated human milk, measures of its purification, effect of diet, lifestyle on quality of human milk and its effect on growth and development of infant. Recent researches have also shown that the variation in human milk composition is associated with the maternal diet, environment, and potentially with genetic factors. Studies have reported that individual mothers within species often demonstrate variation in milk composition and/or yield affecting infant growth and development. Prakriti refers to genetic account of an individual, so there may be variation in human milk composition because of metabolic peculiarity and dietary differences. To analyze the composition of pure and vitiated Human milk as per Prakriti. Total 200 healthy lactating Indian mothers, aged between 20 to 35 years were registered after getting informed written consent from OPD of Department of Kaumarbhritya, I.M.S, BHU, Varanasi, India during year 2015-2017. Analysis of milk for fat, SNF, density, protein, lactose, freezing point, conductivity and pH was done through Eco milk Analyzer. The vitiation of milk was assessed by water test mentioned in Ayurveda text viz. Charak Samhita, Sushruta Samhita, Ashtanghridyam, Kashyap Samhita etc. Assessment of Prakriti (Psychosomatic constitution) of mothers was done by validated Performa.The mean values of different constituents viz. Fat(%),SNF(%),Density( g/cm3),Protein (%) and Freezing Point(°C) were more in vitiated milk as compared to pure human milk. The variation in composition of milk was also found as per Psychosomatic constitution, the mean values of fat percentage in milk varied significantly as per Prakriti (p=0.0287). On applying Post Hoc Test, the significant pairs were for Vata v/s Pitta (<0.05) and Vata v/s Kapha (<0.05) Prakriti.Present study suggests that the composition of human milk varies as per Prakriti and in vitiated state too. Thus by correction of diet and lifestyle as per Prakriti, the quality of breast milk may be improved and better growth of infants may be obtained by using medicines for purification of milk.