Human breast milk plays a vital role in supplementation for infants and is influenced by maternal health, regular diet and regional conditions. Here we hypothesized that the nutritional composition of breast milk was altered based on maternal age, regular diet and regional locations that lead to new-born infant growth and development in early months. In this work, we collected nearly 60 human (22-33 age) breast milk samples within a 150 km radius in Hyderabad. All biochemical (protein, pH, total fat, lactose, iron, calcium and nitrogen) analysis was carried forward into a UV visible spectrophotometer at different wavelengths. Obtained results were showed there alterations in milk compositions are influenced by maternal especially dietary factors. Urban and rural mothers showed variations in all these biochemical analyses based on age, veg and non-veg food intake, area of living and several comorbidities. Overall these findings reveal that emphasizing the role of maternal supplementation and lifestyle in maintaining breast milk quality and effectiveness on infant growth and development provides a clear idea for targeted micro and macro nutritional interferences.