Background: Pasteurization is the most crucial step for donor milk processing in human milk banks and Holder pasteurization is the most widely accepted method. Automated instruments are available for pasteurization but they are costly. Shaker water bath is also used for pasteurization which is a cheaper device. In this study we checked the quality of pasteurization carried out in shaker water bath. Objective: Evaluation of biological safety of pasteurized human Milk by Shaker water bath and to see if pasteurization using shaker water bath alters macro-nutrient values of milk using human milk analyzer. Methods: (a) Bacteriologic quality of post pasteurized milk of 80 pasteurization cycles by shaker water bath. (b) Macro-nutrient values of pre and post pasteurized milk samples from shaker water bath are measured from 20 different cycles using Human Milk Analyzer (MIRIS AB, Sweden). These 20 samples were selected using computer generated random table. Results: Bacteriologic testof pre pasteurized sampleswere comparable and colony count of all samples were within acceptable limit (Gram negative bacilli <104, Gram positive cocci <104). None of the post pasteurized samples showed any growth. Shaker water bath pasteurization did not alter macro nutrient values (protein: 1.56±0.23g/dl, fat: 3.20±0.35g/dl, energy: 64.00±3.08Kcal/dl). Conclusion: Shaker water bath is a low cost pasteurization device for Holder pasteurization of human milk; there is no alteration of milk quality and is bacteriologically safe as well.