Introduction: The purity of the hemodialysis fluids is crucial for hemodialysis patients who are inevitably exposed to a large volume of water during hemodialysis. During this process the semi-permeable artificial membrane comes into direct contact with the bloodstream. Therefore it is important to monitor the purity of dialysis water. Aim and objectives: To compare two different methods for dialysis water analysis. Material and methods: 50 samples of dialysis water were collected from Saveetha Medical College and Hospital for this study. The study was conducted from 2018 December to 2019 March after getting Institutional Human Ethical Committee (IHEC) Clearance. By using the criteria of the AAMI, the present study is taken up to analyse the sensitivity of two different cultures technique i.e., spread plate and membrane filteration technique. Results: The standardization shows that spread plate technique was 80% effective and membrane filtration technique was 70% effective in identifying 100 CFU/mL of bacteria tested. Out of 50 unknown samples tested, 2 were ultra-pure, 21 were between 0.1-50 CFU/mL, 16 were between 50-100 CFU/mL and 11 were >100 CFU/mL by Spread plate technique. Likewise, 6 were ultra-pure, 30 were between 0.1-50 CFU/mL, 10 were between 50-100 CFU/mL and 4 were >100 CFU/mL by Membrane Filtration technique. Conclusion: From this study, spread plate technique proves to be equally sensitive with membrane filtration technique for analyzing dialysis water but when ultrapure water needs to be analyzed, spread plate technique gives much better bacterial recovery. i. e. , on l y 2 samples were proved to be ultra-pure by Spread plate technique.