Biomarker discovery is relying on the sensitivity and specificity of the detected biomolecules in clinical samples. Several potential biomarkers against the particular disease cannot be detected because of the unavailability of either specific procedure or sensitive instrument or both, which hamper the diagnosis. In spite of that some of the potential biomarkers are available in trace amount in biological fluids like serum, urine, buccal swab, sputum etc. and need a sensitive method to detect precisely those biomolecules. Now days Selected Reaction Monitoring (SRM) is Mass Spectrometry based approached who can overcome the problems associated with biomarker discovery. India have adequate and variety of clinical samples but some time due to lack of infrastructure and knowledge we are unable to utilise those samples for human welfare. In this review, we are discussing about the experimental setup and procedure of SRM experiment.