The Department of Biochemistry & Department of Medical Genetics, JSS Medical College and Hospital, JSS Academy of Higher Education & Research, Mysuru a Hands-on Workshop titled “Expanded Newborn Screening Using LC-MS/MS” on 9th May, 2024. During the hands-on workshop, four different sessions including a pre-workshop test, followed by a guest lecture, a hands-on demonstration, and a post-workshop test were conducted. The resource faculty Mr. Kankan Jana (Sciex LC-MS/MS Analyst) provided comprehensive insights into the working principle, hardware, and applications of LC-MS/MS technology. The lectures were followed by a demonstration of the instrumental tuning methodology and the expanded newborn screening (NBS) multiplex assay using the Sciex Triple Quad 4500 LC-MS/MS platform available at the Clinical Biochemistry Laboratory, JSS Hospital. The hands-on workshop was attended by 15 delegates including faculty, undergraduate and postgraduate students. They highly appreciated the informative lectures and efforts provided by the guest speaker, and organizing faculty and staff during the demonstration of the protocols. Results of the pre- and post-workshop evaluations and feedback from the participants indicated that the hands-on workshop was beneficial in providing a clear insight into the emerging, highly sensitive LC-MS/MS technology and its wide utility, especially in the clinical set-up. During the valedictory ceremony, the top two highest scorers of the pre- and post-workshop tests were appreciated with a souvenir by Dr. Akila Prashant. Mr. Hemanth Vikram P R (Research Scholar, JSS College of Pharmacy, Mysuru) secured first place, and Dr. Indumathi T (Postgraduate Student, Hassan Institute of Medical Sciences, Hassan) secured second place.