Background: Blood sample collection is the most frequent intrusive practise that hurts patients in hospital settings. A non-pharmacological and economical way to lessen pain during blood sample collection is the Valsalva Maneuver. Objectives; The aim of this study was to evaluate the effect of Valsalva Maneuver during blood sample collection. Methods: A quasi-experimental design (post-test only control group design). Self-structured questionnaire on Socio demographic information, clinical parameter and Wong baker face pain scale was used to observe the pain score of participants. The research population includes all the adult patients between the ages of 18 and 50 admitted in IPD. The sample size for the study comprises of 500 Participants who met the inclusion criteria. Purposive sampling technique was used to identify adult patients between the ages of 18 and 50 who were having blood sample collection admitted in IPD in IMS & SUM hospital, Bhubaneswar, Odisha. Results: The results revealed that there is significant reduction in pain during blood sample collection in experimental group with (p=0.000). No association was found between level pain and socio-demographic and clinical parameters. Discussion: The Valsalva manoeuvre is a non-invasive, non-pharmacological, and efficient way to lessen pain related to drawing blood samples. During the collection of blood samples, nurses should demonstrate the Valsalva manoeuvre to patients. In-service education programmes for nurses and students should be included by hospital administration in order to promote the non-pharmacological technique of pain alleviation during blood sample collection.