Background The aim of this study was to observe and analyze vascular function in ‘prolonged sitting’, followed by a yoga asana routine and pranayama intervention. Participants in this study include those who work from desks in offices. The study required the participants to attend on three separate days at random, and they had to finish a computerized test on each day. On the first day, participants were required to complete a computer test while sitting still for four hours (with the exception of washroom breaks). The next day, they underwent a computerized test along with a pranayama intervention. Finally, on the last day, they underwent a computerized test along with a yoga asana intervention. At the start of the study and after two and four hours, we measured the diameter and velocity of the common carotid artery (CCA) and superficial femoral artery (SFA). Methods The study was a within-subjects prospective single-center trial conducted in the Department of Radio-Diagnosis and Imaging, Kasturba Medical Hospital, Manipal, India, between September 2022 and January 2023. Participants were asked to do one of the following ‘activities’ over successive weeks: Week 1 – Prolonged sitting; Week 2 – Pranayama intervention; and Week 3 – Yoga asana intervention during prolonged sitting. The baseline and follow-up variables of pulse velocity, endothelial thickness, and shear rate were assessed for normality through a Shapiro-Wilk Test. Results Our sample included 11 participants with moderate physical activity, five with high physical activity and one with low physical activity. Yoga asana intervention comprised participants sitting continuously for four hours, with a yoga asana intervention being provided every hour, lasting for 10 minutes. Conclusions Yoga asana improves vascular functions in prolonged sitting conditions. This routine can promote the concept of interrupted sitting and ways to reduce it with efficient yoga asana practice without changing the work culture and provide better physical relief. Trial registration Clinical Trials Registry – India ( CTRI/2022/09/045628), date of registration: 19/09/2022(CTRI/2022/9/045628)https://ctri.nic.in/Clinicaltrials/main1.php?EncHid=16349.27799,