Introduction: Heart rate variability (HRV) analysis has given a non-invasive way for assessing cardiac autonomic control. Re duced HRV is an independent indicator of poor prognosis in both heart disease patients and the general population. Aim/Objective: To establish reliability and validity for Kubios HRV smart phone application. Materials and methods: Fifty (n =50) office workers volunteered for the study, with 10 females and 40 males ranging in age from 30 to 50 years (Mean age =38.4±5.6). To evaluate Intra and Inter-rater reliability and validity the of HRV parameter, R-R intervals comparing simultaneous recording from ECG and Kubios HRV app, at different three times (day1,day2 andday3) one day apart and by two different trained examiners in the same participants. Each participants rested in supine for 10 minutes prior to the assessment and was instructed to remain relaxed, breathe properly, and refrain from talking and sleeping during the meas urement. The RR intervals (R-R) were recorded during a 5 min period in supine position using ECG and polar H10 monitor with Kubios HRV smartphone application. Results: Kubios HRV smart phone application shows excellent reliability, Intra-rater (α =0.868) and Inter-rater (α =0.890) and Validity (r = 0.94) with R-R intervals calculated from ECG. Conclusion: The Kubios HRV smart phone application provides an accurate and reliable alternative to the ECG for acquiring inter-beat interval time series data. The Kubios HRV app gives an R-R interval that can be used for short term HRV analysis in office workers from steady resting conditions, supporting the selection of this approach of evaluation in research and clinical practise.