Traditionally, the estimation of oxygen consumption (VO2) at work using heart rate (HR) has required the determination of individual HR/VO2 calibration curves in a separate exercise test in a laboratory (VO2-TRAD). Recently, a new neural network-, and heart rate variability-based method has been developed (Firstbeat PRO heartbeat analysis software) for the estimation of VO2 without individual calibration (VO2-HRV). In the present study, the VO2-values by the VO2-HRV were compared with the values by VO2-TRAD in 22 postal workers. Within individuals the correlation between the two methods was high (range 0.80-0.99). The VO2-TRAD gave higher values of VO2 compared to VO2-HRV (19%) especially during low physical activity work when non-metabolic factors may increase HR. When assessed in different HR categories, the smallest difference (11%), and highest correlations (range 0.83-0.99) in VO2 between the methods were observed at higher HR levels. The results indicate that the VO2-HRV is a potentially useful method to estimate VO2 in the field without laboratory calibration.