Pain is a multidimensional experience that combines sensory, affective, and cognitive factors, causing its treatment to be extremely complicated and often ineffective. Mindfulness meditation has been shown to be an effective approach to decreasing pain. While these benefits have largely been seen over extensive, eight-week long training regimens, we have repeatedly shown that just three to four days of mindfulness meditation training can significantly reduce pain. Mindfulness-based analgesia after brief mental training is associated with multiple neural processes involved in the cognitive modulation of pain [i.e. rostral anterior cingulate cortex (rACC), orbitofrontal cortex (OFC)]. Further, the rACC and OFC are implicated in modulating efferent parasympathetic activity. Yet, the role of the parasympathetic nervous system (PNS) in meditative analgesia remains unknown. The primary objective of the present study was to determine if increased high frequency heart rate variability (HF HRV), a marker of higher PNS activity, is associated with mindfulness-based pain relief. 60 healthy volunteers (30 males; 30 females) were randomly assigned to receive a four-session (25 minutes per day) mindfulness meditation or placebo mindfulness (i.e. sham mindfulness meditation) intervention. Before and after the four-session training regimen, HF HRV and behavioral pain intensity and unpleasantness ratings were examined in response to noxious heat (49°C) and innocuous neutral series (35°C) to the right calf. We hypothesized that the analgesic effects of mindfulness meditation, but not sham mindfulness meditation, would be associated with increased HF HRV. After training, mindfulness meditation and sham mindfulness meditation significantly reduced pain intensity and unpleasantness ratings and increased HF HRV when compared to rest ( p < 0.001). Mindfulness-based reductions in pain unpleasantness (p < 0.05) were associated with increases in HF HRV. Sham mindfulness-based analgesia was not significantly associated with HF HRV. These findings suggest that mindfulness-based increases in HF HRV could be optimized to better treat clinical pain.