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This paper reports the preliminary results of a pilot study testing the efficacy of a new experimental protocol to measure the psychophysiological basis of entrepreneurial intuition—that part of entrepreneurial decision and action that is not based on reason or memory, but on an awareness of energetically-encoded information of the future. A multi-methods approach incorporating electrophysiological measures of the autonomic nervous system (ANS) was used to investigate intuition in a sample of 8 serial entrepreneurs with usable data from the Cambridge Technopol. The results are promising: the mean pattern of recordings for all subjects show that informational input was received by the ANS some 6 to 7 seconds before the outcome of the investment choice was known. Also random permutation analysis of individual recordings identified five instances in which the physiological measures had significant or marginally significant predictive power in discriminating future win/loss outcomes. Overall, the physiological measures were able to detect intuitive perception of a future outcome in four of the eight entrepreneurs. However, some improvements to the protocol’s design and data collection procedures (when used in non-laboratory field settings) will be necessary to optimize the protocol’s targeting and measurement of entrepreneurial intuition.