We investigated the effect of the position of electrodes relative to the innervation zone (IZ) of the biceps brachii muscle during isometric elbow flexion using eight-channel surface array electrodes. We estimated the location of the IZ near the centre of the muscle in 20 male subjects. The pulse peaks from electromyogram (EMG) waveforms were detected for each channel and averaged, the triphasic pulse was determined, and the peak values of the first and third phases were compared. The results showed significantly greater pulse values for the first phase when the electrode placement was proximal to the estimated IZ, and for the third phase when the electrode placement was distal to the estimated IZ. Using this method, the positional relationship between electrodes and IZ can be determined using a surface EMG waveform recorded with a pair of bipolar electrodes. This method may be clinically useful in confirming the reliability of a recorded surface EMG.