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Muscular activity in terms of surface electromyography (sEMG) is usually normalised to maximal voluntary isometric contractions (MVICs). This study aims to compare two different MVIC-modes in handcycling and examine the effect of moving average window-size. Twelve able-bodied male competitive triathletes performed ten MVICs against manual resistance and four sport-specific trials against fixed cranks. sEMG of ten muscles [M. trapezius (TD); M. pectoralis major (PM); M. deltoideus, Pars clavicularis (DA); M. deltoideus, Pars spinalis (DP); M. biceps brachii (BB); M. triceps brachii (TB); forearm flexors (FC); forearm extensors (EC); M. latissimus dorsi (LD) and M. rectus abdominis (RA)] was recorded and filtered using moving average window-sizes of 150, 200, 250 and 300 ms. Sport-specific MVICs were higher compared to manual resistance for TB, DA, DP and LD, whereas FC, TD, BB and RA demonstrated lower values. PM and EC demonstrated no significant difference between MVIC-modes. Moving average window-size had no effect on MVIC outcomes. MVIC-mode should be taken into account when normalised sEMG data are illustrated in handcycling. Sport-specific MVICs seem to be suitable for some muscles (TB, DA, DP and LD), but should be augmented by MVICs against manual/mechanical resistance for FC, TD, BB and RA.