Based on the technology of near-infrared spectroscopy, a novel near-infrared moisture content determination instrument has been developed with three laser diodes of different wavelengths as light source, with one InGaAs detector as determination device, and with integrating sphere as sample holding device. Besides, LabVIEW virtual instrument program was employed for development of its operation instructions. This instrument has made full use of the integrating sphere system to remove the spectroscopic parts which are necessary in general spectrometers and has thus facilitated the improvement of system stability and signal noise ratio. Determination tests of moisture in sesame seeds and tea fresh leaves verified high precision of the instrument, showing that it can meet the primary requirement of moisture determination in practical production.