Bioimpedance spectroscopy (BIS) is a detection technology that uses the bioimpedance characteristics and changes law of human tissue to analyze its physiological and pathological states, and is widely used in clinical and scientific research applications. Traditional BIS measurement needs to satisfy the Nyquist sampling theorem, so as to ensure that the measurement signal has no frequency aliasing, but at the same time, the sampling frequency and the number of sampling points will be increased, which will increase the computation and hardware cost. This paper proposes a novel BIS measurement method based on multisine excitation and integer-period undersampling (IPUS) technology. Firstly, the multisine-based IPUS theory was deduced, and the BIS measurement principle based on multisine excitation and IPUS technology was introduced. Secondly, a BIS measurement system based on FPGA+ADC+DAC architecture was designed, and a multisine excitation with 32 pseudo-logarithmically distributed frequency components in the range of 2-997 kHz was generated. The comparative BIS measurement experiments on three RC three-element models were carried out under the Nyquist sampling condition (fs = 2.56 MHz) and under the IPUS condition (fs = 512 kHz), respectively. Experimental results showed that the average amplitude error of BIS measurement under the Nyquist sampling condition is 0.80% (mean) ± 1.19% (std), while the average amplitude error under the IPUS condition is 1.02% (mean) ± 1.13% (std). Moreover, the signal to nosie ratio (SNRz) is calculated in 40 repeated BIS measurements, where the mean SNRz is 63.60dB under IPUS condition, similar to the value of 62.77dB under Nyquist sampling condition. The proposed multisine-based IPUS theory and its implementation method in this paper can complete a BIS measurement with only one fundamental period, and need the sampling frequency and sampling points lower than the requirements of the Nyquist theory, which lay a theoretical and technical foundation for BIS measurement system with reduced hardware requirements and computation cost.