Abstract: The relatively mature horizontal axial measurement technique is difficult to obtain the non-disturbing layered acoustic properties of marine sediment samples in laboratory. The crosswise measurement technology can solve this problem. The research on this measurement technique is carried out by using water as the reference standard for seabed sediment measurement to calibrate the measurement system. The concept of equivalent transducer is proposed, the sound propagation medium other than the sample is integrated into a part of the equivalent transducer, the sound delay of the equivalent transducer is calibrated and the initial energy of sound propagation is measured. By taking water as the standard medium and comparing calibration results in measuring the commonly used Polyvinyl chloride (PVC) pipes of 75, 90, and 110 mm diameters with three acoustic transducers of 33, 80 and 100 kHz, the measured sound velocities are consistent with the standard calculated sound velocity, in which the error of measuring 90 mm diameter PVC pipe at 80 kHz frequency is the smallest. Based on the above calibration method, it is concluded that when the relative distance between the transducer pairs in the water-proof coupling method is 142.02 mm, the calibrated sound delay of the equivalent transducer is 45.88±0.08 μs, and the initial value of the sound propagation energy represented by the receiving voltage of the receiving transducer is 0.86±0.03 V. Based on the sound delay of the equivalent transducer and the initial value of the sound propagation energy, the calculated acoustic velocity and acoustic attenuation of seabed sediments are consistent with the acoustic properties of clay silt.