Fourier-transform infrared (FT-IR) spectroscopy method for measuring small microplastic (SMP) concentration in marine environment is time-consuming and labor-intensive due to sample pre-treatment. In contrast, Raman spectroscopy is less influenced by water and can directly measure SMP samples in water, making it a more efficient method to measure SMP concentration. Therefore, a method that can directly estimate the concentration of SMPs in water was developed, and the relationship between SMP concentration and experimental Raman spectra were established by testing with standard polyethylene (PE) samples. It was found that average spectra acquired in water solution could reflect characteristic peaks of the plastic after baseline correction. Further investigation found that there is a significant functional relationship between correlation coefficient of sample spectra and the concentration of PE particles, and such relationship can be modelled by Langmuir model. The empirical functional relationships can be used to estimate SMP concentrations by measuring average Raman spectra. The developed methodology is helpful for developing rapid SMP identification and monitoring methods in a more complex manner.•A method of directly measuring MP concentration in water is proposed.•Experimental procedures are provided.•Data analysis methods are outlined.