A novel model extended from the Bouguer-Lambert-Beer law can describe the non-linear absorbance of potassium dichromate solutions and microalgae suspensions.

Introduction: The Bouguer-Lambert-Beer law is widely used as the fundamental equation for quantification in absorption spectroscopy. However, deviations from the Bouguer-Lambert-Beer law have also been observed, such as chemical deviation and light scattering effect. While it has been proven and shown that the Bouguer-Lambert-Beer law is valid only under very restricted limitations, there are only a few alternatives of analytical models to this law. Based on the observation in the experiments, we propose a novel model to solve the problem of chemical deviation and light scattering effect. Methods: To test the proposed model, a systematic verification was conducted using potassium dichromate solutions and two types of microalgae suspensions with varying concentrations and path lengths. Results: Our proposed model demonstrated excellent performance, with a correlation coefficient ( R 2 ) exceeding 0.995 for all tested materials, significantly surpassing the Bouguer-Lambert-Beer law, which had an R 2 as low as 0.94. Our results confirm that the absorbance of pure pigment solutions follows the Bouguer-Lambert-Beer law, while the microalgae suspensions do not due to the light scattering effect. We also show that this scattering effect leads to huge deviations for the commonly used linear scaling of the spectra, and we provide a better solution based on the proposed model. Discussion: This work provides a powerful tool for chemical analysis and especially for the quantification of microorganisms, such as the concentration of biomass or intracellular biomolecules. Not only the high accuracy but also the simplicity of the model makes it a practical alternative to the existing Bouguer-Lambert-Beer law.