Comparative study of four matrix effect correction strategies for bile acid quantification in mouse serum

Endogenous substances are components produced by metabolism within the body, and they play a crucial role in disease diagnosis, treatment, and the regulation of physiological functions. However, it is difficult to obtain a blank matrix or substitute analytes, which leads to uncertainties in quantitative analysis. Common analytical strategies for evaluating matrix effects include standard addition, background subtraction, surrogate analyte, and surrogate matrix. It requires experimental studies to comprehensively compare these four strategies. This study employed three different types of serum matrices: authentic serum, artificial serum, and serum treated with activated carbon at different concentrations. Common endogenous compounds (bile acids) were selected as representative analytes, and quality control samples were prepared. calibration standard curves were established and calibrated according to the different analytical strategies. The four strategies were then comprehensively compared by evaluating the accuracy, precision, and matrix effects. The results demonstrated that the treatment method using activated carbon adsorption (at 0.1 g/mL) performed the best. Furthermore, based on the comparison of the four strategies, a complete method validation was performed for fifteen common bile acids, and the results met all the requirements for detection and analysis. This study provides an important reference for the absolute quantitative analysis of endogenous substances.