A simplified high-performance liquid chromatography method to measure skatole and indole in porcine plasma without the use of acetonitrile was developed and validated in this study. The mobile phase consisted of water and methanol used in a gradient programme. Fluorescence detection was performed on the supernatant obtained from plasma after protein precipitation with 100% acetone. Limits of quantification were 0.5 ng∙ml^-1 for skatole and 1.0 ng∙ml^-1 for indole. Accuracy and precision had less than 12% deviation in the linear ranges (0.5–256 ng∙ml^-1 and R² = 0.9999 for skatole, 1.0–256 ng∙ml^-1 and R² = 0.9999 for indole). The correlation between plasma and serum concentrations was strong for skatole (slope = 1.01, R² = 0.999) and moderate for indole (slope = 0.65, R² = 0.95). Analysis of skatole in plasma was in good accordance with our previous acetonitrile-based method (slope = 0.91, R² = 0.988). The proposed method is suited for rapid routine analysis because of its high selectivity, accuracy and precision. Furthermore, it needs only simple sample preparation and the use of methanol instead of acetonitrile in the mobile phase. This method is of practical use to researchers in the field of boar taint.