The effect of a selective estrogen receptor modulator – raloxifene – on the levels of tryptophan and kynurenic acid in the livers of rats as studied via RP-HPLC-FL

Abstract

The levels of tryptophan (TRP) and its main metabolite kynurenic acid (KYNA) were measured in rat livers treated with raloxifene – a selective estrogen receptor modulator. The research was conducted by applying high-performance liquid chromatography on a 5 μm Zorbax Eclipse XDB-C18 column. Selective fluorescence detection (FL) was performed at an excitation of 219 nm and emission of 360 nm for TRP and KYNA. The assays showed good linearity (R2 >0.95) within the tested ranges of 0.045-0.20 µg mL−1, 0.025-0.32 µg mL−1, respectively, for KYNA and TRP. The limits of the detection were found to be 0.1480 ng mL−1 for KYNA and 0.0332 ng mL-1 for TRP. The deproteinization of the liver homogenate samples was accomplished by 80% methanol addition combined with boiling precipitation. The average recovery values were between 94.84% and 99.54% with RSDs no more than 5.5%. The work revealed that raloxifene decreased the mean value of tryptophan, as compared with the control group, while simultaneously leaving kynurenic acid at the same level. For the first time the research suggests that, in the case of raloxifene therapy, tryptophan is not metabolized via the kynurenine pathway.