The objective of this study was to develop a simple and reproducible method for the qualitative and quantitative analysis of betalains in plasma samples, based on Solid Phase Extraction (SPE) and micro-high performance liquid chromatography coupled with mass spectrometry (micro-HPLC-MS/MS). The eight betalain compounds detected and quantified were characterized in the fortified rat blood plasma samples. The developed method showed a good coefficient of determination (R² = 0.999), good recovery, precision, and appropriate limits of detection (LOD) and quantification (LOQ) for these compounds. Application of this method for the treatment of rat plasma samples collected after the betalain preparation administration, for the first time, revealed the presence of native betalains and their metabolites in plasma samples. Moreover, among them, betanin (2.14 ± 0.06 µmol/L) and isobetanin (3.28 ± 0.04 µmol/L) were found at the highest concentration. The results indicated that the combination of an SPE method with a micro-HPLC-MS/MS analysis may be successfully applied for the determination of betalains in the blood plasma.
Keywords: betalains; micro-HPLC-MS/MS; plasma; solid phase extraction (SPE); validation method.