A method for the determination of Rhodamine B in dry chilli, fruit drink and red wine was developed and validated. The analyte was extracted with acetonitrile from foodstuffs. A novel electrospun polymer nanofibers packed micro. column was used for cleaning and concentrating the Rhodamine B in these foodstuffs. High performance liquid chromatography with fluorescence detection( HPLC-Flu ) was used for the determination of Rhodamine B in the samples. The mobile phase was composed of 3 g/ L phosphate buffer and methanol( 3: 7, volume ratio ), the pH was adjusted to 7. 0 with orthophosphoric acid. The results showed that the standard curve was linear over the validated concentrations range of 1-100 ng/ mL( ng/ g), and the limit of detection( LOD ) for Rhodamine B spiked samples was 0. 1 ng/ g in dry chilli, 0. 2 ng/ mL in both fruit drink and red wine. The limit of quantitation( LOQ ) for Rhodamine B spiked into dry chilli was 0. 6 ng/ g, fruit drink 0. 5 ng/ mL and red wine 0. 5 ng/ mL, respectively. The average recoveries of Rhodamine B were 98. 2%-110. 3% for dry chilli, 94. 6%-102. 2% for fruit drink, and 90. 4%-104. 6% for red wine, respectively. The relative standard deviation of the method was 2. 3%-9. 0%. This proposed method was applied to real samples including dry chilli, fruit drink as well as red wine, and there was no Rhodamine B found in fruit drink or red wine, but Rhodamine B was found in chilli powder.