Simultaneous quantification of capsaicinoids and ascorbic acid from pungent peppers

The development of simultaneous extraction and determination of bioactive molecules from natural products is becoming more popular. The present study reports the development of a method for the simultaneous extraction and determination of both capsaicinoids and ascorbic acid in peppers. Capsaicin (341.61 µg/g), dihydrocapsaicin (119.91 µg/g) and ascorbic acid (2,109.60 µg/g) were extracted with 3% metaphosphoric acid-ethanol (2:8) as a solvent. The efficient extraction of capsaicinoids (412.61 µg/g) and ascorbic acid (2,785.93 µg/g) was achieved at a sample-to-solvent ratio of 1:8 after 30 minutes of sonication. Simultaneous separation of capsaicinoids and ascorbic acid was achieved using a Gemini C18 column with a gradient elution of 0.03M phosphoric acid and methanol. Capsaicinoids and ascorbic acid were simultaneously detected at 282 and 254 nm, respectively. The recovery of capsaicinoids ranged from 96.21 to 108.71%, and the recovery of ascorbic acid ranged from 97.01 to 98.83%. The limits of detection for capsaicin, dihydrocapsaicin and ascorbic acid were 0.24, 0.21 and 0.26 µg, respectively. Relative standard deviation for the intra-day and inter-day variability in the results was less than 3%, indicating that the method produced highly reproducible results. Therefore, this method enables the reproducible, simultaneous separation and quantification of capsaicinoids and ascorbic acid from peppers.