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J Chromatogr Sci. 2013 May-Jun;51(5):412-8. doi: 10.1093/chromsci/bms156. Epub 2012 Oct 18.

Simultaneous quantification of capsaicinoids and ascorbic acid from pungent peppers.

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  • 1Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, Texas A&M University, College Station, TX 77845, USA.


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.

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