All samples were first dried, then extracted using the method of Collins et al. [30] with slight
modifications. For capsaicinoid extraction, each dried pepper sample (5 g) was placed in ethanol
(5 mL) in a 120 mL glass bottle equipped with a Teflon lined lid. Bottles were capped and placed in a
water bath at 80 °C for 4 hours, then swirled manually every hour. Samples were removed from the
water bath and cooled to room temperature. The supernatant layer of each sample (5 mL) was filtered
through 0.45 μm filter paper into a HPLC sample vial using a 5 mL disposable syringe (Millipore,
Bedford, MA, USA). The vial was capped and stored at 5 °C in a refrigerator until analysis.