A validated method for the thin-layer chromatography in situ densitometric quantitation of capsaicinoids in Habanero pepper (Capsicum chinense Jacq.)

A thin-layer chromatography method for the separation and in situ densitometric quantitation of total capsaicinoids from dried Habanero pepper pods was developed, following selected guidelines from the International Council for Harmonisation. The method was sensitive enough to detect and quantify 0.5 and 0.75 μg of total capsaicinoids, respectively, from minimal amounts of plant tissue and little sample processing. Method displayed precision, with under 5% variation both at intra- and inter-day analysis; accuracy, as comparable results were obtained using other validated methods; and robustness, estimated by modifying analysis conditions. Due to its simple execution, up to 20 analyses could be performed from beginning to end in less than two hours.