A method was developed and validated for the determination of capsaicin in real chili samples collected from the market by reverse phase HPLC. The recovery data were obtained by spiking blank samples of reagent methanol with capsaicin at concentration levels of 10.0 μg/ml, yielding recoveries in the range of 90 – 110%. Precision values expressed as relative standard deviation (RSD) were in the range of 1.62 – 13.18%. Linearity was studied in the range 10 – 200 μg/ml and the coefficient of correlation was higher than 0.98. for all capsacinoid compounds. Method Detection Limits (MDLs) and Limits of Quantification (LOQs) were established. The overall uncertainty of the method was estimated. About 20 chili samples collected from the market were analysed for capsaicin content. The pungency of these samples expressed in SHU units were also determined. According to the validation data and performance characteristics as well as the high sample throughput, the proposed method is suitable for routine application. The capsaicinoids content and Pungency of the samples were between 0.17 – 0.82% and 25500 -124290 SHU, respectively.