Pepper known as the king of spices is one of the most widely used spice in the world, and it has its fair share of medicinal properties as well. It has been used in traditional medicine for many centuries owing to its medicinal properties apart from its flavour and pungent taste. Piperine is the most important bioactive compound present in different Piper species. Differences among the three Piper species common in South India viz. <i>Piper nigrum</i>, <i>Piper longum</i>, and <i>Piper cubeba</i> were evaluated by estimating the various phytochemicals such as alkaloids, flavonoids, and phenols in them. Further, the quantity of piperine in these three <i>Piper</i> species were determined using HPLC analysis. The piperine content in <i>Piper nigrum</i>, <i>Piper longum</i>, and <i>Piper cubeba</i> were found to be 235.05 μg/mL, 268.50 μg/mL, and 8.56 μg/mL respectively. Since inflammation is the major pathology involved in most of the disease conditions and since pepper is traditionally used in managing respiratory inflammation, we ventured to identify its anti-inflammatory potential. Herein, the anti-inflammatory properties of piperine were checked by in silico docking analysis of piperine with inflammatory proteins. The interaction of piperine with <i>NF</i>-κB, COX-2, COX-1, and <i>TNF</i>-α, as evidenced by the binding scores reveals that piperine interacts with them and modulates inflammation. Further <i>in-vitro</i> and in-vivo studies are warranted to scientifically validate the traditional claims.