Background: Essential oils represent a major class of natural products which are known for their antimicrobial activity. This study aimed to determine the composition of four Piper essential oils by gas chromatography mass spectrometry, attenuated total reflection infrared, and chemometric analysis. Results: Monoterpene was the most predominant class in Piper nigrum and white pepper (87.6 and 80%, respectively) with the dominance of α-pinene, β-pinene, 3-carene, limonene, and βcaryophyllene. Sesquiterpenes represented 50, 19.6, and 12.3% of the essential oils of Piper longum, white pepper, and P. nigrum, respectively. Unlike other species, Piper cubeba oil was found to be rich in aromatics (59%), with eugenol (10.7%) and methyl eugenol (47.4%) representing the major components along with β-myrcene (21.2%) and 1,8-cineole (6.4%). Only P. longum essential oil comprised about 18.2% of alkanes and 13.6% of alkenes. Application of chemometric analysis utilizing GC/MS and ATR-IR data displayed the same segregation pattern where both principal component analysis and hierarchal cluster analysis revealed that white pepper was most closely related to P. nigrum while being completely discriminated from other Piper species. The Piper oils showed promising inhibitory effects on Helicobacter pylori. P. longum oil recorded the most efficient anti-Helicobacter activity [minimum inhibitory concentration (MIC) value of 1.95 μg/ml, which is the same as the MIC of clarithromycin], followed by the oil of white pepper (MIC = 3.90 μg/ml), while P. cubeba and P. nigrum produced the lowest activity (MIC value of 7.81 μg/ ml). Conclusion: Piper essential oils can be used as nutritional supplements or therapeutic drugs to protect against H. pylori infection.