Chemical communication plays an integral part of conserving tigers. Tigers, elephants, lions, and many other mammals use marking fluid (MF) and other excrements as means to communicate with each other and their ecosystem.

This research focuses on understanding which compounds eluted in tiger (MF) contribute to the overall odor of MF. Specifically, which compounds are responsible for behavioral responses. This study collected MF samples from four, sixteen year old Panthera tigris tigris from South Khayerbari Tiger Rescue Center in West Bengal, India. In this work, MF has been studied to gain a perspective on how tigers utilize MF for territorial and reproductive communication. Specifically, this study expanded upon previous research methodologies that characterized the odor of MF, of Panthera tigris tigris, based on thin-layer chromatography (TLC). TLC was utilized for the separation and identification of lipid compounds. Alkali was added to the TLC paper for the identification of compounds, specifically 2-acetyl-1-pyrroline (2-AP). This research study compared the concepts of TLC against solid-phase microextraction (SPME) and multidimensional gas chromatography mass spectrometry-olfactometry (mdGC-MS-O) for detection of odorous volatile organic compounds. TLC analysis of the samples was performed at the University of Calcutta and mdGC-MS-O work was performed at Iowa State University.

The mdGC-MS-O technique uncovered 19 odorous compounds including 2-AP, suggesting that 2-AP is not the only contributing odor to the smell of Panthera tigris tigris MF