Analysis of Odorants in Marking Fluid of Siberian Tiger (Panthera tigris altaica) Using Simultaneous Sensory and Chemical Analysis with Headspace Solid-Phase Microextraction and Multidimensional Gas Chromatography-Mass Spectrometry-Olfactometry

Scent-marking is the most effective method of communication in the presence or absence of a signaler. These complex mixtures result in a multifaceted interaction triggered by the sense of smell. The objective was to identify volatile organic compound (VOC) composition and odors emitted by total marking fluid (MF) associated with Siberian tigers (Panthera tigris altaica). Siberian tiger, an endangered species, was chosen because its MF had never been analyzed. Solid phase microextraction (SPME) for headspace volatile collection combined with multidimensional gas chromatography-mass spectrometry-olfactometry for simultaneous chemical and sensory analyses were used. Thirty-two VOCs emitted from MF were identified. 2-acetyl-1-pyrroline, the sole previously identified compound responsible for the “characteristic” odor of P. tigris MF, was identified along with two additional compounds confirmed with standards (urea, furfural) and four tentatively identified compounds (3-methylbutanamine, (R)-3-methylcyclopentanone, propanedioic acid, and 3-hydroxybutanal) as being responsible for the characteristic aroma of Siberian tiger MF. Simultaneous chemical and sensory analyses improved characterization of scent-markings and identified compounds not previously reported in MF of other tiger species. This research will assist animal ecologists, behaviorists, and zookeepers in understanding how scents from specific MF compounds impact tiger and wildlife communication and improve management practices related to animal behavior. Simultaneous chemical and sensory analyses is applicable to unlocking scent-marking information for other species

Determining an optimal method for the detection of odorous volatile organic compounds in tiger marking fluid in an effort to aid conservation

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 M