Profiling the Triacylglyceride Contents in Bat Integumentary Lipids by Preparative Thin Layer Chromatography and MALDI-TOF Mass Spectrometry

Abstract

The mammalian integument includes sebaceous glands that secrete an oily material onto the skin surface. Sebum production is part of the innate immune system that is protective against pathogenic microbes. Abnormal sebum production and chemical composition are also a clinical symptom of specific skin diseases. Sebum contains a complex mixture of lipids, including triacylglycerides, which is species-specific. The broad chemical properties exhibited by diverse lipid classes hinder the specific determination of sebum composition. Analytical techniques for lipids typically require chemical derivatizations that are labor-intensive and increase sample preparation costs. This paper describes how to extract lipids from mammalian integument, separate broad lipid classes by thin-layer chromatography, and profile the triacylglyceride contents using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This robust method enables a direct determination of the triacylglyceride profiles among species and individuals, and it can be readily applied to any taxonomic group of mammals.

Glycerophospholipid Analysis of Eastern Red Bat (Lasiurus Borealis) Hair by Electrospray Ionization Tandem Mass Spectrometry

“…With over 100 phospholipids known from mammals (Yamashita et al 2014), cataloging GPs present on fur, combined with sebum composition from other body regions (Pannkuk et al 2012, 2013a, 2013b), is a critical first step before making and testing predictions about the role of these lipid molecules in the ecology of free-living animals. L. borealis is a particularly interesting species for investigation due to a number of ecological and life history factors.…”

“…Hair of L. borealis (1.0 g) was removed with scissors and lipid was immediately isolated with chloroform:methanol (C:M) as previously described (Pannkuk et al 2012; 2013b) (Arkansas State University’s Institutional Biosafety Committee approval # 135349-1). This relatively non-destructive method minimized integral hair lipids rather than sebaceous secretions present in the sample.…”

High-Throughput Lipidomic and Transcriptomic Analysis To Compare SP2/0, CHO, and HEK-293 Mammalian Cell Lines

“…The samples were separated with a CHCl 3 –MeOH–H 2 O (71:25:2.5, v/v/v) solvent system and then rotated 90° and developed on a second dimension separation with a CHCl 3 –MeOH–acetic acid–H 2 O (76:9:12:2, v/v/v/v) solvent system . Another method based on 1D-HPTLC was implemented using hexane–diethyl ether–acetic acid (80:20:1.5, v/v/v) for nonpolar lipids separation . The lipid bands were visualized under the UV lamp with a 0.2% ethanol solution of 2,7-dichlorofluorescein dye .…”

Glycerophospholipid Profiles of Bats with White-Nose Syndrome

“…2a,c,e) or necrotic (inoculated bats; fig. 2b,d,f was used for targeting necrotic tissue) tissue sections were excised, total lipid was extracted with chloroform:methanol as previously described (Pannkuk et al 2013), then solvent was evaporated under a stream of N 2 and stored at −0°C. All storage solvents contained 0.05% butylated hydroxytoluene (Law et al 1995) and were of HPLC grade (Fisher Scientific, U.S.A.).…”