Effects of Echinostoma caproni infection on the neutral and polar lipids of intestinal and non-intestinal organs in the BALB/c mouse as determined by high-performance thin-layer chromatography

Abstract

The goal of this study was to characterize and quantify the various neutral and polar lipid classes in the BALB/c mouse that are associated with Echinostoma caproni infection. Ten infected mice and 10 uninfected control mice were used for this study (five infected and five uninfected were used for each of the neutral lipid and polar lipid studies). After 3 weeks postinfection, the mice were necropsied and various organs were removed and prepared for lipid class analysis. The organs used were liver, kidney, spleen, colon, cecum, anterior portion of the small intestine (SI), middle portion of the SI, and posterior portion of the SI. Lipids were determined by high-performance thin-layer chromatography (HPTLC) with Analtech 10 × 20 cm HPTLC-HLF silica gel plates. For neutral lipids, petroleum ether-diethyl ether-glacial acetic acid (80:20:1) mobile phase and 5% ethanolic phosphomolybdic acid detection reagent were used to determine the neutral lipids in each organ. Chloroform-methanol-deionized water (65:25:4) mobile phase and 10% cupric sulfate in 8% phosphoric acid detection reagent were used to determine the polar lipids in each organ. The analyzed polar lipids in all organs were phosphatidylcholine (PC), phosphatidylethanolamine (PE), and sphingomyelin (SM). Using HPTLC-densitometry for quantification, PC was found in the greatest amount and SM the smallest of all organs analyzed. The PE in the anterior portion of the SI was determined to be significantly greater (using the Student’s t test with P < 0.05), with about twice the amount of PE in mice infected with E. caproni relative to the uninfected mice. No significant differences in any of the neutral lipid classes were found between infected and uninfected samples.