In Vitro anthelmintic evaluation of Gliricidia Sepium, Leucaena Leucocephala, and Pithecellobium dulce: Fingerprint analysis of extracts by UHPLC-Orbitrap mass spectrometry

Abstract

In the present work, the anthelmintic activity (AA) of ethanolic extracts obtained fromGliricidia sepium,Leucaena leucocephala, andPithecellobium dulcewas evaluated using the third-stage-larval (L-3) exsheathment inhibition test (LEIT) and egg hatch test (EHT) onHaemonchus contortus. Extracts were tested at concentrations of 0.3, 0.6, 1.2, 2.5, 5.0, 10, 20, and 40 mg/mL. The larval exsheathment inhibition (LEI) results showed thatG. sepiumachieved the highest average inhibition of 91.2%, compared with 44.6% forP. dulceand 41.0% forL. leucocephalaat a concentration of 40 mg/mL; the corresponding IC(50)values were 22.4, 41.7, and 43.3 mg/mL, respectively. The rates of egg hatching inhibition (EHI) at a concentration of 5 mg/mL were 99.5% forG. sepium, 64.2% forP. dulce, and 54% forL. leucocephala; the corresponding IC(50)values were 1.9 mg/mL forG. sepium, 3.9 mg/mL forP. dulce, and 4.3 mg/mL forL. leucocephala. The species extracts studied here were also analyzed by ultra-high performance liquid chromatography and Orbitrap high resolution mass spectrometry (UHPLC-Q/Orbitrap/MS/MS), resulting in the compounds' identification associated with AA. Glycosylated flavonoids and methoxyphenols were observed in all three species: fatty acids inG. sepiumandP. dulce; phenylpropanoids, anthraquinone glycosides, amino acids and glycosylated phenolic acids inG. sepium; and flavonoids inL. leucocephala.Comparatively,G. sepiumpresented a greater diversity of compounds potentially active against the control of gastrointestinal nematodes, which was associated with the results obtained in the applied tests.