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 from Gliricidia sepium, Leucaena leucocephala, and Pithecellobium dulce was evaluated using the third-stage-larval (L3) exsheathment inhibition test (LEIT) and egg hatch test (EHT) on Haemonchus 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 that G. sepium achieved the highest average inhibition of 91.2%, compared with 44.6% for P. dulce and 41.0% for L. leucocephala at a concentration of 40 mg/mL; the corresponding IC50 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% for G. sepium, 64.2% for P. dulce, and 54% for L. leucocephala; the corresponding IC50 values were 1.9 mg/mL for G. sepium, 3.9 mg/mL for P. dulce, and 4.3 mg/mL for L. 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 in G. sepium and P. dulce; phenylpropanoids, anthraquinone glycosides, amino acids and glycosylated phenolic acids in G. sepium; and flavonoids in L. leucocephala. Comparatively, G. sepium presented a greater diversity of compounds potentially active against the control of gastrointestinal nematodes, which was associated with the results obtained in the applied tests.

Anthelmintic Effect of Leucaena leucocephala Extract and Its Active Compound, Mimosine, on Vital Behavioral Activities in Caenorhabditis elegans

“…The egg-laying activity in this study was also found to be significantly reduced after exposure to LL extract and mimosine, as demonstrated by the decrease in the mean number of eggs laid. Similar research findings concordantly showed the inhibition of egg-laying and -hatching post-treatment with LL extracts in different nematode species [ 30 , 32 , 74 , 75 , 76 , 77 , 78 , 79 , 80 ]. For instance, a recent study by Romero et al (2020) [ 76 ] reported that the ethanolic extract of LL leaves significantly inhibited egg-hatching activity up to 54% in Haemonchus contortus.…”

“…Similar research findings concordantly showed the inhibition of egg-laying and -hatching post-treatment with LL extracts in different nematode species [30,32,[74][75][76][77][78][79][80]. For instance, a recent study by Romero et al (2020) [76] reported that the ethanolic extract of LL leaves significantly inhibited egg-hatching activity up to 54% in Haemonchus contortus. These studies strongly support the ability of LL plant extracts to inhibit the laying and hatching of eggs in nematodes.…”

LC-QTOF-MS Characterization, Antioxidant Activity, and In Vitro Toxicity of Medicinal Plants from the Tri-Than-Thip Remedy

“…The last herb component, P. dulce , contained many chemical groups with antioxidant effects that were similar to those of the two previous herbs, such as glycosylated compounds and flavonoids (quercetin-3-glucoside, luteolin-7-O-glucoside, and kaempferol-3-O-rhamnoside) as well as fatty acids (azelaic acid, etc.) [ 45 ]. The chemicals found in the herbal components and Tri-TT remedy belonged to similar chemical groups as the primary and secondary metabolite constituents of plant parts, with an emphasis on phenolic compounds and flavonoids with potent antioxidant activities ( Figure 3 ).…”

Impact of Seasonal and Organ-Related Fluctuations on the Anthelmintic Properties and Chemical Profile of Cladium mariscus (L.) Pohl Extracts

“…Still, the considerably lower amount of flavonoid glycosides in the inflorescences extract perhaps may have unmasked the interactions with other metabolites present (e.g., proanthocyanidins and luteolin) leading to improved anthelmintic effects. Flavonoid glycosides have been identified in the chemical profile of several plant species with proven anthelmintic effects ( Barrau et al, 2005 ; Mengistu et al, 2017 ; Araújo et al, 2019 ; Romero et al, 2020 ), and some authors have posed the possibility of occurring interactions of this type of compounds with GIN ( Barrau et al, 2005 ; Alonso-Díaz et al, 2008 ). Barrau et al (2005) determined that 3 flavonol glycosides (quercetin-3- O -rutinoside or rutin, kaempferol-3-rutinoside or nicotiflorin and isorhamnetin-3-rutinoside or narcissin) reduced the migration of H. contortus L3 larvae in 25–35% when applied at 1,200 μg mL –1 .…”