An Efficient Method for the Purification and Characterization of Nematicidal Azadirachtins A, B, and H, Using MPLC and ESIMS

Abstract

Azadirachtin A enriched concentrate containing 60% active ingredient (a.i.) was prepared from the methanolic extract of the de-fatted neem (Azadirachta indica A. Juss) seed kernels. Azadirachtins A, B, and H, the three major bioactive constituents of neem seed kernel, were purified from this methanolic concentrate by employing reverse phase medium-pressure liquid chromatography (MPLC), using methanol-water solvent system as an eluant. The three pure azadirachtin congeners thus obtained were characterized by their unique mass spectral fragmentation, using electrospray probe in positive ion mode (ESI). All three azadirachtins exhibited nematicidal and antifungal activities. Azadirachtin B was the most effective against the reniform nematode Rotylenchulus reniformis (EC(50) 96.6 ppm), followed by Azadirachtin A (119.1 ppm) and H (141.2 ppm). At 200-ppm concentration, the test compounds caused 50-65% mortality of Caenorhabditis elegans nematode. Azadirachtin H showed the highest activity against the phytophagous fungi Rhizoctonia solani (EC(50) 63.7 ppm) and Sclerotium rolfsii (EC(50) 43.9 ppm), followed by B and A. The isolation of pure azadirachtins A, B, and H directly by MPLC purification from its concentrate and their characterization by ESIMS are unique and less time-consuming.

Novel Anti-inflammatory Activity of Epoxyazadiradione against Macrophage Migration Inhibitory Factor

“…Extraction of Neem Limonoids-Neem limonoids (salannin, nimbin, and azadirachtins A and B) from the neem seed kernel were extracted as described earlier (49,50) with slight modification. In brief, 1 kg of neem seed kernel was extracted with petroleum ether (4 ϫ 5 liters) with stirring for at least 3 h in every turn.…”

Comparative efficacy of different herbal and modern anthelmintics against gastrointestinal nematodiasis in fowl

“…In case of L 3 Korolla and papaya were significantly effective (table 3). (Sharma et al, 2003). Besides, neem leaves contain quercetin, a polyphenolic flavonoid, acts as anti-oxidant (Ghani, 2003) and vitamin C which also an anti-oxidant (Kayser, 2002).…”

In Vitro Efficacy of Medicinal Plant Material on the Inhibition of Development of Egg of Ascaridia galli

“…Number of researchers reported the anthelmintic activity of turmeric (C. domestica) against adult A. galli (Ahktar and Riffat, 1985;Ali, 2006) and other helminths (Rahman, 2002;Hordegen et al, 2003;Sharma et al, 2003;Mishra et al, 2004;Githiori et al, 2004;Chandrawathani et al, 2006;Szewczuk et al, 2006). In this experiment, highest efficacy of turmeric leaves was found in 20% juice (78.9%), followed by 10% juice (73.9%) and dust at 20% concentration (73.3%).…”