Ovicidal and larvicidal effects of extracts from leaves of Andrographis paniculata (Burm. f.) Wall.ex Nees against field isolates of human hookworm (Ancylostoma duodenale).

ETHNOPHARMACOLOGICAL RELEVANCE: The whole plant of Andrographis paniculata (Burm. f.) Wall.ex Nees is used traditionally in different forms by the local people of Asian countries owing to its myriad medicinal properties. Its use as an anthelmintic has been mentioned in literature but has not been well elucidated. AIM OF THE STUDY: To determine anthelmintic effects of extracts from leaves of A.paniculata against human hookworm species based on a standard assay system and to establish the effects of major active compounds responsible for the effects. MATERIALS AND METHODS: Ovicidal and larvicidal activities of extracts of leaves of A.paniculata in different solvents ethanol (Et), methanol (Met), ethyl acetate (EA) and petroleum ether (PE) was studied against field isolates of Ancylostoma duodenale collected and cultivated from hookworm infected human stool samples by egg hatch and larval motility assays. Major active compounds namely andrographolide (AP1), neoandrographolide (AP2) and andrograpanin (AP3) were estimated quantitatively in all the extracts by high-performance liquid chromatography (HPLC) and mass spectrometry (MS) analysis. Anthelmintic effects (ED50, LC50) and presence of the marker compounds in each extract was statistically analyzed by principal component analysis (PCA). Further, biological activities of pure compounds of AP1, AP2, AP3 were assessed to validate the results of the study. RESULTS: Extracts in ethanol and methanol showed highest activity in inhibition of egg hatching with lowest ED50 values (0.017 and 0.02 mg/mL respectively) while ethyl acetate extract had the highest activity against larval motility (0.001 mg/mL) followed by ethanol (0.019 mg/mL). On HPLC analysis, andrographolide content (%), the major diterpene compound, in Met and Et was 0.85 and 1.43 respectively. On PCA, andrographolide component in the extracts was associated with significant inhibitory effects both on egg hatching and larval motility. Pure compound AP1 also showed significant ovicidal and larvicidal activities at concentrations 0.125 µg/mL and 0.019 mg/mL respectively. CONCLUSION: Andrographolide is one of the main phytochemical responsible for significant ovicidal and larvicidal activity against field isolates of A.duodenale from human infections and can be developed as a potential therapeutic choice.

Phytochemical and antioxidative potential of orange, red, yellow, rainbow and black coloured tropical carrots (Daucus carota subsp. sativus Schubl. & Martens).

This study was executed to determine phytochemical content i.e. total carotenoids, phenolics and flavonoids, and antioxidant ability expressed in the form of FRAP, CUPRAC and ABTS activity among different coloured tropical carrots (orange, red, yellow, rainbow and black carrot) developed at ICAR-IIVR, Varanasi, Uttar Pradesh, India. Overall, within different colour group, the extent of variation for various phytochemical content and antioxidant potentiality is narrow i.e. ranged from 1.04- to 3.21-fold; but at the same time, the genotypic variability across genotypes is too wide which varied 20.90- to 57.92-fold for phytochemical and antioxidants is an indication of broad genetic base of carrot germplasm. Among all the carrots, black carrot had an exceptionally high content of total phenolics and flavonoids, and thereby led to the highest antioxidant ability in the terms of FRAP, CUPRAC and ABTS activity expressing about 76-83% relative potentiality followed by rainbow carrot, and least in orange, red and yellow carrot (black carrot > rainbow carrot > red carrot ≈ orange carrot ≈ yellow carrot). The content of phenolics and flavonoids were highly correlated with antioxidant activity (0.955** to 0.992**). However, the most cultivated and consumed carrots, orange and red one, possessed higher amount of carotenoids. The content of carotenoids negatively correlated with total phenolics, flavonoids and antioxidants activity (- 0.612** to - 0.627**). Broad genetic base and selection based on total phenolics content could be pivotal in the future breeding to harness the genetic wealth of carrot efficiently.

Anti-diabetic Phenolic Compounds of Black Carrot (Daucus carota Subspecies sativus var. atrorubens Alef.) Inhibit Enzymes of Glucose Metabolism: An in silico and in vitro Validation.

BACKGROUND: Black carrot is known to be effective against Type 2 diabetes. The phenolic compounds present in black carrot are responsible for this property, but limited information was available about the mechanism of action and target enzymes. OBJECTIVE: The present study aims at understanding molecular interactions of phenolic compounds of black carrot with enzymes involved in glucose metabolism in human to identify the potential inhibitor that can be used as candidate drug molecule to control diabetes. METHOD: In vitro assay for inhibition of α-amylase, α-glucosidase and DPP-IV was carried out using black carrot purified extract and the standard inhibitor acarbose and vildagliptin, recpectively. The inhibition activity of selected phenolic compounds was also studied by in silico docking with all these three enzymes for the proper understanding of interactions. Encapsulation of purified black carrot extract was also carried out. RESULTS: In vitro IC50 value of purified extract was found to be better than the standard inhibitor acarbose for α-amylase and α-glucosidase, and vildagliptin for DPP-IV. Similarly, docking scores of few anthocyanin molecules were found to be higher than their respective inhibitors, suggesting more effective inhibition. Among anthocyanin molecules of black carrot, cyanidin 3-xylosyl galactoside was found to be the potential drug to inhibit these enzymes, whereas dipeptidyl peptidase IV was identified as the best target to control diabetes with anthocyanins of black carrot. CONCLUSION: Anthocyanins from black carrot were found to be effective to control diabetes and very first time we propose that cyanidin 3-xylosyl galactoside is the best potential molecule for inhibiting enzymes involved in glucose metabolism. The study also shows the encapsulation of anthocyanin compounds using ß-cyclodextrin.