PHYTOCHEMICAL AND ANTIVENOM EVALUATIONS OF METHANOL LEAF EXTRACT OF CLERODENDRUM CAPITATUM (WILLD) SCHUM. AND THONN. (VERBENACEAE)
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ABSTRACT
Clerodendrum capitatum (Willd) Schumach and Thonn. is a common medicinal plant found mainly in the tropical and sub-tropical regions of the world where it is used in traditional medicine for the treatment of various conditions including erectile dysfunction, wounds, diabetes mellitus, hypertension. The investigations started with the qualitative microscopic and macroscopic evaluation of the leaf material and establishment of its quality parameters, including physicochemical and phytochemical evaluation as no detailed pharmacognostic study or establishment of quality parameters has been done on this plant to date. The powdered leaf was then macerated using 70% aqueous methanol to obtain a brownish gummy extract. The extract was partitioned into n-Hexane fraction (HF), Ethylacetate fraction (EF) and n-Butanol fraction (NF) and the EF was further subjected to phytochemical analysis. In addition, the extract was then evaluated for its acute toxicity and anti-venom properties. Prominent characters include numerous multicellular covering trichomes, anomocytic stomata, vascular bundles and prisms, sheath and cluster of calcium oxalate crystals were recorded. Various physicochemical parameters were also established. Phytochemical screening revealed the presence of many therapeutically important classes of phytoconstituents such as alkaloids, flavonoids, phenolics, sterols, triterpenoids, saponins, proteins and carbohydrates. Analysis of EF through column and thin layer chromatography led to the isolation and identification of the known compound Quercetin3-O-α-arabinoside. The structure of this compound was determined by spectroscopic methods including 1D NMR as well as comparison with published data. The LD50 of 3.8 and 4.4 gKg-1 body weight for Lorke’s and Karber’s arithmetic methods via i.p and oral routes respectively reveals the relative safety of the extract. Investigations carried out on isolated frog rectus abdominis muscle showed that C. capitatum extract did not demonstrate any 8 contracting or blocking effects on frog rectus abdominis muscle when varying doses of 0.1 mg/ml, 1 mg/ml, 10 mg/ml were used. The optimum contraction of ACh and median inhibitory concentration (IC50) for the venom is 0.0004 and 0.00083 mgmL-1 organ bath concentrations respectively. In the in vitro neutralization studies, the extract at concentration of 10 mgmL-1 and 20 mgmL-1 showed 60% and 100% survival respectively. The extract at 20 mgmL-1 showed significant level of detoxification where all the mice survived. The phospholipase A2 assay showed that the crude extract inhibited the enzyme (crude phospholipase) in a non competitive manner as revealed by the double reciprocal plots. The Km was calculated as 2.17 mgmL-1 and Vmax as 1, 1.13 and 1.28 mg/mL. Such a study would serve as a useful tool in standardization of the leaf material, isolation of medicinally important phytoconstituents, performing pharmacological investigations and ensuring quality formulations. The activity of the extract and the presence of Quercetin-3-O-αarabinoside in the extract may provide a scientific support to the use of the plant C. capitatum for the treatment of snake bites.
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