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Pharmacognostic Evaluation of the Leaves of Secamone Afzelii (Schult) K Schum (Asclepiadaceae)

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Pharmacognostic Evaluation of the Leaves of Secamone Afzelii (Schult) K Schum (Asclepiadaceae) Abere & Onwukaeme Tropical Journal of Pharmaceutical Research February 2012; 11 (1): 125-131 © Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, 300001 Nigeria. All rights reserved . Available online at http://www.tjpr.org http://dx.doi.org/10.4314/tjpr.v11i1.16 Research Article Pharmacognostic Evaluation of the Leaves of Secamone afzelii (Schult) K Schum (Asclepiadaceae) Tavs A Abere and Doris N Onwukaeme Department of Pharmacognosy, Faculty of Pharmacy, University of Benin, Benin City 300001, Nigeria . Abstract Purpose: Establishment of the pharmacognostic profile of the leaves of Secamone afzelii (Schult) K. Schum, known for its antimicrobial, antioxidant and free radical scavenging properties, will assist in standardization, quality assurance, purity and sample identification. Methods: Evaluation of the fresh, powdered and anatomical sections of the leaves were carried out to determine the macromorphological, micromorphological, chemomicroscopic, numerical (palisade ratio, stomata number, stomata index, vein–islet number and veinlet termination number, moisture content, total ash, acid–insoluble ash, water–soluble ash, alcohol and water soluble extractive values) and phytochemical profiles. Results: Macro-and microscopical studies indicated the presence of pinnately compound leaf, an entire margin with lanceolate shape, acute base, accumulate apex and reticulate venation. Epidermal walls were straight with numerous calcium oxalate crystals. Stomata arrangement was paracytic, with numerous unicellular uniseriate covering trichomes on both surfaces. Chemomicroscopic characters present included lignin, cellulose, mucilage, suberin and cutin, while phytochemical evaluation revealed the presence of alkaloids, tannins, cardiac glycosides and saponins. The findings also included numerical and quantitative leaf microscopy. Conclusion: These findings could serve as a basis for proper identification, collection and investigation of Secamone afzelii. Keywords: Secamone afzelii , Pharmacognostic evaluation, Sample identification. Received: 10 June 2011 Revised accepted: 18 December 2011 *Corresponding author: Email: [email protected]; Tel: +2348023395616 Trop J Pharm Res, February 2012;11 (1): 125 Abere & Onwukaeme INTRODUCTION from the plant having an IC 50 of 15 µg (sol.)/ml in the same system, thus The family Asclepiadaceae is of the order demonstrating the presence of other Rubiales, formerly called Gentianales. The antioxidants [5]. family consists of about 130 genera and 2,000 species distributed all over the world. The extracts of Secamone afzelii have also Some of them are tropical and subtropical been shown to have anti–inflammatory and shrubs, often twining, or perennial herbs. The antioxidant properties due to flavonoids, latex cells usually contain a latex rich in triterpenoids, diterpenoids and caffeic acid triterpenes. Other constituents include: derivatives [6]. The plant also possesses cyanogenetic glycosides, saponins, tannins antibacterial and antifungal activities. and cyclitols [1]. Some drugs of plant origin in conventional Secamone afzelii , known in three Nigerian medical practice are not pure compounds but languages as arilu, ailu or alu in Yoruba, direct extracts or plant materials that have utunta (Ibo) and Ewuonkwonegie (Bini) [2] is been suitably prepared and standardized [7]. a familiar creeping woody climber found on The parameters obtained from the fences and on trees and grows to a very long pharmacognostic evaluation of medicinal length of about 2-3 cm. It is a creeping woody plants have been used in their identification climber with pinnately compound leaves. It is and differentiation of the various species. The often seen as a nuisance to other plants or Apocynaceae family is closely allied to the crops because of its domineering spread Asclepiadaceae family [1]. Establishment of wherever it grows. the pharmacognostic profile of the leaves of Secamone afzelii will assist in Secamone afzelii is used in traditional standardization, which can guarantee quality, medicine for stomach problems, diabetes, purity and identification of samples. colic, dysentery and also for kidney problems. The whole plant boiled with rice is used as EXPERIMENTAL purgative for children. The decoction of the entire plant is prescribed for cough, catarrhal Collection, identification and preparation conditions and as galactogogue. For the of plant material treatment of gonorrhoea, the whole plant is crushed with fresh palm nuts and oil [2]. Its Fresh leaves of Secamone afzelii were synonym, Secamone myrtifolia is claimed to collected in Ugbowo area of Benin City, be used for the management of cough, sore Nigeria. Identification and confirmation were throat, backache, purge and galactogogue done by Usang Felix of Forest Research [3]. The root is said to be poisonous but is Institute of Nigeria (FRIN), Ibadan, Nigeria, used, more the less, by the Zulu medicine where a voucher specimen was deposited man as a remedy for spinal disease [4]. The with the number, FIH 107158. The fresh antioxidant activity of a methanolic extract of leaves were air-dried, powdered in an electric Secamone afzelii stems was tested using the mill (Moulinex), and then stored in a dry and Diphenyl-1-picrylhydrazyl (DPPH) assay and well-sealed bottle. the active compound was identified as α– tocopherol. High Performance Liquid Macroscopy Chromatography (HPLC) determination showed that 0.12% w (sol.)/w α–tocopherol The following macroscopic characters for the was present in the plant. The total extract fresh leaves were noted: size and shape, also showed effective free radical scavenging colour, surfaces, venation, presence or activity in the assay for non–enzymatic lipid absence of petiole, the apex, margin, base, peroxidation in liposomes with an IC value 50 lamina, texture, odour and taste [1,8]. of 90 µg (sol.)/ml, with α–tocopherol isolated Trop J Pharm Res, February 2012;11 (1): 126 Abere & Onwukaeme Microscopy RESULTS The outer epidermal membranous layer (in Macroscopic description of the leaves of fragments) were cleared in chloral hydrate, S. afzelii mounted with glycerin and observed under a compound microscope. The presence as well The leaf was dark green in the upper surface as absence of the epidermal cells, stomata and light green in the lower surface. It had a (type and distribution) and epidermal hairs stipule and a short petiole. Pulvinus was (types of trichomes and distribution) was absent. It was pinnately compound, with a observed. The transverse sections of the lanceolate shape. The margin was entire, fresh leaves through the lamina and the apex was accumulate, base acute and midrib as well as a small quantity of the venation was reticulate. Average leaf size powdered leaves were also cleared, mounted was 4.4±0.8 cm (length) and 2.4±0.7 cm and observed [9]. (breadth). The fresh leaf exudes a white gummy substance when cut, odourless and Chemomicroscopic examination slightly acrid in taste. Examination of the powder for starch grains, Microscopic description of the leaves of lignin, mucilage, calcium oxalate crystals, S. afzelii cutin and suberin were carried out using standard techniques [1]. Micromorphological features revealed that anticlinal walls were thick and straight. Phytochemical investigation Stomata were present in both lower and upper epidermi. The stoma was surrounded Chemical tests were employed in the by two epidermal cells whose axis was preliminary phytochemical screening for parallel to the axis of the guard cells various secondary metabolites such as (Paracytic arrangement) (Fig 1). There were tannins, glycosides and alkaloids using numerous uniseriate covering trichomes standard techniques [1,10-12]. present on both surfaces, mostly unicellular (Fig 2) and multicellular (Fig 3). Quantitative investigation Quantitative leaf microscopy to determine palisade ratio, stomata number, stomata Straight walled index, vein–islet number and veinlet epidermal cells termination number were carried out on epidermal strips. Other parameters determined for the powdered leaves were moisture content, total ash, acid–insoluble ash, water–soluble ash, alcohol (90 % ethanol) and water soluble extractive values (X 400) [13]. Fig 1: Photomicrograph of S. afzelii with straight walled epidermal cells and paracytic stomata Statistical analysis arrangement The data obtained were expressed as mean A transverse section of the leaf across the ± SEM (standard error of mean), and n mid–rib showed an upper and lower represents the number of replicates in an epidermis consisting of cells of similar sizes experiment. (Fig 4). There were uniseriate covering trichomes on both surfaces. It had a bifacial Trop J Pharm Res, February 2012;11 (1): 127 Abere & Onwukaeme mucilage, calcium oxalate crystals and cellulose. Unicellular covering Phytochemical analysis of the leaves of S. trichomes afzelii Phytochemical evaluation (Table 1) revealed the presence of alkaloids, tannins, cardiac glycosides and saponins. (X 400) Fig 2: Photomicrograph of S. afzelii with Table 1: Preliminary phytochemical unicellular covering trichomes. screening of Secamone afzelii leaves Secondary Secamone afzelii metabolites Alkaloids Multicellular Dragendorff’s reagent + covering Wagner’s reagent + trichomes Mayer’s reagent + 1% picric acid reagent + (X 400). Fig 3: Photomicrograph of S. afzelii with Anthracene derivatives multicellular covering trichomes Combined anthraquinones - Free anthraquinones - Tannins Phenazone test + Iron complex test + Formaldehyde
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