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Pharmacognostic and Phytochemical Investigation of Pongamia pinnata

Research Article Open Access
Pramod Kumar1, Munesh Kumar2* and Jaime A Teixeira da Silva3
1Department of Pharmaceutical Sciences, H. N. B. Garhwal University, Srinagar, Garhwal, Uttarakhand – 246174, India
2Department of Forestry, H. N. B. Garhwal University, Srinagar, Garhwal, Uttarakhand – 246174, India
3Faculty of Agriculture and Graduate School of Agriculture, Kagawa University, Miki-cho, Kagawa, Japan
*Corresponding author: Munesh Kumar
Department of Forestry
H. N. B. Garhwal University
Srinagar, Garhwal, Uttarakhand – 246174, India
E-mail: muneshmzu@yahoo.com
 
Received January 08, 2013; Published February 25, 2013
 
Citation: Kumar P, Kumar M, da Silva JAT (2013) Pharmacognostic and Phytochemical Investigation of Pongamia pinnata. 2:634 doi:10.4172/scientificreports.634
 
Copyright: © 2013 Kumar P, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
 
Abstract
 
Pongamia pinnata Linn. (Papilionaceae family) shows potent antidiabetic activity. The plant is also used for the treatment of hypertension and hyperlipidemia, as well as mycobacterium and skin infections. This paper is a detailed pharmacognostic evaluation of the crude drug of P. pinnata pods. The detailed pharmacognostic study uses physico-chemical; morphological and histological parameters recommended by the WHO and may be used for the authenticity of P. pinnata plants. Total ash, acid insoluble and water soluble ash value were 3.377, 0.876 and 1.141% respectively. Crude drugs showed a hot extractive value in petroleum ether, chloroform, methanol and water of 0.1765, 0.4651, 0.2182 and 0.2155% respectively while in normal (unheated) petroleum ether, chloroform, methanol and water, the values were 0.2101, 0.1130, 0.1025 and 0.5937% respectively. Resin content and foaming index were 0.73% (w/w) and <100 respectively.
 
Keywords
 
Pongamia pinnata; Papilionaceae; HPTLC; Pharmacognosy
 
Introduction
 
Pongamia (Papilionaceae) is a monospecific genus, namely Pongamia pinnata. P. pinnata, commonly known as karanja, is distributed throughout India in tidal and beach forests, often as a mangrove plant. It is used medicinally in India, China, Australia and the Philippines. In the Indian traditional system of medicine Ayurveda, P. pinnata has been used in the treatment of bronchitis, whooping cough, rheumatic joints and quench dipsia in diabetes [1]. The flower furnishes an aliphatic waxy matter kaempferol, pongamin (C15H12O5), γ-sitosterol glucoside, quercertin, neoglabrin (A complex amino acids) resembling glabrin and galbrosaponin (C50H84O23) [2]. A furanoflavone i.e., pongone has been isolated from flowers [2]. P. pinnata contains flavonoids and related compounds including flavones, furanoflavonoids, chromenoflavone, chromenocalchones, coumarins, flavone glycosides sterol, terpenes and modified phenylalanine dipeptide [3]. The seeds contain 13.5% mucilage, traces of essential oil and complex amino acids, termed glabrin. Four furanoflavones karanjin, pongapin (C19H12O6), kanjone (C18H12O4) and pongaglabrone (C18H10O5), identified as 3',4'-methylenedioxy furano [2',3',7,8] flavone, have been isolated from Indian Karanja seed [4]. Three furanoflavonoids (Pongamosides A, B and C) and a flavonol, glucoside Pongamoside D, have been reported from the n-butanol-soluble fraction of the ethanolic extract P. pinnata fruit [5]. Pongaglabol, a hydroxyfuranoflavone, and aurantiamide acetate, a rarely occurring modified phenylalanine dipeptide, have been isolated together with four furanoflavones (karanjin, lancheolatin B, kanjone and pinnatin) [6]. Two hydroxychalcones – onganones I and II – have been isolated from bark and characterized. Moreover, two phenylpropanoids – Pongapinone A and B – have been isolated from bark of Indonesian Karanja plants [7]. Five flavonoids (Pongamone A, B, C, D and E) have been isolated from P. pinnata [8]. Seeds are bitter and acrid, carminative, and purify and enrich the blood, relieve inflammation, cure earache, lumbago, chest complaints and chronic fever. Seed is considered useful in the treatment of scabies, leprosy, piles, ulcers, bronchitis and whooping cough [9]. Seeds are mainly valued for their oil, in cosmetic industry and Ayurvedic herbal medicine [10]. Different parts of P. pinnata show different activity anti-inflammatory [ 11], antihyperglycemic and antilipidperoxidative [12] antiulcer [13], analgesic [14], antimycobacterial [15] and antifilarial activity [16].
 
Materials and Methods
 
Plant material
 
The pods of P. pinnata were collected from Jamia Hamdard campus in October 2006 and were identified in the Department of Botany, Jamia Hamdard Hamdard University New Delhi. A specimen for further reference has been retained (Voucher No. PRL-001-06).
 
Morphological properties
 
P. pinnata is a medium-sized glabrous tree with short bole and spreading crown up to 18 m high or sometimes even more and 1.5 m in girth. Pods are compressed, woody, indehiscent, yellowish gray when ripe, varying in size and shape, elliptic to obliquely oblong, 4.0-7.5 cm long and 1.7-3.2 cm broad with short curved beak. Seed usually 1 rarely 2, elliptical or reniform 1.7-2.0 cm long and 1.2-1.8 cm broad, wrinkled with reddish brown leathery testa. The specimens consisted of pods/ fruits of P. pinnata with the following morphological characters. The sample consists of 5% shell and 95% oleaginous seed kernels [1].
 
Powdered microscopic
 
Pods were ground into fine powder with help of grinding machine and treated with phuloroglucinol and conc. hydrochloric acid and mounted on a glass slide with glycerin. A cover slide was added and samples were examined under a confocal microscope (Fluoview FV- 1000; Olympus).
 
Extraction
 
Different extracts of P. pinnata pods were made with a Soxhlet apparatus. Extracts were evaporated and dried to a powder. Dried extracts were kept in desiccators with calcium carbonate adsorbent to prevent moisture absorption by the dried extract [17].
 
Preliminary phytochemical and physiochemical analysis
 
The methanolic extract was subjected to preliminary phytochemical screening to detect major phytoconstitutents using a standard procedure [18,19] and to generate some physiochemical parameters that can be utilized for the identification of plant materials (Table 1). P. pinnata pods can be evaluated morphologically but this study will really help in the authentification of P. pinnata when supplied in powdered form.
 
Table 1: Presence or absence of Phytoconstituents in the P. pinnata.
 
Phytochemical evaluation by thin-layer chromatography
 
Different extracts like petroleum ether, chloroform and methanolic extract of P. pinnata pods were subjected for HPTLC fingerprinting analysis and TLC profile ) in different solvent systems (Table 4) and visualized with the help of a UV chamber (254 and 266 nm) and iodine vapor. Moreover detailed physicochemical parameters have been studied (Table 5).
 
Table 2: Effect of different chemical reagents on P. pinnata powder.
 
Table 3: Effect of different chemical reagents on the fluorescence behavior of P. pinnata powder.
 
Table 4: TLC profile of different extracts of Pongamia pinnata.
 
Table 5: Physicochemical Analysis of P. pinnata pods Powder.
 
Results and Discussion
 
Organoleptic properties
 
A brown to yellowish brown powder with a faint characteristic odour and bitter unpleasant taste.
 
Powdered microscopic studies of pods
 
 
Pods and seeds were powdered. Microscopy revealed the character of pericarp, seeds and attached pedicel.
 
Epicarp: It consists of a layer of cells that were diametric, nonpigmented, and polygonal in a surface view with a moderately thick wall (Figures 1 and 11) and a thick cuticle (seen on fragments in sectional view).
 
Figure 1: Epicarp( Inside face view).
 
Testa: The epidermis is composed of a layer of conical and thickwalled palisade cells with a thick cuticle (Figure 2). In surface view, the epidermal cells appeared more regular, polygonal and yellowish-brown ( Figure 3).
 
Figure 2: Testa in sectional view.
 
Figure 3: Testa (Surface view).
 
Bearer cells: Adherent to the palisade epidermis of the testa, a few of stone cells in patches and a layer of bearer cells could be found. The cells were parenchymatous, polygonal with thickened radial walls and slightly contracted in the middle region. When viewed from below the lumen of the central constriction and those on the either side appeared as rings (Figures 2, 4 and 5).
 
Figure 4: Bearer cells of testa.
 
Figure 5: Bearer cells (When seen from below).
 
Trichomes: Unicellular, conical, thick, and distinctly warty and bent near the base, trichomes were scattered and rare (Figure 6).
 
Figure 6: Trichomes.
 
Calcium oxalate crystal: Several clusters and prismatic calcium oxalate crystals were found in the powder (Figure 7).
 
Figure 7: Calcium Oxalate crystals (Cluster & Prism type).
 
Fibres of measocarp: Appeared as two crossed layers and covered on one side with thin-walled parenchymatous cells containing prism calcium oxalate crystal. The individual fibers were narrow with moderately thickened walls, occasional pits and a distinct lumen ( Figure 8).
 
Figure 8: Fibres of Mesocarp.
 
Sclereids: Single or cluster or Stone cells with an irregular ovoid outline and moderately lignified cell walls with pits were abundantly seen (Figures 9, 12 and 13).
 
Figure 9: Sclereids.
 
Figure 10: Vessels.
 
Figure 11: T.S of pod.
 
Figure 12: Single sclereid.
 
Figure 13: Sclereids in clusters.
 
Vessels: Narrow lignified with annular thickenings were also seen ( Figures 10 and 14).
 
Figure 14: Vessels.
 
Endosperm: Abundant fragments of stratified food material containing cells were observed. Moreover transverse sections of seeds of P. pinnata were performed (Figure 15). The cells were polygonal in outline and with a thick cell wall.
 
Figure 15: Transverse section of seed.
 
Portion of pedicel
 
The fibers and sclereids of the pedicel were similar to those found in the pods but were larger. The parenchymatous cell that associated with the fibers was more thick walled and contained bigger cluster-type calcium oxalate crystals. Preliminary phytochemical screening of different extracts showed the presence of alkaloids, carbohydrates, glycosides, phenolic compounds, tannins, flavonoids, proteins and free amino acids, saponins, sterols, acidic compounds, mucilage, lipids, fats and resin (Table 1). The crude powder of P. pinnata pods was treated with different chemical reagents and evaluated for fluorescence behavior with the naked eye and UV light (Tables 2 and 3). TLC profiles of different extracts of this powder were obtained in different solvent system. The methanolic extract in solvent system toluene: ethyl acetate: methanol in a 5:1:0.3 ratio showed five clear spots at Rf = 0.15, 0.34, 0.47, 0.69 and 0.85 (Table 4). Similarly, HPTLC (Figure 16) was performed in the same solvent system (Table 5). The different physicochemical parameters of P. pinnata pods assessed in this study would be useful in the authentication of plant material (Table 5). Moisture content of the crude drug is not too high therefore the possibility of bacterial and yeast growth is minimal. The ash value was determined to evaluate the purity of the crude drug. Total ash, acid insoluble and water soluble ash value were 3.377, 0.876 and 1.141% respectively. Crude drugs showed a hot extractive value in petroleum ether, chloroform, methanol and water of 0.1765, 0.4651, 0.2182 and 0.2155% respectively while in normal (unheated) petroleum ether, chloroform, methanol and water, the values were 0.2101, 0.1130, 0.1025 and 0.5937% respectively. Resin content and foaming index were 0.73% (w/w) and <100 respectively.
 
Figure 16: HPTLC fingerprinting of different extracts of P. pinnata.
 
Acknowledgment
 
The authors are thankful to Jamia Hamdard (Hamdard University) New Delhi for providing the necessary facility to carry out this study.
 
 
References