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Neuropharmacological Screening and Isolation of Cinnamoyl and Coumaroyl- Glucosides from Leaf Fraction of Carpolobia Lutea G

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Neuropharmacological Screening and Isolation of Cinnamoyl and Coumaroyl- Glucosides from Leaf Fraction of Carpolobia Lutea G Indian Journal of Novel Drug delivery 4(1), Jan-Mar, 2012, 28-37 IJNDD Indian Journal of Novel Drug Delivery An Official Publication of Karnataka Education and Scientific Society Research Article Neuropharmacological Screening and isolation of cinnamoyl and coumaroyl- glucosides from leaf fraction of Carpolobia lutea G. Don (Polygalaceae) LUCKY LEBGOSI NWIDU* 1, PAUL ALOZIE NWAFOR 2; WAGNER VILEGAS 3 1 Department of Pharmacology and Toxicology, Faculty of Pharmacy, Niger Delta University, Wilberforce Island, Bayelsa State, NIGERIA 2 Department of Pharmacology and Toxicology, Faculty of Pharmacy, University of Uyo, Uyo, Akwa Ibom State, NIGERIA. 3 UNESP – Univ Estadual Paulista - Departamento de Química Orgânica, Instituto de Química, cp 355, CEP 14801-970, Araraquara, SP, BRAZIL. A R T I C L E D E T A I L S A B S T R A C T Article history: The leaf of Carpolobia lutea G. Don (Polygalaceae) is widely used among the Ibibio Received on 05 August 2011 and Efik in Nigeria to control psychotic patient by sedation. This antipsychotic Modified on 20 November 2011 effect among other uses of the plant is yet to be investigated pharmacologically. Accepted on 29 November 2011 This is a first report of neuropharmacological screening of the leaf. Gradient Keywords: extraction was executed after maceration of air-dried leaf of C. lutea in n-hexane, Carpolobia lutea, chloroform, ethyl acetate and ethanol solvent for 72 hours to obtain the four Psychopharmacology, fractions. The effects of these four fractions of the leaf (192.5-770mg/kg) were Neuropharmacology, investigated on thiopentol-induced sleeping time, locomotor activity, Anticonvulsant. pentylenetetrazole (PTZ) - and strychnine (STN)-induced convulsion in mice. The most active fraction, ethyl acetate fraction, was submitted to semi- preparative HPLC to structurally isolate and characterized active compounds. The ethyl acetate fraction reveal a dose dependent significant (P < 0.01) prolongation of sleeping time duration but no effect on sleeping time latency when compared with control group. The ethyl acetate fraction reveals a dose dependent significant (p < 0.05- 0.001) decrease on locomotor activity. In the anticonvulsant assay, it demonstrates 60% and 40% protection in the PTZ- and strychnine-induced convulsion in mice respectively. The effects of other fractions were not as significant compared to the ethyl acetate fraction. Five polyphenols were isolated, the first four molecules is reported for the first time. The neuropharmacological screening results and isolated compounds could in part lend credence to antipsychotic ethnomedical uses of C. lutea in management of mad people in Akwa Ibom State of Nigeria. © KESS All rights reserved INTRODUCTION Carpolobia lutea G. Don is reputable and widely Polygalaceae belongs to the order Fabales. It has acknowledge among the Efik, Ibibio, Ibo and about 800 species in 12-20 genera which are of Yoruba speaking ethnic groups in Nigeria for its cosmopolitan distribution except in New anti-inflammatory, anti-arthritic and aphrodisiac Zealand, Polynesia and the Arctic zone. Polygala potential. This specie is a small tree distributed is the most important genus among these in West and Central tropical Africa. C. lutea is species. Carpolobia, Atroxima, Muratia , and called cattle stick (English), Ikpafum (Ibibio), Securidaca belong to Africa. Carpolobia has Agba or Angalagala (Igbo) and Egbo Oshunshun fourteen species of which ten is native to Africa (Yoruba) in Nigeria [2] . It has androgenic [1] . The species are: Carpolobia afzeliana, C. alba, properties, used to cure insanity, rheumatism, C. caudate, C. conradsiana, C. delvauxii, C. dubia, C. fever, analgesic, dermal infection, venereal goetzei, C. glabrescens, C. leandriana, C. lutea, C. diseases; used to combat sterility and promote macrostachya, C. suaveolens, C. versicolo and C. child birth; used as taeniafuge and vermifuge [2-5] . zenkeri [1] . The stem bark is dried and taken as snuff to cure migraine headache [6] . The leaf is reported to have anti-inflammatory and anti-arthritic *Author for Correspondence: properties [7] ; diabetes mellitus (Etefia, Personal Email: [email protected] communication); evacuant fever, headache, leprosy, snake bite, venereal disease, wounds 28 Lucky Lebgosi Nwidu et al / Indian Journal of Novel Drug Delivery 4(1) Jan-Mar, 2012, 28-37 healing [8, 9] . Polygalaceae is known to contain and mortar. The pulverized leaves were stored at species with a variety polyphenolic compounds room temperature. such as xanthones, flavonoids and biphenyl derivatives, which exhibit significant biological Animals activities [10] . Adult albino mice and rats were used. All the animals were housed in standard cages under Many plants with neuropharmacological laboratory conditions in the University of Uyo, properties have been investigated [11,12] . However Pharmacology department. The animals used there is no scientific research verifying the were fed with pellet feeds (Vital Feed and Flour neuropharmacological property of the leaf of C. Mill Limited, Edo State, Nigeria) and water ad lutea as the plant infusion has been used by libitum . All animals used have free access to tap herbalist to treat insanity among the Effik people water under standard conditions of 12 h dark–12 of Akwa Ibom State, Nigeria. Some h light and temperature (21±1%). The protocols pharmacological studies have been executed on were approved by the University of Uyo the plant leaf such as evaluations of the anti- Institutional animal Care and Use Committee diarrhoeal and anti-ulcerogenic potential in (UUAEC) which follows the guidelines of CPSCEA rodents [13] and preliminary gastro-protective (Committee for the purpose of control and evaluation of various fractions [14] . supervision of experimental animals) (UUAEC C. lutea leaf enjoyed wide patronage traditionally No. 2004/013)). for the management of various diseases, there has been no active compounds isolated and Extraction and Fractionation of C. lutea characterized with biological activities to our Procedure of gradient solvent extraction is as knowledge. The ethyl acetate leaf fraction described in earlier report [14] . The chemical demonstrate neuropharmacological and constituents present in C. lutea were analyzed anticonvulsant potential. Therefore, this most according to Wagner et al . [15] . The active fraction, which also demonstrate good chromatographic analyses were performed by antiulcerogenic activity in earlier report [14] was thin layer chromatography (TLC) (Fluka, silica submitted to semi- preparative HPLC to gel plates on glass, 20 × 20 cm × 0.25 mm) eluted structurally isolate and characterized active with: CHCl 3/MeOH/H 2O (43:37:20, v/v/v), compounds. HPLC fingerprint of the polar extract CHCl 3/MeOH/H 2O (80:18:2, v/v/v), revealed the presence of polyphenolics with the CHCl 3/MeOH/H 2O/Acetic acid (43:37:30:1, ethyl acetate fraction pre-concentrating the v/v/v/v) and CHCl 3/MeOH/ n-PrOH/H 2O phenols of the leaf fraction. This is the first (310:380:60:250, v/v/v/). report of isolation and characterization of 750 g of the dry leaf powder yielded 3.66, 3.03, biological active compounds from 3.63, and 15.07% for n-hexane, chloroform, ethyl neuropharmacological active fraction. acetate and ethanol respectively [14] . An aliquot of 4.00 g of the ethyl acetate fraction (EAF) was MATERIALS AND METHODS dissolved in 10 ml methanol and centrifuged at Chemicals and Drugs 2,500g for 15 minutes. This procedure was The chemicals used were all of analytical grade: performed 3 times. The supernatant was filtered absolute ethanol, ethyl acetate, chloroform, and submitted to gel permeation acetic acid (Reidel-de Haem, Germany), methanol chromatography using Sephadex LH-20 (Synth, Brazil). Drugs like PTZ, strychnine, (Pharmacia) in a glass column (700 × 35 mm, thiopental sodium, diazepam i.d.) using methanol 98.98%. Fractions were obtained using automatic fraction collector set at Plant Material 9 ml per tube at a flow rate of 3.0 ml/min. Carpolobia lutea leaves were collected and Eighteen fractions (C1-C18) were obtained after supplied by Mr. Etefia, the traditional herbalist, combining into different groups based on the attached to the Pharmacognosy Department, chromatographic profile on TLC and revealed University of Uyo. The plant was identified and either with Natural Product Polyethylene glycol authenticated by Dr. (Mrs.) Margaret Bassey of reagent (NP/PEG) reagent or Department of Botany, University of Uyo, Akwa anisaldehyde/sulfuric acid solution. Fraction C7 Ibom State, Nigeria. A voucher specimen (UUH (1.5 g) obtained from 4.0 g of EAF were analyzed 998) was deposited at the University Herbarium, by HPLC and after submitted to the isolation of University of Uyo, Uyo, Akwa Ibom State, Nigeria. compounds. The leaves were air dried, powdered with pestle 29 Lucky Lebgosi Nwidu et al / Indian Journal of Novel Drug Delivery 4(1) Jan-Mar, 2012, 28-37 HPLC analysis and isolation electrospray interface. The data obtained were The EAF and fraction C7 aliquots were dissolved compared with those found in literature [17, 18] . in 80% aqueous MeOH and filtered through a 0.50 µm Teflon syringe filter, then the filtrate (30 Neuropharmacological studies µl) was injected into the HPLC. HPLC analysis Prolongation effect on thiopental sodium - was conducted on an HPLC system using a Jasco induced sleeping time (Tokyo, Japan) liquid chromatography equipped The latency and duration of loss and gain of with a PU-2089 quaternary solvent pump, a MD- righting reflex were investigated according to 2010 PAD and an AS-2055 autosampler injector methods of De Paris et al., [19] and De Almeida et with a 20 μL sample loop. The analytical column al., [20] . Test samples suspended in 20% Tween was a Phenomenex Synergi Hydro RP18 (250 × 80 were administered to male mice (5/group) 4.6 mm i.d.; 4 μm) equipped with a Phenomenex were divided into 6 groups. Group 1 (served as security guard column (4.0 × 2.0 mm i.d.). The control) receives 10ml/kg of 20% Tween 80, mobile phase composition was: water (eluent A) vehicle only.
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