See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/257476284 Anti-acne activity of tannin-related compounds isolated from Terminalia laxiflora ARTICLE in JOURNAL OF WOOD SCIENCE · OCTOBER 2013 Impact Factor: 0.94 · DOI: 10.1007/s10086-013-1344-4 CITATIONS READS 4 159 3 AUTHORS, INCLUDING: Ali Mahmoud Muddathir University of Khartoum 5 PUBLICATIONS 13 CITATIONS SEE PROFILE Available from: Ali Mahmoud Muddathir Retrieved on: 06 February 2016 J Wood Sci (2013) 59:426–431 DOI 10.1007/s10086-013-1344-4 ORIGINAL ARTICLE Anti-acne activity of tannin-related compounds isolated from Terminalia laxiflora Ali Mahmoud Muddathir • Kosei Yamauchi • Tohru Mitsunaga Received: 21 February 2013 / Accepted: 17 April 2013 / Published online: 17 May 2013 Ó The Japan Wood Research Society 2013 Abstract In our investigation to find out new anti-acne Keywords Terminalia laxiflora Engl & Diels Á agent, we focused on Terminalia laxiflora Engl & Diels Propionibacterium acnes Á Lipase inhibitor Á Terchebulin Á (Combretaceae) methanolic wood extract, which has been Flavogallonic acid dilactone selected during previous screening experiments for anti- acne agents, which included 29 species of Sudanese medicinal plants. Based on the biologically guided Introduction fractionation using an antibacterial assay against Propion- ibacterium acnes, a lipase inhibitory assay and 1,1-diphe- Acne vulgaris is the most common skin disease that affects nyl-2-picrylhydrazyl (DPPH) radical scavenging activity areas containing the largest oil glands including face, back assay; five tannin-related compounds were isolated, such as and trunk [1]. Propionibacterium acnes have been impli- ellagic acid, flavogallonic acid dilactone, terchebulin and cated over other cutaneous microflora in contributing to the gallic acid. Terchebulin showed good antibacterial activity; inflammatory response of acne. It acts as an immunosti- minimum inhibitory concentration (MIC) = 125 lg/ml and mulator through producing enzymes such as lipases and minimum bactericidal concentration (MBC) = 250 lg/ml. proteases which are involved in the development of Gallic acid exhibited lipase inhibitory activity with IC50 inflammatory process [2]. Several treatments have been value of 149.3 lM, which showed strong inhibition com- introduced to decrease the esthetic and psychological pared with terchebulin, IC50 260.7 lM. However, all problems caused by acne. The topical application of ther- compounds exhibited better or equal DPPH radical scav- apeutic agents has been found to be more feasible than enging activity to (?)-catechin as positive control. Ellagic hormonal treatment and laser therapy. The ingredients in acid and terchebulin showed the best DPPH radical scav- topical acne treatments, particularly herbs and naturally enging activities, IC50 4.86 and 4.90 lM, respectively. This derived compounds, have received considerable interest as study demonstrated that terchebulin has potentiality as an they show fewer adverse effects than synthetic agents [3]. anti-acne agent. Based on our previous screening experiments for anti-acne activity, the methanolic and 50 % ethanolic extracts of Terminalia laxiflora Engl & Diels wood showed potent activity among the 29 species of Sudanese medicinal plants examined [4]. The genus Terminalia is the second largest genus of A. M. Muddathir Á K. Yamauchi Combretaceae family consisting of 200 species, distributed United Graduate School of Agricultural Science, Gifu University, Gifu 501-1193, Japan in the tropics and subtropics. About 30 species of Termi- nalia are found in Africa [5]. About 200 woody species of T. Mitsunaga (&) Terminalia are used as resources in the timber, pharma- Department of Applied Biological Science, Faculty of Applied ceutical, and leather industries [6]. Although traditional Biological Science, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan healers throughout Africa have used species of the Com- e-mail: [email protected] bretaceae for the treatment of a wide range of disorders, 123 J Wood Sci (2013) 59:426–431 427 only a few species have been subjected to scientific studies column (YMC-DispoPack AT ODS-25:120 g).The column [7]. Several biological activities of Terminalia sp. have was conditioned with the first eluent used for separation for been reported such antibacterial, antifungal [8], anticancer 30 min with flow rate 0.5 ml/min to ensure that no other [9], anti-inflammatory [10], antidiabetic [11], hypocholes- contaminations were present on the column. MPLC sepa- terolemic [12], anti-ulcer activity [13], anticaries agent ration was performed using a chromatography pump (540 [14], anti-HIV-1, anti-malarial [15], antioxidant and mel- Yamazen, Japan), UV detector at 280 nm wavelength (UV- anin inhibitory activity [16]. These pharmacological stud- 10V Yamazen, Japan) and fraction collector (SF-2120, ies in a number of Terminalia species have led to isolation Advantec Toyo Ltd., Japan). of several compounds such as gallic acid, ellagic acid, Elution with H2O/MeOH = 95/5 (v/v) and 20/80 (v/v) punicalagin, terchebulin, isoterchebulin, chebulagic acid, gave two fractions (F1 and F2). Fraction one (F1) (200 mg) chebulinic acid and others. was subjected to column chromatography on a Sephadex T. laxiflora is a common indigenous tree species in LH-20 using MeOH and 70 % acetone in water (v/v) as the woodland and semi-humid Savannah of the Sudan. It has eluent to give four fractions (F1-1–F1-4). Separation of multipurpose uses with a high potential of timber produc- these sub-fractions (F1-1, F1-2 and F1-4) were performed tion and traditional medicinal uses. In Sudan decoction of using preparative high performance liquid chromatography T. laxiflora stem bark is used for malaria and cough (HPLC) with reversed phase Inertsil OD-3 column (GL treatments [17]. Women also use heartwood for fumigant Sciences Inc. 10 mm i.d. 9 250 mm) monitored at ‘‘smoke bath’’. Exposure to the smoke bath is believed to 280 nm. The solvent system used was as follows: a gra- relieve rheumatic pain, smoothen skin and achieve general dient program for 60 min from 10 to 100 % methanol in body relaxation besides other cosmetic and medicinal water with 0.05 % TFA at a flow rate 5 ml/min. This beautification [18]. Earlier work done on the root bark of separation gave gallic acid (1) 10 mg, flavogallonic acid T. laxiflora has led to isolation of several compounds like dilactone (2) 27.5 mg and terchebulin (3) 16.9 mg. Frac- laxiflorin, ellagic acid, trimethylellagic acid, tetramethyl- tion two (F2) was also subjected to preparative HPLC under ellagic acid and terminolic acid [19]. the same condition to isolate ellagic acid (4) 35 mg. In this study five compounds from methanolic wood Chemical structures of the isolated compounds (1)–(4) are extract of T. laxiflora have been isolated and evaluated for shown in Fig. 1. their anti-acne activities using antibacterial assay against P. acnes, a lipase inhibitory assay and 1,1-diphenyl-2-picryl- Identification of isolated compounds from T. laxiflora hydrazyl (DPPH) radical scavenging activity assay. Compounds were identified by 1H, 13C nuclear magnetic resonance (NMR) and liquid chromatography–mass Materials and methods spectrometry (LC–MS). Methanol-d4 was used as the NMR solvent. NMR measurements were obtained by Plant materials using JEOL ECP 600 MHz NMR. LC–MS (Waters WatersÒXevoTM QTof MS) measurements were per- T. laxiflora wood was purchased from Khartoum, Sudan, in formed using column C18 (2.1 mm i.d. 9 100 mm) with March 2011. The specimen was authenticated by Dr. MeOH/H2O = 5/95 (v/v) (30 min), 100/0 (10 min) with a Ashraf Mohamed (Faculty of Forest, University of Khar- linear gradient as eluent. The data were collected in toum). Voucher specimen (SD-KH-03) was deposited at negative ionization mode. Spectroscopic data of flavo- the Department of Horticulture, Faculty of Agriculture, gallonic acid dilactone (2) and terchebulin (3) are known University of Khartoum. from the literature. Gallic acid (1) and ellagic acid (4) were identified by comparing the spectroscopic data of Extraction and isolation commercial reagents. Compound 2 Flavogallonic acid dilactone, a tan powder. T. laxiflora wood (300 g) was air-dried at room tempera- LC–MS (negative ion mode) m/z: 469(M-H); 1H-NMR (in 13 ture, powdered and extracted with 3 L of methanol for 12 h CD3OD): d (ppm) 7.26 (s), 7.50 (s). C-NMR (in 0 0 three times. The extract was filtered then the solvent was CD3OD): d (ppm) 108.1 (C-1, 1 ), 110.1–114.4 (C-6, 6 ), removed under vacuum using rotary evaporator at 30 °C. 112.8 (C-5), 113.3 (C-600), 117.5–120.2 (C-50,200), 124.9 The yield of methanol extract was 42.9 g (14.3 %). (C-100), 135.7 (C-2), 136.3 (C-3), 136.5 (C-4), 137.8 (C-20), Part of the extract (10 g) was separated by medium 139.2 (C-30), 143.2 (C-40), 144.1 (C-300), 145.9 (C-400), pressure liquid chromatography (MPLC) using ODS 147.8 (C-500), 158.9-160.4 (C-7, 70), 168.9(C-700). 123 428 J Wood Sci (2013) 59:426–431 Fig. 1 Structures of compound O OH OH (1) gallic acid, compound (2) flavogallonic acid, Compound O O OH 2' 3' O OH (3) terchebulin and compound 7 4' 7'' 1' 5' (4) ellagic acid 6' 6 5 1 1'' 2'' 4 2 7' 6'' HO OH 3 3'' 5'' HO O O 4'' OH HO OH OH OH Compound ( Compound ( OH HO 4 5 6 3 D HO 1 2 O OH 7 O O OH O O O 5 4 6 7 3 OH 1 2 6'' O O OH 5'' 1'' HO O O 2' O 1' 7' 4'' 2'' O HO 3' 6' 3'' 4' 5' O OH O 7 O O Compound ( HO 7 O 7 1 2 HO 6 3 OH 6 O A 5 1 5 4 C 6 1 4 2 2 3 B 5 3 OH HO 4 HO HO OH OH OH Compound ( 1H-NMR and 13C-NMR spectral data coincided to that Bioassay tests of published report [20].