Molecular characteristics of anti-inflammatory activities in wood extractives of Quercus Aliena.
Q. aliena, which was native to Korea, Japan and China, had straight trunk, beautiful patterns, large diameter at breast height and high hardness. The wood of Q. aliena was very fine and beautiful, then often used for boat building and wood flooring in house for a long time. At present, the utilization of Q. aliena biomass was synthetically studied more and more. The seed was firstly crushed into a powder, and then used as soup thickener, cereals, breads, substitute for coffee, and so on. The bark, which had a rich source of tannin, was used as a powerful astringent (1,2,4). Yue-Ken et al found that the ethanol extracts of leaf, bark and xylem of Q. aliena Blume had antimicrobial effect against Gram-positive and Gram-negative bacteria, and the ethyl acetate was the strongest antimicrobial activity (4). Zhu-Ping et al studied the synthetic drugs (5-8). Especially, the researchers in our university had analyzed many wooden products, pyrolysis products (9-15), and woody extractives (12-23), and some researchers also explored a variety of biological active ingredients and wood biomass (16-30). Therefore, the wood extractives of Q. aliena was obtained by kinds of extraction methods, and the molecular characteristics were investigated in order to better utilize this wood resource.
MATERIALS AND METHODS
Fresh wood of Q. aliena was collected from the Tongbai Mountain, Henan Province, China. The fresh wood was dried at room temperature, pound to pieces, and then kept in vacuum. Acetic ether, methanol, benzene, petroleum ether and ethanol were chromatographic grade for the subsequent experiments. Cotton thread and cotton bag were both extracted in benzene/ethanol solution for 12 h.
Experiment methods Single extraction
Weighed 54 pieces of wood, each was about 5g (0.1mg accuracy) and finally parceled by cotton bag tied by cotton thread. Extraction was carried out in 350ml solvents by the Foss method for 1, 3, 4, 5, 6, 7 hours, respectively. Solvents were ethanol/methanol ([V.sub.ethanol]/[V.sub.ethanol] = 2), petroleum ether/acetic ether ([V.sub.petroleum ether]/[V.sub.acetic ether] = 2), and benzene/ethanol solution ([V.sub.ethanol]/[V.sub.benzene] = 2). Ethanol/methanol extraction, petroleum ether/ acetic ether extraction, and benzene/ethanol extraction were carried out at temperature of 75[degrees]C, 90[degrees]C and 95[degrees]C, respectively. After extraction, one piece was took out, dried in 105[degrees]C to oven dry, and weighed. The extractives was obtained by evaporation at the temperature of 60~70[degrees]C.
Weighed 27 pieces of wood, each was 20g (1.0mg accuracy), and finally parceled by cotton bag tied by cotton thread. Three-step extraction was carried out by large-caliber Soxhlet according to different orders combined by EMPA-BE (ethanol/methanol '! petroleum ether/acetic ether' !benzene/ ethanol), PA-BE-EM (petroleum ether/ acetic ether '! benzene/ethanol'! ethanol/methanol), BE-EM-PA (benzene/ethanol'! ethanol/methanol'! petroleum ether/acetic ether), respectively. After each step extraction, one piece was took out, dried in 105[degrees]C to oven dry, and weighed. The extractives was obtained by evaporation at the temperature of 60~70[degrees]C.
Among the above extractives, the BE extractives, EM extractives, EM extractives in EMPA-BE method, empaBE extractives in EM-PA-BE extraction, BE extractives in BE-EM-PA method, beEM extractives in BE-EM-PA method, paBE extractives in PA-BE-EM method, pabeEM extractives in PA-BE-EM method were analyzed, respectively. Each 0.5 mg extractives was analyzed by online linked GC/MS (gas chromatograph/ mass spectrometer), respectively. The GC/MS analysis was done as the same as the documents (16-23).
The leaching rates of single extractions and three-step extractions were listed in Table-1 and Table-2. The EB, ME, EM, empaBE, BE, beEM, paBE, pabeEM wood extractives were obtained, respectively. The total ion chromatograms of 8 extractives by GC/MS were shown in Figure-1. Based on the MS data, NIST standard MS map, open-published books and papers, the components and their contents were identified (16-29).
Leaching rule of wood extractives of Q. aliena
The leaching rate trend of Q. aliena wood extractives in different solvents was described in Table-1. It was observed that during ethanol/methanol extraction, the leaching rate of stem extractives fluctuated, and reached the maximum (3.91%) when extraction time was 6h. During petroleum ether/acetic ether extraction, the leaching rate of stem extractives first increased and then decreased, and reached the maximum (2.94.70%) when extraction time was 7h. During benzene/ alcohol extraction, the leaching rate of stem extractives fluctuated, and reached the maximum (4.20%) when extraction time was 7h. The optimal extraction time of ethanol/methanol extraction, petroleum ether/acetic ether extraction, and benzene/alcohol extraction were 6h, 7h, and 7h, respectively.
During three-step extraction, the ethanol/methanol extraction, petroleum ether/acetic ether extraction, and benzene/alcohol extraction were done for 6h, 7h, and 7h, respectively. The statistical results showed that the leaching rates of Q. aliena wood extractives by EM-PA-BE method were 8.17%, 5.74% by BE-EM-PA method, and 10.61% by PA-BE-EM method, resprectively. And it was observed that the leaching rate of each single extraction was less than that of three-step extractions. Table-2 also showed that the threestep extractions gradually increased leaching rates of wood extractives, which were larger than that of any single extraction. During three-step extraction, PA-BE-EM method was the optimum extraction mode for the leaching rate was 10.61%.
Molecular Properties of wood Extractives of Q. aliena
According to GC/MS result, 3 components were identified from EB extractives of Q. aliena wood in single extraction as: 1,5-hexadien-3-yne (97.95%), phthalic acid, butyl hexyl ester (0.61%), 2-p- nitrophenyl-oxadiazol-1,3,4-one-5 (1.43%).
Only 1 component was identified from ME extractives of Q. aliena wood in single extraction. The result showed that the 1 component was 1,5-hexadien-3-yne.
21 components were identified from EM extractives of Q. aliena wood. The result showed that the main components were phthalic acid, butyl hexyl ester (22.86%), 4-cyclohexene-1,2-dicarboximide, N-butyl-, cis- (15.79%), myo- Inositol (9.49%), 9,10-methanoanthracen- 11-ol, 9,10-dihydro-9,10, 11-trimethyl-(8.94%), oleic acid (4.52%), n-hexadecanoic acid (4.47%), 9,12-octadecadienoic acid (Z,Z)- (3.79%), indolizine, 2(4-methylphenyl)-(3.69%), [1,2,4]triazolo [1,5-a] pyrimidine-6- carboxylic acid, 4,7-dihydro-7-imino, ethyl ester (3.62%), benzo[h]quinoline, 2,4-dimethyl-(3.41%), inositol, 1-deoxy-(3.33%), 1heptacosanol (3.05%), phthalic acid, dodecyl octyl ester (2.97%), 1H-indole, 1-methyl-2-phenyl-(2.10%), (all-E)-2,6,10,15,19,23-hexamethyl-2,6, 10,14,18,22-tetracosahexaene (1.84%), 6,13-diazadispiro [188.8.131.52] tetradecan-14-one (1.63%), hexanedioic acid, bis(2-ethylhexyl) ester (1.28%), 3(2H)-furanone, 4-methoxy-2,5-dimethyl-(1.22%), 2-naphthalenemethanol,1,2,3,4,4a,5,6,7-octahydro-[+ or -], [+ or -],4a,8-tetramethyl-, (2R-cis)-(0.73%), agarospirol (0.70%), octadecanoic acid (0.54%).
16 components were identified from BE extractives of Q. aliena wood. The result showed that the main components were dibutyl phthalate (40.40%), phthalic acid, 2-ethylhexyl hexyl ester (13.26%), 7-heptadecyne, 17-chloro- (11.44%), 3,3,7,11-tetramethyltricyclo[184.108.40.206(4,11)]undecan-1-ol(9.47%), [1,2,4]triazolo[1,5-a] pyrimidine-6-carboxylic acid, 7-amino-, ethyl ester (3.56%), 9,12octadecadienoic acid (Z,Z)- (3.48%), benz[b]-1,4oxazepine-4(5H)-thione, 2,3-dihydro- 2,8-dimethyl(3.07%), 1,2,5- oxadiazol-3-amine, 4-(4-methoxy phenoxy)-(2.95%), 1H-indole, 1-methyl-2-phenyl-(2.30%), n-hexadecanoic acid (2.25%), hexanedioic acid, bis(2-ethylhexyl) ester (2.03%), 19,23-hexamethyl-, (all-E)- (1.87%), cyclopentadecane (1.32%), 1- eicosene (1.30%), phthalic acid, butyl tetradecyl ester (0.64%), benzo [h]quinoline, 2,4-dimethyl-(0.64%).
10 components were identified from beEM extractives of Q. aliena wood. The result showed that the main components were phthalic acid, butyl hexyl ester (42.85%), hexanedioic acid, bis(2-ethylhexyl) ester (5.22%), phthalic acid, 2ethylhexyl hexyl ester (13.96%), 2,6,10,14, 18,22tetracosahexaene, 2,6,10, 15, 19,23-hexamethyl-, (allE)-(3.13%), 1H-indole, 1-methyl-2- phenyl(1.07%), 2-amino-4-hydroxy-6,8-dimethyl-7 (8H)-pteridinone (11.69%), benzo[h] quinoline, 2,4dimethyl-(1.43%), 2,4,6-cyclo-heptatrien-1-one, 3,5-bis-trimethylsilyl- (2.50%), 3,3,7,11-tetramethyltricyclo [220.127.116.11(4,11)]undecan-1-ol-(1.43%), cyclohexane, 1,1,2-trimethyl-3,5- bis(1-methylethenyl)-, (2[+ or -],32,52)-(16.73%).
7 components were identified from bcJY extractives of Q. aliena wood. The result showed that the main components were phthalic acid, butyl isohexyl ester (2.06%), dibutyl phthalate (53.84%), 9,17-octadecadienal, (Z)- (0.73%), hexanedioic acid, bis(2-ethylhexyl) ester (3.92%), phthalic acid, 2-ethylhexyl isobutyl ester (24.35%), 1H-indole, 1-methyl-2-phenyl-(6.12%), 1,2,5-oxadiazol-3-amine, 4-(4-methoxyphenoxy)-(8.99%).
8 components were identified from pabeEM extractives of Q. aliena wood. The result showed that the main components were dibutyl phthalate (25.15%), hexanedioic acid, bis(2-ethylhexyl) ester (1.43%), phthalic acid, neopentyl 2-propyl ester (5.55%), 2H bisoxireno[2,3:8,8a]azuleno [4,5-b] furan-7(3aH)-one, octahydro-3a, 8c-dimethyl-6- methylene(15.96%), 1,2,5-oxadiazol-3-amine, 4-(4methoxyphenoxy)-(11.50%), 1H-pyrrole-2,5-dione, 1-(4-chlorophenyl)-(25.04%), 2,4,6 cycloheptatrien-1-one, 3,5-bis-trimethylsilyl-(12.08%), 2,4-cyclo- hexadien- 1-one, 3,5-bis(1,1-dimethylethyl)-4-hydroxy-(3.29%).
3 components were identified from empaBE extractives of Q. aliena wood. The result showed that the main components were dibutyl phthalate (50.84%), phthalic acid, 2-methoxyethyl undecyl ester (49.16%), 3,3,7,11 tetramethyltricyclo[5.4. 0.0 (4,11)]undecan-1-ol.
Resource Properties of wood extractives of Q. aliena
There were many biomedical components in the wood extractives of Q. aliena. Because of its officinal value, 3,3,7,11-tetramethyltricyclo [18.104.22.168(4,11)] undecan- 1-ol was the one volatile alcohol of Picea crassifolia needle and branch which could lure Ips typographus Linnaeus (24). Phthalic acid, isobutyl nonyl ester could potentially cure chronic cardiovascular and cerebrovascular diseases and had anti-tumor, anti-inflammatory, antibacterial functions (25). In 1996, Okugawa et al found that agarospirol could be considered to be neuroleptic. (all-E)-2,6,10,15,19,23-hexamethyl-2,6,10,14,18,22-tetracosahexaene, which could protect liver, resist fatigue and strengthen the body's resistance, and improve human immunity, was considered as important substances in practical and clinical uses with a huge potential in nutraceutical and pharmaceutical industries (15-23). 2,4,6-cycloheptatrien-1-one, 3,5-bistrimethylsilyl-was one of the bioactive components of Microcosmus exasperatus which might heal some diseases[20-25]. 9,12-octadecadienoic acid, methyl ester, and 9,12-octadecadienoic acid (Z,Z)-had been identified as the main medical component of dried worms, and has diuretic, swelling and detoxification properties (2-24). According to the relative content , the extractives was suitable to extract phthalic acid derivatives, dibutyl phthalate, and (all-E)-2,6,10,15, 19,23- hexamethyl-2,6,10,14, 18,22-tetracosahexaene. And there were some drug activities in wood extractives of Q. aliena.
The leaching rule of wood extractives from Q. aliena was obvious. The optimal extraction time of ethanol/methanol extraction, petroleum ether/acetic ether extraction, and benzene/alcohol extraction were 6h, 7h, and 7h, respectively. The leaching rate of each single extraction was less than that of three-step extractions among which PA-BE-EM method was the optimum extraction mode for the leaching rate was 10.61%. What's more, wood extractives of Q. aliena was rich in drug activities, such as 3,3,7,11- tetramethyltricyclo [22.214.171.124(4,11)]undecan-1-ol, 2,6,10,14,18,22-tetracosahexaene, 2,6,10,15,19,23-hexamethyl-, (all-E)-. And the wood extractives of Q. aliena contained many rare drug activities.
This work was financially supported by Invitation Fellowship Programs for Research in Japan of Japan Society for the Promotion of Science (ID No. L14713), and a Project Supported by Special Fund for Forest Scientific Research in the Public Welfare (201504507).
(1.) Mong-Huai S, Sheng-Chieh W, Chang-Fu H, SinI C, Kuoh-Cheng Y. Rediscovery of Quercus aliena Blume (Fagaceae) in Taiwan. Taiwania 2003, 48(2): 112-117
(2.) Kang YX, Xia GW, Liu JJ, Zhou W, Chen GP. Soil respiration characteristics in the clear-cutting site of Quercus aliena var. acuteserrata forest in Xiaolong Mountain in Qinling Mountains. Ying Yong Sheng Tai Xue Bao 2014, 25(2): 342-350
(3.) ChMzaburM T. Tanaka's Cyclopedia of Edible Plants of the World. Tokyo : Yugaku-sha : distributed by Keigaku Pub. Co., 1976, 1-924
(4.) Yue-Ken L, Miao-Ching C. Micropropagation of Quercus aliena Blume var. aliena from Explants of Mature Trees. Journal Taiwan Journal of Forest Science 2014, 29(2): 117-31
(5.) Zhu-Ping X, Zhi-Yun P, Jing-Jun D, Rui-Cheng D, Xu-Dong W, Hui O, et al. Synthesis, molecular docking and kinetic properties of 2-hydroxy-2-phenylpropionyl-hydroxamic acids as Helicobacter pylori urease inhibitors. European Journal of Medicinal Chemistry 2013, 68: 212-221
(6.) Wanxi P, Lansheng W, Fengjuan W, Qiu X. 3-(4Bromophenyl)-4-(4-hydroxyanilino) furan-2(5H)-one. Acta Crystallographica Section EStructure Reports Online 2011, 67(9): O2329U206
(7.) Wanxi P, Fengjuan W, Lansheng W, Qiu X. Crystal structure of 3-(4-bromophenyl)-4-(4chloropheny lamino) furan-2(5H)-one, C16H11BrClNO2. Zeitschrift Fur Kristallographie-New Crystal Structures 2012, 227(1): 61-62
(8.) Wan-Xi P, Le C. Crystal structure of 3-(3-bromophenyl)-4-(3,5-dichloro-phenylamino) furan-2(5H)-one, C16H10BrCl2NO2. Zeitschrift Fur Kristallographie-New Crystal Structures 2012, 227(2): 267-268
(9.) Wanxi P, Lansheng W, Qiu X, Qingding W, Shilong X. TD-GC-MS Analysis on Thermal Release Behavior of Poplar Composite Biomaterial Under High Temperature. Journal of Computational and Theoretical Nanoscience 2012, 9(9): 1431-1433
(10.) Lansheng W, Wanxi P, Zhi L, Minglong Z. Molecule Characteristics of Eucalyptus Hemicelluloses for Medical Microbiology. Journal of Pure and Applied Microbiology 2013, 7(2): 1345-1349
(11.) Lansheng W, Wanxi P, Minglong Z, Zhi L. Separation Characteristics of Lignin from Eucalyptus Lignincellulose for Medicinal Biocellulose Preparation. Journal of Pure and Applied Microbiology 2013, 7: 59-66
(12.) Qiu X, Wanxi P, Makoto O. Molecular Bonding Characteristics Of Self-Plasticized Bamboo Composites. Pakistan Journal of Pharmaceutical Sciences 2014, 27: 975-982
(13.) Le C, Wanxi P, Zhengjun S, Lili S, Guoning C (2014a). Weibull Statistical Analysis Of Tensile Strength Of Vascular Bundle In Inner Layer Of Moso Bamboo Culm In Molecular Parasitology And Vector Biology. Pakistan Journal of Pharmaceutical Sciences 2014, 27: 1083-1087
(14.) Yong-Chang S, Zhi L, Wan-Xi P, Tong-Qi Y, Feng X, Yi-Qiang W, et al. Chemical changes of raw materials and manufactured binderless boards during hot pressing: Lignin isolation and characterization. BioResources 2014, 9(1): 1055-1071
(15.) Wanxi P, Lansheng W, Minglong Z, Zhi L. Separation characteristics of lignin from Eucalyptus camaldulensis lignincelluloses for biomedical cellulose. Pakistan Journal of Pharmaceutical Sciences 2014, 27:723-728
(16.) Wanxi P, Shengbo G, Dongli L, Bo M, Daochun Q, Makoto O. Molecular Basis Of Antibacterial Activities In Extracts Of Eucommia Ulmoides Wood. Pakistan Journal of Pharmaceutical Sciences 2014, 27: 2133-2138
(17.) Dongli L, Wanxi P, Shengbo G, Bo M, Zhongfeng Z, Daochun Q. Analysis on active molecules in Populus nigra wood extractives by GC-MS. Pakistan Journal of Pharmaceutical Sciences 2014, 27: 2061-2065
(18.) Wanxi P, Qiu X, Makoto O. Immune Effects Of Extractives On Bamboo Biomass SelfPlasticization. Pakistan Journal of Pharmaceutical Sciences 2014, 27: 991-999
(19.) Hongchen Q, Wanxi P, Yiqiang W, Shubin W, Ganjun X. Effects of Alkaline Extraction on Micro/Nano Particles of Eucalyptus Camaldulensis Biology. Journal of Computational and Theoretical Nanoscience 2012, 9(9): 1525-1528
(20.) Wanxi P, Lansheng W, Zhi L, Minglong, Minglong Z. Identification and Chemical Bond Characterization of Wood Extractives in Three Species of Eucalyptus Biomass. Journal of Pure and Applied Microbiology 2013, 7: 67-73
(21.) Wanxi P, Zhi L, Junbo C, Fangliang G, Xiangwei Z, Zhongfeng Z. Immunology Molecular Characteristics of JYBS Extractives from Illicium verum Biomass. Journal of Pure and Applied Microbiology 2013, 7(2): 1237-1243
(22.) Wanxi P, Zhongfeng Z, Zhi L, Ohkoshi M, Junbo C, Fangliang G, et al. Molecular Characteristics of Biomedical and Bacteriostasis Extractives of Illicium verum Fruit. Journal of Pure and Applied Microbiology 2013, 7(3): 2017-2024
(23.) Wanxi P, Zhi L, Junbo C, Fangliang G, Xiangwei Z. Biomedical molecular characteristics of YBSJ extractives from Illicium Verum fruit. Biotechnology and Biotechnological Equipment 2013, 27(6): 4311-4316
(24.) Shi R J, Xie S A, Zhao W, Lv S J, Song X B, Guo X R. Analysis of Volatile Components in the Needle and Branch of Picea crassifolia by SPME/GC/MS., Journal of Northwest Forestry University 2011, 6: 95-99
(25.) Hao H, Xin Z, Biao J. Survey in study on chemical constituents from plants of Elaeagnaceae. Chinese Traditional and Herbal Drugs 2006, 37(2): 307-309
(26.) Lin Z, Ge SB, Li DL, Peng WX. Structure Characteristics of Acidic Pretreated Fiber and Self-bind Bio-boards for Public Health. Journal of Pure and Applied Microbiology, 2015, 9: 221-226
(27.) Peng WX, Lin Z, Chen H, Wu JG. Biochemical Group Characteristics of Self-Bonded Boards During Acidic Oxidation for Public Health. Journal of Pure and Applied Microbiology, 2015, 9: 307-311
(28.) Cui L, Peng WX, Sun ZJ, Lu HF, Chen GN. Variability of macroscopic dimensions of Moso bamboo. Pakistan Journal of Pharmaceutical Sciences, 2015, 28: 675-679
(29.) Sun YC, Lin Z, Peng WX, Yuan TQ, Xu F, Wu YQ, Yang J, Wang YS , Sun RC. Chemical Changes of Raw Materials and Manufactured Binderless Boards during Hot Pressing: Lignin Isolation and Characterization. Bioresources, 2014, 9(1): 1055-1071
(30.) DL Li, SB Ge, WX Peng, QD Wu, JG Wu. Chemical structure characteristics of wood/lignin composites during mold pressing. Polymer Composites, Article first published online, 2015 DOI: 10.1002/pc.23658.
M.A. Qingzhi [1,2], M.O. Bo , Hong Chen , Zhang Dangquan  *, Yuzo Furuta  *
 Central South University of Forestry and Technology, Changsha, China.
 Laboratory of Biomaterials Science, Kyoto Prefectural University, Kyoto, Japan.
(Received: 01 June 2015; accepted: 25 July 2015)
* To whom all correspondence should be addressed. E-mail: email@example.com
Caption: Fig. 1. Total ion chromatogram of 8 wood extractives of Q. aliena by GC/MS
Table 1. Leaching rate trend of each single extraction [%] Extraction ethanol petroleum ether benzene time [h] /methanol /acetic ether /ethanol 1 0.43 2.53 2.79 3 3.33 2.16 2.17 4 2.82 1.58 4.13 5 3.51 2.03 2.16 6 3.91 2.69 3.83 7 2.33 2.94 4.20 Table 2. Leaching rate of three-step extraction [%] Extraction EM-PA-BE BE-EM-PA Time[h] 6 7 7 7 6 7 Step 1st 2nd 3rd 1st 2nd 3rd Leaching rate 3.33 0.83 4.01 1.97 0.89 2.88 Extraction PA-BE-EM Time[h] 7 7 6 Step 1st 2nd 3rd Leaching rate 4.47 4.72 1.42
|Printer friendly Cite/link Email Feedback|
|Author:||Qingzhi, M.A.; Bo, M.O.; Chen, Hong; Dangquan, Zhang; Furuta, Yuzo|
|Publication:||Journal of Pure and Applied Microbiology|
|Date:||Dec 1, 2015|
|Previous Article:||Isolation of an effective nitrogen-fixing strain N1115 from rice rhizosphere by rice germ lectin.|
|Next Article:||Molecular characteristics of three extractives of Cinnamomum camphora leaves.|