この論文は国立情報学研究所の学術雑誌公開支援事業により電子化されました。Chemical analyses were carried out on a graft-wood of hinoki (Chamaecyparis obtusa) and sawara cypress (C. pisifera), and the results were compared with the data from normal woods of hinoki and sawara. The amount of boiling water soluble matter were much higher in the graft-wood than in normal woods (1.5-3.6 times), and also the amount of alcohol-benzene soluble matter in the graft-woods were two times higher than those of the normal woods. Lignin content varied little among the samples. α-Terpineol which is the most characteristic component to identify hinoki and sawara woods, borneol, DL-limonene and bornyl acetate were identified, respectively. Some group peaks which are supposed to be cadinenes and cadinols were also found on g.l.c. of steam volatile fraction. Sawaranin [2,3-desoxy-2-(p-hydroxy phenyl hexonic lactol)] was first isolated from methanol extracts of normal woods of hinoki and identified with the results of u.v.. i.r. and n.m.r

HIGUCHI, Takayoshi

KITAMURA, Teruko

SATO, Akira

Kyoto University Research Information Repository

TitleSome Chemical Observation on a Natural Graft-Wood ofJapanese Cypress (Chamaecyparis obtusa ENDL.) and SawaraCypress (Chamaecyparis pisifera SIEB. et ZUCC.)Author(s)SATO, Akira; KITAMURA, Teruko; HIGUCHI, TakayoshiCitationWood research : bulletin of the Wood Research Institute KyotoUniversity (1970), 50: 12-18Issue Date1970-12-19URL http://hdl.handle.net/2433/53424RightType Departmental Bulletin PaperTextversionpublisherKyoto UniversitySome Chemical Observation on a Natural Graft-Wood of Japanese Cypress (Chamaecyparis obtusaENDL.) and Sawara Cypress (Chamaecyparispisifera SIEB. et Zucc.)*Akira SATO**, Teruko KITAMURA** and Takayoshi HIGUCHI**Abstract--Chemical analyses were carried out on a graft-wood of hinoki (Chamaecy-paris obtusa) and sawara cypress (C. pisijera) , and the results were compared with thedata from normal woods of hinoki and sawara. The amount of boiling water solublematter were much higher in the graft-wood thar. in normal woods (1.5-3.6 times), andalso the amount of alcohol-benzene soluble matter in the graft-woods were two timeshigher than those of the normal woods. Lignin content varied little among the samples.a-Terpineol which is the most characteristic component to identify hinoki and sawarawoods, borneol, D1.-limonene and bornyl acetate were identified, respectively. Some grouppeaks which are supposed to be cadinenes and cadinols were also found on g.l.c. of steamvolatile fraction. Sawaranin [2,3-desoxy-2-(p-hydroxy phenyl hexonic lactol)J was firstisolated from methanol extracts of normal woods of hinoki and identified with the resultsof u.v., i.r. and n.m.r.IntroductionHinoki (Chamaecyparis obtusa ENDL.) and sawara (Chamaecyparis pisifera SIEB.et Zucc.) cypresses are closely related in species and a natural graft of both woodshas been discovered at the National Forest, Tsukechi District, Gifu prefecture.The authors have recently obtained such a graft-wood of hinoki and sawara shownin Fig. 1 from the National Forest.In the wood of hinoki (Japanese cypress), the following chemical compoundshave been found; a-pinene, thujopsene, cadinene, cadinol, hinokiol, hinokione,hinokinine (cubein)ll and hinokiresinoFl. On the wood of sawara cypress theoccurrence of the following compounds as well as some of the above terpenoidshas been reported; sawaranin (2,3-desoxy-2-(p-hydroxy-phenyl hexonic lactol)],arabinose, potassium hydrogen oxalate3l and savinin (hibalactone)4l.It is interesting at the standpoint of chemotaxonomy and comparative bio-chemistry to know the distribution of chemical constituents in such a graft-woodin relation to the possible variation of the constituents of each species.* Presented at the 19th Annual Meeting of the Japan Wood Research Society, Sapporo, July1969.** Division of Lignin Chemistry.- 12-SATO,;KITAMURA, HIGUCHI: Chemical Observation on a Natural Graft-WoodFig. 1. A Sample Board of Natural Graft-Wood. Left hand side shows partof Sawara, and right hand side does Hinoki. Size of a scale is 30 em long. 'In the present experiment, ordinary chemical analysis of the hinoki and sawaraparts of the graft-wood and normal woods of both cypresses have been carriedout. Although hinokinin, savinin, ~inokiresinol and sawaranin which were con-sidered as suitable indices for the above described purpose, none of them wereidentified through this experiment except sawaranin. However, sawaranin wasfound in normal wood of hinoki, unexpectedly.Results and DiscussionThe mean values qf dual or triple analyses of moisture, boiling water solublematter, alcohol-benzene soluble matter and lignin are given in Table 1. Chemicalcompositions of normal woods were generally reasonable comparing with thoseTable 1. Chemical composition of Chamaecyparis sp.Sawara HinokiNormaiIGraft. NormalIGraft.Moisture (%) 18.3 14.2 10.5 13.8Boiling water soluble (%) 1.7 6.2 4.5 6.3Alcohol-benzene soluble (%) 8.3 17.3 7.6 14.5Lignin (%) 31. 0 29.7 30.7 29.5- 13-WOOD RESEARCH No. 50 (1970)reported in old literatures5,6), except the quantities"of alcohol-benzene soluble matterwere about two times higher. On the other hand, contents of boiling water solublematter and alcohol benzene soluble matter differed c~siderablybetween the normaland the graft-wood. For instance, the contents of boiling water soluble matter in thegraft-wood showed 3.6 and 1.4 times higher than those of sawara and hinoki normalwoods, respectively, whereas the difference in the graft-wood between hinoki andsawara parts was minute. The contents of the alcohol-benzene soluble matter fromeach part of the graft-wood were almost two times higher than those of the normalwoods, and that from the p~rt of sawara wood was slightly higher than that fromhinoki. Such higher value is seemed to be ascribed to an abnormal conjunctivetissue. However, lignin contents were quite similar among the samples.Gas-liquid chromatograms of the steam volatile fraction from each normal andgraft-wood are shown in Fig. 2 and 3, .respectively. Sensitivity range of the in-strument was switched over to half aL the point of mark S on the chromatograms.As shown in Fig. 2, two groups of peaks which appeared over 200°C were ~etected.It is suggested that the former at lower temperature contains cadinenes and thelatter of higher temperature does cadinols. Other peaks at lower temperaturefrom 80°C to 200°C, were established to be monoterpenes and the peaks of a-terpineoland borneol were identified in both woods. Minor peaks of DL-limonene and bornylacetate were also identified in sawara. Main peaks due to a-terpinol and anotherunidentified peak which appeared around 190°C were characteristic in sawara.(S)(5)2m /0% ApiezonNon Chr, AWTeO. 4°C/m. up, He/50/64(J)SAWARA/80200220(6)236Fig. 2. Gas-liquid Chromatograms of the Volatile Oil from Normal Woodof Hinoki and Sawara.(1) borneol (2) DL-limonene (3) a-terpineol (4) bornyl acetate(5) sesquiterpenes (cadinenes) (6) sesquiterpenealcohols (cadinols)-14 -SATO, KITAMURA, HIGUCHI: Chemical Observation on a Natural Graft-WoodGenerally, the group of cadinenes was more contained than cadinols in both woods.In the case of the graft-wood, chromatograms of both hinoki and sawara partsseparated kept characteristics of each original wood as shown in Fig. 3, althoughthe pattern of chromatograms was similar to each other.( 6)(5)2m /0% ApiezonN on ChrAWTCD, 4°c/min. up, He(3)SAWARA(3)HINOKIl2 )235 210 200 190 180 170 160 150°CFig. 3. Gas-liquid Chromatograms of the Voltatile Oil from aGraft-Wood of Hinoki and Sawara.Acetone extracts were analyzed by Ll.c. Each sample solution was preparedfrom either n-hexane insoluble or soluble fraction after removing steam volatilematter, and the fractions from normal woods thus obtained were fractionated by1 % sodium hydroxide solution. The results are summarized in Fig. 4. Thechromatogram developed with chloroform-acetic acid and colored with diazotized1.0 SiLicageL G CHCL3'AcOH (9: IV/V)0.9 0 0 \]HD 0 00.8 0 8G-0.7 0 CJ 0 00.6 0 0 0OF oERf 0 00.5i 04 'I:' G0.. Q0.3 d 0 OC0.2 ~ 0 0 0 0 o B0./ ~ 0 il 0 R A0.0 ~ 0 0 6-0ST. NoOH Nor. GR. GR. Nor. ST.NaOH- Hinoki- -Sawara-Fig. 4. Thin-layer Chromatograms of the Ether Soluble Matters.-15 -WOOD RESEARBH No. 50 (1970)benzidine is shown in Fig. 4. By this solvent, the spot of sawaranin did not movefrom the original point. The spots of hinokinin, savinin and a few kinds offlavonoids were supposed to be present on the chromatogram, although each spotwas not yet fully identified. Only main difference between the normal woods ofsawara and hinoki was of the spot marked C and D in Fig. 4. For instance, spotC which gives Rf value of 0.30 appeared only in hinoki, but spot D of Rf value0040 did only in sawara wood. Differences between the graft-wood and the normalwoods were generally quite small. Spot H showed a little difference in hinoki, spotD in the normal sawara disappeared and spot F newly appe'ared in the sawarapart of the graft-wood. Such similarity is not surprised as both species belong tothe same genus. Thus, the special pattern of the extractives was not found inthe graft-wood, and the hinoki and sawara parts of the graft-wood generally gavechemical components of the respective normal woods.Pale yellow crystals were isolated as flakes from the methanol extract of the====:=:::Ii===:c::..-=~--r-~~--_ ,------ ---I~~ I I ::-=r== i !40003600 3200 28002400200(; 1900 /800 1700800 /roo 1400 1300 120C 1/00 /000 300 800 700 600em-IFig. 5. LR. Charts of Sawaranin.iii Iii 1 iii I Iii I I , Iii iii iii ,-~ -r-r . -I~r-,-,-.--r-r.,-r-.,..--,-,--,-"540 480 420 360 300 240 f80 f20 60 OHz. , 'IiUl-Fig. 6. N.M.R. Chart of Sawaranin.-16 -TMSI I I ISATO, KITAMURA, HIGUCHI: Chemical Observation on a Natural Graft-Woodnormal hinoki wood. This compound was eluted with solvents containing acetonefrom silicic acid column, recrystalized with boiling water and melted at 212-214°C.Elementary analysis was coincident with formura C12H140 6 and spectrum of U.v.absorption showed Max. at 277 nm in 95 % EtOH. The absorption spectra of Lr.and n.m.r. measured with the solution of deutero DMSO were also obtained and"-the results were coincident with those of authentic sawaranin. The Lr. and n.m.r.charts are shown in Fig. 5 and 6, respectiviely. Thus it was established thatsawaranin was not only distributed in the wood of sawara, but also in the woodof hinoki, and the fact was very interesting from the view point of chemotaxonomyof genus, Chamaecyparis.ExperirnentalSamples of a natural graft-wood between hinoki and sawara, and respectivenormal woods as standard were collected from Tsukechi District, Gifu Prefecture.Wood meal named grafting hinoki or sawara were prepared from the each partof the wood using Willey's mill, respectively.Wood meal from the normal wood of both species which mainly composed ofheartwood was also prepared with Willey's mill and named as normal hinoki annsawara.Each sample wood was used for chemical analyses to determine, moisture.boiling water extract, alcohol-benzene extract and lignin by the methods of JISstandard; namely, ]IS P-8002, P-8005, P-8010 and P-8008, in order.Steam Distillation Two hundreds grams of wood meal were subjected to steamdistillation for two hours. The distillate caught in ice cold acetone was transferedto ether layer as usual, and the concentrated ether solutions were applied for g.l.c.Gas Liquid Chromatography (g.l.c.) The instrument, lEOL 750 fixed with 2meters of 10 % Apiezon N on chromosorb W (3 mm¢ stainless steel) was operatedwith temperature range 80°C to 240°C, in the atmosphere of helium, and compoundswere detected with TeD. Monoterpenes were identified comparing with retentiontimes of the authentic ones.Thin-Layer Chromatography (t.l.c.) Glass plates coated with Silica gel G (Merck;were used for ether soluble fractions and developed with chroloform-acetic acid(9; 1 v/v) as shown in Fig. 4. Diazotized benzidine was used as a coloringreagent. The fraction named St. NaOH was once extracted with 1 % sodiumhydroxide.Samples for the Ll.c. were prepared from acetone extract by using a Soxhlet'sapparatus.The i.r. charts were obtained using KBr pellets with JASCO IR-S. U.v. chart- 17-WOOD RESEARCH No. 50 (1970)was recorded by using Hitachi 120 double beam spectrophotometer in 95 % ethanolicsolution. Charts of n.m.r. were taken in the 10 % solution of D6-DMSO at 22°Cusing }EOL, Minimer (MH-60, 60 MHz).As extractives for isolating sawaranin the ether soluble parts of methanolsoluble matters of normal hinoki were chosen.For instance, five grams of methanol soluble and ether soluble fractions fromnormal hinoki were placed on the top of silicic acid column (Merck, 0.05-0.2 mm;10 mm¢, 30 cm long) packed with chloroform, and following solvent systems wereapplied for elution, in order; ether-chroloform (1: 1 vlv, 300 ml), chloroform(150 ml), chloroform-acetic acid (9: 1 vlv, 500 ml) and acetone-chloroform-aceticacid (50: 45 : 5 vlv, 500 ml). Crude crystals (217 mg) were collected from a red-brownish solution of the eluate with the above last solvent system. The solventsystem of ethylacetate-acetone (1: 1 vIv), instead of the above acetone-chloroform-acetic acid system, was also available to obtain the eluate in which sawaranin wascontained. The above described crystals were established to be sawaranin incomparison with the data of Amax. in u.v. (277 nm in 95 % ethanol) and theidentity of Lr. chart with authentic one as shown in Fig. 5. Recrystalized sawara-nin was insoluble in ether and following results were obtained from the elementaryanalysis,Found: C., 56.48 %; H., 5.49 %Calcd. for C12H140 6 : C., 56.68 %; H., 5.56 %AcknowledgementThe authors express their thanks to Mr. T. Higuchi for the gift of samplewoods at Goverment Forestry Station at Tsukechi, Gifu, and also thanks to Dr.H. IMAMURA for giving us the authentic compound of sawaranin.Literature1) YOSHIKI, Y. and T. ISHIGURO, j. Pharm. Soc. japan, 53, 1 (1933).2) HIROSE, Y., N. OISHI, H. NAGAKI and T. NAKATSUKA, Tetrahedron Letters, 3665 (1965).3) IMAMURA, H., Bull. of the Gover. Forest Experi. Station, japan, No. 138, 1 (1962).4) TAKASHIMA, T., M. OGUSHI, Y. KIUCHI and F. AKABANE, J, japan Wood Res. Soc.,14, 391 (1968).5) NISHIDA, K., "Wood Industry" Vol. 1, p. 74, Asakura Publishing Co. (1946), (in japanese).6) MIGITA, N., "Wood Chemistry" Vol. 1, p. 72, Kyoritsu Publishing Co. (1968), (in japanese).-18 -

Some Chemical Observation on a Natural Graft-Wood of Japanese Cypress (Chamaecyparis obtusa ENDL.) and Sawara Cypress (Chamaecyparis pisifera SIEB. et ZUCC.)

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Some Chemical Observation on a Natural Graft-Wood of Japanese Cypress (Chamaecyparis obtusa ENDL.) and Sawara Cypress (Chamaecyparis pisifera SIEB. et ZUCC.)

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