eat ye rie Net ANALYSIS OF NUTMEG OIL USING CHROMATOGRAPHIC METHODS Gaon Rrch Comgay Li, CHI Diente (Sets) Various chromatographic methods were ied for he alse of utero In audition tote fourten known monoterpene hydrocarbons, opimethyistyrene hs been identied togeier with seen monotepenealeohol even eters and eight sesquiterpene hydrocarbons. The osartence ofthe mins mors iomer of pment enc and a bifenstionl derivative ofthe same pe Uranepmenth- antl fi are of spel interes. Bess the terpenoid constants two unstated al Dhatic compounds, sme (E)-Smetylstdecen--l and is actat, were detected forthe fit time na natural soba “The characteris odours of natural substrates (pies, essen lls, foods in senera) are mainly coud by volatle components. Gas iud chromatography (GLC) ste most important 90 forthe analysis ofthese compounds. Oler ana ‘al methods worked well nl forthe main coasttuets, but were unstable forthe tac components which so often ae responsibie or the overall favour or odour. The tow methods developed forall Kinds of chromatograph techniques slow eich sr guaitatve and quantitative montorng of compontons. The rapid dveop- ‘ment of expiry column in gat chromatography fas facitated the avestization of Complex mistres suchas een oi, The fei of spicy eset oil, nutmeg cil and mae ol play an important ‘ein the favour ad fragrance nduty, Owing to thei known content of noous Shanes suchas saflabd myrsic it was cestabl to obtain more information ‘boot the qualitative and quantitative compositions im ration tothe odour and ‘avour charters of there ol. Based on the compton of Van Staaten we ‘hve now confirmed ll ofthe components aes) knows and adde to thelist of ute onsitent the olowing compounds: one onotrpene hydrocarbon; ast {uierpene hydrocarbons six monoterpee alcool thee exer; vanilin and two ‘light compounds detected forthe fst time in nature, (E)-methyl--decen--l ‘and its asa Ge Table).‘uine olof commercial grade (Roure-Bertand, Grasse, Frans) was fist ised In acon, Cetin fractions mere combined snd then chromstoerapied on ‘lca pel #0 (E- Mere, Damsint, CER) using hese diy er fa various ato (10:0 (00:10) inorder to soit specie components ofthe i, Thus, 20 of ‘dilation fraction (op. 34-39°C1DO4 mmlsy 1.87. of the tla of) were Bet ‘hromatogaphed on 150g of sca gl 6D. Aller clinton ofthe hydrocarbon, ‘esane-dty ter (10:1) wa ed fo lute 43g ofa complex mice which wat {cchromatoprapbe on 130g of sen sl 6. Eution with hexane eter (101) ‘hen afoded + mature (0.6) consisting mail ofthe known rausabinee Byte, ‘ai and ranepmeath-Pen--l. The pronously unknown ransssomer Was olated {n pure fom (64 mg) by means of preparative GLC. ‘Another stilton Fraction (op 49-&5°C/003 maby; 34.19% ofthe toul co) was teat the same ay to it 200 mg of (E)- meth -+ece 0 (purity $02) ing exaneetyl ether (Se) a eluent Final elation with et ete forded tone menth-2eneI-hiol 70% puny. Both constituents were farther ured by preparative GLC. ‘AI GLC reaurement were conducted on 2 Carlo Erba Gl appara. gas sluma (8m > 11am LD) packed with 10-59, Crowns 20M (ong Ssresing Steps with creasing dsanee fom column entrance) on Supelcoport treated with terephthalic ae, assed for preparative work Temperate othemal, 100 of 20°C. Cari pu nitrogen, 200 mimi. A lass clan (30> mum .D.) packs with 3% Carbowax 20M 0a Chromosore G HP (80-100. mest, teted wth Teephtalic ai, war and or the quantiaive determination of mytsticin. Tem Patz: programmed fom to 29°C at 6Clnin. Cave gas: um, 5 min Cepia columis (Om > 0.1 mm ID.) wee cated with UCON. HB 3100 or (Crbowa ID epesvely.Tenperstue: programmes fom $010 185°Cat Cj (Carrier ps: Blt, 2 Sli. ‘Specioicopic data of ote compounds were recorded on Perin-Emer Model 187 (nates), Broker WH60 (11 muta magecic resonance, NMR) and Yaran XLIOIA (°C NMR) spectophotometes. A Vatan-MAT CHS or 212 ambition wth Varian SS 10 or Pinan Inco 2400 compter site ate Formas spectrometry (MS) nd GLC-MS coupling experimen Monoterpene hydrocarbons represent about 70% of nutmeg oi In GLC caperinents, only Carboway 1000 revoked al een monoterpene hyerorabons ‘Wi i stashonary phase even 1inol swell sparted fom the adjacent yaro= carbons limonene and pphelandreae) ap Dinethystyrene, already known 0 oeear In other oar sures, war newly eteied a ace component in ute ol By GLE-MS couping expense "Te eal columns ud showed ice’ separation of sesquiterpene hydro: carbons from mooterpene alcohols, ecopuene from iranosabiaene hydate of Styopiylene Irom tanrpmenth-leni-ol They asp enabled the detection of & ‘isquburpene hydrocarbon which seams to dccomposs during the pectesiment via sii gel column chromatography. We ssume that this s @ grmacrenesype ses ‘terpenes hyeocrbon in ve ofthe other Meta compounds which reread {Oa common intredat (Pip 1). Gemacrenes are noha tobe very seni and poe to seltal ecrangerens unde ace condtions™ Two pair of ciranitomere monoterpene alcohol ual ose togeter ia ‘zen sof several spes (specially in Larnacee derived oy for instance fosemary i), names, et aneaabnene Hydrate and evanspsenth-2en-- ‘Thesof te compounds ae known occur inating The ming fourth compound (Gram pil nen bas now also een ented iis oation nd compan ‘fect than authentic pene ‘Another interesting finding waste ideatifston of trompmenth-2ene- 4. it in tne. Tks hl, slay Known a5 conten of Mitcham peppermint {ste ft bfeneonal monoteene desta detected in nuteg of sucure twas asspned by comparaon wth sntte specimen of known sereochems ‘curl was redvod ith ipenyipoephine’™ and give aftr hycofss he ran Ftomer ident inal rests withthe soled mate Icons, reduction of ‘tearl with ium aluminum hdd lads tothe ceiomer, 8 Set undetected Four atural subst, “The ocsrrence of (5) -methy!-tdecenol([aF*— 3205) ¢ = 1085, CHC) ina natural material bere eepored fr te fet tne, The statue of the isolated alcobl flows fom specrscopc data and comparison with n autbenti, sate ample! This represents 2 good example of the successful isolation and structureslucdtion by combination of tedious chromatographic sep with modern spec teosopieans The natural lebol seems to be opal pute, sn no addon lines were observed when an optically active Eu} reagent war added. [a ontras the sate specinen shows oulingo the character signals tothe presne of bol posible omer ‘Addionaly, dmth--cecelyl seetate was detested a trace component by GLC-MS where the mass specometer was operted in the chemical fomaton| (i) moves the CT spectrum sing obutane as eacton ga exhib intense nals a CcomPosimion oF NUTMEG on. et aoe eta ares ‘beans nab ete erent ‘Gone ast ‘Gerace BF Sore Fe 3 remtass rom Apa ‘rowing "EMetidn .t Nees os SMe ae ims ~ 153 and 213, whereas he noemaly recorded lesteon impact peta com pletely lacks the molclar fon a mje = 212 “The soli ofthe roca mace othe oof he ded alo of mutes revels only quantitative bu no lgicnt ulate Jiereaces when compe with the eluting ol The higher percentage of eatyopylene andthe shah lower Amousts of safol and myrisicin are to be noted. For she le, 4 noxious oustunt with prepoatve, tore ropertis, a quaniatve GLC method wat Sesloped sing dc phthalate a nteral standard. Our intrest in developing {80h a method nas aroused by the fact that the iterate ales obaed by GLE ‘measurements show some dserepancy. The esl ftom ou method artespond well {o the values obtained by conventional workup (13-1820: nutmeg 139 05% mass 1015 08% 1 Table the approximate composition of nue lis given forthe min component, ths undeining the prodomiance (107) of moeterpene hydowar one 1 B.A. atten, agen, he 8809 92, 28 Stn (ey, it Compound, Catal ae or Nu and Feed 3 eVen Oar Saal ewe, Tee at, 1968) 26, {1 Mormon and ce, Teron Le 90) 199 SANs WW une, Was X,8. Kaa LS. Chee, Terns (998 6 Dagunay aid He hen, 1. tag hr Varkomnen nt Ari rile Pisin Boma, WS Maton, Nab, GR Paes and FV. Rabon, et Tar fad sae (9 8.6.0 Sees, Kin, 0 Buhl S. Serer nd GONE, Sn Lis A, Chey 9 kta, Kt K. Hashimoto and M, Koby, Kye ala Zah, 6198) 10.6.0. avon a0, Rau, J Or. Cha 34058 8 11 4a te Raga ate ta C3 Shy Bch, 14097 2987. HE Benoland PEs a Chon Ao 52 (9099