Interntionl Journl of Applied Reserh in Nturl Produts Vol. 9 (1), pp. 9-13. Diretory of Open Aess Journls 2008-2016. IJARNP-HS Pulition Originl Reserh Chemil Composition of Lef Essentil Oils of Syzygium smrngense (BL.) Merr. et Perry v. Pink t Three Mturity Stges Pei-Chi Lee 1, Hong-Yi Guo 1, Chi-Cho Hung 2, Chin-Feng Chn 1* 1 Deprtment of Applied Cosmetology, Hungkung University, No.1018, Se. 6, Tiwn Boulevrd, Shlu Distrit, Tihung, Tiwn, R.O.C. 2 Fengshn Tropil Hortiulturl Experimentl Brnh, Tiwn Agriulturl Reserh Institute, Tiwn, R.O.C. Summry. The hemil omposition of the lef essentil oils of Syzygium smrngense (BL.) Merr. et Perry v. Pink t young, mture, nd old stges ws nlyzed through gs hromtogrphy mss spetrometry (GC-MS). There were 33 ompounds identified. The min omponents of 3 stges of lef essentil oils were α-pinene, γ-terpinene, β-ryophyllene, α-selinene, β-selinene, nd selin-6-en-4-ol. The results demonstrted tht levels of β-elemene, γ-terpinene, nd β-ryophyllene deresed during mturtion. However, ryophyllene oxide nd p-ymene grdully inresed. The mture stge oil exhiited higher level of monoterpene whih my e ssoited with the potent superoxide svenging effet ut exhiited lower level of sesquiterpene ompre to other stges of essentil oils. The old stge essentil oil exhiited more plesnt fruity rom whih my e relted to inreses of p-ymene, ryophyllene oxide, nd trns-3-ren-2-ol. Industril relevne. The remrkle rdils svenging effet nd rom exhiited y the lef oil n e inorported into the food, drug or osmeti formultions to redue the risk of ner, rdiovsulr diseses or skin geing nd to inrese flvor of foods or osmetis. We re urrently fous on investigting the pplition of extrts of Syzygium smrngense on skin re nd on ntimiroil tivity in osmeti industry. Keywords. Syzygium smrngense (BL.) Merr. et Perry v. Pink; essentil oil; gs hromtogrphy mss spetrometry; superoxide INTRODUCTION Syzygium smrngense (BL.) Merr. et Perry v. Pink is pink vriety of wx pple, whih is one of the most frequently plnted fruit rops in Tiwn (Figure 1). 1 The fruit extrts of S. smrngense were reported to hve potent ntirdil tivity euse of high levels of totl phenols nd nthoynins. 2 In ddition, the methnoli extrts of the pulp nd seeds of S. smrngense lso exhiited ntiner tivity. 3 The flower ud of S. smrngense exhiited potent ntioxidnt tivity, nd 40 hemil omponents in it hve een identified, inluding ryophyllene (7.95%), 1,2,4,5- tetrmethyl enzene (7.187%), nd ryophyllene oxide (6.319%). 4 Essentil oil from the lef of Syzygium smrngense n inhiit Esherihi oli growth y reduing its extrellulr protese. 5 However, little or no study onduted on the hemil omposition of the lef essentil oils of S. smrngense during mturtion. Figure 1. Piture of Syzygium smrngense (BL.) Merr. et Perry v. Pink *Corresponding Author. fhnjmes@hotmil.om 886-4-2631-8652 ext. 5314 Aville online http.//www.ijrnp.org
Lee et l. Chemil ompositions nd iologil tivities t vrious stges of mturtion plnt tissues my hve signifint differene. 6 Artemisi nnu essentil oils exhiited high levels of monoterpene t preflowering phse (89.11%). By ontrst, high levels of sesquiterpenes were otined t postflowering phse (58.73%). 6 It hs een reported tht vrious flowering stges of Artemisi nnu essentil oils exhiited distint hemil ompositions nd ntietylholinesterse tivities. 6 MATERIALS AND METHODS Plnt mteril nd essentil oil isoltion. Fresh plnt leves of Syzygium smrngense were olleted t vrious stges of mturtion from the Fengshn Tropil Hortiulturl Experiment Brnh, Agriulturl Reserh Institute, Counil of Agriulture in Tiwn, R.O.C. All leves were olleted from August 2012 to Deemer 2012. Young leves were hrtreuse or ple green in olor nd developed in 2 months. They further grew for pproximtely 2 to 4 months nd the resulting olor ws green ssoited with mture leves. The olor of the old leves ws drk green, nd ppered fter longer growth phse of pproximtely 4 to 6 months. One kilogrm of fresh shredded leves ws ir dried in the shde. The leves were sujeted to stem distilltion, using Clevenger-type pprtus for 3 h. 4 The essentil oil ws evported together with wter vpor nd olleted fter denttion. Cupri sulphte nhydrous ws used to remove the wter fter extrtion. The essentil oil ws stored t 4 C until tested nd nlyzed. Gs hromtogrphy MS nlysis. Voltile omponents were nlyzed using GC-MS QP 2010 (Shimzu; Seiskusho, Jpn) equipped with BP-X5 pillry olumn (28.8 m 0.25 mm; oting thikness 0.25 μm). Crrier gs helium t 1 ml/min; volume injeted, 1 μl of solution in methnol of the oil; injetor temperture: 220 C; oven temperture progrm: 70 C to 220 C (5 C/min); splitting rtio: 1:10. The MS sn onditions inluded trnsfer line temperture of 250 C, n interfe temperture of 250 C, n ion soure temperture of 200 C, n ioniztion tehnique, n eletroni impt (EI) of 70 ev, nd n quisition rnge of 40 to 400 m/z. 5 Identifition of the onstituents ws sed on omprison of retention indies nd on omputer mthing ginst ommeril (NIST 05) nd homemde lirries, nd on omponents of known oils nd GC-MS dt provided in the literture. Sttistil Anlysis. All tests were performed in triplite. Sttistil evlution ws performed y running the Dunn s multiple rnge test nd n nlysis of vrine ws ompleted using SigmSttR (Version 3.11.0, Systt Softwre In., Sn Jose, CA, USA). The nlysis signifine level ws α =.05, nd differene ws onsidered sttistilly signifint for p vlue less thn 0.05. RESULTS AND DISCUSSION Extrtion yields. The essentil oils otined from young, mture, nd old leves of S. smrngense produed 0.02%, 0.03%, nd 0.02% yield (v/w), respetively. Chemil omposition of essentil oils. The identified ompound, retention indies (RI), nd the perentge of the pek re of S. smrngense lef essentil oils nlyzed using GC-MS re summrized in Tle 1. Thirty-three omponents were identified t eh development stge of the lef essentil oils. The min omponents in the three lef essentil oils were α-pinene (21.85% 26.40%), γ-terpinene (9.51% 11.17%), β- ryophyllene (4.67% 7.72%), α-selinene (4.67% 5.60%), β-selinene (4.94% 6.00%), nd selin-6-en-4-ol (5.07% 5.21%) (Figure 2). Trns-3-ren-2-ol ws only found in mture nd old lef essentil oils. However, germrene D ws only found in young lef essentil oil. Germrene D is onsidered to e preursor to mny sesquiterpene hydrorons nd tht my e ssoited with lower levels of sesquiteterpene in mture nd old leves. 7 Vrious hnges to the hemil ompositions of lef essentil oils during mturtion were oserved in this study. Monoterpene ws inresed in young leves from 42.85% to 50.34% in mture leves, nd my e relted to the inrese of α-pinene from 21.85% to 26.40%. Sesquiterpene ws deresed from 32.06% to 22.06%, whih my e relted to redution of β-elemene nd β-ryophyllene. Surprisingly, the results were not onsistent with the flowering stge of A. nnu. In this study, the β-ryophyllene ontent in young lef essentil oil ws higher thn in mture nd old lef essentil oils (Figure 3). In previous study, β-ryophyllene ws oxidized into ryophyllene oxide over time. 8 Furthermore, β- ryophyllene indued hlorosis in leves. 9 Therefore, the mount of norml growth β-ryophyllene in leves ws deresed nd trnsformed into ryophyllene oxide over time, 9 whih ws onsistent with our results. Cryophyllene oxide is frequently used in osmetis nd s flvoring gent in foods. 10 Therefore, inresing ryophyllene oxide in old lef essentil oil my ontriute to its more mild nd plesnt odor thn in young lef essentil oil. The result is onsistent with the vrious mturtion stges of flowering essentil oils of A. nnu. 11 The ryophyllene of A. nnu oils deresed from preflowering to postflowering phse. By ontrst, β-ryophyllene oxide inresed drmtilly during flowing stges. 11 The other monoterpene γ-terpinene n romtize into p-ymene, 12 whih is widely used s n intermedite in the synthesis of fine hemils for flvorings, frgrnes, perfumes, fungiides, nd pestiides. 13 In this study, γ-terpinene deresed during mturtion nd the mount of p-ymene grdully inresed, whih my indite tht γ-terpinene ws trnsformed to p-ymene t the mture nd old lef stges (Figure 3). Trns-3-ren-2-ol ws only found in mture nd old lef essentil oils (Figure 4). Cr-3-ene hs een reported to ontriute to the hrteristi mngo odor of the rhizome of Curum md Rox nd the Venezueln mngo. 14, 15 Trns- 3-ren-2-ol my lso use the fruity rom of mture nd old leves. Therefore, the plesnt fruity rom of old lef essentil oil my e relted to p-ymene, ryophyllene oxide, nd trns-3-ren-2-ol. 10
Lef essentil oils of Syzygium smrngense Tle 1. Min omponents of Syzygium smrngense lef essentil oils t 3 different mturity stges No. RI Compound Pek re (%) of lef essentil oils Young Mture Old 1 902 α-thujene 1.64 ± 0.37 1.84 ± 0.05 0.76 ± 0.04 2 943 β-pinene 1.97 ± 0.28 2.47 ± 0.12 2.21 ± 0.07 3 948 α-pinene 21.85 ± 2.77 26.40 ± 1.72 26.35 ± 2.47 4 969 α-phellndrene 0.66 ± 0.13 0.74 ± 0.11 0.40 ± 0.04 5 976 β-is-oimene 3.24 ± 0.15 3.95 ± 0.07 0.60 ± 0.03 6 998 α-terpinene 1.00 ± 0.17 1.40 ± 0.03 0.99 ± 0.15 7 998 γ-terpinene 11.17 ± 0.78 11.13 ± 0.32 9.51 ± 0.61 8 1042 p-cymene 1.33 ± 0.07 2.41 ± 0.02 3.22 ± 0.31 9 1052 α- Terpineol 3.69 ± 0.17 3.79 ± 0.06 2.65 ± 0.21 10 1136 trns-3-cren-2-ol - 3.93 ± 0.46 5.70 ± 0.36 11 1344 α-copene 1.06 ± 0.08 0.49 ± 0.04 0.93 ± 0.09 12 1398 β-elemene 3.23 ± 0.07 0.72 ± 0.06 0.36 ± 0.05 13 1410 α-elemene 1.00 ± 0.11 0.89 ± 0.05 1.10 ± 0.07 14 1435 γ-cdinene 0.52 ± 0.06 0.39 ± 0.05 0.51 ± 0.07 15 1469 β-selinene 6.00 ± 0.19 4.94 ± 0.03 5.52 ± 0.48 16 1469 δ-cdinene 1.68 ± 0.12 1.22 ± 0.08 1.29 ± 0.03 17 1474 α-selinene 5.60 ± 0.28 4.67 ± 0.01 4.71 ± 0.24 18 1494 β-cryophyllene 7.72 ± 0.23 5.29 ± 0.01 4.21 ± 0.26 19 1502 γ-selinene 3.13 ± 0.18 2.43 ± 0.09 2.42 ± 0.14 20 1507 Cryophyllene oxide 0.56 ± 0.04 1.58 ± 0.09 3.51 ± 0.06 21 1515 Germrene D 0.65 ± 0.31 - - 22 1530 Ledol 0.15 ± 0.55 0.39 ± 0.02 0.45 ± 0.01 23 1530 (-)-Gloulol 2.55 ± 0.15 2.74 ± 0.03 2.97 ± 0.05 24 1564 Nerolidol 0.48 ± 0.08 0.37 ± 0.02 0.59 ± 0.08 25 1579 α-cryophyllene 1.46 ± 0.09 1.01 ± 0.06 0.90 ± 0.09 26 1580 Cuenol 0.32 ± 0.46 0.97 ± 0.33 0.78 ± 0.09 27 1580 τ-cdinol 1.27 ± 0.13 1.11 ± 0.05 1.86 ± 0.34 28 1580 α-cdinol 1.08 ± 0.30 0.66 ± 0.06 0.68 ± 0.16 29 1580 δ-cdinol 0.28 ± 0.37 1.00 ± 0.06 0.58 ± 0.16 30 1593 Selin-6-en-4-ol 5.18 ± 0.29 5.07 ± 0.09 5.21 ± 0.33 31 1593 Crotol 1.86 ± 0.25 1.52 ± 0.06 2.01 ± 0.13 32 1598 Hinesol 1.96 ± 0.19 1.96 ± 0.14 2.43 ± 0.05 33 1647 Juniper mphor 0.65 ± 0.09 0.61 ± 0.07 0.81 ± 0.07 34 1710 Frnesol 2.85 ± 0.33 1.9 ± 0.18 3.33 ± 0.14 Totl 97.79 100 100 Monterpenes 42.85 50.34 44.03 Oxygented monterpenes 3.69 7.72 8.35 Sesquiterpenes 32.06 22.06 22.41 Oxygented sesquiterpenes 19.19 19.88 25.21 Retention indies reltive to MS dtse. Reltive quntittive dt otined y GC-FID nlysis. Vlues re mens of triplite nlyses. Figure 2. The min omponents of old Syzygium smrngense lef essentil oils y GC-MS nlysis 11
Lee et l. () () Figure 3. Content of the different omponents of Syzygium smrngense lef essentil oils t 3 different mturity stges. () Content of the β-ryophyllene nd ryophyllene oxide of young, mture nd old oils. () Content of γ-terpinene nd p-ymene of young, mture nd old oils. The sme omponent of three different mturity oils with different lower se letters indites signifint differene (p < 0.05). Figure 4. The ontent of trns-3-ren-2-ol of Syzygium smrngense lef essentil oils t 3 different mturity stges. The omponent of three different mturity oils with different lower se letters indites signifint differene (p < 0.05). 12
Lef essentil oils of Syzygium smrngense ACKNOWLEDGEMENTS The uthors thnk the Fengshn Tropil Hortiulturl Experiment Brnh, Agriulturl Reserh Institute, Counil of Agriulture, Exeutive Yun, Tiwn, R.O.C. for kindly providing fresh leves of S. smrngense t vrious mturtion stges. The uthors lso thnk the Ntionl Siene Counil, Tiwn, R.O.C. for finnil support (NSC102-2622-E-241-006-CC3). REFERENCES 1. Annul report : Counil of Agriulture (COA). Tiwn: Agriulture nd Food Ageny, Exeutive Yun (Ed.); 2009. 2. Reynertson KA, Yng H, Jing B, Bsile MJ, Kennelly EJ. Quntittive nlysis of ntirdil phenoli onstituents from fourteen edile Myrtee fruits. Food Chem. 2008;109(4):883-890. 3. Simirgiotis MJ, Adhi S, To S, Yng H, Reynertson KA, Bsile MJ, et l. Cytotoxi hlones nd ntioxidnts from the fruits of Syzygium smrngense (Wx Jmu). Food Chem. 2008;107(2):813-819. 4. Go Y, Hu Q, Li X. Chemil omposition nd ntioxidnt tivity of essentil oil from Syzygium smrngense (BL.) Merr. et Perry flower-ud. Sptul DD. 2012;2:23-33. 5. Reddy LJ, Jose B. Chemil omposition nd ntiteril tivity of the voltile oil from the lef of Syzygium smrngense (Blume) Merr. & L.M. Perry. Asin J Biohem Phrm Res. 2011;3(1):263-269. 6. Yu Z, Wng B, Yng F, Sun Q, Yng Z, Zhu L. Chemil ompositionnd nti-etyl holinesterse tivity of flower essentil oils of rtemisinnut different flowering stge. Irn J Phrm Res. 2011;10(2):265-271. 7. Bulow N, Konig WA. The role of germrene D s preursor in sesquiterpene iosynthesis: investigtions of id tlyzed, photohemilly nd thermlly indued rerrngements. Phytohemistry. 2000;55(2):141-168. 8. Skold M, Krlerg AT, Mtur M, Borje A. The frgrne hemil et-ryophyllene-ir oxidtion nd skin sensitiztion. Food Chem Toxiol. 2006;44(4):538-545. 9. Snhez-Munoz BA, Aguilr MI, King-Diz B, Rivero JF, Lotin-Hennsen B. The sesquiterpenes et-ryophyllene nd ryophyllene oxide isolted from Seneio slignus t s phytogrowth nd photosynthesis inhiitors. Moleules. 2012;17(2):1437-1447. 10. Di Sotto A, Mffei F, Hreli P, Cstelli F, Srpietro MG, Mzznti G. Genotoxiity ssessment of et-ryophyllene oxide. Regul Toxiol Phrmol. 2013;66(3):264-268. 11. Lio WC, Li YC, Yun MC, Hsu YL, Chn CF. Antioxidtive tivity of wter extrt of sweet potto leves in Tiwn. Food Chem. 2011;127(3):1224-1228. 12. Botnik I, Xue W, Br E, Idh M, Shwrtz A, Joel DM, et l. Distriution of primry nd speilized metolites in Nigell stiv seeds, spie with vst trditionl nd historil uses. Moleules. 2012;17(9):10159-10177. 13. Kmitsou M, Pngiotou GD, Trintfyllidis KS, Bouriks K, Lyourghiotis A, Kordulis C. Trnsformtion of α-limonene into p- ymene over oxide over oxide tlysts: A green hemistry pproh. Appl Ctl A : Gen. 2014;474:224-229. 14. Gholp AS, Bndyopdhyy C. Chrteriztion of mngo-like rom in Curum md Rox. J Agri Food Chem. 1984;32:57-59. 15. MLeod AJ, Pieris NM. Comprison of the voltile omponents of some mngo ultivrs. Phytohemistry. 1984;23:361-366. 13