Variation in Chemical Composition of Essential Oil Extracted from the Fruits and Leaves of Cinnamomum Tenuipile Kosterm (Sugandhakokila) of Nepal

2019J. Pl. Res. Vol. 17, No. 1, pp 86-93, 2019 Journal of Plant Resources Vol.17, No. 1

Variation in Chemical Composition of Essential Oil Extracted From the Fruits and Leaves of Cinnamomum tenuipile Kosterm (Sugandhakokila) of Nepal

Tara Datt Bhatt*, Amit Dhungana and Jyoti Joshi Department of Plant Resources, Thapathali, Kathmandu, Nepal *Email: [email protected]

Abstract

The purpose of this study was to find out the variation of major chemical constituents present in the essential oil of leaves and fruits of Cinnamomum tenuipile Kosterm analyzed by GCMS instrument. Leaves and Fruits of Cinnamomum tenuipile Kosterm were collected from Brindaban Botanical Garden of Makawanpur district of Nepal. The extraction of essential oil was performed by hydro-distillation using Clevenger apparatus and then their chemical composition was identified by gas chromatography coupled with mass spectrometry (GC-MS). The results of chromatographic analysis have shown somehow similar compounds except camphor which was found in fruits whereas it was absent in leaf oil. By GCMS analysis 13 and 15 compounds were identified respectively in which eucalyptol (24.17%) and methyl cinnamate (52.18%) were found as major compound in leaf oil while eucalyptol (38.23%), camphor (19.57%) and methyl cinnamate (22.53%) were found as major compound in fruit oil.

Keywords: Camphor, Clevenger , Eucalyptol, GC-MS, Methyl cinnamate

Introduction The different parts of Sugandhakokila tree contains essential oil in different percentages which is used for The evergreen, Cinnamomum tenuipile Kosterm the formulation of perfume as well as uesd in the form (syn. Cinnamomum cecidodaphne), part of the of scent (Adhikary, 2018). The nematicidal, termiticidal, Lauraceae family, is native to Nepal and grows wild mosquito larvicidal (Satyal et al., 2013), insecticidal, in the districts of Dang, Rolpa and Sallyan in antifungal, antiaflatoxin, antioxidant (Prakash et al., the Rapti Zone (Rema et al., 2002, Adhikari , 2018). 2013) and antibacterial (Rajendra et al., 2013) This species is a diploid and can grow to an altitude activities of essential oils have been also reported. of 1300 meters (Ravindran et al., 2003). Cinnamomum tenuipile Kosterm is recognized as an aromatic plant, In the present study, essential oil is extracted from meaning it has an elevated level of essential oil the fruits and leaves of Cinnamomum tenuipile (Gurung, 2015). Using steam distillation, the dried Kosterm (Sugandhakokila) collected from berries of Cinnamomum tenuipile Kosterm produce Brindaban Botanical Garden, Makawanpur Nepal the essential oil commonly known as sugandha and oil was analysed by Schimadzu GC-MS QP 2010 kokila oil (Ravindran et al., 2003), which is yellow Plus. The purpose of this study was to compare the in color and has a camphoraceous, spicy aroma chemical constituents present in the both oil samples. (HPPCL, 2015). This product can be used as a fragrance in soaps, detergents, cosmetics, perfumes Materials and Methods and industrial fragrances (Gurung, 2015). Sugandha kokila oil is also used in indigenous medicine as a Collection of plant materials demulcent and stimulant (Rema et al., 2002). The The fruits and leaves of Cinnamomum tenuipile Nepal Trade Integration Strategy 2010, identified Kosterm were collected from Brindaban Botanical Medicinal and Aromatic Plants (MAPs) as one of Garden, Makawanpur district of Nepal. The fresh Nepals top twenty goods and services with export fruits and leaves were collected and dried in shed potential (Sharma, 2015). before extraction of essential oil.

86 The fruits and leaves of Cinnamomum tenuipile Kosterm were collected from Brindaban Botanical Garden, Makawanpur district of Nepal. The fresh fruits and leaves were collected and dried in shed before extraction of essential oil. 2019J. Pl. Res. Vol. 17, No. 1, pp 86-93, 2019 Journal of Plant Resources Vol.17, No. 1 2019 Journal of Plant Resources Vol.17, No. 1

Extraction of essential oil Variation in Chemical Composition of Essential Oil Extracted From Extraction of essential oil column oven temperature was set at 40°C and the the Fruits and Leaves of Cinnamomum tenuipile Kosterm A Clevenger apparatus was used for the extractioninjection of essential temperature oil from was 250°C. the fruits and leaves of ACinnamomum Clevenger apparatus tenuipile was Kosterm used for through the extraction hydro distillation (Waheed et al., 2011). The fruits and leaves (Sugandhakokila) of Nepal ofwere essential thoroughly oil fromwashed the and fruits placed and in leavesClevenger of apparatusThe qualitative and subjected analysis to hydro of the distillation essential foroil aboutwas Cinnamomum8 hours. The steamtenuipile and vaporizedKosterm throughoil were condensedhydro further into liquid continued by a vertical in a Shimadzu condenser GCMS-QP2010 and collected in Tara Datt Bhatt*, Amit Dhungana and Jyoti Joshi distillationmeasuring (Waheedtube. Being et al.,immiscible 2011). Theand lighterfruits and than water,Plus. Duringthe volatile the analysis, oil separated the ion out source as an temperature upper layer. Department of Plant Resources, Thapathali, Kathmandu, Nepal leavesThe oil were was thenthoroughly separated washed from water and andplaced collected in andin small the interface glass bottles, temperature dried with was anhydrousset at 250°C sodium and *Email: [email protected] Clevengersulphate, sealed, apparatus labelled and and subjected stored in glassto hydro vials. 200°C respectively. The detector scanning start time distillation for about 8 hours. The steam and Abstract was 4 min and end time was 68 min; scan speed vaporizedGas chromatography- oil were condensed Mass into spectrometry liquid by a vertical (GC-MS) was 666 with scanning range of m/z 40.00-350.00. The purpose of this study was to find out the variation of major chemical constituents present in condenser and collected in measuring tube. Being Identification of compounds was done by comparing the essential oil of leaves and fruits of Cinnamomum tenuipile Kosterm analyzed by GCMS immiscibleThe chemical and constituentslighter than inwater, the essentialthe volatile oils oilwere separated by using a Shimadzu Gas chromatograph instrument. Leaves and Fruits of Cinnamomum tenuipile Kosterm were collected from Brindaban the Mass spectral data present in the mass spectral separatedMass Spectrophotometer out as an upper (GCMSlayer. The QP oil 2010 was Plus) then withlibrary Rtx-5MS NIST column 2017 and (30mX0.25mmX0.25 FFNSC 1.3. µm). 1 µL Botanical Garden of Makawanpur district of Nepal. The extraction of essential oil was performed of the essential oil diluted with spectroscopic grade hexane (10:1) was injected into the GC inlet by hydro-distillation using Clevenger apparatus and then their chemical composition was separated from water and collected in small glass bottles,maintaining dried column with anhydrous flow rate ofsodium 0.68 mL/minsulphate, and purge flow 3 mL/min in the split mode. The initial identified by gas chromatography coupled with mass spectrometry (GC-MS). The results of 0 Results and Discussion0 chromatographic analysis have shown somehow similar compounds except camphor which was sealed,column labelled oven temperature and stored inwas glass set vials.at 40 C and the injection temperature was 250 C. found in fruits whereas it was absent in leaf oil. By GCMS analysis 13 and 15 compounds were The oil extracted from fruits and leaves of identified respectively in which eucalyptol (24.17%) and methyl cinnamate (52.18%) were found GasThe chromatography- qualitative analysis Mass of spectrometry the essential (GC-MS) oil was furtherCinnamomum continued tenuipilein a Shimadzu Kosterm GCMS-QP2010 (Sugandhakokila) Plus. as major compound in leaf oil while eucalyptol (38.23%), camphor (19.57%) and methyl cinnamate During the analysis, the ion source temperature and the interface temperature was set at 250 0C and 200 (22.53%) were found as major compound in fruit oil. The0 chemical constituents in the essential oils were was analyzed by GCMS instrument and the C respectively. The detector scanning start time wascomposition 4 min and ofend various time was constituents 68 min; scanpresent speed in thewas separated666 with by scanning using a Shimadzurange of m/z Gas 40.00-350.00. chromatograph Identification of compounds was done by comparing the Keywords: Camphor, Clevenger , Eucalyptol, GC-MS, Methyl cinnamate respective oil is tabulated below in Table 1 & Table MassMass Spectrophotometer spectral data present (GCMS in the massQP 2010spectral Plus) library NIST 2017 and FFNSC 1.3. 2 respectively which is nearly similar to the analysis with Rtx-5MS column (30mX0.25mmX0.25µm). Introduction The different parts of Sugandhakokila tree contains 1 µL of the essential oil diluted with spectroscopic of essential oil of fruits of sugandhakokila (Adhikary essential oil in different percentages which is used for gradeResults hexane and (10:1)Discussion was injected into the GC inlet et al., 2011) which shows 1,8-cineole, methyl The evergreen, Cinnamomum tenuipile Kosterm the formulation of perfume as well as uesd in the form maintaining column flow rate of 0.68 mL/min and cinnamate, alpha-terpineol as major constituents. (syn. Cinnamomum cecidodaphne), part of the of scent (Adhikary, 2018). The nematicidal, termiticidal, purgeThe flow oil extracted 3 mL/min from in the fruits split andmode. leaves The initial of Cinnamomum tenuipile Kosterm (Sugandhakokila) was Lauraceae family, is native to Nepal and grows wild mosquito larvicidal (Satyal et al., 2013), insecticidal, analyzed by GCMS instrument and the composition of various constituents present in the respective oil in the districts of Dang, Rolpa and Sallyan in antifungal, antiaflatoxin, antioxidant (Prakash et al., is tabulated below in Table 1 & table 2 respectively which is nearly similar to the analysis of essential the Rapti Zone (Rema et al., 2002, Adhikari , 2018). 2013) and antibacterial (Rajendra et al., 2013) oil of fruits of sugandhakokila (Adhikary et al., 2011) which shows 1,8-cineole, methyl cinnamate, This species is a diploid and can grow to an altitude activities of essential oils have been also reported. alpha-terpineol as major constituents. of 1300 meters (Ravindran et al., 2003). Cinnamomum tenuipile Kosterm is recognized as an aromatic plant, In the present study, essential oil is extracted from Table 1: Chemical constituents present in the essential oil extracted from fruits of Cinnamomum tenuipile Kosterm meaning it has an elevated level of essential oil the fruits and leaves of Cinnamomum tenuipile Table(Sugandhakokila) 1: Chemical based constituents on GCMS present analysis. in the essential oil extracted from fruits of Cinnamomum tenuipile Kosterm (Sugandhakokila) based on GCMS analysis. (Gurung, 2015). Using steam distillation, the dried Kosterm (Sugandhakokila) collected from berries of Cinnamomum tenuipile Kosterm produce Brindaban Botanical Garden, Makawanpur Nepal Peak R. Time Area Area% Name of the Compounds 1 11.668 539761 1.93 Pinene the essential oil commonly known as sugandha and oil was analysed by Schimadzu GC-MS QP 2010 2 13.505 909094 3.26 Sabinene kokila oil (Ravindran et al., 2003), which is yellow Plus. The purpose of this study was to compare the 3 13.64 530067 1.9 Pinene in color and has a camphoraceous, spicy aroma chemical constituents present in the both oil samples. 4 15.937 304184 1.09 Cymene (HPPCL, 2015). This product can be used as a 5 16.287 10674502 38.23 Eucalyptol fragrance in soaps, detergents, cosmetics, perfumes Materials and Methods 6 17.61 168681 0.6 Terpinene 7 19.61 355533 1.27 Linalool and industrial fragrances (Gurung, 2015). Sugandha 8 21.826 5463814 19.57 Camphor kokila oil is also used in indigenous medicine as a Collection of plant materials 9 22.882 173829 0.62 Terpineol demulcent and stimulant (Rema et al., 2002). The The fruits and leaves of Cinnamomum tenuipile 10 23.379 1014454 3.63 Terpinen-4-ol 11 24.02 1194704 4.28 Terpineol Nepal Trade Integration Strategy 2010, identified Kosterm were collected from Brindaban Botanical Medicinal and Aromatic Plants (MAPs) as one of 12 29.339 288113 1.03 Cinnamate Garden, Makawanpur district of Nepal. The fresh 13 32.807 6002721 21.5 Cinnamate <(E)-, methyl-> Nepals top twenty goods and services with export fruits and leaves were collected and dried in shed 14 38.319 151850 0.54 2-Acetylbenzoic acid potential (Sharma, 2015). before extraction of essential oil. 15 40.995 152191 0.55 Caryophyllene oxide

86 87 2019 Journal of Plant Resources Vol.17, No. 1

(x1,000,000)

TIC (1.00)(x1,000,000) 5 2.5 TIC (1.00) 5 2.5

2.0 2.0

1.5 1.5 3 8 3 8 1 1 1.0 1.0

0.5 1

0.5 0 1 2 1 1 1 0 3 1 2 2 1 4 5 7 1 4 6 9 1 1 1 3 2 4 7 5 1 4 6 9 1 10 20 30 1 40 50 60 70 10 20 30 40 50 60 70 Figure 1: GCMS chromatogram of essential oil of fruits of Sugandhakokila collected from Brindaban, MakawanpurFigure 1: GCMS chromatogram of essential oil of fruits of Sugandhakokila collected from Brindaban, Makawanpur

:C].:R]1JVVJ :G1JVJV GV :R]1JVJV ]:`:RH7IVJV %H:C7] QC 1J:CQQC :I].Q` V`]1JVJRRQC :C].:R V`]1JVQC IV .7CH1JJ:I: V\]

IV .7CH1JJ:I: V\] `V:Q

Figure 2: Graphical representation for the major chemical constituents present in the essential oilFigure of fruits 2: Graphical of Cinnamomum representation tenuipile for the Kosterm major chemical analyzed constituents by GCMS. present in the essential oil of fruits of Cinnamomum Table 2: Chemicaltenuipile constituents Kosterm present analyzed in by the GCMS. essential oil extracted from leaves of Cinnamomum tenuipile Kosterm Table 2: Chemical constituents present in the essential oil extracted from leaves of Cinnamomum tenuipile Kosterm (Sugandhakokila) based on GCMS analysis. (Sugandhakokila) based on GCMS analysis. Peak R. Time Area Area% Name of the Compounds 1 11.689 732795 2.49 Pinene 2 13.529 2061207 6.99 Sabinene 3 13.664 985045 3.34 Pinene 4 14.362 216021 0.73 Myrcene 5 16.166 205549 0.7 Limonene 6 16.298 7126612 24.17 Eucalyptol 7 23.396 430624 1.46 Terpinen-4-ol 8 24.038 1286930 4.36 Terpineol 9 29.361 563838 1.91 Cinnamate 10 32.89 14824373 50.27 Cinnamate <(E)-, methyl-> 11 34.354 505255 1.71 trans-.alpha.-Bergamotene 12 38.588 231513 0.79 Cadinene 13 43.681 317663 1.08 Cadin-4-en-10-ol

88 (x1,000,000) 0 TIC (1.00) 1

2.0 6

1.5

1.0 2 0.5 8 3 1 1 9 3 2 1 7 1 4 5 1

10 20 30 40 50 60 70

Figure 3 : GCMS chromatogram of essential oil of leaves of Sugandhakokila collected from Brindaban, Makawanpur 2019 Journal of Plant Resources Vol.17, No. 1 2019 Journal of Plant Resources Vol.17, No. 1

(x1,000,000) (x1,000,000) 0 5 TIC (1.00)(x1,000,000) TIC (1.00) 1 2.5 TIC (1.00) 5 2.5 2.0

2.0 6 2.0 1.5 1.5 1.5 3 8 3 8 1 1 1.0 1.0 1.0 0.5 2 8

0.5 3 1 1

0.5 0 1 1 2 9 1 3 1 2 7 1 1 4 1 5 1 0 3 1 2 2 1 4 5 7 1 4 6 9 1 1 1 3 2 4 7 5 1 4 6 9 1 10 20 30 1 40 50 60 70 10 20 30 40 50 60 70 10 20 30 40 50 60 70 Figure 3 : GCMS chromatogram of essential oil of leaves of Sugandhakokila collected from Brindaban, Figure 1: GCMS chromatogram of essential oil of fruits of Sugandhakokila collected from Brindaban, Makawanpur MakawanpurFigure 1: GCMS chromatogram of essential oil of fruits of Sugandhakokila collected from Brindaban, Makawanpur

:C].:R]1JVVJ :G1JVJV :C].:R1JVJV GV :R]1JVJV :G1J1JV ]:`:RH7IVJV %H:C7] QC GV :R1JVJV 1J:CQQC %H:C7] QC :I].Q` :C].:RV`]1JVQC V`]1JVJRRQC IV .7C 1JJ:I: V \] :C].:R V`]1JVQC IV .7CH1JJ:I: V\] IV .7CH1JJ:I: V\] `V: Q `V:Q Figure 4: Graphical representation for the major chemical constituents present in the essential oil of Figure 2: Graphical representation for the major chemical constituents present in the essential leaves of Cinnamomum tenuipile Kosterm analyzed by GCMS. oilFigure of fruits 2: Graphical of Cinnamomum representation tenuipile for the Kosterm major chemical analyzed constituents by GCMS. present in the essential oil of fruits of Cinnamomum Figure 4: Graphical representation for the major chemical constituents present in the essential oil of leaves of Cinnamomum Table 2: Chemicaltenuipile constituents Kosterm present analyzed in by the GCMS. essential oil extracted from leaves of Cinnamomum tenuipile Kosterm tenuipile Kosterm analyzed by GCMS. Table 2: Chemical constituents present in the essential oil extracted from leaves of Cinnamomum tenuipile Kosterm (Sugandhakokila) based on GCMS analysis. Mass fragmentation pattern of various compounds that are identified by GCMS analysis (Sugandhakokila) based on GCMS analysis. (x10,000) 1.00 Peak R. Time Area Area% Name of the Compounds 93 0.75 1 11.689 732795 2.49 Pinene Mass fragmentation pattern of various compounds that are identified by GCMS analysis 2 13.529 2061207 6.99 Sabinene 0.50 41 77 0.25 3 13.664 985045 3.34 Pinene (x10,000) 1.00 67 105 121 93 136 0.00 155 178 194 203 215 237 256 263 289 303 317 331 346 4 14.362 216021 0.73 Myrcene 50.0 0.75 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 5 16.166 205549 0.7 Limonene Figure 5:0.50 Mass Fragmentation of Pinene 6 16.298 7126612 24.17 Eucalyptol 41 77 0.25 (x10,000) 105 7 23.396 430624 1.46 Terpinen-4-ol 1.00 67 121 93 136 0.00 155 178 194 203 215 237 256 263 289 303 317 331 346 8 24.038 1286930 4.36 Terpineol 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 0.75 9 29.361 563838 1.91 Cinnamate 0.50 41 10 32.89 14824373 50.27 Cinnamate <(E)-, methyl-> Figure 5:77 Mass Fragmentation of Pinene 0.25 69 136 11 34.354 505255 1.71 trans-.alpha.-Bergamotene 121 107 158 172 198 217 233 248 277 299 312 332 343 0.00 (x10,000) 1.00 12 38.588 231513 0.79 Cadinene 50.0 75.0 100.0 93 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 13 43.681 317663 1.08 Cadin-4-en-10-ol Figure 6:0.75 Mass Fragmentation of Sabinene 0.50 41 77

0.25 88 69 136 89 (x1,000,000) 121 107 158 172 198 217 233 248 277 299 312 332 343

0 0.00 TIC (1.00) 1 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0

2.0 Figure 7: Mass Fragmentation of Sabinene 6

(x10,000) 1.00 93 1.5 41

0.75

1.0 0.50 69 0.25 79

107 121

2 136 0.00 154 170 188 204 216 243 256 263 282 308 322 333 346 0.5 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 8 3 1 1 9 3 2 1 7 1 4 5 1 Figure 8: Mass Fragmentation of Pinene 10 20 30 40 50 60 70

(x10,000) 1.00 41 Figure 3 : GCMS chromatogram of essential oil of leaves of Sugandhakokila collected from Brindaban, Makawanpur 0.75 0.50 93

69 0.25 79 121 0.00 107 136 152 171 186 210 216 238 252 265 282 309 320 329 346 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 2019 Journal of Plant Resources Vol.17, No. 1

(x10,000) 1.00 93 41

0.75

0.50

69 0.25 79 121 107 136 0.00 154 170 188 204 216 243 256 263 282 308 322 333 346 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 Figure 7: Mass Fragmentation of Pinene

(x10,000) 1.00 41

0.75

0.50 93

69 0.25 79 107 121 0.00 136 152 171 186 210 216 238 252 265 282 309 320 329 346 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 8: Mass Fragmentation of Myrcene

(x10,000) 1.00 93

0.75 68

0.50 41 79

107 0.25 121 136 349 232 314 330 0.00 157 177 188 199 257 270 280 305 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 9: Mass Fragmentation of Limonene

(x10,000) 1.00 43

0.75

0.50

71 81 0.25 108 93 139 154 125 0.00 168 183 199 215 233 256 274 281 295 314 327 344 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 Figure 10 : Mass Fragmentation of Eucalyptol

(x10,000) 1.00 43 71

0.75 93 0.50 111 55 0.25 136 154 121 0.00 173 190 202 230 241 261 268 288 308 322 333 344 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 11: Mass Fragmentation of Terpinen-4-ol

(x10,000) 1.00 59

0.75 43 0.50 93 121 81 0.25 136 107 0.00 154 170 184 199 219 235 253 265 279 311 331 344 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 12: Mass Fragmentation of Terpineol

(x10,000) 1.00 103

131 0.75

0.50 51 77 162 0.25 63 347 117 144 0.00 176 188 206 220 247 254 279 294 307 317 337 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 Figure 13: Mass Fragmentation of Cinnamate

90 2019 Journal of Plant Resources Vol.17, No. 1 2019 Journal of Plant Resources Vol.17, No. 1

(x10,000) (x10,000) 1.00 1.00 93 131 41

0.75 0.75

77 103 0.50 0.50 162 51 69 0.25 79 0.25 102 107 121 63 136 144 0.00 154 170 188 204 216 243 256 263 282 308 322 333 346 0.00 168 183 199 215 233 247 263 281 295 311 327 343 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 7: Mass Fragmentation of Pinene Figure 14: Mass Fragmentation of Cinnamate <(E)-, methyl->

(x10,000) (x10,000) 1.00 1.00 41 41

0.75 0.75

0.50 93 0.50 93 79 105 69 69 0.25 0.25 133 79 147 161 349 107 121 175 189 204 0.00 136 152 171 186 210 216 238 252 265 282 309 320 329 346 0.00 223 236 258 266 291 304 323 339 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 8: Mass Fragmentation of Myrcene Figure 15: Mass Fragmentation of trans-.alpha.-Bergamotene

(x10,000) (x10,000) 1.00 1.00 93 105

41 161 0.75 0.75 68 91

0.50 41 79 0.50 134 204 81 107 0.25 0.25 121 136 349 69 189 232 314 330 147 176 290 0.00 157 177 188 199 257 270 280 305 0.00 223 238 256 275 310 317 336 345 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 9: Mass Fragmentation of Limonene Figure 16: Mass Fragmentation of Cadinene

(x10,000) (x10,000) 1.00 1.00 43 43

0.75 0.75 95

0.50 0.50 121 71 81 79 0.25 108 0.25 105 55 161 93 139 154 204 137 125 189 222 0.00 168 183 199 215 233 256 274 281 295 314 327 344 0.00 179 240 255 264 288 298 326 338343 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 Figure 10 : Mass Fragmentation of Eucalyptol Figure 17: Mass Fragmentation of Cadin-4-en-10-ol

(x10,000) (x10,000) 1.00 1.00 43 71 119

0.75 0.75 93 0.50 111 0.50 91 55 0.25 0.25 41 117 136 154 65 77 121 135 0.00 173 190 202 230 241 261 268 288 308 322 333 344 0.00 158 184 210 217 243 267 283 299 321 330 347 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 11: Mass Fragmentation of Terpinen-4-ol Figure 18: Mass Fragmentation of Cymene

(x10,000) (x10,000) 1.00 1.00 59 93

0.75 0.75 43 0.50 93 0.50 121 43 77 81 0.25 0.25 136 121 136 105 107 65 0.00 154 170 184 199 219 235 253 265 279 311 331 344 0.00 160 170 188 215 234 257 263 293 307 323 333 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 Figure 12: Mass Fragmentation of Terpineol Figure 19: Mass Fragmentation of Terpinene

(x10,000) (x10,000) 1.00 1.00 103 43

131 0.75 0.75

71 0.50 51 77 0.50 55 93 162 0.25 0.25 63 347 121 117 144 107 136 0.00 176 188 206 220 247 254 279 294 307 317 337 0.00 156 167 195 223 234 254 263 287 307 321 331 343 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 13: Mass Fragmentation of Cinnamate Figure 20: Mass Fragmentation of Linalool

90 91 2019 Journal of Plant Resources Vol.17, No. 1

(x10,000) 1.00 41 95 0.75 55 81 0.50 108

0.25 152 137 0.00 123 170 186 199 216 238 256 270 280 298 317 327 344 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 21: Mass Fragmentation of Camphor

(x10,000) 1.00 59

0.75 43

0.50 81

95 0.25 136 110 121 0.00 154 167 193 199 219 237 259 271 281 299 323 333 346 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 22: Mass Fragmentation of Terpineol

(x10,000) 1.00 149

0.75

0.50

177 0.25 105 50 65 76 93 121 164 204 0.00 185 222 245 258 265 290 306 317 333 345 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 23: Mass Fragmentation of 2-Acetylbenzoic acid

(x10,000) 1.00 41

0.75

0.50 79 91 55 0.25 107 121 349 138 159 177 189 235 0.00 204 220 253 275 292 300 324 330 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 24: Mass Fragmentation of Caryophyllene oxide

Conclusion providing resources and encourages for research work. We would like to thank Plant Research Centre, The results of chromatographic analysis of essential Makawanpur especially Mr. Raj Kishor Pandit, oil of fruits and leaves have shown that they are Assisstant Scientific Officer for his co-operation somewhat similar but some compounds were present during sample collection. The authors would like to only in fruit oil and some were present in leaf oil thank Department of Plant Resources for giving an only. Camphor is present in fruit oil while in leaf oil opportunity for research work in a very much friendly it was absent. By GCMS analysis 13 compounds and sound environment and for financial support were identified in leaf oil where as 15 compounds without which the research work was not possible. were identified in fruit oil. The major compounds were Eucalyptol (38.23%), Camphor (19.57%) and References Methyl cinnamate (22.53%) in fruit oil and Eucalyptol (24.17%) and Methyl cinnamate Rema, J., Krishnamoorthy, B., Sasikumar, B., Saji, (52.18%) in leaf oil. K.V. & Mathew, P.A. (2002). Cinnamomum Cecidodaphne Meissn. Indian Journal of Arecanut, Acknowledgments Spices & Medicinal Plants. 4 (1), 5961. The authors would like to thank Mr. Sanjeev Kumar Adhikari, S. (2018). Essential oils of Nepal. Rai, Director General and Mr. Mohan Dev Joshi Deputy Thapathali, Kathmandu, Nepal: Department of Director General, Department of Plant Resources for Plant Resources.

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(x10,000) Ravindran, P.N., Nirmal-Babu, K.& Shylaja, M. camphora, C. tamala and C. glaucescens. Natural 1.00 41 95 0.75 (2003). Cinnamon and cassia: the genus Product Communications, 12, 1777-1784. 55 81 0.50 Cinnamomum. : CRC press. pp. 337 (Retrieved 108 Bhanu, P., Singh, P., Yadav, S., Singh, S.C.& Dubey, 0.25 152 23/11/ 2015). 137 N.K. (2013). Safety Profile Assessment a n d 0.00 123 170 186 199 216 238 256 270 280 298 317 327 344 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Gurung, K.(n.d.). Study on Quality Issues of Efficacy of Chemically Characterized Figure 21: Mass Fragmentation of Camphor Medicinal and Aromatic Plants (MAPs) Sector Cinnamomum glaucescens Essential Oil against

(x10,000) 1.00 in Nepal. Kathmandu, Nepal: Jadibuti Storage Fungi, Insect, Aflatoxin Secretion and as 59

0.75 Association of Nepal. Retrieved 23 November Antioxidant. Food and Chemical Toxicology, 53, 43

0.50 81 2015. 160-167. 95 0.25 136 110 121 0.00 154 167 193 199 219 237 259 271 281 299 323 333 346 MOFSC.(2011).Essential Oils. Kathmandu, Gyawali, R., Bhandari J., Amatya, S., Piya, E., 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Nepal: Herbs Production & Processing Co. Ltd. Pradhan, U.L., Paudyal, R., Shrestha, R., & Figure 22: Mass Fragmentation of Terpineol Shrestha, T.M., (2013). Antibacterial and

(x10,000) Sharma, P. & Shrestha, N.(2011) Promoting 1.00 Cytotoxic Activities of High Altitude Essential 149 Exports of Medicinal and Aromatic Plants 0.75 Oils from Nepalese Himalaya. Journal of (MAPs) and Essential Oils from Nepal. South 0.50 Medicinal Plant Research, 7, 738-743. 177 0.25 105 Asia Watch on Trade, Economics and 50 65 76 93 121 164 204 0.00 185 222 245 258 265 290 306 317 333 345 Environment (SAWTEE). Adhikary, S.R., Tuladhar, B.S., Sheak, A., Tens, A., 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 & Gerrit P. L. (1992). Investigation of Figure 23: Mass Fragmentation of 2-Acetylbenzoic acid Satyal, P., Paudel, P., Poudel, A., Dosoky, N.S., Nepalese Essential Oils. I. The Oil of (x10,000) 1.00 Pokharel, K.K. & Setzer, W.N. (2013). 41 Cinnamomum glaucescens (Sugandha Kokila) Bioactivities and Compositional Analyses of 0.75 Journal of Essential Oil Research, 4(2), 151-159. 0.50 79 Cinnamomum Essential Oils from Nepal: C. 91 55 0.25 107 121 349 138 159 177 189 235 0.00 204 220 253 275 292 300 324 330 50.0 75.0 100.0 125.0 150.0 175.0 200.0 225.0 250.0 275.0 300.0 325.0 350.0 Figure 24: Mass Fragmentation of Caryophyllene oxide

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