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Composition of leaf oil of Gelam (M. leucadendra (L.)L.) growing in various peat swamp regions of Central Kalimantan Indonesia.

INTRODUCTION

Gelam (tea tree) is tree species belong to Myrtaceae family. In Indonesia, Gelam found at Sumatera, Java, Kalimantan, Molucca, Seram, Buru, an d Nusa Tenggara Timur. In Central Kalimantan, large population of Gelam grows naturally at various types of peat swamp regions. This tree known for their ability to grow in varied soil conditions even on peat land. Therefore these trees also planted for restoration of peat swamp forests. However, there is increasing demand of Gelam wood as building material. This condition hampered restoration program and endangered wild population of Gelam trees. Thus, it is necessary to utilize non timber product of Gelam trees to prevent excessive logging.

Potential non-timber product of this tress is Cajuput oil, which use as traditional medicine for pain, burns, cold, influenza, dyspepsia. Indonesia is one of major producer of this oil with Java as main source of raw material for oil production (Budiadi et al., 2005). In Java, Melaleuca leucadendra L.(L.) grow on soil dominated volcanic ash with organic material content less than 5% while organic material content of swamp peat forest could reach upper 65% (Anwar et al., 1984). Some studies showed that soil condition and geographic variation produces significant variation of chemical composition of Cajuput oil (Brophy et al., 1989; Osaki et al., 1998; Oyen and Dung, 1999). Unfortunately, study of Cajuput oil of Central Kalimantan is lacking which prevent any development of this oil as non-timber product of Central Kalimantan. Therefore, a study on chemical composition of Cajuput oil of Gelam trees of Central Kalimantan grow at various swamp peat forest was carried out.

Methodology:

Plant Material:

Fresh leaves of M. leucadendra L. (L.) samples were harvested by hand at study area. Study area was located in Block C, Pulang Pisau District, Central Kalimantan. Study area previously designed as One Million Hectare Rice Field Project which abandoned since 1999 and prior to project predominantly occupied by Gelam Tree. Leaf samples were collected from nine sampling sites (Table 1)

Active Compound Separation and Analysis: Oil Extraction:

The air dried (water content 30-50%) leaf biomass (200g) of M. leucadendra L. (L.) subjected to steam distillation process (Clark, 2005). The essential oil produced from steam distillation process, collected in vial bottle and weighed to determine its production.

Oil Active Compound Analysis:

Analysis of active compound in the essential oil samples were conducted using GC/MS (Gas Chromatography Mass Spectrometry) (Skoog et al, 2007). GC/MS analysis was carried out in EI mode using Shimadzu gC-MS (Mode QP-5000, Shimadzu Co., Kyoto, Japan). The sample inlet of GC 7890A, injection source of GCALS 7690, and Mass Spectrometry of 5970 MSD with Triple Axis Detector. Oven temperature was programmed at 40[degrees]C (Isothermal for 2 minutes) was ramped to 280[degrees]C at 10[degrees]C/min followed by to 299[degrees]C at 30[degrees]C/min. Helium was used as the carrier gas at a flow rate of 1ml/min, with using 1:20 split ratio. Mass spectral data were compared with those of published data.

RESULTS AND DISCUSSION

The oil yield was 0.003% (equal to 0.63 mg) on the basis of air-dried leaf weight which considered low compared with oil yield of same and similar species (Budiadi et al., 2005). Fifty seven chemical compound were identified from leaf oil of M. leucadendra L. (L.) grow in various types of peatlands. Oil from shallow peatland region more complex than others which show some effects of peatlands character to chemical content of oil (Table 2).

Chemical content of leaf oil originated from shallow peatland dominated by one unknown substance (average 11.69%) followed by [alpha]-terpinolene (average 10.95%), caryophyllene (average 10.77%), and [gamma]-terpinene (average 10.55%). Specific chemical content of oil leaf from thin peatland were [alpha]-amorphene, Cadina3, 9-diene, Germacerene-B, Guaiol, [beta]-guaiene, Cubenol, [alpha]-cadinol, [alpha]-eudesmol, Bulnesol, 1,4-dimethoxy-6, 7, 8, 9, and 1, 4-naphtalenedione which account for 3.35% of total oil. Chemical content of leaf oil originated from medium depth peatland dominated by caryophyllene (average 15.62%) and [gamma]-terpinene (average 13.02%). Specific chemical content of oil leaf from medium depth peatland were 2-isopropenyl-4a, 8-, Globulol, 12-oxabicyclo (9.1.0), and Isospathulenol which account for 1.57% of total oil. Chemical content of leaf oil originated from deep peatland dominated by [alpha]-terpinolene (average 18.71%) and [gamma]-terpinene (average 15.98%) and specific chemical content of the oil only [beta]-linalool which account for 0.04% of total oil.

On the other hand, content of 1, 8-cineole, the most economic important chemical of oil leaf of this species, ranged from non-existed to 20.09% (Table 2). Result on chemical content from this research was different compare with similar oil from Java region of Vietnam (Cuong et al., 1994; Pino et al., 2002; Kumar et al., 2005) where 1, 8-cineole (Eucalyptol) and viridiflorol considered as major component. Based on this, it is not promising for commercialized leaf oil from as cajuput oil.

Conclusion:

Most oil contained significantly higher monoterpenes than sesquiterpene, except oil from medium depth peatland where amount of sesquiterpenes slightly higher (38.94% compare with 38.36% monoterpene) (Table 3). High level of monoterpenes is common feature of plant grow in excessive stress due to low-nutrient soil. Monoterpenes and sequisterpen are found in essential oils. Monoterpenes is an essence compound and it has characteristic odor. Monoterpens known widely used as an antiseptic, axpectorant, spasmolytic, anesthetic, and sedatives. Utilization of this compound is also known as a food flavor and perfume concentrates and it is heavily trafficked commercial compounds. sesquiterpene compounds have large enough bioactivity, such as antifeedant, hormones, antimicrobials, antibiotics and toxins and the regulat or plant growth and sweeteners (Lenny, 2006). some of the secondary metabolites (limonene, [alpha]-pinene, [beta]-pinene, [delta]-3 carene, sabinene, caryophyllene, and [delta]-cadinene) belonging to these terpenes, are in the form of resins (Langenheim, 2003). In conclusion, this report on the chemical composition of leaf oil of M. leucadendra L. (L.), growing at various peat swamp region types is the first report from any part of the world. As the essential oil of this species contain appreciable amount of [gamma]-terpinene, [alpha]-terpinolene, caryophyllene, [alpha]-humulene which have high demand in pharmaceutical industry and other related industries, this oil may benefited restoration program of peatland in Central Kalimantan.

ARTICLE INFO

Article history:

Received 4 December 2014

Received in revised form 10 January 2015

Accepted 8 February 2015

Available online 20 February 2015

REFFERENCES

Anwar, J., S.J.D amanik, N. Hisyam and A.J. Whitten, 1984. Ekologi ekosistem Sumatera. Gadjah Mada Univ. Press. Jogyakarta.

Brophy, J.J., D.J. Boland and E.V. Lassak, 1989. "Leaf essentail oils of Melaleuca and Leptospermum species from tropical Australia". p.193-203. In: D.J. Boland (ed.), Tree for the Tropics: Growing Australia multipurpose trees and shrubs in developing countries, Monograph No. 10, Australian Centre for International Agricultural Research, Canberra.

Budiadi, Hiroaki, I.T., S. Sigit, K. Yoichi, 2005. "Variation in Kayu Putih (Melaleuca leucadendron UNN) oil quality under different farming system in Java, Indonesia". Eurasian J. For. Res, 8(1): 15-20.

Clark, J., 2005. Immiscible liquids and steam distillation, Chemguide. www.chemguide.co.uk/physical/phaseeqia/immiscible.html

Cuong, N.D., T.T.X uyen, O. Motl, K. Stanky, J. Presslova, Jedlikova and V. Sery, 1994. "Antibacterial properties of Vietnamese cajuput oil". JEOR, 6(1): 63-67.

Kumar, A., S. Tandon and A. Yadaf, 2005. "Chemical composition of the essential oil from fresh leaves of M. leucadendron L. from North India". J. of Ess Oil Bear Plants. 8(1): 19-22.

Langenheim, J.H., 2003. Plant Resins: Chemistry, Evolution, Ecology, and Ethnobotany. Portland, Cambridge. Timber Press.

Lenny, S., 2006. "Terpenoid dan steroid'. Karya Ilmiah. Departemen Kmia. Fakultas MIPA. Universitas Sumatera Utara. Medan. Repository USU@2006: 12-14.

Osaki, M., T. Watanabe, T. Ishizawa, C. Nilnond, T. Nuyim, C. Sittibush and T. Tadano, 1998. "Nutritional characteristics in leaves of native plants grown in acid sulfate, peat, sandy podzolic and saline soils distributed in Peninsular Thailand". Plant and Soil., 201: 175-182.

Oyen, L.P.A. and N.X. Dung, (ed.)., 1999. Plant resources of south-east Asia. No.19. Essential-oil plants, Prosea Foundation, Bogor.

Pino, J., A. Bello, A. Urquiola, J. Aguero. and R. Marbot, 2002. "Chemical composition of cajuput oil (M. leucadendra L.) from Cuba". JEOR. 14(1): 10-11.

Skoog, D.A., H. James, R.C. Stanley, 2007. Principles of instrumental analysis, 6th Edition.United States: Thomson Brooks/Cole.

(1) Robert Manurung, (2, 3) Ana Widiana, (3) Taufikurahman, (4) Suwido H. Limin

(1) Department of Bio Engineering, School of Life Science and Technology, Bandung Institute of Technology

(2) Department of Biologi, Faculty of Science and Technology, UIN Sunan Gunung Djati Bandung

(3) Department of Biology, School of Life Science and Technology, Bandung Institute of Technology

(4) Department of Agricultural, Paalngka Raya University

Corresponding Author: Robert Manurung, Department of Bio Engineering, School of Life Science and Technology, Bandung Institute of Technology. Indonesia. Mobile

Ph: 062-811248240. E-mail: manurung@sith.itb.ac.id.
Table 1: Feature of Sampling areas

Area                      Land      Water     Soil      EC ([micro]s/
                          History   Acidity   Acidity   cm)

Deep Peatlands
Jabiren                   Burned    4.70      4.80      53.00
Tumbang Nusa              Wetland   3.80      3.80      65.00
Taruna I                  Cut off   3.50      6.20      75.00
Medium Depth Peatlands
Kelampangan               Burned    3.70      4.20      82.00
Taruna II                 Wetland   3.90      3.75      55.00
TunjungNyaho              Cut off   4.90      2.00      46.00
Shallow Petlands
Buntoi                    Burned    4.50      4.30      38.00
Bukit Keminting           Wetland   5.20      5.50      26.00
Maliku                    Cut off   4.10      4.90      20.00

Area                      Peat        Temperature    Humidity
                          Thickness   ([degrees]C)   (%)
                          (m)

Deep Peatlands
Jabiren                   3.00        31.70          71.20
Tumbang Nusa              3.00        33.00          54.00
Taruna I                  3.00        38.00          54.00
Medium Depth Peatlands
Kelampangan               1.50        31.00          65.00
Taruna II                 1.50        33.00          61.00
TunjungNyaho              1.40        31.00          69.00
Shallow Petlands
Buntoi                    0.80        32.20          68.80
Bukit Keminting           0.70        31.00          66.00
Maliku                    0.50        32.00          38.00

Table 2: Chemical composition of Meulaleuca leucadendra L. (L.)
Oil Leaves

Name                       RI               Percentage

                                          Shalow Peatland

                                    Buntoi   Bukit       Maliku
                                             Keminting

[alpha]-thujene            932.6    1.94     2.27        1.99
[alpha]-pinene             940.1    1.17     5.07        1.74
Sabinene                   977.5    0.00     0.00        0.00
[beta]-pinene              980.8    0.00     3.14        0.00
[beta]-myrcene             995.4    0.00     0.88        0.12
[alpha]-phellandrene       1008.4   2.02     0.24        1.88
[delta]-3 carene           1014.6   0.00     0.00        0.00
[alpha]-terpinene          1021.2   2.56     1.67        2.21
o-cymene                   1029.6   0.00     5.76        5.28
[beta]-cymene              1036.0   4.06     0.00        0.00
Limonene                   1033.5   1.28     7.77        1.04
1,8-cineole                1043.9   0.46     20.09       0.00
[gamma]-terpinene          1063.0   11.84    10.38       9.43
[alpha]-terpinolene        1091.7   14.54    6.39        11.91
[beta]-linalool            1101.8   0.00     0.00        0.00
4-terpineol                1182.8   0.53     0.75        0.69
[alpha]-terpineol          1196.1   0.00     3.40        0.00
[alpha]-terpinenvlacetat   1355.6   0.00     3.13        0.00
[alpha]-copaene            1384.4   0.46     0.00        0.87
[beta]-elemene             1399.6   1.08     0.12        1.80
Caryophyllene              1431.3   12.23    5.89        14.19
[alpha]-humulene           1465.8   5.41     2.08        0.00
[alpha]-caryophyllene      1465.7   0.00     0.94        6.50
2-isopropenyl-4a.8-..      1486.1   0.00     0.00        0.00
Aromadendrene              1487.3   0.31     0.24        0.00
[alpha]-amorphene          1490.3   0.00     0.23        0.00
Germacrene-D               1493.2   2.27     0.00        4.07
[beta]-selinene            1498.5   0.70     1.98        0.00
[alpha]-selinene           1507.2   0.00     1.79        0.00
Bicyclosermacrene          1508.1   0.00     0.00        3.44
Germacrene-A               1518.2   0.00     0.00        2.26
[gamma]-elemene            1512.0   2.54     0.00        0.00
[delta]-guaiene            1523.1   0.75     0.00        0.00
Cadin[alpha]-3,9-diene     1517.7   0.00     0.16        0.00
[delta]-cadinene           1533.4   0.65     0.18        1.23
Germacerene-B              1571.7   0.00     0.36        0.00
(-)-spathulenol            1591.8   0.79     0.24        1.58
Caryophyllene oxide        1598.8   1.13     3.25        1.67
Globulol                   1598.6   0.00     0.00        0.00
Veridiflorol               1606.0   0.00     1.48        0.00
Guaiol                     1609.8   0.00     3.34        0.00
Ledol                      1619.1   0.00     0.18        0.00
Humulene oxide             1625.8   0.00     0.66        0.00
12-oxabicyclo(9.1.0)..     1625.5   0.00     0.00        0.00
Unknown-1                  1631.0   24.85    0.00        10.23
.tau.-cadinol              1639.9   0.00     0.24        0.00
[beta]-auaiene             1642.0   0.00     0.41        0.00
Cubenol                    1651.0   0.00     1.44        0.00
Isospathulenol             1652.9   0.00     0.00        0.00
[alpha]-cadinol            1666.6   0.00     0.99        0.00
[alpha]-eudesmol           1668.9   0.00     1.68        0.00
Juniper camphor            1670.0   0.57     0.29        0.00
Bulnesol                   1681.4   0.00     0.61        0.00
Unknown-2                  1730.3   5.85     0.00        12.90
Unknown-3                  ?        0.00     0.00        2.98
1,4-dimethoxy- 6,7,8,9     1994.9   0.00     0.00        0.38
1,4-naphtalenedione,       2014.0   0.00     0.00        0.43

Total                               100.00   99.70       100.00

Name                                  Percentage

                                Medium depth Peatland

                           Kalampangan   Taruna   Tanjung
                                         II       Nyaho

[alpha]-thujene            1.12          6.52     0.72
[alpha]-pinene             1.19          3.97     0.52
Sabinene                   0.00          0.55     0.00
[beta]-pinene              0.00          0.47     0.00
[beta]-myrcene             0.18          0.82     0.12
[alpha]-phellandrene       1.16          2.87     0.82
[delta]-3 carene           0.00          0.13     0.00
[alpha]-terpinene          1.50          4.40     0.94
o-cymene                   1.46          12.32    0.91
[beta]-cymene              2.11          4.06     0.42
Limonene                   2.89          2.92     0.00
1,8-cineole                7.70          14.70    4.43
[gamma]-terpinene          9.11          24.50    5.44
[alpha]-terpinolene        0.00          11.54    0.27
[beta]-linalool            0.00          0.00     0.00
4-terpineol                2.06          0.78     0.00
[alpha]-terpineol          0.00          1.28     0.00
[alpha]-terpinenvlacetat   0.00          0.16     0.00
[alpha]-copaene            0.89          0.00     0.61
[beta]-elemene             0.88          1.24     2.00
Caryophyllene              17.92         1.71     27.24
[alpha]-humulene           8.43          1.80     13.17
[alpha]-caryophyllene      0.00          0.00     0.00
2-isopropenyl-4a.8-..      0.00          0.00     0.32
Aromadendrene              0.00          0.00     0.00
[alpha]-amorphene          0.00          0.00     0.00
Germacrene-D               2.82          0.66     3.39
[beta]-selinene            6.53          0.00     0.77
[alpha]-selinene           6.70          0.00     0.00
Bicyclosermacrene          1.02          0.00     0.68
Germacrene-A               0.00          0.82     0.76
[gamma]-elemene            0.00          0.00     2.91
[delta]-guaiene            0.00          0.00     0.00
Cadin[alpha]-3,9-diene     0.00          0.00     0.00
[delta]-cadinene           0.87          0.00     0.63
Germacerene-B              0.00          0.00     0.00
(-)-spathulenol            0.67          0.47     0.78
Caryophyllene oxide        9.12          0.31     2.57
Globulol                   0.17          0.00     0.00
Veridiflorol               0.00          0.00     0.00
Guaiol                     0.00          0.00     0.00
Ledol                      0.00          0.00     0.00
Humulene oxide             0.00          0.00     0.00
12-oxabicyclo(9.1.0)..     3.78          0.00     0.00
Unknown-1                  3.19          0.79     15.88
.tau.-cadinol              0.00          0.00     0.00
[beta]-auaiene             0.00          0.00     0.00
Cubenol                    0.00          0.00     0.00
Isospathulenol             0.00          0.00     0.45
[alpha]-cadinol            0.00          0.00     0.00
[alpha]-eudesmol           0.00          0.00     0.00
Juniper camphor            0.00          0.00     0.38
Bulnesol                   0.00          0.00     0.00
Unknown-2                  5.06          0.00     10.36
Unknown-3                  0.00          0.00     1.60
1,4-dimethoxy- 6,7,8,9     0.00          0.00     0.00
1,4-naphtalenedione,       0.00          0.00     0.00

Total                      100.00        100.00   100.00

Name                                Percentage

                                  Deep Peatland

                           Jabiren   Tumbang   Taruna
                                     Nusa      I

[alpha]-thujene            1.81      4.28      6.21
[alpha]-pinene             1.45      3.31      4.06
Sabinene                   0.00      0.23      0.09
[beta]-pinene              0.00      0.15      0.20
[beta]-myrcene             0.00      0.25      0.75
[alpha]-phellandrene       1.82      3.71      2.26
[delta]-3 carene           0.00      0.00      0.14
[alpha]-terpinene          2.29      4.27      2.46
o-cymene                   0.00      6.34      20.40
[beta]-cymene              3.16      0.00      0.00
Limonene                   1.16      1.86      2.94
1,8-cineole                0.75      0.00      3.56
[gamma]-terpinene          10.58     18.56     18.80
[alpha]-terpinolene        12.82     22.79     20.52
[beta]-linalool            0.00      0.00      0.11
4-terpineol                0.58      1.24      2.76
[alpha]-terpineol          0.00      0.15      1.18
[alpha]-terpinenvlacetat   0.00      0.00      0.00
[alpha]-copaene            0.45      0.00      0.00
[beta]-elemene             1.90      1.47      0.61
Caryophyllene              11.88     9.16      3.32
[alpha]-humulene           5.10      0.85      0.92
[alpha]-caryophyllene      0.00      3.26      0.63
2-isopropenyl-4a.8-..      0.00      0.00      0.00
Aromadendrene              0.70      0.00      0.18
[alpha]-amorphene          0.00      0.00      0.00
Germacrene-D               3.04      2.14      0.00
[beta]-selinene            1.23      0.00      0.99
[alpha]-selinene           0.00      0.00      1.31
Bicyclosermacrene          0.00      2.20      0.00
Germacrene-A               0.00      1.05      0.00
[gamma]-elemene            3.87      0.00      0.00
[delta]-guaiene            1.29      0.00      0.00
Cadin[alpha]-3,9-diene     0.00      0.00      0.00
[delta]-cadinene           0.66      0.23      0.15
Germacerene-B              0.00      0.00      0.00
(-)-spathulenol            0.91      0.00      0.88
Caryophyllene oxide        0.97      0.47      2.87
Globulol                   0.00      0.00      0.00
Veridiflorol               0.00      0.00      1.45
Guaiol                     0.00      0.00      0.00
Ledol                      0.00      0.00      0.13
Humulene oxide             0.00      0.00      0.13
12-oxabicyclo(9.1.0)..     0.00      0.00      0.00
Unknown-1                  16.92     8.43      0.00
.tau.-cadinol              0.00      0.00      0.00
[beta]-auaiene             0.00      0.00      0.00
Cubenol                    0.00      0.00      0.00
Isospathulenol             0.00      0.00      0.00
[alpha]-cadinol            0.00      0.00      0.00
[alpha]-eudesmol           0.00      0.00      0.00
Juniper camphor            1.04      0.00      0.00
Bulnesol                   0.00      0.00      0.00
Unknown-2                  13.61     3.35      0.00
Unknown-3                  0.00      0.26      0.00
1,4-dimethoxy- 6,7,8,9     0.00      0.00      0.00
1,4-naphtalenedione,       0.00      0.00      0.00

Total                      100.00    100.00    100.00

Table 3: Classification of Secondary Metabolite

Type of chemical        Shallow Peatland
and amount (%)
                   Buntoi   Bukit       Maliku
                            Keminting

Monoterpene        39.94    41.96       31.01
  Hydrocarbon
Oxyganted-         1.25     20.33       1.58
  Monoterpene
Sesquiterpene      28.10    18.58       36.03
Oxygenated         0.00     9.96        0.00
  Sesquiterpene
Others             30.70    8.89        31.38
Total              99.99    99.72       100.00

Type of chemical         Medium Depth Peatland
and amount (%)
                   Kelampangan   Taruna   TanjungNyaho

Monoterpene        26.13         75.28    13.68
  Hydrocarbon
Oxyganted-         3.56          3.39     0.78
  Monoterpene
Sesquiterpene      55.18         6.54     55.11
Oxygenated         0.17          0.00     0.45
  Sesquiterpene
Others             13.49         14.55    29.88
Total              98.53         99.76    99.90

Type of chemical            Deep Peatland
and amount (%)
                   Jabiren   Tumbang Nusa   Taruna

Monoterpene        35.67     60.80          62.37
  Hydrocarbon
Oxyganted-         1.66      0.00           4.55
  Monoterpene
Sesquiterpene      31.16     20.36          8.24
Oxygenated         0.00      0.00           1.58
  Sesquiterpene
Others             30.53     18.38          20.40
Total              99.02     99.54          97.14
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Author:Manurung, Robert; Widiana, Ana; Taufikurahman; Limin, Suwido H.
Publication:Advances in Natural and Applied Sciences
Article Type:Report
Geographic Code:9INDO
Date:May 1, 2015
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