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Properties of Wax Extracted from Banana Leaves

Conference Paper · January 2003

DOI: 10.13031/2013.14121

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Paper Number: 036026

An ASAE Meeting Presentation

Properties of Wax Extracted from Banana Leaves

Takashi Yanagida
Graduate School of Life and Environmental Science, University of Tsukuba, Tsukuba
305-8572, Japan, e-mail: yanataka@livedoor.com

Naoto Shimizu, Assistant Professor

Inst. of Agriculture & Forest Engineering, University of Tsukuba, Tsukuba
305-8572, Japan

Toshinori Kimura, Professor

Graduate School of Life and Environmental Science, University of Tsukuba, Tsukuba
305-8572, Japan

Written for presentation at the

2003 ASAE Annual International Meeting
Sponsored by ASAE
Riviera Hotel and Convention Center
Las Vegas, Nevada, USA
27- 30 July 2003

Abstract. Three banana varieties were used to recover waxes from banana leaves. Wax yields of
Musa liukiuensis, M. acuminata and M. chilicarpa were 0.58%, 1.05% and 1.41% per dry weight,
respectively. The melting point of banana waxes were 78 - 82 °C, were higher than commercial
natural waxes except carunauba wax. The components of M. chilicarpa were similar to carunauba
wax by thin layer chromatography analysis. Fatty acids with carbon numbers ranging from C14 to
C30 were detected in M. chilicarpa leaf wax after saponification. C22 fatty acid was most abundant in
these fatty acids.
Keywords. banana leaf, tropical countries, waste utilization, wax

The authors are solely responsible for the content of this technical presentation. The technical presentation does not necessarily
reflect the official position of the American Society of Agricultural Engineers (ASAE), and its printing and distribution does not
constitute an endorsement of views which may be expressed. Technical presentations are not subject to the formal peer review
process by ASAE editorial committees; therefore, they are not to be presented as refereed publications. Citation of this work should
state that it is from an ASAE meeting paper. EXAMPLE: Author's Last Name, Initials. 2003. Title of Presentation. ASAE Meeting
Paper No. 03xxxx. St. Joseph, Mich.: ASAE. For information about securing permission to reprint or reproduce a technical
presentation, please contact ASAE at hq@asae.org or 69-429-0300 (2950 Niles Road, St. Joseph, MI 49085-9659 USA).
Introduction
Developing countries in the tropical region are faced with financial difficulties. In order to
overcome such difficulties, obtaining high value products to be exported for the world market is
useful. Banana is well known as an export crop cultivated in agricultural plantations of most
parts of the tropical region. After harvesting the banana fruit, the stems and leaves are regarded
as waste and left in the field or incinerated (Phirke et al., 2001). This residual from agricultural
plantations is an unutilized biomass resource. The putative amount of the banana waste
exceeds one billion tons per year in the world (FAO, 2001). The huge amount of the waste
generates some advantages because of its potential use in the conversion of agricultural wastes
into industrial products as by-product usages. Some studies on banana waste utilization have
been reported (Deivanai et al., 1995, Krishna, 1999, and Kalia et al., 2000).
Banana is a perennial herbaceous crop. The surface of the banana leaf is covered with a
natural wax. Natural waxes are used for various fields in the developed countries for a number
of purposes such as food coating, cosmetics, medicines and chemical bases. Japan imported
3300 tons of carnauba wax, 332 tons of natural plant wax and 227 tons of bees wax in
2001(The Chemical Daily Co., Ltd., 2002). Carnauba wax is the hardest, highest-melting,
natural commercial wax. Carnauba wax has amorphous, hard, tough and lustrous properties.
Carnauba wax is added to other waxes to increase their melting point, hardness, toughness and
luster and decrease stickiness, plasticity and crystallizing tendencies. If the properties of the
banana wax are similar to natural wax such as carnauba and candelilla wax, banana wax may
become an alternative material. Utilization of banana wax provides a great benefit by creating a
high value product from the unutilized biomass resources of banana cultivation in the tropical
developing countries.The leaf surface waxes from several plants have been reported (Smith et
al., 1966, Tulloch, 1973a, Baker, 1979, Genderen et al., 1988, Prasad et al., 1990a, Prasad et
al., 1990b, and Vermeer et al., 2003). In contrast, to the best of our knowledge, there are few
studies on banana wax.
The objectives of this research are to investigate the properties of banana leaf wax. Three
varieties of banana, Musa acuminata, M.liukiuensis and M.chilicarpa were used in this study.
Banana wax was extracted from banana leaves by an organic solvent reflux method. The
melting point and color value of the waxes were examined. Wax components were compared by
thin layer chromatography (TLC). The fatty acid composition of M.chilicarpa wax was identified
by gas liquid chromatography (GLC).

Safety Emphasis
Diazomethane was used for preparing fatty acid methyl ester derivatives in this work. As well as
being extremely toxic and a suspected carcinogen, diazomethane is dangerously explosive in
contact with rough surfaces. Diazomethane synthesis was carried out in a fume cupboard and
behind a safety shield with grate care. Furthermore, most of the organic solvents are highly
toxic, accumulating in the liver and kidneys to produce very serious irreversible effects including
cancer. Organic solvents also were used in the fume cupboard.

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Materials and Methods

Materials
Three varieties of banana leaf materials used in the experiment were Musa acuminata,
M.liukiuensis and M.chilicarpa and also their leaves were fully developed. All banana leaves
were collected in the tropical green house of ‘TSUKUBA BOTANICAL GARDEN , Japan (60 km
north from Tokyo)’ in August 2002.
Natural wax samples, carnouba, candelilla, rice, japan and bees wax were provided by
CERARICA NODA Co.,Ltd., Japan. These natural waxes were used to compare with banana
leaf wax.

Extraction of banana wax

Banana wax was extracted with a reflux method (Schmidt et al., 2002). Banana leaf materials
were cut into small pieces and placed in a round bottom flask for each variety. Hexane (Wako,
Japan) solvent was added into the flask until the leaf material was covered. The banana leaf
and solvent mixture were set on a water bath and maintained at a constant temperature at 60°C
for 10 minutes. A glass condenser was directly connected to the top of the flask.
After refluxing, the solvent mixture was collected into the conical flask through the filter. The
flask was allowed to cool down at room temperature for 2 hours and placed in a freezer for 6
hours.
The precipitated wax was collected using a filter paper (No.1; Advantec, Japan). A pre-
weighed filter paper was placed in the glass funnel and the solvent and wax mixture were
poured into the funnel. The precipitated wax was retained on the surface of the filter paper. The
filter paper was placed in a desiccant chamber for 48 hours. After drying, the filter paper was
weighed to determine wax weight.

Melting Point and Color Value Determinations

The melting point was determined on the micro melting point apparatus Yanaco MP
(Yanagimoto Manufacturing Co., Japan). Color value parameters, L* (lightness), a* (from green
to red) and b* (from blue to yellow) were determined on the color meter MINOLTA CR-20
(MINOLTA, Japan). C* (chroma) parameter is calculated from the former. The results are
means of 10 determinations. The instrument was calibrated against a standard white reference
plate.

TLC Analysis
Wax components of samples were compared by TLC. Silicagel plates (60 F254; Merck,
Germany) were used in this experiment. The wax samples were dissolved in warm chloroform
(Wako, Japan) and applied on the start lines on the plate. TLC development was used two
solvent systems. Solvent system I consisted of hexane-ether-acetic acid (90:15:1 v/v) for
investigation of non-polar component. Solvent system II consisted of hexane-ether-acetic acid
(50:50:1 v/v) for investigation of polar component. After development, the plates were sprayed
with 50% H2SO4 and heated at 105°C until colored spots appeared.

3
Saponification and Separation
Banana wax (M. chilicarpa, 20mg) and 0.5N KOH/ethanol solution (30ml) were mixed and
stirred in a flask. A glass condenser was directly connected to the top of the flask.
Saponification was carried out for 2 h at 95 °C. Then distilled water (10ml) was added to the
saponificated solution. The saponificated mixture was extracted repeatedly with hexane
(100ml). The combined hexane extracts were washed with 0.1N KOH-ethanol (2:3) and dried
over anhydrous Na2SO4. The water layer was combined and added 1N H2SO4 (30ml). The water
layer was extracted repeatedly with ether (100ml). The combined ether extracts were washed
with distilled water and dried over anhydrous Na2SO4.
Fatty alcohols (5.0mg) and fatty acids (12.2mg) were recovered after evaporation of the solvent
from the hexane extract and the ether extract respectively.

GLC Analysis
Fatty acid methyl ester derivatives were prepared using diazomethane and analyzed using a
Shimazu GC-14B fitted with a URBON HR-52 capillary column (0.25mm x 25m). The operating
temperatures were as follows: injector, 330°C; detector (flame ionization detector), 330°C; initial
oven temperature, 200°C, with a ramp rate of 5°C /min. to 290°C, and maintained for 10 min.
He was used as carrier gas.

Results and Discussion

Wax yields
Table 1 shows the yields of wax extracted from banana leaves. Wax yields of Musa liukiuensis,
M. acuminata and M. chilicarpa were 0.58%, 1.05% and 1.41% per dry weight, respectively.
Wax yield of M. chilicarpa showed the highest percentage compared to the other two varieties.

Table 1. Yields of wax from leaves of bananas

Banana varieties
Musa liukiuensis Musa acuminata Musa chilicarpa
No of leaves 2 2 2
Leaf weight (g) 240 405 324
Leaf moisture content (%) 85.95 85.79 84.90
Wax yield (mg) 196.0 604.7 689.6
Wax yield (%; to dry weight) 0.58 1.05 1.41

Melting point and color value

Table 2 shows the melting points and the color values of natural waxes. The melting point of
banana waxes ranged from 78 °C to 82 °C, which was higher than other natural waxes except
carunauba wax. The banana waxes showed the highest degree of lightness, ranging from 72.23
to 86.08.

4
Table 2. The melting points and the color values of natural waxes
Melting point Color values
(°C) L* (lightness) a* (green-red) b*(blue-yellow) C* (chroma)
Musa liukiuensis 78-81 79.70 -0.48 15.45 15.45
Musa acuminata 79-80 86.08 -0.89 8.36 8.41
Musa chilicarpa 78-82 72.23 -1.15 12.74 12.80
Carunauba wax 82-83 63.88 3.04 32.65 32.79
Candelilla wax 67-69 64.41 5.71 41.27 41.66
Rice wax 74-78 68.02 -3.83 28.09 28.35
Japan wax 44-46 65.08 -2.44 8.23 8.59
Bees wax 56-60 56.50 -2.37 9.67 9.95

TLC Analysis
Figure 1 shows a TLC chromatogram of banana waxes and natural waxes. The component
profiles of Musa chilicarpa were similar to carunauba wax. In the results of the TLC
chromatogram using non-polar solvent system I, four spots with Rf values ranging from 0.1 to
0.75 in M. chilicarpa wax showed similar to carunauba wax. In addition, spots with Rf value
ranging from 0 to 0.3 of M. chilicarpa and carunauba wax were similar to each other in the polar
solvent system II.

Figure 1. TLC chromatogram of banana waxes and natural waxes

System I: hexane-ether-acetic acid (90:15:1 v/v)
System II: hexane-ether-acetic acid (50:50:1 v/v)

5
Fatty acid
Wax of M. chilicarpa showed the highest yield and similar chemical components with carnauba
wax was chosen to investigate the fatty acids composition. Figure 2 shows the GLC
chromatogram of fatty acid methyl esters in epicuticular wax extracted from M. chilicarpa leaves.
Fatty acids with carbon numbers ranging from C14 to C30 were detected in the wax after
saponification. C22 fatty acid was most abundant in these fatty acids. The fatty acid
components of banana wax were identified by GLC in comparison with the fatty acid methyl
ester standards or the calibration curve obtained from the standard reference mixture of fatty
acids.

Figure 2. GLC chromatogram of fatty acid methyl esters in epicuticular wax extracted from
banana (Musa chilicarpa) leaves.

Conclusion
Three banana varieties were used to recover waxes from banana leaves. Wax yields of Musa
liukiuensis, M. acuminata and M. chilicarpa were 0.58%, 1.05% and 1.41% per dry weight,
respectively. Hemmers et al. (1986) reported epicuticular waxes extracted from leaves of five
Euphorbia species grown in Europe. There were also some differences in the wax yields
ranging from 0.6 to 2.1 % per dry weight. Melting point is one of the main parameters of the
quality of wax. The melting point of banana waxes was 78 - 82 °C, which is higher than
commercial natural waxes except carunauba wax. Color value is also one of the main
parameters of wax. L* is used for darkness evaluation (100= white and 0= Black). The banana
waxes showed the highest degree of lightness in the natural commercial waxes. These
properties of banana waxes were an expected advantage used industrial material. The
components of M. chilicarpa were similar to carunauba wax by TLC analysis. Tulloch (1973b)
reported that the main components of carnauba wax are monoesters. Therefore M. chilicarpa
wax could be a monoester type wax. Banana waxes were suggested as an alternative material
for carunauba wax. However, there were some differences in the fatty acids composition

6
between M. chilicarpa wax and carunauba wax. Carunauba wax contains fatty acids ranging in
carbon number from C16 to C34 and their predominant compounds are C24 and C28 (Japan Oil
Chemists’ Society, 2001). Fatty acids with carbon numbers ranging from C14 to C30 were
detected in M. chilicarpa leaf wax after saponification. C22 fatty acid was most abundant in
these fatty acids.

Acknowledgements
We are grateful to CERARICA NODA Co.,Ltd., Japan for samples of natural waxes; to
TSUKUBA BOTANICAL GARDEN, Japan for samples of banana leaves used in this work.

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