BUKU E PROSIDING EFAMB

e-Prosiding Festival Agro Makanan Dan Bioteknologi 2020 (E-FAMB2020)

3. RESEARCH METHODOLOGY

3.1 Preparation of Pumpkin Powder

Pumpkin or the Cucurbita moshata species are brought from local supermarket located at Pagoh,
Johor Darul Takzim. The size of the pumpkin is in the range or 2 – 3 kg per fruit. After cleaning and
peeling, the pumpkin flesh was chopped into big uniform strips. The pumpkin strips were then cut
into uniform slabs with a thickness of 2 - 4mm using automated slicer (INEO FV-LVC1). Pumpkin
powder is made by dehydration techniques with temperature of drying was set at 60°C for 6 hours.
Then the dried pumpkin was grounded into powder and sieved using sieve separator and
collected powder with particle size 250 µm. The pumpkin powder then kept in cold, dark and
vacuum packed condition to prevent it from color changed before further used.

3.2 Premix Flour Production

Premix flour (stage 2) is done by mixing all the dry ingredient together such as pumpkin powder,
wheat flour, salt, sugar, grounded chia seed and baking powder and then the premix flour was
packed in aluminum plastic sealer air tight packaging with premix weight around 250 g per pack.
The benefits of this premix flour is to reduce the weighing and preparation time during production
of puri bread as well as can lower the weighing error risk which commonly happen during weighing
the ingredients.

3.3 Puri Bread Production

The pumpkin powder was incorporated at levels of 50% (1:1), 60% (3:2), and 70% (7:3) in puri bread
by replacing the wheat flour. The remaining ingredient which is warm water and oil are added
after that. As the dough is mixing, it will form a mass with nothing left sticking to the sides of the
mixing bowl. This is referred to as clean-up and the kneading process can be stop. Then the puri
bread was fried with deep fried method until it light golden brown or bright orange.

3.4 Sensory Evaluation by Trained Panelist

Descriptive test was used for detection and description of bread of sensory properties quality.
Descriptors for evaluation of bread sensory properties (appearance, taste, color and overall
acceptance) were determined by consensus of all panelists. A sensory evaluation with nine Likert
scale was used to see the overall acceptance by mean of consumption compared to control puri
bread, which are not added with pumpkin and chia seed. The data then being analysis with
ANOVA method using Statistical Package for The Social Sciences (SPSS) software.

4. RESULT AND DISCUSSION

Sensory evaluation is a powerful tool with a broad range of applications in the bakery industry. An
in-house sensory panel consisting of eight trained panelists was used for the sensory evaluations.
Sensory analysis by panelist was carried out using a 9-point hedonic scale, scoring method with 1
is minimum to 9 maximums. Four attributes were identified, which were used with most consistency
among the panelist. They are appearance, color, taste and overall acceptance as shown in table
below.

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Table 2: Sensory Evaluation Attributes with Hedonic Scales

Minimum Maximum

(1) (9)

Appearance Physical structure of puri Weak Strong
bread
Colour Light Dark
Yellowish colour after the
Taste bread fried Weak Strong
Sweet taste ; a basic factor
Overall produced by pumpkin Low High
acceptance Impression of frying bread
following consumption

Figure 2: The Mean Value of Sensory Quality of Frozen Cucurbita

Four attributes were identified, which were used with most consistency among the panelists as
tabulated in Table 2 and Table 3. They are appearance, color, taste and overall acceptance as
shown in tables above. Appearance is a physical observation in frozen Cucurbita puri bread after
frying. Appearance of bread with 50% pumpkin powder give a very soft, crispy and puffier than to
60% and 70% fried Cucurbita puri bread. This is because when addition of too much pumpkin
powder in the formulation it will also increase the fiber content that make the bread heavier and
hard.

The higher the point is the best and preferable but the lower color is better as it is referring to
the yellowness of puri bread. Color of bread with 70% bread are more dark orange in color which
make a significant difference to the control and other formulations. The intensity of color for has a
score of 4.70 ± 1.02 which means that the respondents like them moderately and still consider them
as still acceptable.

On the other hand, taste is a sweet taste that is produced by pumpkin. Puri bread with 50%
and 60% pumpkin powder have a same mean value (7.50 b ± 0.54) thus show that there is no
significant difference between both formulations but give a significant different to control puri
bread and puri bread with 70% pumpkin powder. Nevertheless, overall acceptance attributes are
referred to the overall impression of fried puri bread. All the attributes give a detailed aspect in
terms of quality of puri bread.

From the above discussions, it is obvious that panels favor the 50% formulation more
compared to others. Even though statically for overall acceptance, it shows significant
difference to the others, puri breads are assumed to be acceptable if their mean scores were
above 5 (Lazaridou et al., 2007).

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5. CONCLUSION

From the above discussions, it is obvious that panels favor the 50% formulation more compared to
others as shown to the highest mean value for almost all attributes. Hence, the controlled
formulation too can be assumed as acceptable. Even though statically, it shows significant
difference to the others, but the difference is not very significant. In terms of sensory evaluation, all
the formulations are acceptable to the respondents. Four characteristics was conducted, the
results show that frozen Cucurbita puri bread with chia seed has significant differences in the
sensory evaluation such as appearance, color, taste and overall liking scores. The finding of this
research indicate that consumer preferred and accepted the new product of Cucurbita Puri
Bread with Chia Seed as their new food when using Formulation 1 with only 50% of pumpkin powder
in the formulation. The results also indicate that, by adding Cucurbita or pumpkin powder and chia
seed into the puri bread was affected the consumers’ sensory evaluation of this frozen product.

REFERENCES

Ayerza, R., & Coates, W. (2001). Chia seeds: New source of omega-3 fatty acids, natural
antioxidants and dietetic fiber. AZ, USA: Southwest Center for Natural Products Research &
Commercialization Office of Arid Lands Studies, The University of Arizona.

Bahare, S., Esra, C., Nabil, A., & et, a. (2019). A Review Cucurbits Plants: A Key Emphasis to Its
Pharmacological Potential. MDPI Journal Molecules, 24, 1854: 1-23.

Dr. Szabo P. Balazs, (2012). Sensory Evaluation in Food Industry. 233-253.

Eckardt, J., Ohgren, C., Alp, A., Ekman, S., Astrom, A., Chen, G.,Langton, M. (2013). Long-term
frozen storage of wheat bread and dough - Effect of time, temperature and fiber on sensory
quality, microstructure and state of water. Journal of Cereal Science, 57(1): 125–133.

Ersedo, T. L., (2019). Chemical and Sensory Evaluations of Wheat (Triticumaestivum L.) Bread
Enriched with Pumpkin (Cucurbita maxima L.) Flour. International Journal of Food Science
and Nutrition Engineering. 24-30

Fu, C., Huan, S., Li, Q. (2006). A review on pharmacological activities and utilization technologies
of pumpkin. Plant Foods for Human Nutrition, 61, 73–80.

Jedidah, W. K., Dorcas, K. I. and Peninah, N. N., (2017). Effect of Drying Method on Nutrient Integrity
of Selected Components of Pumpkin (Cucurbita Moschata Duch.) Fruit Flour. ARPN Journal
of Agricultural and Biological Science. 110-116.

Jedidaj Wanjiku Kiharason, Dorcas Khasungu Isutsa. (2019). Shelf-life of Pumpkin Fruit Slices, Flour
and Blended Products. International Journal of Food Science and Biotechnology, 4(1): 14-
25.

Kiharason, J.W., Isutsa, D.K. and Ngoda, P.N. (2017). Nutritive Value of Bakery Products from Wheat
and Pumpkin Composite Flour. Global Journal of Bioscience and Biotechnology. 96-102.

K.M., Masood, S.B., Sharif, H.R. and Nasir, M. (2017). Sensory Evaluation and Consumer
Acceptability. 362-385.

Loreto A. Muñoz ,Angel Cobos,Olga Diaz &José Miguel Aguilera (2013). Chia Seed (Salvia
hispanica): An Ancient Grain and a New Functional Food. Food Reviews International
Volume 29.

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Lazaridou A, Duta D., Papagergiou M., Belc N., Biliaderis C.G (2007). Effects of hydrocolloids on

dough rheology and bread quality parameters in gluten-free formulations. Journal of Food
Engineering 79(3):1033-1047.
Murzaini, N. M. N., Taip, F. S., Aziz, N. A., Rahman, N. A. A. (2020). Effect of Pre-treatment in
Producing Pumpkin Powder Using Air Fryer and Its Application in “Bingka” Baking. Current
Research in Nutrition and Food Science Journal, 8(1): 48–64.
Munoz, L. A., Cobos, A., Diaz, O., & Aguilera, J. M. (2012). Chia seeds: Microstructure, mucilage
extraction and hydration. Journal of Food Engineering, 108: 216–224.
Spruce (2014). Pumpkin. Octopus Publishing Group. 42-60.

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THE PHYSICOCHEMICAL ANALYSIS OF DRINKING WATER QUALITY
CONSUMED BY PAGOH, JOHOR RESIDENTS

Azierah Zawiyyah binti Azmi, Muhammad Habibullah bin Yahaya, Ilakkia A/P Thandaithabany

Politeknik Tun Syed Nasir Syeb Ismail, KM1 Jalan Pachor ,
Hub Pendidikan Tinggi Pagoh,Johor Malaysia
*[email protected]

Abstract. The source of domestic water through pipeline system is susceptible to biological
pollutants, chemical contaminants and other problems that affect the quality of the water,
especially for drinking water. This study was carried out to evaluate the physical and chemical
properties of drinking water in Pagoh, Johor. There were 8 numbers of water sample has been
collected on 4 different types of drinking water which were tap water, boiled water, filtered water
(labelled as A and B), and vending machine water (labelled as C, D, E and F). Several parameters
such as pH, Biological Oxygen Demand (BOD), chlorine, Total Suspended Solid (TSS) and heavy
metal contents in each type of drinking water were analyzed in this study. Based on the result, for
pH, total soluble solid and BOD values were within the permissible range of both WHO and NDWQS.
However, the chlorine content in all water samples were below the standard limit, this is somehow
alarming as it may contribute to the growth of microbes in drinking water. Meanwhile, the major
problems identified were that the heavy metal remaining in the water even though treatment of
boiled and filtration had been done. This condition may cause by the efficiency of water
treatment method for each drinking water in order to attain good quality water.

Keywords: pH, Biological Oxygen Demand (BOD), Chlorine, Total Suspended Solid (TSS), Heavy Metal

1. INTRODUCTION

Water is essential for life. All living things in the world like humans, animals and plants need water,
without water there would be no life on earth. Diarrhea is one of the major water-borne diseases
that still give health burden in many parts of the world. According to World Health Organization
(WHO), (2002) reported that 5.7% of the global disease were cause by water, which about 4 billion
clinical cases of diarrhea per year. Even though there are other sources for contamination with
pathogen of gastrointestinal tract like poor sanitation and food, drinking water is the major source
of microbial pathogens in developing regions (Ashbolt, 2004). Failure to supply safe water will
place a serious problem to humanity. The source of domestic water through pipeline system is
susceptible to biological pollutants, chemical contaminants and other problems that affect the
quality of the water, especially for drinking water. The contamination of water may cause from
landfills and septic systems, through careless disposal of hazardous household products,
agricultural chemicals, and leaking of underground storage tanks (Yasin et al., 2015).

Tap water quality is identified as one of the major factors on the increase in demand from
consumers who are mentally prepared to pay for the price of better quality drinking water. They
thought the quality of filtered drinking water and vending machine water are much better in term
of hygiene, safety and mineral content. Nowadays, in Malaysia there are increasing population
that installing the water filter at their home. This is because consumers are concern on the health
risk regarding the safety of potable water. There are various brands of water filter that comes into
existence such as Coway, Cuckoo, SK Magic and others. The water filters also have very high
market potential, and it is being a contributor to the national economy. Although, the water
utilities from the pipline systems had strong assurance that water been delivered is safe for direct
human consumption as the water had treated according to the Malaysian National Standards for
Drinking Water Quality (NSDWQ), Malaysian still seek to other measures such as boiling water,

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installing a water filtration system, use vending machine water and buying bottled water claimed
by Othman et.al, 2013. When installing the filtered water or buying bottled water or buying water
from the vending machine, the consumer confident that the water is safe to drink without further
process of boiling it, as that has been practiced previously. It is believed that, by filtering tap water
before consumption are the best way to ensure the water is safe for drinking as filtering removes
sediments in the water. Apart from thinking about personal health, consumers like to use this kind
of water because it makes easier for consumers to drink water without having to cook and do not
need to use containers to store the water. These are the research questions, i. Is it true that the
water is safe to drink without further processing? and whether Is it true that the process of water
filtration can filter out all the harmful contents in the water?

Based on the questions, this study was designed to examine the current safety status of
different water sources that was used for drinking. The water samples were collected in Pagoh,
Johor which is located at the Southern Peninsular Malaysia. The drinking water sources included
in this study were tap water, boiling water, filtered and vending machine water. These samples
were chosen because Pagoh residents primarily consuming this type of drinking water, based on
the survey that has been done to 50 respondents who live around Pagoh. A number of
parameters such as pH, Biological Oxygen Demand (BOD), chlorine, Total Suspended Solid (TSS)
and heavy metal contents like Magnesium (Mg), Calcium (Ca), Ferum (Fe), Zinc (Zn), Nikel (Ni),
Copper (Cu), Arsenic (As), Chromium (Cr), Cadmium (Cd), and lead (Pb) in each drinking water
were analyzed in this study. The results of each parameter were compared to the guidelines and
standards set by the WHO and local standards such as NDWQS.

2. LITERATURE REVIEW

The entire population in Malaysia nearly has accessed to improve water supply. Although, the
water supply system is generally satisfactory, there are still problems remain with the quality of
potable water. In the news of Utusan Malaysia April 4, 2005 reported that the Malaysian Water
Association has pointed out 90% of Malaysians receive low quality potable water. This statement
also has been supported by Law (2005) and Aini et al. (2007). Law (2005) reported that around
75% of Kuala Lumpur residents were not satisfied with the quality of piped water in their homes.
Meanwhile Aini et al. (2007) stated that majority of the respondents in Seremban City rated the
quality of water supplied is very poor. The respondents stated that the colour, taste and odour of
piped water is the main problem. So that, the study found most of the residents took an additional
method to improve the water quality by boiling the water, installing water filters or using vending
machine water.

Access to safe drinking water is a universal need, there were many researches had been
conducted to evaluate the quality of the drinking water in physical, chemical and biological
aspects around the world. In Malaysia, research was also being done in specific area. The quality
of the drinking water shown both positive and negative results. Positive results indicate that the
parameter was not exceeding the maximum limit of standard regulation while negative results
represent that the value of the parameter were not according to the standard regulation. The
data obtained were vary depending on the specific location of the research.

According to Chan et al. (2007) on the quality of filtered water collected randomly from a
total of five houses in Klang Valley, Malaysia. The water samples were collected before (labelled
as A1, B1, C1, D1, E1) and after filtration treatment process (labelled as A2, B2, C2, D2, E2). This
research mainly done to observe the impact of tap water before and after the filtration
treatment. In this result, total plate count analysis and the pH value for all the water samples gave
negative results. While for the total suspended solids and the turbidity for all the samples gave
positive results, which mean that it complies with regulations.

Another research has been done by Hashim & Yusop in 2012. The vending machine water
quality was investigated from eight locations in Parit Raja, Johor in terms of pH, total dissolve
solids (TDS), turbidity and heavy metal content (chromium, arsenic, cadmium, lead and nickel).
Results for TDS

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and heavy metals show positive results. On the other hand, the pH value gave negative results
while the turbidity was found to be 45 to 95 times higher than 0.1 as required by the Malaysian
Food Act Regulation. The data obtained in that study would suggest the important of routine
maintenance and inspection of vending machine water in order to maintain a good quality,
hygienic and the safety level of drinking water.

There is also research that reported positive data for all the physicochemical parameters
conducted on the drinking water quality in the state of Perak, Malaysia (Rahmanian et al., 2015).
This indicated that all the parameter within the safe limit. The drinking water samples was carried
out in different residential and commercial areas of the state while the parameter analyzed were
pH, turbidity, conductivity, total suspended solids (TSS), total dissolved solids (TDS), and heavy
metals such as Cu, Zn, Mg, Fe, Cd, Pb, Cr, As, Hg, and Sn. Researcher conclude that the water
from all the locations in perak state was found to be safe as drinking water.

Based on the previous studies, it can be assumed that the quality of drinking water from
various treatment approaches in Malaysia will give different results according to the location of
the pipeline system. Different locations or state in Malaysia provide different data in terms of
physical, chemical and microbiological. Even though the water treatment plants are following
the same standard, each state has a different governing body that responsible for the treatment
of household water supply for each state in Malaysia. It could be hypothesized that it is important
to conduct research in every location in Malaysia for evaluating the quality of the drinkingwater.

3. RESEARCH METHODOLOGY

3.1 Study site and period

The study was conducted at Pagoh, Johor which is located at the southern of Malaysia. Pagoh is
estimated about 154 km from Johor Bahru and 75.6 km from Melaka via the North-South
Expressway. The authority who responsible for the treatment of raw water in this area is Ranhill SAJ
(Syarikat air Johor) Muar. The study was conducted from August 2019 to October 2019.

3.2 Survey research

Questionnaires were used to gather pertinent information on the source of drinking water
consumed by the Pagoh residents. Fifty questionnaire form were distributed to Pagoh residents in
July 2019 where it comprises the area of Pagoh Jaya, Kampung Sari Baru and Pekan Pagoh. The
background of the respondents was selected randomly to represent the population of Pagoh
residents. Then, the data were analyzed using Microsoft Excel 2010.

3.3 Water sampling

The drinking water samples were collected from tap water, boiled water, two popular brands of
filter water (labelled as A and B) and four different brand of vending machine water (labelled as
C, D, E and F) which are accessible in Pagoh, Johor. Samples were aseptically collected from
each sampling site in sterile glass bottles analysed within 12 hours of sample collection.

3.4 pH

30 ml of each type of drinking water was collected and measured each the sample thrice using
Jenway Model 3510 pH meter, UK.

3.5 BOD

Sterilized small empty bottle with distilled water. Then, 250 ml of water sample was filled in the small
empty bottle. Next, the YSI Model 5905/5010/200-BOD/Pro-BOD probe, USA was inserted into the

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water sample to measure the reading of DO value. The reading was recorded. The measurement
of each water sample was triplicated. After that, the testing continues for up to 24 hours with the
testers being incubated in the incubator at 28 ˚C. The next day, the final reading of DO value was
recorded. Finally, the BOD value was calculated according to the formula given below stated by
Hassan et.al (2014).

Eq 1: BOD mg/L = D2 - D1

Where:

D1: Dissolved oxygen of diluted sample immediately after preparation, mg/L
D2: Dissolved oxygen of diluted sample immediately after 24 hours at 28 ˚C, mg/L

3.6 Chlorine

Potassium chromate solution (5%) was prepared by dissolving 5 g of potassium chloride in 100 ml
of distilled water. Then, 17.5 g of sliver nitrate was dissolved in 1000ml of distilled water. Next, 10 ml
of water sample was measured accurately. After that, 1 ml of 5% w/v potassium chromate solution
was added in 10 ml of water sample. Later, 0.1 M silver nitrate was titrated until a permanent faint
reddish brown colour is obtained. The titration was performed in triplicate. Lastly, the chlorine
residual calculated according to the formula given.

Eq 2: Normality of silver nitrate = sodium chloride in g
solution, AgNO3 silver nitrate titrate value in ml x 58.44

Eq 3: Residual chlorine (Cl2 g/L) = Titration value (ml) x normality of AgNo3 x 35.5 x 100
Sample size in ml x 1000

3.7 Total Suspended Solid

The Total Suspended Solid (TSS) was according to Mazlin et al, 2003. A filter paper was weighed
and later wet with sterile distilled water. The sample was stirred homogenously to ensure no
precipitation. The sample was measured accurately and poured through the filter paper. The
filtering was done using a filter funnel. Next, the filter paper was transfer into the oven for drying at
103°C for 24 hours. The filter paper was left to cool before being weighed. The TSS was counted
according to the formula below:

eq 4: mg of total suspended particles/ L = (A – B) x 1000
ml sample

3.8 Presence of heavy metal

Samples (30mL) was measured using measuring cylinder from the collection. Afterwards, 10 drops
of 1:1 HNO3 is added to each sample to prevent the adsorption of soluble metal ions to inner wall
of the bottle and to reduce the microbial activity after sampling. Next, the samples are stored
immediately in the refrigerator at 4˚C until analysis. Finally, the concentration of heavy metal Mg,
Ca, Fe, Zn, Ni, Cu, As, Cr, Cd, and Pb in the water samples were analyzed using Agilent 7700e ICP-

MS, USA according to standard method GB/T 2050.6-2006 cited by Lu & Yuan (2017).

4. RESULTS AND DISCUSSION

The drinking water consumed by Pagoh residents were represented in Figure 1. From the data, the
highest drinking water consume was from the filter water. This result proved that the increasing

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trend of installing the filter water as the source of drinking water in the house. The data also proved,
there were still a big number of households that still use the normal practice which is boiling the
tap water that coming from the pipeline. This research investigated the physical, biological and
chemical properties of the drinking water in term of pH, TSS, BOD, chlorine, and heavy metal
contents. Only water that come directly from the pipeline with or without further treatment process
were selected as the samples.

42% 6% Bottled drinking water
9% Bottled mineral water
Vending machine water
11% Boiled water
Filter water
32%

Figure 1: Drinking water consume by Pagoh residents

Table 1 show the results for pH, TSS, BOD and chlorine content for all the water samples. pH
and TSS were categories as physical properties. pH is classed as one of the most important water
quality parameters, it represents whether the drinking water is acidic or alkaline. According to
standard of WHO and NDWQS the normal pH of drinking water range between 6.5 to 8.5. Among
the 8 water sample, filter water A is more effective in producing 7.48 pH value of neutral water.
This followed by filter water B 6.98, vending machine water C 6.79, boiled water 6.62, tap water
6.58 and vending machine water D 6.56. But the E and F vending machine water was lower than
the standard value which is 6.39 and 6.24 respectively. The similar result also obtained by Hashim,
& Yusop (2012) that vending machine water was founded in between pH range 5.5 – 6.4. But in
this analysis the taste perceptions of all water samples with minimum and maximum pH level were
all considered satisfactory among the consumers.

Table 1: The results for pH, TSS, BOD and chlorine content for all the water samples

Water sample pH TSS (g) BOD (ppm) Chlorine (mg/L)

Tap water 6.58 0.006 0.14 3.5560 x 10-5
Boiled water 6.62 0.002 1.49 3.5465 x 10-5
7.48 0.001 0.09 3.5486 x 10-5
A 6.98 0.001 0.10 3.5500 x 10-5
B 6.79 0.009 0.11 3.5465 x 10-5
C 6.56 0.003 0.08 3.5536 x 10-5
D 6.39 0.015 0.27 3.5528 x 10-5
E 6.24 0.014 0.82 3.5518 x 10-5
F

Meanwhile, for TSS, the maximum allowable value in drinking water is 25 mg/L according to
NDWQS. The total suspended solid is formed when there is mineral content in water which means
the higher the mineral content in water sample will form more total suspended solid. The finding
of total suspended solid in this analysis obtain are in range of 0.001 to 0.015 g. The highest value of
total suspended solid in drinking water is vending machine water E 0.015 g whereas the lowest
value is filter water A and B 0.001 g. This indicates that filter cartridge of filter water A and B are
able to remove the suspended materials finely to produce pure water. The amount of total
suspended solid for other water sample such as tap water 0.006 g, boiled water 0.002 g, vending
machine water C 0.009 g, vending machine water D 0.003 g, and vending machine water F 0.014
g. This shows that all the water sample does not exceed the limit of NDWQS standard.

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On the other hand, BOD is represented for the biological properties of the drinking water.
According to BOD results in table 1, the highest BOD level in drinking water is boiled water, 1.49
ppm whereas the lowest value of BOD level among the water sample is D, which is coming from
vending machine, 0.08 ppm. Water sample A and B were filtered water where its range between
0.09 to 0.10 ppm which means that the filtration system was able to filter the organic waste in tap
water, 0.14 ppm which directly come from pipe holds and make it safer to drink. Moreover, water
sample C, D, E, F were vending machine water, the range of BOD level is between 0.11 to 0.82
ppm. All the water samples are in the range of BOD level of 1 – 3 ppm which indicate the quality
of water is more clean and there is no much organic waste present in the water supply based on
NDWQS stated by Nur Diyana (2011).

Last but not least, the chemical properties were investigated in term of the chlorine and
heavy metal content. Residual chlorine in water distribution system is maintained to prevent from
waterborne diseases throughout the distribution. From the results, chlorine content in water sample
is range from 3.5465 x 10 -5 to 3.5560 x 10 -5 mg/L. The highest value of chlorine content is tap water
3.5560 × 10 -5 mg/L whereas the lowest value is boiled water and vending machine water C 3.5465
× 10 -5 mg/L. After the tap water undergoes boiling and filtration process the chlorine content was
reduced because the chlorine compound was breakdown or filtered. For filter water A and B the
chlorine content is 3. 5486 × 10-5 mg/L and 3.5500 × 10-5 mg/L respectively whereas vending
machine D, E, F is between 3.5518 × 10-5 to 3.5536 × 10-5 mg/L. According to Malaysian Standard
of Drinking Water, the concentration of chlorine in disinfected drinking water should be within 0.2
to 1.0 mg/L. The chlorine content in all water sample are below the standard limit which may
causes the microbial growth in water. In previous research by Codony et al. (2005) stated that the
microbial count in drinking water can increase by 10 fold in the low concentration of chlorine
residual.

Table 2: The average value of the concentration of heavy metals in each water sample

Water Mg Ca Fe Heavy metal analysis (mg/L) Cd Pd
sample 30 400 0.06 Zn Ni Cu As Cr 0.0006 0.002
Allowable 3.36 0.59 0.14 0.6 0.004 0.2 0.001 0.01 0.00657 0.019
Tap water 3.42 1.65 0.10 2.57 0.0105 2.80 0.0045 0.80 0.00036
Boiled water 2.50 21.1 0.21 0.012 0.0002 14.3 0.0002 Tr 0.00001 Tr
3.07 25.7 0.04 Tr 0.0701 ND 0.0358 ND 0.00003 ND
A 3.08 3.32 0.03 0.019 0.0058 ND 0.0420 0.30 0.00036 Tr
B 3.02 0.7 0.009 2.2 0.0007 12.3 0.0127 Tr 0.00049 Tr
C 2.43 4.66 0.6 0.0005 4.30 ND ND 0.00039 0.003
D 3.16 4.56 Tr 0.008 0.0355 15.1 0.0026 Tr 0.00045 0.001
E 0.073 24.3 0.0068 12.2 0.00009 Tr Tr
F

Analysis on the heavy metal concentration of Mg, Ca, Al, Zn, Ni, Cu, As, Cr, Cd, and Pb
content (table 2) showed that the heavy metals were present in all water sample. The allowable
amounts in table 2 were referred to Food Act 1983 (Act 281) and Regulations, 2018. The most
contaminated sample was tap water, similar result was obtained by Azlan, et.al (2012). After
filtration process, the results showed that the concentration of the heavy metals were decreasing
although some of the heavy metal were exceeded the permissible value. This indicates that the
filtration process of tap water is important in reducing the potential of health effect that cause
from heavy metal that become toxic to human body stated by Karavoltsos et al. (2008). The
content of heavy metal in the water sample may be due to corrosion of pipeline cause from the
leaching of heavy metals due from ageing and pitting as cited by WHO (2006). Pin–like formation
is called pitting in the metal surface cause by corrosion of pipe when water directly contact.
Meanwhile improper filtration and poor maintenance service similarly cause the heavy metals
accumulation at the filter cartridge that can reduce the efficiency of filtration. Furthermore, all
the water treatment methods need maintenance on regular basis to maintain optimum water
quality.

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5. CONCLUSION

Water samples which consist of boiled water, filtered water and vending machine water showed
that almost all of the physicochemical parameters like pH, TSS and BOD amounts were within the
permissible range of both WHO and NDWQS, expressing its suitability for drinking purpose.
However, the chlorine content in all water sample is below the standard limit, this is somehow
alarming as it may contribute to the growth of microbial organisms in drinking water. Further study
should be emphasized on this matter to ensure the drinking water safe to be consumed.
Meanwhile, the major problems identified were that the heavy metal remaining in the water even
though treatment of boiling and filtration had been done. The importance of regular service to
maintain optimum drinking water quality is critical. Poor water sources from the water treatment
plant due to the corrosion and pitting of pipeline system contribute to the heavy metal
contaminations. Therefore, there is a need to study and design an efficient raw water treatment
process, waste disposal system and a catchment area management system around the Pagoh
water sources. In addition to ensure that the water is safe for human consumption, regular water
quality assessment of the source, main distribution tanks, distribution systems, and pipelines should
be carried out. This present study was limited in evaluating the physical and chemical parameters
from household tap water supply that had been boiled and filtered by in-house water filters and
vending machines. Thus, beside study in microbial aspects, further study should consider the
addition of other potential water contaminations such as others chemical parameter and
radiological materials for a longer period, in order to assess the overall water quality in Pagoh.

REFERENCE

Aini, M. S., Fakhrul-Razi, A., Mumtazah, O., & Meow Chen, J. M. (2007). Malaysian households’
drinking water practices: A case study. International Journal of Sustainable Development &
World Ecology, 14, 503– 510.

Ashbolt, NJ. (2004). Microbial contamination of drinking water and disease outcomes in
developing regions. Toxicology. 38,198-229

Azlan, A., Khoo, H. E., Idris, M. A., Ismail, A., & Razman, M. R. (2012). Evaluation of Minerals Content
of Drinking Water in Malaysia. The Scientific World Journal, 1–10. doi:10.1100/2012/403574

C.L., Chan, Zalifah, M.K. & Norrakiah, A.S. (2007). Microbiological And Physicochemical Quality Of
Drinking Water. The Malaysian Journal of Analytical Sciences, 11(2), 414 – 420

Codony, F., Marato, J. and Mas, J. (2005). Role of discontinuous chlorination on microbial
production by drinking water biofilms. Water Res., 39, 1896 – 1906.

Food Act 1983 (Act 281) & Regulations. 2018. Regulations 394 (1) and 360B (3), Twenty-Fifth
Schedule Standard for Water and Packaged Drinking Water. pp.316. Malaysia: International
Law Book Services.

Hashim N.H and Yusop H.M. (2012). Drinking Water Quality of Water Vending Machines in Parit
Raja, Batu Pahat, Johor. doi:10.1088/1757-899X/136/1/012053.

Hassan R.P, et.al. (2014). Determination of Biochemical Oxygen Demand (BOD) Without
Nitrification and Mineral Oxidant Bacteria Interferences by Carbonate Turbidimetry.

Jamal Othman, Goh Hong Lip and Yaghoob Jafari. (2013). Benefits valuation of potable water
quality improvement in Malaysia: the case of Kajang Municipality. Vol. 30, No. 4, 621–634,
Retrieved from http://dx.doi.org/10.1080/07900627.2013.876851.

Karavoltsos S., Sakellari A., Mihopoulos N., Dassenakis M., Scoullos M.J. (2008). Evaluation of the

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e-Prosiding Festival Agro Makanan Dan Bioteknologi 2020 (E-FAMB2020)

quality of drinking water in regions of Greece. Desalination; 224:317–29.
Law BB (2005). The Usage of Domestic Water Filtration Systems in Malaysia A dissertation

unpublished. University of Southern Queensland Faculty of Engineering and Surveying.
Pp.132
Lu J & Yuan F. (2017). The effect of drinking water quality on the health and longevity of people -
A case study in Mayang, Hunan Province, China. IOP Conf. Series: Earth and Environmental
Science 82 (2017) 012005. doi :10.1088/1755- 1315/82/1/012005.
Mazlin, M., Mohd Talib, L. and Lee, Y. H. (2003). Water Chemistry. Kuala Lumpur: Utusan Publications
and Distributions Sdn. Bhd.
Ministry of Health Malaysia, NDWQS: National Drinking Water Quality Standard, Engineering of
Services Division, Ministry of Health Malaysia, 2nd edition, 2004
N. Rahmanian, Siti Hajar Bt Ali, M. Homayoonfard, N. J. Ali, M. Rehan, Y. Sadef, A. S. Nizami (2015)
Analysis of Physiochemical Parameters to Evaluate the Drinking Water Quality in the State of
Perak, Malaysia. Journal of Chemistry, vol. 2015.doi:10.1155/2015/716125
Nur Diyana M.S. (2011). Community Water Quality Monitoring. Unpublished degree’s thesis,
University Malaysia, Pahang.
Utusan Malaysia. (2005, April 12). 90% penduduk dapat air bersih kurang baik (90% of population
received low quality clean water). Retrieved from
http://www.utusan.com.my/utusan/info.asp
WHO. The World Health Report. Switzerland: World Health Organization; 2002.
World Health Organization (WHO). (2011). Guidelines for DrinkingWater Quality, WHO Press,
Geneva, Switzerland, 4th edition.
World Health Organization (WHO). (2006). Health aspect of plumbing.
Yasin, M., Ketema, T., & Bacha, K. (2015). Physico-chemical and bacteriological quality of drinking
water of different sources, Jimma zone, Southwest Ethiopia. BMC Reseacrh Notes, 8, 54

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THE PRODUCTION OF NATURAL DANDRUFF OIL BY USING
ORANGE PEEL WASTE

Noorizyan Binti Asimat, Ganjanan A/L Gunalan, Nurul ‘Izzati Binti Borhan
Politeknik Tun Syed Nasir Syed Ismail, Jorak,
84500 Bukit Pasir, Johor.
*[email protected]

Abstract. Anoteworthy insufficiency of numerous investigations intothe idea of dandruff has
been an inability to characterize the turmoil unequivocally. We characterize dandruff as
incessant non fiery scaling of the scalp. Albeit incessant, the fluctuating character of
dandruff has since quite a while ago entangled its investigation. Dandruff is the basic
grievancesandisenduredbythesame number of ashalf of thenumber of inhabitantsinworld
sooner or later during theirlife. Thebasic topicalreadinessusedtotreat dandruff incorporates
ketoconazloe, flucanazole, zinc-pyrthrone, selenium sulphide, coal tar and so on. Both
engineered and normal medications are administered in various plan like shampoos,
cream, moisturizers, emulsions, hair oils and other restorative detailing. Alongside
manufactured treatment, natural components are increasingly favoured for reducing the
dandruff. This purposed product is to produce natural product, the fundamental oil from
orange peel for dandruff problem. Orange peel has a lot of benefit and has a potential to
become a great product. As we know, orange peel content a d-limonene and carotenoid
which is a beneficial antioxidant and antibacterial substance. A method used is using
extraction or water distillation to create oil. Other than that, to explore the adequacy, the
correlation will be done between utilizing this normal basic oil from orange strip waste and
dandruff itemsin market.

Keywords: dandruff, orange peel, natural dandruff oil, extraction, waste.

1. INTRODUCTION

Dandruff is a typical scalp issue influencing practically 50% of the post pubertal populace of
anyethnicityandthetwosexual orientations.Dandruffistheregularprotestsandisenduredby
the same number of as half of the number of inhabitants in world sooner or later during their
life. The condition is for the most part described by the nearness of chips on scalp and in the
hair. The indications can fluctuate and the seriousness can extend from mellow scaling to
extreme scaling. Its pervasiveness and seriousness is most noteworthy in youngsters and kids.
Themoreseasoned peopleendure lessasoften aspossible to dry scalp and dandruff. Among
800 male, 521 (65.1%) endure with dandruff, though 279 (34.9%) don't endure, especiallythe
individualofagebunch21- 40 endure most with dandruff than other age gatherings. There
are number of variables that in charge of the dandruff like parasitic diseases, hormonal
irregularity, cool, dry climate, poor sterile propensities, long haul pressure and nervousness,
inconsistent shampooing ordeficient washing of the scalp and hair, horrible eating routine
and so on (Hati, Bhavnagar, 2013).

Regular fundamental oil is a characteristic item that can be utilized to decrease
dandruff issues. Fundamental oil is gotten from plant material which held inside certain piece
of the plant or explicit piece of the plant cells; it might be from leaves, seeds, strips, or stalks
relying uponthe species. Citrus natural product have a place with six general (Fortunnella,
Eromocitrus, Clymmedia, Poncirus, Microcitrus, and Citrus), which are local to the tropical
and sub-tropical districts of Asia, yet the real business incorporates a few significant organic
products,for example, oranges, mandarin, lime, lemon, and grape natural products. The
basic oil is available in the natural product's strip in extraordinaryamounts. Essential oil from

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citrus is a large type of natural flavours and fragrances which is popularly used in food
industries, daily chemical products and health care field. The citrus species are potential
sources of variable oil which might be utilized for edible and other industrial purposes. The
exact function of essential oils in a plant is unknown, it maybetoattractinsectsfor pollination,
ortorepelharmfulinsects oritmaybe simplyametabolic intermediate (Giwa,2018).

The main component in citrus oil or orange peel essential oil is limonene. Its
concentration intheoil differ between 30% and 99%, be depending on the variety: 30 -40% in
bergamot,40-75% inlemonand68-98%insweetorange(Moufida,2003).Limonenemayhave
severalhealthbenefits that have been shown to possess anti-inflammatory, antioxidant, anti-
stress and many more. Other than that, Limonene enhanced good mood-elevating effect.
Relaxing indicas high inlimonene counteract the potential for lower moods from increased
sedative terpenes such as myrcene. Mycrene are known for producing joyful and euphoric
effects, in addition to an overall feeling of relaxation which can be found in natural
fragrance. In this project, we also discover the presence of carotenoid in orange peel oil.
Carotenoid always be found in variety of plant, fruits, algae and bacteria. It was important
substance which is the part of antioxidant defense system.

In this project, we used extraction by water distillation to obtain the orange peel oil.
The use of appropriate extraction technology, plant material, manufacturing equipment,
extraction method and solvent and the adherence to good manufacturing practices
certainly help to producea goodquality extract(Tsegaye,2019).Forthisproject,wechoose
using water distillation because this method protects the oil to be extracted till a certain
degree since surrounding water prevents it from overheating. Water distillation is the most
favored method of production of citronella oil from plant material (Majeed,2013). Then we
obtained the presence of limonene, carotenoids and mycrene by analyze using UV-VIS
Spectroscopy.

2. BACKGROUND STUDY AND OBJECTIVE

Nowadays, people become more natural conscious and try to recycle or reuse the waste
they generated. Mostly, people more prefer to natural products which do not contain
chemical substance thatcancause long-termhealtheffect.Besides, thenaturalproduct will
notbeharmful to the sensitive part especially skin.

There are several benefits of the “essential oil’ solution in daily life. One of the
advantages is to save money by convert kitchen waste such as orange peel waste into
multipurpose natural essential oil that can help against dandruff, hair fall and scalps. Other
than that, “essential oil” solution produced from orange peel waste can help reducing the
kitchen waste of orange peel causes from dailyuses.

The objective of this project is:

1. To produce natural dandruff oil from orange peel waste.

2. To detect the presence of limonene, carotenoid and mycrene in the orange peel
waste by analyze using UV-VIS Spectroscopy.

3. METHODOLOGY

The research methodology is the most important part of a study to be used to ensure that
studies are carried out according to the methods, procedures and the correct way. By
following our main objective which is to produce natural dandruff oil, we carried out a
research to find the best method to produce this product so, the process to complete this

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project willrun smoothlyand the product can be obtained.
3.1 Methodology flowchart

Gathering Information:-
- Research on solvent extraction
- Interview
- Questionnaire

Selection and preparation of Raw
Material:-
- Orange peel
- Fractional coconut oil

Preparation of sample of essential oil
from water distillation

No
Analysis

End

Figure 1: Methodology Flowchart

3.2 Procedure of water distillation process
150 g of the orange peel waste sample is put in the round bottom flask with huge measure of
water added to cover the peel. The flask was connected with the still section which was
connected with the condenser, as shown in Figure 2. The steam is produced from the

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heated water and then extractedtheessentialoilwhich wascondensed aspart of thesteam
and it waspassing through the condenser. A mixture of water and oil was then collected
and the other flask where it was separated into two layers. Later, the mixture was separated
by using separating funnel to obtain the oil. The summary of the procedure has been shown
in flowchart at figure 3.

Figure 2: Water distillationapparatus

Prepare sample ( orange peel +
water) in round bottom flask

figure 2) until themixture of oil
and water obtained

Separation process using
separating funnel to separate the
oil-water mixture

Figure 3: Water DistillationProcedure

3.3 Analysis method using uv-vis spectroscopy
Analysis method are used to describe the analysis process that uses for our method. This is to
determine the contamination needed for our product. This can be done through reading
past project journal that related to our project.

UV/VIS/NIR spectroscopy is an amazing diagnostic strategy to decide the optical
properties (transmittance, reflectance and absorbance) of fluids and solids. It very well may
be applied to portray semiconductor materials, coatings, glass and numerous other
research and assembling materials. UV/VIS/NIRworksintheopticalrangebetween 175 nmto
3300 nm.

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UV-Vis spectroscopy is commonly used to decide analyte focuses or the synthetic
change of a part in arrangement. The method estimates the retention of light over the ideal
opticalrange. An example is administered into a cuvette and set in the way between the
optical light source and an identifier. As per the Beer-Lambert law, with a steady light way
length and known assimilation coefficient (subordinate upon wavelength), the
centralization of a compound being referred to can be resolved from the light ingested at
that wavelength.

3.2.1 uv-vis spectroscopy analysis to detect the presence of carotenoid, limonene and mycrene

From the previous study, the majority of carotenoid exhibit absorption in visible region of
spectrum between 400 to 500 nm.(Scott, 2001) or 470 nm (Borello, 2019). Thelimonenecan be
detected atmaximumabsorption 220nm(Lide,1993). Formycrene,itcanbedetected byUV-
Vis at maximum absorption 230 nm (Victor,2018).

4. RESULT AND ANALYSIS

For analysis, we use two sample. Sample 1 is pure orange peel oil which has been distillated.
Intheotherhand,sample 2isthereferencesample wheretheoilisdilutedwithfractional coconut
oil as the oil carrier

4.1 Physical properties

Table 1: Physical properties

Odor Sample 1 Sample 2
Color (orange peel oil +
Solubility (orange peel oil by fractional coconut oil)
water distillation)
Orange smell moderate
colorless yellowish
Not soluble
Soluble

4.2 Chemical properties

4.2.1 Carotenoids wavelength at 470 nm

Carotenoids act as antioxidants in the human body. They have strong cancer-fighting
properties, according to the Physicians Committee for Responsible Medicine. Some
carotenoids are converted by the body to vitamin A, which is essential to vision and normal
growth and development.

For this project, the wavelength of the carotenoid at UV-VIS Spectrometer is to be set
at 470 nm( Borello, 2019). The absorbance of the carotenoid is shown at table 2.

Table 2: Absorbance reading of carotenoids at 470 nm.

Abs reading 1 2 3 Average
Sample 1 0.4540 0.4302 0.4314 0.4385

Sample 2 0.0633 0.0637 0.0643 0.0638

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Abs Reading Carotenoids at 470 nm Sample 1
Sample 2
0.5
0.45

0.4
0.35

0.3
0.25

0.2
0.15

0.1
0.05

123

Figure 4: Carotenoids at 470nm

According to Beer-Lambert Law, the absorbance will be proportional to the
concentration in the solution. The law is usually obeyed for absorbance between 0.1 to 2.0,
especially for the wavelength at the absorbance peak (Tang, 2018).

From the results, the carotenoid is detected at both sample. Sample 1, is pure orange
peel oil has detected at higher absorbance than sample 2 which is the orange peel oil which
is diluted with coconut oil as oil carrier. Usually coconut oil contains carotenoid in range 0.34
± 0.1 (ppm). Therefore, from this results we know that pure orange peel oil is better as the
absorbanceishigher and the total amount of carotenoid is also higher. As we know, coconut
oil is also used widely at market as a good oil for dandruff treatment.

4.2.2 Limonene wavelength at 220 nm

Limonene is one of the most widely recognized terpenes found in nature and may offer a few
medical advantages. It has been appeared to have calming, cancer prevention agent,
hostile to stretch, and perhaps infection averting properties. At wavelength 220 nm, the
absorbance will detect at the higher absorbance.

Table 3: Absorbance reading of limonene at 230 nm.

Abs reading 1 2 3 Average
Sample 1 1.7508 1.7409 1.7361 1.7426

Sample 2 1.8158 1.8304 1.8458 1.8307

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Figure 5; Limonene at 470nm

From the reading at Table 3, limonene can be detected for both sample
wavelength 230nm with Sample 2 have higher absorbance reading than Sample 1. But
from the result the different is not too vary. Therefore, we can have concluded that
limonene can be detected at both sample.

4.2.3 ß-myrcene wavelength at 230 nm

Beta myrcene has additionally been advanced for its enemy of oxidant properties, for
example, in a recent report where it was looked at close by curcumin and demonstrated a
solid connection with a diminishing in oxidative worries in rodents.

Table 4: Absorbance reading of b-mycrene at 230 nm.

Abs reading 1 2 3 Average
Sample 1 1.0977 1.1721 1.1721 1.1473

Sample 2 0.5860 0.6582 0.5850 0.6097

Figure 6: Beta mycrene at 230 nm

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From table 4, At wavelength 230, the absorbance of beta myrcene in Sample 1
was detected at higher absorbance than Sample 2. Therefore, weknow that Sample 1 which
is orange peel oil have more mycrene content than Sample 2 which is diluted with carrier oil.

5. CONCLUSION

From this project, we can conclude that carotenoid, limonene and beta mycrene can be
detected in both sample. As for the result, the pure orange peel oil gives better results than
the orange oil which is diluted with coconut oil. Even the results for limonene is more
detected at sample 2 but the different is not too much. In the other hand the pure orange oil
has more beta mycrene content which can be as anti-stress and joyful agent. The smell of
orange is also strong for orange oil sample. For the result of our product, we can say that the
product has a lot of benefits and is safe for daily usage. As it contains carotenoid and
limonene that are good for hair care and health. It also has good smell and beta mycrene
which can give a joyful feeling and anti-stress agent. Forfurther study, it couldbebetterifthe
analysis can be done using HPLC to detect all the substance in the orange peel oil. It is also
important in next study to get the user feedback.

REFERENCES

Engineering, F. (2017). Citrus peel essential oils: a review on composition and antimicrobial
activities, 9(5), 38–44.

Giwa, S. O., Muhammad, M., & Giwa, A. (2018). Utilizing Orange Peels for Essential Oil
Production, 13(1), 17–27.

Hati, D., Bhatnagar, S. P., & Sethi, K. K. (2013). Development and Evaluation of Polyherbal
Antidandruff Hair Oil. Pharmacognosy Journal, 2(10), 328–334.

Saidani Maufida, Brahim Mauzuq (2003). Biochemical characterization of blood orange,
sweet orange, lemon, bergamot and bitter orange. Elsevier Science Ltd, 1283-
1289.

Abdul-Majeed, B.A., Hassan, A.A. and Kurji, B.M. (2013) Extraction of Oil from Eucalyptus
Camadulensis Using Water Distillation Method. Iraqi Journal of Chemical and
Petroleum Engineering, 14(2), 7-12.

Eleonora Borello and Valentina Domenici (2019), Determination of Pigments in Virgin and
Extra- Virgin Olive Oils: A Comparison between Two Near UV-Vis Spectroscopic
Techniques, Journal of Foods, MDPI.

Lide, D.R. (ed.). (1993). CRC Handbook of Chemistry and Physics. 73rd ed. Boca Raton, FL:
CRC Press Inc., 1992-1993., p. 3-308

Yeung Chiu Dennis Tang (2018), Nano and Advanced Materials Institute
Victor Talrose, Eugeny B. Stern, Antonina A. Goncharova, Natalia A. Messineva, Natalia V.

Trusova, MargaritaV.Efimkina. (2018). NIST Chemistry WebBook, SRD 69, U.S. Secretary of
Commerce on behalf of the United States of America
Piérard-Franchimont, C., Xhauflaire-Uhoda, E., & Piérard, G. E. (2006). Revisiting dandruff.
International Journal of Cosmetic Science, 28(5),311–318.
Tshilenge, J. (2009). Dissertation-Distillation, (February).
Wikandari, R., Nguyen, H., Millati, R., Niklasson, C.,& Taherzadeh, M. J. (2015). Improvement of
biogas production from orange peel waste by leaching of limonene. BioMed
Research International, 2015.

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CHARACTERIZATION OF LEMONGRASS (Cymbopogon citratus)
OIL

Nor Aziyan binti Mohd Nasir, Dewi Nilam Rupa’a binti Razali
Politeknik Tun Syed Nasir Syed Ismail, Hab Pendidikan Tinggi Pagoh, KM 1, Jalan Panchor, 84600

Pagoh, Muar, Johor
*[email protected]

Abstract. Cymbopogon citratus or lemongrass is a plant that is widely grown in the region of
India, South Africa, America and Southeast Asia. In this study, the characterization of
lemongrass oil that has been carried out includes the determination of moisture content,
iodine value and acid value. Lemongrass oil is first extracted by Soxhlet extraction method.
The moisture content determination is carried out by oven drying method. The measurement
of Iodine value of the lemongrass oil is determined by the Wijs method while the acid value
is determined by neutralization of potassium hydroxide. The average data of moisture
content of the lemongrass oil is 21.805%. The average number of iodine value in g is 13.215%
while the average number of the acid value is 8.470%. The characterization study is
important to regulate the handling and storage of the lemongrass oil.

Keywords: lemongrass oil, moisture content, Iodine value, acid value

1. INTRODUCTION

Cymbopogon citratus or lemongrass are a plant that widely grown mostly in the region of
India, Indonesia, South Africa, America, Madagascar and many parts of Asia. This plant is
described to have a 3-meter height with the rootlets that connect with the soil at the
bottom. Lemongrass have known in many names; different in other country. The name
lemongrass came from the lemon-like odour that is present from the essential oil odour itself.

Lemongrass is often extracted to be an essential oil. The essential oil is extracted by
stim-distilling the lemongrass biomass. Lemongrass essential oil has a lot of applications
especially in food and pharmaceutical industries, perfumery and cosmetics, and eco-
friendly pesticides (Zheljazkov et al., 2011). It also has antibacterial and antifungal properties
(Zheljazkov et al., 2011). Essential oil is a concentrated hydrophobic liquid extracted from
plants that has a characteristic akin to other vegetable oil, and is high in volatile compound
that gives lemongrass its characteristic smell. However, there is very little research
investigating the characteristics of the triglyceride form of lemongrass oil.

In this research, the lemongrass is extracted to produce the crude lemongrass oil,
and several characterization experiments has been carried out.

2. RESEARCH BACKGROUND

Cymbopogon citratus is the scientific name for lemongrass. Lemongrass oil is high in
phytochemical compounds such as geranial, neral and myrcene (Mohamed Hanaa et al.,
2012). In addition to that, the essential oil of lemongrass consists of small quantities of
geraniol, geranyl acetate and monoterpene olefins, such as limonene (Mohamed Hanaa
et al., 2012). This contributes to the smell, texture, and appearance of the oil. Additionally,
lemongrass essential oil is used as a raw material for the production of ionone, vitamin A and

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β-carotene because of its high content of citral (constituted by the isomers neral and
geranial) (Nur Ain et al., 2012).

Lemongrass essential oil is widely used in perfumery industry since it blends well with
a great variety of essential oil (Nur Ain et al., 2012). Other than that, it is used as a scent in
cleaning agents and also widely used as insect repellents especially mosquitoes and even
as aromatherapy. Other uses of lemongrass oil as folk medicine for several treatments;
usually of nervous and gastrointestinal disturbances, and as antispasmodic, analgesic, anti-
inflammatory, anti-pyretic, diuretic and sedative (Mohamed Hanaa et al., 2012).

There is not many information that is available about the characteristics of the
triglyceride form of lemongrass oil. Lemongrass oil is first extracted by Soxhlet extraction
method and then is proceeded to be characterized by conducting several experiments to
determine the moisture content, iodine value and acid value. Mostly, lemongrass is being
extracted to produce its essential oil. In this research, the data provided is important to
determine the quality parameters of the lemongrass oil which help regulate the handling
and storing of the oil. This is because every oil type need to have a specific storage and
handling procedure to prevent oil deterioration that can lower the quality of oil.

3. METHODOLOGY

3.1 Sample preparation
The fresh lemongrass stalks are collected from a farm in Taman Kelemak Jaya, Alor Gajah,
Melaka in June 2019. Firstly, the lemongrass stalks are washed well to remove the solid
particles like soil. The lemongrass stalks then were cut into small pieces to provide a greater
surface area. Then it is oven-dried for 12 hours 30 minutes at 45 °C.

3.2 Oil extraction
Lemongrass oil is extracted out of the dried lemongrass sample by Soxhlet extraction
method. The Soxhlet extraction was held for 12 hours 30 minutes at boiling temperature of
methanol which is 64 °C. The oil is then separated from the solvent by rotary evaporation.
Then petroleum ether is added to the gathered oil and is further mixed by vortex mixer. The
oil is separated again using separating funnel.

3.3 Moisture content
1 g of the crude lemongrass oil sample was weighed on a watch glass using the weighing
scale. Then, it was oven dried at 45℃ for 6 hours. After 6 hours, the oil was weighed again to
obtain the final weight after drying.

3.4 Iodine value determination
0.5 g of crude lemongrass oil is weighed in a conical flask. Then, 20 ml of cyclohexane and
25 ml of the Wij’s reagent were added to the oil sample. It was shaken gently and been
placed in the dark for 30 minutes at room temperature. After standing, 20 ml of potassium
iodide solution and 100 ml of distilled water were added. The sample was titrated with
sodium thiosulphate solution until the yellow colour due to iodine has almost disappeared.
Then, 1 ml of starch indicator solution is added and titrated until the blue colour just
disappears after very vigorous shaking. A blank test is carried out simultaneously under the
same condition.

3.5 Acid value determination
The analysis was carried out by weighing 5 g of crude lemongrass oil sample into a 250 ml
conical flask. Then, 50 ml of ethanol was added and the mixture was heated for 6 minutes
(until it started to boil) by using the hot plate. After that, 2 drops of phenolphthalein indicator
were added to the solution. It was then titrated against KOH solution. The appearance of

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the pink colour indicates the end point. The final volume of the KOH in the burette was
recorded.

4. RESULTS AND DISCUSSIONS

Table 1 presents the value of moisture content of lemongrass oil.

Table 1: Moisture Content of Lemongrass Oil

Sample Final weight (g) Initial weight (g) Moisture
content (%)
1 0.795 1.016
2 0.790 1.011 21.750
3 0.792 0.964
21.860
21.805

The moisture content of the lemongrass before extract been calculated by using
this formula:

Moisture content (%) = Initial weight - final weight × 100
Initial weight

The average moisture content of the lemongrass oil is high which is 21.805%. The
higher the moisture content, the lower the shelf stability of the lemongrass oil. Moisture is also
undesirable because it will support microbial growth and facilitate lipid hydrolysis. This may
be due to the samples that was not been dried completely during drying. Other parameters
such as iodine value and acid value will also be affected by the high moisture content. High
moisture content will lead to hydrolysis process due to the presence of water and lipase
enzyme in the oil. This hydrolysis process will produce free fatty acid which will degrade the
oil quality. Table 2 presents the iodine value of lemongrass oil.

Table 2: Iodine Value of Lemongrass Oil

Sample Weight of Vb (ml) Vs (ml) N Iodine

sample (g) value

1 0.516 7.40 5.10 0.1 5.656

2 0.535 7.40 2.60 0.1 11.385

3 0.523 7.40 1.20 0.1 15.044

The average iodine value of the sample 2 and 3 of lemongrass oil is 13.215 (gI2/100g)
The iodine value was calculated by the formula below:

Iodine value = 12.69× N× (Vb - Vs)
W

Vb = volume (ml) of sodium thiosulphate used for blank
Vs = volume (ml) of sodium thiosulphate used for the determination
N = normality of sodium thiosulphate solution used
W = weight of sample

Table 2 showed the result of iodine value of lemongrass oil. An average of 13.215
(gI2/100g) iodine value was observed which is considered as the lowest value among
various vegetable oils (Chebet et al., 2016). To get a precise average value, only sample 2

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and 3 are considered for the calculation since the value of sample 1 is far more different
than that of the sample 2 and 3.

Iodine value is an important analytic method characteristic of the oil since it
measures the degree of unsaturation or the double bond of fatty acid that can react with
the iodine. It is also important because the oxidative stability and the melting points of the
oil are related to the degree of unsaturation. So that, the iodine value provides some
estimation of these quality factors. Since iodine value of the lemongrass oil is low, it has low
susceptibility to oxidation. Table 3 presents the acid value of lemongrass oil.

Table 3: Acid Value of Lemongrass Oil

Sample Weight of Volume of Normality Acid value
sample(g) KOH (ml) (N) (mg KOH/g)
1 0.2
2 5.010 2.50 0.2 5.600
3 5.070 3.70 0.2 8.188
5.000 3.90 8.752

The acid value was calculated using the formula below:

Acid value = (Volume of KOH × Normality of KOH × 56.1)
Weight of oil sample

The average acid value of the lemongrass oil is 8.470 mg KOH/g which is considered
as high compared to other vegetable oils (O’Brien, 2008). In this analysis, titration method
was used.

The acid value is a common parameter in the specification of oils. The acid value
measures the free fatty acid (FFA) in the lemongrass oil which indicate the hydrolysis of
triglyceride. Thus, according to literature review the acid value increases when the moisture
content of the oil increases. The reaction process of oxidation and hydrolysis in the oil will
reduce the amount of unsaturated fatty acid which led to an increase in the acid value. In
this research the acid value and the moisture content of the lemongrass oil are both high.
In this study, higher acid value may have resulted from the high value of moisture content,
since high moisture content leads to hydrolysis process in the lemongrass oil. Acid value or
FFA can be used to measure hydrolytic rancidity if the fatty acids are volatile.

5. CONCLUSION

In this study, three characterization analysis has been conducted on lemongrass oil, which
is moisture content, iodine value and acid value. The average moisture content value of
lemongrass oil is 21.805%. The average number of iodine value in g is 13.215% while the
average number of the acid value is 8.470%. The data obtained from this characterization
study is important to determine the quality parameters of the lemongrass oil which help
regulate the handling and storing of the oil.

REFERENCES

Chebet, J., Kinyanjui, T., Cheplogoi, P. K. (2016). Impact of frying on iodine value of
vegetable oils before and after deep frying in different types of food in Kenya. Journal
of Scientific and Innovative Research 2016; 5(5): 193-196

O’Brien, R. D. (2008). Fats and Oils Formulating and Processing for Application. CRC Press
LLC.

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Mohamed Hanaa, A. R., Sallam, Y. I., El-Leithy, A. S., Safaa E. Aly. (2012). Lemongrass

(Cymbopogon citratus) essential oil as affected by drying methods. Annals of
Agrcultural Science, 57(2), 113–116.
Nur Ain, A. H., Zaibunnisa, A. H., Halimahton Zahrah, M. S. and Norashikin, S. (2013). An
Experimental Design Approach for The Extraction of Lemongrass (Cymbopogon
Citratus) Oleoresin Using Pressurised Liquid Extraction (PLE). International Food
Research Journal 20(1): 451-455.
Zheljazkov, V. D., Cantrell C. L., Astatkie, T., Cannon, J.B. (2011). Lemongrass Productivity, Oil
Content, and Composition as a Function of Nitrogen, Sulfur, and Harvest Time.
Agronomy Journal 103:805–812.

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TRANSESTERIFICATION ON WASTE COOKING OIL AS
SOIL AMENDMENT

Rahizana bt Mohd Ibrahim, Kamal Firdausi bin Abd Razak, Fazliana bt Abdullah

Politeknik Tun Syed Nasir Syeb Ismail, Hab Pendidikan Tinggi Pagoh,
KM 1 Jalan Panchor,84600 Panchor Johor, Malaysia
*[email protected]

Abstrak. Nowadays, the waste cooking oil (WCO) was collected, processed and converted
into a biodiesel product. However, the biodiesel was produced about 10% (w/w) of crude
glycerol and become a waste product insignificant economic value. This research reported
the addition of crude glycerol into the soil could increase the moisture content of the soil
due to its hygroscopic properties. Firstly, the waste cooking oil collected base on the color
of the oil which depends on how many it was use. Black oil is CG4, dark brown oil is CG3,
light brown oil is CG2 and yellow oil is CG1. Pre-treatment is done to the oil by filtering its
impurities, heated with ginger to remove the bad odor and filtered with activated carbon
to reduce the amount of free fatty acids in the WCO. Then, the transesterification process is
done using Sodium Hydroxide dissolved in ethanol as catalyst to produce glycerin. The yield,
density, viscosity and pH value of the crude glycerol was measured. FTIR and UV-Vis tests
are also done to identify the functional group and absorbance of the crude glycerin (CG)
produced to analysis the impurity content. From the analysis, CG4 have showed the highest
yield of crude glycerin (90%) with the density at 1.15g/ml and 12.52 of pH value that suitable
for soil amendment. The CG was added to soil to analyses the effect of CG to the soil. It has
shows that the CG has increased the moisture content of the soil up to 5% for 5 days.

Kata Kunci: Waste cooking oil, crude glycerol, soil

1. INTRODUCTION

Biodiesel (fatty acid methyl esters) was created using transesterification method from
vegetable oils has been studied in recent literature reviews (Gashaw et al. 2014). However
due to higher cost and limitated sources using the fresh vegetable oil, the feedstock’s are
always a critical issues. Therefore, waste cooking oil (WCO) become the most favorable
feed stock for biodiesel production since the price is 2–3 times cheaper than the fresh
vegetable oils. However. the fast development of the biodiesel industry produced 10%(w/w)
of glycerol as a by product. The number of excess glycerol has generated an environmental
concerns associated with contaminated glycerol for disposal. To date, most of the crude
glycerol is burned and discharged the carbon to the atmosphere. Subsequently, treatment
of this low-value substance to higher added-value by the implies of chemical or
biotechnological is much interested. However, the increasing the purity of glycerol is
required higher cost, especially for intermediate and small biodiesel production. Recently,
there is a number of research has been studied to improve the economic feasibility of the
crude glycerol (Vassilev et al. 2016 ). Conceivable outcomes such as combustion,
composting, animal feed, thermochemical conversion, and biological conversion have
been applied to the crude glycerol processing. Chen et al. (2018) reported that value of
crude glycerol from any feedstock is generally between 60 and 70% (wt) glycerol. Chai et
al. (2015) reported that waste vegetable oil from waste cooking oil had a higher level (76.6
%) of glycerol than the soy oil feedstock (67.8%). The crude glycerol contained a variety of
elements originating from the primary oil, such as K (0 to 217 ppm), P (12 to 37 ppm), S (14
to 128 ppm), Na (1.06 to 1.40), C (24 to 37%), N (0.04 to 0.12%), and protein (0.05 to
0.44%)(Nanda et al. 2014). Direct application of crude glycerol in the soil has been
investigated in Western Australia by Bonnardeaux (2006). Primary results from the research

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showed that the crude glycerol could offer assistance rectify water repellency of the non-
wetting sandy soils by covering the non-wetting coatings on soil particles without including
a critical fetched. To date, the information on the effects of direct application of crude
glycerol to soils is still limited. The objective of this paper is to study the impact of addition of
glycerol on plant growth and soil characteristics for Centella Asiatica plant.

2. SCOPE OF RESEARCH

This research aims to study the potential of crude glycerol from biodiesel by-products as a
soil amendment to enhance the agricultural field. In this research, waste palm cooking oil
from various restaurants at Taman Pagoh Jaya and Polytechnic canteen have been used
as raw material to produce biodiesel and crude glycerol. Pre-treatment, transesterification,
and neutralization process have been choosing as a method to produce biodiesel and
crude glycerol. The result would be compared with the standard data of biodiesel and
crude glycerol for verification before being used as a soil amendment.

3. METHODOLOGY

3.1 Pre-Treatment process
Variety sources of waste cooking oil (WCO) was collected from Taman Pagoh Jaya area,
restaurants and cafeteria PTSN. Then, the samples were graded according to the physical
properties (color) such as CG1 for yellow, CG2 for light brown, CG3 for dark brown and CG4
for black color. The WCO was strained with cloth filter for the several times (2-3) to remove
coarse particles, then was to heat at 50°C with ginger slice to remove odor from the oil. Then
the WCO was continue filtered for the second time by using an activated carbon to absorb
any impurities from the waste cooking oil (WCO).

3.2 Titration process
Titration process is to determine the concentration of acid in waste cooking oil (WCO) using
a reference solution. About 10ml of sample for each WCO was mixed with 100ml of isopropyl
alcohol. Then, 10 drops of phenolphthalein indicator were added into the samples and
isopropyl alcohol solution. About 0.10%(w/v) NaOH solution was prepared and was added
slowly to the solution until the solution change from colourless to pink color. The volume of
0.10% NaOH solution was recorded. Those steps were repeated for three times to get the
average amount of NaOH needed.

3.3 Transesterification process
All the sample of waste cooking oil which were collected at different sources have been
graded by CG1, CG2, CG3 and CG4 according to the physical properties. The molar ratio
of Ethanol: WCO used is 4:1, 5:1, 6:1 and 7:1. Then, the process is carried out by adding
NaOH. The mass of NaOH pallet used was measured and put it into ethanol solution. The
ethanol solution and NaOH pallet was mixed using a magnetic stirred bar at 150 rpm. After
the NaOH pallet and ethanol solution was mixed, the WCO was heated to 45°C. Once the
WCO achieved at the setting temperature, the mixed ethanol and NaOH solution was
combined with the WCO solution. The WCO and mixed solution was stirred and heated for
30 minutes then the mixed solution was put in a room temperature for natural separation
process. The solution would be separated into two layers, the top layer was biodiesel and
the bottom layer is crude glycerol. The steps were repeated for the following sample; CG2,
CG3 and CG4, respectively.

3.4 Neutralization process
About 5ml of crude glycerol samples of each CG1, CG2, CG3 and CG4 were prepared in
the test tubes. Each sample was titrated with 0.1M hydrochloric acid (HCl and was

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monitored by pH meter until it reached pH 7 using Addison method (Addison et al. 2006).
The reading of the pH shown in the pH meter was recorded.

3.5 Soil Amendment
The soil amendment was tested using 200g soil and 10g of organic fertilizer to the growth of
Centella Asiatica plant. Each pot was varied by 2g of different type of crude glycerol
produced with ratio of 7:1 (1.0% of the soil). Crude glycerol with ratio 7:1 was chosen
because it produced the most yield and has the highest purity among all ratio. The first pot
did not contain any glycerol for experiment control data. Second pot contained of CG1,
third pot contained CG2, fourth pot contained CG3 while CG4 was poured into the fifth
pot. The pots were labelled accordingly. About 50mL of tap water was watered to the all
pots every morning for 5 days. The growth of the plants was observed every day while the
moisture content of soil was analysis.

4. RESULTS AND DISCUSSION

4.1 Crude glycerol Yield (%) Analyses
Four experiments were carried out by varied the molar ratio of ethanol/waste cooking oil
between 4:1 and 7:1. Table 1 has shown the percentage of yield of crude glycerol (CG) with
the ratio of ethanol to waste cooking oil. From the table, the percentage of yield showns
the highest value from CG1 to CG4 which are 76.67%. 83%, 87%, and 90%. The percentage
of yield increased as the molar ratio increased, with the best results (90%) being for a molar
ratio 7:1 for CG4. This results indicated that the excess of alcohol could improve the solubility
of glycerol, a reaction product that physically blocks the entrance of catalyst pores.
Talebian et al. (2013) in their research also showed that high ethanol/waste cooking oil
molar ratios would cause lipases deactivation. This is because proteins are usually unstable
in the presence of short-chain alcohols, such as ethanol and methanol. Thus, an excess of
alcohol causes an opposite effects on the lipase reaction thus decreased biocatalyst
stability and increased the formation of a homogeneous suspension of the reactants and
biocatalyst. In consequence, the alcohol/oil molar ratio is one of the most important
variables affecting the esters yield, and although the stoichiometric ratio for
transesterification required three moles of alcohol and one mole of triglyceride, an excess
of alcohol is used in practice. Hence, the alcohol molar/oil ratio was a crucial variable.

Table 1: Percentage of yield against ratio of ethanol to WCO

Ratio Yield %

(Ethanol:WCO) CG1 CG2 CG3 CG4

4:1 43.00 60.00 73.33 76.67

5:1 55.00 70.00 80.00 83.00

6:1 66.30 70.00 83.67 87.00

7:1 75.00 76.67 89.33 90.00

4.2 Physical properties
The parameter of physical properties of crude glycerol had been observed for the value of
pH and density. Table 2 shows the result of the pH value for crude glycerol to the ratio of
ethanol and waste cooking oil. From the table, the pH value for all samples have been
found alkaline with the value range from 9 to 13. These results were closed to the crude
glycerol standard data as obtained from Wan Nor Roslam et al. (2015).

Table 2: PH value against ratio of ethanol to WCO

Ratio pH
(Ethanol:WCO)
CG1 CG2 CG3 CG4

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4:1 10.70 12.32 9.84 11.84

5:1 12.31 9.72 9.50 10.64

6:1 10.04 11.39 11.71 12.03

7:1 12.18 12.06 10.12 12.52

Figure 1 was plotted in line graph of the density of crude glycerol. The density of the
glycerol samples was determined between 1.08 to highest value of 1.147 g/ml. This result
showed that this parameter was varied with the level of impurities in the sample. The
impurities were included alcohol, water, fatty acids, catalyst residues, and other
compounds originating from the oil used in the sample of WCO. The results presented in
Figure 1 showed an increasing in the density of the crude glycerol with the increased in the
ratio of ethanol to waste cooking oil. The increasing in density is most pronounced while
comparing the distilled and crude glycerol samples in the initial purification steps. The crude
glycerol in ratio 7:1 of CG4 had the highest density of 1.147 g/mL while the lowest density of
crude glycerol obtained was ratio 4:1, CG1 which obtained the density of 1.083 g/mL

Figure 1 : Graph of density crude glycerol against ratio Ethanol:WCO

4.3 Viscosity
Table 3 shows the percentage of viscosity against ratio ethanol to waste cooking oil. The
crude glycerol was tested using Brookfield DV1 Digital Viscometer with constant spindle LV2-
62, speed at 100rpm, in a container of 10mL, at 25 C. Each sample was run in 5 minutes and
the data was recorded in the table and was plot in line graph. Based on Figure 2, ratio 7:1
of each CG give the highest value compared to the ratio 4:1. However, compared to each
CG in ratio 7:1, CG4 shows the highest percentage of 21.9%. This is because, CG4 of ratio
7:1 contain high impurities and component inside the solution which lead to high viscosity
compared to other CG in ratio 7:1. While in ratio 4:1, the lowest percentage is 5.5%, CG1 as
it consists of low impurities and less component inside the solution.

Table 3: Percentage of viscosity against ratio of ethanol to WCO

Ratio Viscosity(%)
(Ethanol:WCO)
CG1 CG2 CG3 CG4
4:1
5:1 5.50 10.50 14.70 20.30
6:1 7.70 12.20 16.40 20.80
11.80 13.20 17.90 21.60

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7:1 12.00 14.40 19.10 21.90

Figure 2: Graph of crude glycerol against ratio Ethanol:WCO

4.4 pH of Soil Amendment
The pH of soil amendment was around 5.0 to 6.5 while adding the crude glycerol. The pH of
soil without glycerol is 5.5 as a control sample. In this research, the Centella Asiatica was
selected since it has suitability to growth at the pH value within a range of 4 to 6.5. After 5
days, the pH value was dropped and became more acidic. From the table, the plant that
using CG4 is more lush than other plant using different samples of CG.

Table 4 : pH of CG1, CG2, CG3 and CG4

Glycerol pH

Constant Initial Final
CG1
CG2 5.5 5.5
CG3
CG4 5.0 4.0

5.9 4.9

6.0 6.0

6.5 5.0

4.5 Moisture content on soil amendment
Figure 3 was plotted to compare the percentage of moisture content of the soil amendment
while all the resulted are tabulated in table 5. This experiment was carried out to investigate
the effect of crude glycerol in the ecosystem by monitoring the moisture content of the soil
and observed the condition of the plant within 5 days. Container A is a sample of plant
without crude glycerol and B samples with crude glycerol. The result shows that the moisture
content in B with crude glycerin was increased as compared to the container A without
inserted the crude glycerin. This is because crude glycerin has hygroscopic properties which
means it attracts and holds water molecules from the surrounding environment. This could
have made the water unavailable to the plants and demonstrating a clear maximum
concentration in this non-nutrient-limited soil. In nutrient limited soils such as found in the
humid subtropical region of Northwest Arkansas, more pronounced inhibition effects are
seen as demonstrated in the runoff testing and in studies by Schoenau et al. (2009) and Qian
et al. (2011) due to nitrogen tie-up. Hence, this result led to the survival of the plants and
animals.

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Figure 3: Percentage of Moisture Content of Soil Amendment

Table 5: Moisture content of soil for 5 days

Soil Container Moisture content (%)

Day 1 Day 2 Day 3 Day 4 Day 5
55.68
Without crude glycerin A 19.09 30.45 47.76 50.99 58.62

With crude glycerin B 19.15 33.67 51.50 55.35

5. CONCLUSION

This primary research could be a pioneer research for the usage of crude glycerol as a
benefit by product. The results showed that crude glycerol may have a huge benefit
towards the soil and also to the growth of the plants. The crude glycerol as a soil
amendment, thus could enhanced the properties of the soil by retaining the moisture
content of the soil. This research has been proven that the utilization of glycerol should be
investigated since the biodiesel generation has demonstrated to be a decent option in
contrast to oil-based fuel.

REFERENCE

Addison, K. (2006). Separating Glycerine/FFAs. Homemades Project, Journey to Forever.

Alemayehu Gashaw, Abile Teshita (2014). Production of biodiesel from waste cooking oil
and factors affecting its formation: A review. International Journal of Renewable and
Sustainable Energy; 3(5): 92-98.

Amin Talebian-Kiakalaieh, Nor Aishah Saidina Amin, Hossein Mazaheri. (2013). A review on
novel processes of biodiesel production from waste cooking oil. Journal of Applied
Energy 104 (2013) 683–710.

Bonnardeaux, J. (2006). Agricultural Uses of Glycerine, in page 12 of Glycerine Overview,
Department of Agriculture and Food, Government of Western Australia.

Jiaxin Chen, Song Yan, Xiaolei Zhang, Rajeshwar Dayal Tyagi, Rao Y. Surampalli, J.R. Valéro
.(2018). Chemical and biological conversion of crude glycerol derived from waste
cooking oil to biodiesel. Journal of Waste Management; 71: 164-175,

Nanda MR, Yuan Z, Qin W, Poirier MA and Chunbao X. (2014). Purification of Crude Glycerol
using Acidification: Effects of Acid Types and Product Characterization. Austin Journal
of Chemical Engineering.

Nikolay Vassilev, Eligio Malus Antonia Reyes Requena, Vanessa Martos, AnaLópez, Ivana
Maksimovic, MariaVassilev. (2016). Potential application of glycerol in the production
of plant beneficial microorganisms. Journal of Industrial Microbiology and
Biotechnology.

Peiyuan Qian, Jeff Schoenau, and Ron Urton. (2011) . Effect of soil amendment with thin

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stillage and glycerol on plant growth and soil properties. Journal of Plant Nutrition,
34:2206–2221.
Schoenau J, Qian P, Alotaibi K, Urton R. (2009) Potential for using biofuel, crop and animal
processing by-products as soil amendments to increase fertility: effect of soil
amendment with dehydrated alfalfa, dry and wet distillers grains, thin stillage and
glycerol on growth of canola, soil properties, and microbial activity final report,
Department of Soil Science, College of Agriculture, University of Saskatchewan,
Saskatoon, Saskatchewan, Canada.
Wan Nor Roslam Wan Isahak, Zatil Amali Che Ramli, Manal Ismail, Jamaliah Mohd
Jahim,Mohd Ambar Yarmo (2015). Recovery and Purification of Crude Glycerol
fromVegetable Oil Transesterification. Separation & Purification Reviews, 44: 250–267.
Zi-Zhe Cai, Yong Wan, Ying-LaiTen, Ka-Man Chong, Jia-WeiWang, Jie-Wen Zhang, De-Po
Yang (2015). A two-step biodiesel production process from waste cooking oil
viarecycling crude glycerol esterification catalyzed by alkali catalys. Fuel Processing
Technology; 137: 186-193.

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ANTIMICROBIAL PROPERTIES OF SELECTED LOCAL FRUITS AND
SPICES AGAINST ORAL BACTERIA

Nurul Shafiqah Binti Sulaiman , Munirah Mihat, Nur ‘Assyakirin Mohamed Paid
Politeknik Nilai, Kompleks Pendidikan Nilai, Bandar Enstek, 71760 Labu, Negeri Sembilan

*[email protected]

Abstract. Oral bacteria are bacteria commonly found in the mouth. Oral bacteria contributed
to become a major health issue worldwide; which has led to many diseases that included oral
and pharyngeal cancers, oral soft and hard tissue lesions, orofacial disorders, tooth loss, dental
caries and periodontal diseases. Periodontitis is an infectious disease of microbial origin with
over 700 different known strains of bacteria detected in the human mouth including the most
common strains; Staphylococcus and Streptococcus namely Porphyromonas gingivalis,
Prevotella intermedia, Bacteroides forsythus, Campylobacter rectus, Fusobacterium
nucleatum and Aggregatibacter actinomycetemcomitans. These microorganisms play crucial
role in pathogenesis of periodontal disease. Successful periodontal therapy is aimed at
eliminating these pathogenic organisms. Despite the abundance of oral hygiene and
healthcare products offered in the market, caries and cavities cases are still increasing in
children. An oral mouthwash made from natural derivatives will be less harmful for children to
swallow. This project investigated the feasibility of using plant and spices extracts namely Piper
betle(Betel), Murraya koenigii (Curry leaves), Illicium verum (Star anise), Areca catechu(Areca),
Zingiber officinale (Ginger flower), Syzygium aromaticum (Clove), Averrhoa carambola
(Starfruit), Hylocereus undatus (Dragon fruit, Cucumis sativus (Cucumber), Citrofortunella
microcarpa (Calamansi) and Annona muricata (Soursop) as raw materials for mouthwash by
investigating their antimicrobial properties towards oral bacteria. The extracts were tested
against samples of oral bacteria obtained from students of Taska Politeknik Nilai. From the data
obtained, Star anise, Curry leaves, Bilimbi, Calamansi, and Starfruit exhibited the highest
antimicrobial properties against the oral bacteria with different types of solvents used for the
extracts of these plants and spices have different antibacterial properties, due to the different
active compounds extracted.

Keywords: Oral Bacteria, Antimicrobial, Herbs, Fruits.

1. INTRODUCTION

Oral health is important to ensure a higher quality of life. Good oral health is a condition free
of mouth and facial pain, oral and throat cancer, oral infection and sores, periodontal disease,
tooth decay, tooth loss, and other diseases and disorders that limit the ability of a person to
bite, chew, smile, speak, and psychosocial well-being. There are two main dental caries that
may affect oral health known as tooth decay and periodontal the gum disease. Dental caries
happens when tooth surfaces were demineralized by bacterial processes that produced
cavities or holes in the teeth (Gilsenan-O'Neill, 2015). Bacterial infection produced biofilm on
the surfaces of gum and tooth and the immune system cause inflammatory responses to reacts
against the infection. These responses may damage the tissues and bones that surround and
anchor teeth, which when persisted leads to periodontal gum disease. It has been reported
that the numbers of people who continue to suffer high levels of tooth decay has been
increasing (Gilsenan-O'Neill, 2015). There are a range of other serious oral disorders in addition
to dental caries and periodontal disease that affect oral health and well-being, including oral
cancer, dental trauma, teeth fracture due to injury, tooth wear, dental decay, attrition and

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abrasion, dry mouth, mouth ulcers, cold sores, tooth sensitivity and halitosis (Gilsenan-O'Neill,
2015).

Since 1984, oral health staff have performed oral health programmes for preschool
children. These programmes concentrate on the promotion and prevention of oral health with
the goal of increasing awareness of oral health at an early age and exposing children to oral
health personnel and oral healthcare in a comfortable and familiar environment. In Malaysia,
dental epidemiological surveys have shown a high prevalence of caries among 5 to 6-year-
olds conducted in Malaysia, despite a decreasing trend over the last three decades. In
Peninsular Malaysia in 1970, the proportion of 6-year-olds with one or more carious teeth in
deciduous dentition was 95.4 percent (Malaysia, 1970). This decreased to 88.6% in 1988 and by
1997, a further decline to 80.6% was noted (Malaysia M. o., 1997). In a comparable Sarawak
report in 1982, the prevalence of caries in the same age group was 91.7 percent. By 1994, the
incidence of caries had decreased to 88.2 percent, and three years later this further decreased
to 79.6 percent (Malaysia M. o., Dental epidemiological survey of school children in Sarawak ,
1994). The prevalence of caries among 5-year-old children in Sarawak was found to be 85.8
percent in a regional pre-school survey in 1995, with 49.6 percent deemed high risk of 5 or more
carious teeth (Malaysia M. o., 1995). Just 10.7% of damaged teeth have been recovered,
indicating the extremely unmet needs of care seen elsewhere in Malaysia. Sabah reported the
highest prevalence of caries among 6-year-olds among the regions at 96.9 percent in 1985
and 94.7 percent in 1997. This suggests a very slight decline over the 12-year period of 2.2 per
cent (Malaysia M. o., Dental epidemiological survey of school children in Sabah , 1985).
Despite the decrease in the incidence of caries; despite the continuous efforts of the Ministry
of Health Malaysia, the disease is still prevalent in Malaysia. This pattern is observed not only in
Malaysia, but also globally.

In America, approximately 21 percent of children have untreated dental caries (tooth
decay) in their primary teeth. Non-Hispanic black and Mexican-American children (27 percent
and 31 percent, respectively) have a higher prevalence of untreated tooth decay than non-
Hispanic white children (18 percent) (Eugenio D. Beltrán-Aguilar, 2002). Children and
adolescents from families with low incomes are more than twice as likely to have untreated
tooth decay in their permanent teeth as those from families with higher incomes (Griffin SO1 &
CDC Dental Sealant Systematic Review Work Group, 2008). A variety of population and
individual methods, such as community water fluoridation, topical fluorides (e.g., fluoride
toothpaste, fluoride varnish, and fluoride mouth rinse), dental sealants, education, and dietary
approaches, include the prevention of tooth decay in children and adolescents (Tomar SL,
2010). The statistical analysis shown in both countries showed that despite the awareness and
the increased income, incidence of cases is still high. This may be due to the diet and lifestyles
that may contribute to poor oral hygiene.

The need for oral health care is the most prevalent unmet health care need among
children and adolescents. (Newacheck PW, 2000). Oral health care is the most prevalent
unmet health care need for children and adolescents with special health care needs,
compared with all other health care services. Unmet needs for oral health services impact
almost twice the number of children and adolescents as unmet needs for mental health care.
According to parents, children and adolescents with special health care needs without
insurance and from families with low incomes are more likely not to receive the health care
services they need. Those from families living below the federal poverty level (FPL) are three
times more likely not to receive the services they need as their counterparts from families with
incomes of 400 percent FPL or more (25 percent vs. 7 percent) (Rockville, 2008). For children
and adolescents with special health care needs, more than half of dental schools provide
students with fewer than 5 hours of classroom instruction and less than 5 percent of clinical time.
Hands-on dental school education experiences have a profound effect on the attitudes of
dentists regarding barriers to treatment for children and adolescents with special health care
needs (Romer M, 1999). This indicated that more consideration should be given to the children
as well as adolescents with special needs.

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Treating oral diseases is costly (Petersen PE, 2005), even in high income countries where
5– 10% of public health spending is used for dental care (Baelum V, 2007). While no comparable
data are available for low-income countries, it has been estimated that the treatment of
childhood caries would cost between US$ 1618 and 3513 per 1000 mixed-age children
between 6 and 18 years of age, a sum that exceeds the funding available for the provision of
an essential public health care package for children in most low-income countries (Yee R,
2002). Despite the abundance of oral hygiene and healthcare products offered in the market,
caries and cavities cases are still increasingly reported. This is possibly because children dental
care is hard to be taken care of, as they tend to resent the brushing and flossing required twice
daily. Aside from that, parents tend to feel uneasy when young children incline to swallow their
toothpaste when brushing. Thus, specially made products for children’s teeth, which is safe for
them to consume is a necessity to curb the increasing problem of children’s teeth health
problem.

An oral mouthwash made from natural derivatives will be less harmful for children to

swallow. Thus, this study investigated the feasibility of the extracts of some plants and spices

that is normally used in folklore to combat toothaches namely Piper betle(Betel), Murraya

koenigii (Curry leaves), Illicium verum (Star anise), Areca catechu(Areca), Zingiber officinale

(Ginger flower), Syzygium aromaticum (Clove), Averrhoa carambola (Starfruit),

Hylocereus undatus (Dragon fruit, Cucumis sativus (Cucumber), Citrofortunella microcarpa

(Calamansi) and Annona muricata (Soursop) to be used as raw materials for oral mouthwash.

Some of these plants has been reported to have antimicrobial properties namely clove, bilimbi,

and calamansi, however a study of antibacterial properties against wide ranges of oral

bacteria has not been done. The extracts of these plants were tested against a wide range of

common oral bacterial strains to evaluate their potential as the main ingredients in the oral

mouthwash.

2. LITERATURE REVIEW

Oral bacteria have developed mechanisms for sensing their environment and avoiding the
host or altering it. Both the tooth surface and gingival epithelium are ecologically influenced
by bacteria. However, bacterial colonization is continuously controlled by a highly efficient
innate host defense mechanism and bacterial invasion of local tissues is prevented. There is a
dynamic equilibrium between bacteria from the dental plaque and the innate host defense
system. These diseases are among the most important oral health problems. These included
oral and pharyngeal cancers and oral soft and hard tissue lesions, or facial disorders, tooth loss,
dental cavities and periodontal diseases.

World Human Organization (WHO), which states that worldwide about 60-90% of school
children have dental cavities, often leading to pain and discomfort. According to Oral Health
Divison (1998), studies shown that more than 70% of children of twelve years of age suffered
from dental decay. Then, dental cavities can be prevented by maintaining a constant low
level of fluoride in the oral cavity (Noguti, 2012).

Current oral products in the market do not really contribute to the healthcare of
toddlers. Oral health of toddlers should be heeded properly because it is easily exposed to
bacteria. With the production of ready to eat toddler food nowadays, the need has become
more imminent than before. Therefore, actions need to be carried out to control the number
of oral bacteria in toddlers so it could reduce the diseases related to the oral bacteria.
Currently, oral healthcare in Malaysia is provided by both the public and private sectors
(Ministry of Health, Malaysia, 1997). Oral healthcare is provided to the population in the public
sector through a comprehensive network of decentralized dental facilities developed through
five-year development plans in the country. There were a total of 2,043 dental facilities in

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Malaysia in the year 2005. Private dental clinics are mostly spread throughout the country in
well-populated metropolitan areas (Ministry of Health, Malaysia, 1997). Reduced cases of
dental problem may have an indirect improved impact towards the nations’ economy.

The use of chemical preservatives and additives in ready to eat food may result in
health hazards when consumed regularly for example harmful bacterial production in teeth.
Researches have been carried out to prevent or decrease these problems. In addition to
having less harmful effects compared to chemical or synthetic additives, natural preservatives
may be more effective in preserving food products against chemical degradation and
microbial spoilage (Rababah et al., 2004). Plants have gained a great deal of attention as the
most effective source of biologically active compounds as a natural antimicrobial agent, such
as medicinal plants, herbs and spices. Polyphenolic compounds, alkaloids, tannins, vitamins,
essential oils and terpenoids constitute the most common of these bioactive compounds.
Bioactive compounds can be extracted and isolated using water or ethanol from plant raw
materials (Khoobchandani et al., 2011).

One of the easier ways to overcome this problem is to extract the antimicrobial
properties from common plants. Different extraction solvents such as water, ethanol and
essential oil can be used to extract the plant. Alcoholic extract of Areca catechu husk fibers
showed dose dependent positive inhibitory effect against Candida albicans with zone of
inhibition of 5-9 mm (Verma, et al., 2012). No important antimicrobial activity against other
studied species was seen in alcoholic and aqueous extracts (Verma, et al., 2012). The findings,
however, show that areca husk contains chemical components with an antifungal effect. This
plant material may therefore be a potential source of natural antifungal agents to cultivate
against C. Albicans, a prevalent oral pathogen (Verma, et al., 2012).

Aside from that, Averrhoa bilimbi fruit can be used for removing stains from clothing and
for washing hands (Zakaria, et al, 2007). By preventing the growth of bacteria, antibacterial
agents are of great benefit to society. Since commercialised antibacterial agents are
expensive, people prefer to find alternative ways to prevent these diseases through the use of
natural and safe, but cheaper, antibacterial agents (Zakaria, et al, 2007). Staphylococcus
aureus, a gram-positive bacterium, frequently found in a healthy person’s nose and skin, is also
present in raw food and in certain food with high osmotic pressure that commonly causes food
poisoning and other skin (Zakaria, et al, 2007). Thus, by the use of common plant parts such as
Averrhoa bilimbi leaves and fruits extract as an antibacterial agent against Staphylococcus
aureus, will help in lessening the rampant spreading of disease in the society (Zakaria, et al,
2007).

These plants have been shown to exhibit some antimicrobial properties against certain
species of bacteria, and are edible. Despite showing antibacterial properties against certain
strain of oral bacteria, a study that showed antibacterial properties against a wide range of
oral bacteria is not available. Thus, their extracts, if proven to be able to inhibit the growth of
oral bacteria, can be used to formulate a natural oral mouth wash that may be safe to
consume. This formula will be all natural, with no chemicals added, and safe for the
consumptions of young children. This will in turn, improve the oral health statistics in young
children.

3. RESEARCH METHODOLOGY

3.1 Oral Bacteria Culture
Bacteria samples were obtained from swab samples of children in Taska Politeknik Nilai with the
age range form 2-4 years old. The bacteria were grown in LB (Luria broth) and peptone water.
These two media were used as they were non selective; thus, all the strains of oral bacteria will
be preserved.

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3.2 Water Extraction
The plants were washed under running water and then with sterile distilled water. The parts of
plants were cut into thin pieces. The plants were weighed. Later, the plants were air dried at
room temperature (370C for 4 days). The fruits were weighed. The dry plants were grinded to a
fine powder using a blender or pestle and mortar and was stored it in air tight bottles. 10g of
grind plants were soaked in 100ml of sterile distilled water and grinded using blender. Each
preparation was filtered through a sterilized Whatman No 1 filter paper. The filtrate was
transferred to the rotatory evaporator and the extraction process run for 3hours at 40 0 C at
37psi. Each extraction was stored in sterile bottles with label in a freezer at 40C for further use.
The plants were washed under running water and then with sterile distilled water. 10g of fresh
plants were soaked in 100ml of sterile distilled water and grinded using blender. Each
preparation was filtered through a sterilized Whatman No 1 filter paper. The filtrate was
transferred to the rotary evaporator and the extraction process run for 3hours at 40 0 C at 37psi.
Each extraction was stored in sterile bottles with label in a freezer at 40 0 C for further use .

3.3 Methanol Extraction
In methanol extraction, the plants were washed under running water and then with sterile
distilled water. The plants were cut into thin pieces. The plants were weighed. Later, the plants
were air dried at room temperature (370C for 4 days). The plants were weighed. The dry plants
were grinded to a fine powder using a blender or pestle and mortar and was stored it in air
tight bottles.10g of grind plants was soaked in 100ml of 96% methanol and grinded using
blender. Each preparation was filtered through a sterilized Whatman No 1 filter paper. The
filtrate was then transferred to the rotatory evaporator and the extraction process run for 3
hours at 40 0C at 332psi. The extracts were stored in sterile bottles was stored with label in a
freezer at 4 0C for further use.

The plants were washed under running water and then with sterile distilled water. 10g of
fresh plants was soaked in 100ml of 96% methanol and grinded using blender. Each
preparation was filtered through a sterilized Whatman No 1 filter paper. The filtrate was then
transferred to the rotary evaporator and the extraction process run for 3 hours at 40 0C at
332psi. The extracts were stored in sterile bottles was stored with label in a freezer at 4 0C for
further use.

3.4 Disk Diffusion Assay
Oral bacteria from stock culture were spread on nutrient agar plate and LB agar plate and
incubated for 24 hours at room temperature. Up to 12 commercially prepared, fixed
concentrations were mounted on the inoculated agar surface, paper discs dipped into
samples and controls. Prior to evaluating the results, plates were incubated for 16-24 h at 35 °
C. The growth inhibition zones were measured up to the nearest millimetre around each of the
antibiotic discs. The diameter of the inhibition zone was linked to the isolate 's susceptibility and
the rate of drug diffusion through the agar medium. The zone diameters of each drug were
interpreted using the criteria published for the discs by the Clinical and Laboratory Standards
Institute (CLSI, formerly the National Committee for Clinical Laboratory Standards or NCCLS) or
by those used in the product inserts approved by the US Food and Drug Administration (FDA).

4. RESULT AND ANALYSIS

When the oral bacteria were grown on the LB agar, the inhibition zone for both water and
methanol plant extracts were shown as in Table 1.

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Table 1: The inhibition zone of oral bacteria on LB Agar

Types of Inhibition zone of juice Inhibition zone of water Inhibition zone of
extracts extraction (mm) extraction (mm)
methanol extraction
Fresh fruit Dry fruit
(mm)
76
Fresh juice Pasteurized 66 Fresh fruit Dry fruit
juice --
Calamansi 8 11 67 10 7
Bilimbi 11 11 67 79
- - -- -7
Dragon fruit 7 9 --
Starfruit 9 7 10 5
- - 8-
Cucumber
Soursop

When the oral bacteria were grown on the nutrient agar, the inhibition zone for both
water and methanol plant extracts were shown as in Table 2.

Table 2: The inhibition zone of oral bacteria on nutrient Agar

Types of Inhibition zone of juice Inhibition zone of water Inhibition zone of
extracts extraction (mm) extraction (mm)
methanol extraction

(mm)

Fresh juice Pasteurized Fresh fruit Dry fruit Fresh fruit Dry fruit
juice
Calamansi 8 11 76 10 7
Bilimbi 11 11 66 79
- - -- -7
Dragon
fruit 7 9 67 --
9 7 67 10 5
Starfruit - - -- 8-
Cucumber

Soursop

For the herb extracts, the results shown for the LB agar and for nutrient agar was shown in Table
3.

Table 3: The inhibition zone of oral bacteria for herb extracts

Inhibition Zone of inhibition Zone of inhibition Zone of inhibition
After 6 against common
Hours against oral bacteria on against oral bacteria on bacteria on LB agar

LB agar (mm) nutrient agar (mm) (mm)

Betel Water Methanol Water Methanol Water Methanol
Star anise extract extract extract extract extract extract
13 10 3
Areca 11 15 9 11 8 5
18 11 10 8 6 4
Ginger 13 13 15 11 5 5
flower 9 9 7
Clove 12 10 8
10 14 8 11 9 4
Curry 8 12 8
leaves

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.
Table 1,2 and 3 shown different bacterial inhibition zone for both bacteria grown in LB
and nutrient agar. This was due to the sample of bacteria were not one type of oral bacteria,
but contained different strains of bacteria. Thus, different types of media gave rise to different
strains, hence the bacteria grown for both media, will be in different composition, with
difference in growth rate, hence the difference in inhibition.

Aside from that, the results also shown different bacterial inhibition ability for water
extracts and methanol extracts. This is due to the active compound that is extracted from each
method is different due to the compounds’ solubility. Some compounds were water soluble
thus was extracted in water extracts, while compound that were not water soluble were
extracted in methanol extracts. Thus, each types of extracts have different composition that
exhibit antimicrobial properties. To investigate the active compound for the antimicrobial
properties, further investigation is needed, but form all samples that were used Star anise, Curry
leaves, Bilimbi, Calamansi and Starfruit exhibit the highest bacterial inhibition properties that
may warrant for further investigation.

5. CONCLUSION

In conclusion, different types of solvents used for the extracts will have different antibacterial
properties, due to the different active compounds extracted. Different raw materials will also
have different antimicrobial properties, but the ones that exhibit the highest antimicrobial
properties were Star anise, Curry leaves, Bilimbi, Calamansi and Starfruit. However, further
analysis should be done to investigate the exact active compound in these extracts that is
responsible for the antimicrobial properties towards the oral bacteria. Aside from that, an
investigation on how to stabilized these active compounds should also be done as it is shown
that the inhibition properties of the samples were reduced when the samples were stored as
compared to when they were freshly extracted.

REFERENCES

Baelum V, v. P. (2007). A global perspective on changes in the burden of caries and
periodontitis: implications for dentistry. J Oral Rehabil , 872-906.

Eugenio D. Beltrán-Aguilar, D. L. (2002). Centers for Disease Control and Prevention. Surveillance
for Dental Caries, Dental Sealants, Tooth Retention, Edentulism, and Enamel Fluorosis ---
United States, 1-44.

Gilsenan-O'Neill, P. (2015). Dental Caries. Dental Health Foundation Ireland , 6.

Griffin SO1, O. E., & CDC Dental Sealant Systematic Review Work Group, B. J. (2008). The
effectiveness of sealants in managing caries lesions. US National Library of Medicine
National Institutes of Health , 74.

Khoobchandani, M., Ganesh, N., Gabbanini, S., Valgimigli, L., & Srivastava, M. M. (2011).
Phytochemical potential of Eruca sativa for inhibition of melanoma tumor growth.
Fitoterapia, 82(4), 647-653.

Malaysia, M. o. (1995). Dental epidemiological survey of pre-school children in Sarawak . Dental
Services Division, Ministry of Health Malaysia , 3.

Malaysia, M. o. (1985). Dental epidemiological survey of school children in Sabah . Dental
Services Division, Ministry of Health Malaysia , 3.

Malaysia, M. o. (1994). Dental epidemiological survey of school children in Sarawak . Dental
Services Division, Ministry of Health Malaysia , 3.

Malaysia, M. o. (1970). Dental Services Division, Ministry of Health Malaysia. Dental

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e-Prosiding Festival Agro Makanan Dan Bioteknologi 2020 (E-FAMB2020)

epidemiological survey of school children in West Malaysia , 3.
Malaysia, M. o. (1997). National Oral Health Of School. Oral Health Division, Ministry Of Health

Malaysia , 3.
Newacheck PW, M. M. (2000). Access to health care for children with special health care.

Pediatrics , 760-766.
Noguti, J., de Oliveira, F., Peres, R. C., Renno, A. C. M., & Ribeiro, D. A. (2012). The role of fluoride

on the process of titanium corrosion in oral cavity. Biometals, 25(5), 859-862.
Oral Health Division, Ministry of Health, Malaysia. (1998). A Lifetime of Healthy Smiles. National

Oral Health Plan for Malaysia 2011-2020. National Oral Health Survey of School Children.
Petersen PE, B. D.-D. (2005). The global burden of oral diseases and risks to oral health. Bull World

Health Organ , 661-669.
Rababah, T. M., Hettiarachchy, N. S., & Horax, R. (2004). Total phenolics and antioxidant

activities of fenugreek, green tea, black tea, grape seed, ginger, rosemary, gotu kola,
and ginkgo extracts, vitamin E, and tert-butylhydroquinone. Journal of Agricultural and
Food Chemistry, 52(16), 5183-5186.
Rockville, M. (2008). The National Survey of Children with Special Health Care Needs. Maternal
and Child Health Bureau .
Romer M, D. N.-L. (1999). Predoctoral education in special care dentistry. Journal of Dentistry
for Children , 132-135.
Tomar SL, R. A. (2010). Changes in the oral health of U.S. children and adolescents and dental
public health infrastructure. Academic Pediatrics , 388-395.
Verma, D. K., Bharat, M., Nayak, D., Shanbhag, T., Shanbhag, V., & Rajput, R. S. (2012). Areca
catechu: Effect of topical ethanolic extract on burn wound healing in albino rats. Int J
Pharmacol and Clin Sci, 1, 74-8.
Yee R, S. A. (2002). The burden of restorative dental treatment for children in Third World
countries. Int Dent J , 1-9.
Zakaria, Z. A., Zaiton, H., Henie, E. F. P., Jais, A. M., & Zainuddin, E. N. (2007). In vitro antibacterial
activity of Averrhoa bilimbi L. leaves and fruits extracts. International Journal of Tropical
Medicine, 2(3), 96-100.

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KEBERKESANAN PELET LITINI CAMPURAN DAUN KELADI (Colocasia
esculenta L) DAN MENGKUDU (Morinda citrifolia) TERHADAP
TUMBESARAN IKAN KELI AFRIKA (C. gariepinus)

Hisyam bin Che Utama*, Amirul Farhan bin Othman@Mustaffa, Nurhafizam Shah bin Husin

Politeknik Jeli Kelantan
Jalan Raya Timur-Barat,

17600 Jeli, Kelantan.
*[email protected]

Abstrak. Kos pemberian makanan menyumbang kepada 60% daripada keseluruhan kos
operasi dalam akuakultur. Kos makanan yang tinggi disebabkan sumber bahan mentah yang
mahal dan susah didapati. Kajian ini dijalankan sebagai alternatif kepada sumber bahan
mentah dalam penghasilan pellet LITINI yang lebih jimat dengan menggunakan daun keladi
(Colocasia eculenta L) dan daun mengkudu (Morinda citrifolia). Daun keladi dan mengkudu
dipilih dalam kajian ini kerana mempunyai kandungan protein yang agak tinggi. Objektif
kajian ini adalah untuk menunjukkan keberkesanan pellet LITINI terhadap tumbesaran ikan keli
Afrika (C. gariepinus). Kajian ini dijalankan selama 5 minggu untuk mengkaji perbezaan
tumbesaran antara ikan keli Afrika yang diberi makan menggunakan pellet LITINI dan ikan keli
Afrika yang diberi makan menggunakan pellet biasa. Sebanyak 120 ekor ikan keli Afrika bersaiz
3 inci digunakan dalam kajian ini. Ikan tersebut dibahagikan kepada 4 tangki (A, B, C dan D),
dimana tangki A dan B diberi makan menggunakan pellet LITINI dan tangki C dan D diberi
makan menggunakan pellet biasa. Berat dan panjang ikan keli dalam setiap tangki diukur
setiap minggu bagi tujuan pengumpulan data dalam kajian ini. Ikan tersebut diberi makan
pada kadar dan kuantiti yang sama bagi kesemua tangki supaya dapat menunjukkan hasil
yang lebih tepat. Hasil kajian mendapati ikan keli dalam tangki yang diberi makan
menggunakan pellet LITINI (a dan B) menunjukkan kadar tumbesaran yang lebih tinggi
berbanding ikan keli dalam tangki C dan D. Walaubagaimanapun, kadar perbezaan yang
ditunjukkan dalam keempat-empat tangki tersebut tidak begitu ketara. Kesimpulan dalam
kajian ini menunjukkan bahawa daun keladi dan daun mengkudu berupaya untuk
menggantikan bahan yang mentah yang lebih mahal dalam pembuatan pellet makanan
ikan air tawar. Kajian ini boleh ditambahbaik dengan menjalankan kajian lebih lama supaya
dapat melihat kesan panjang penggunaan pellet LITINI. Selain itu, ia juga boleh dicubapakai
pada spesies ikan air tawar yang lain.

Kata Kunci: Daun Keladi, Daun Mengkudu, Ikan keli Afrika, Pelet LITINI

1. PENGENALAN

Pokok mengkudu merupakan sejenis tumbuhan yang terdapat di hutan Malaysia serta di negara –
negara Asia yang lain (Syahruddin E, 2011). Selain itu, daun keladi pula berasal dari india
selatan dan Asia tenggara. Iklim dan Tanah untuk pokok keladi memerlukan keadaan panas
dan lembap dengan purata suhu harian diantara 219 ⁰C - 30 ⁰C. Ia ditanam di kawasan-
kawasan rendah dan juga di kawasan yang tingginya sehingga 1,800 a.p.l. Purata hujan yang
diperlukan ialah lebih kurang 250 mm setahun. Ia juga sesuai hidup di kebanyakan jenis tanah
yang kaya dengan bahan organik. Kebanyakannya boleh didapati di tanah yang subur dan
sentiasa berkeadaan lembap atau mempunyai kandungan air yang tinggi (Setyowati,2007).

Pada umumnya, makanan ikan menyumbang dari 60% hingga 70% kepada kos
operasi bagi sesuatu spesis ikan yang diternak. Secara tidak langsung, keuntungan amat
bergantung kepada kos makanan ternakan selain daripada keberkesanan nisbah penukaran

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makanan tersebut oleh ikan yang diternak. Oleh itu, kesan utama kepada penggunaan
makanan berformula kepada para penternak kecil dan sederhana adalah harga yang mahal
dan semakin meningkat. Justeru, tujuan penghasilan pelet daripada daun keladi, dan
mengkudu adalah untuk mengurangkan kos pembelian pelet yang mahal (Jabatan
Perikanan Malaysia, 2009).

Peningkatan harga makanan ikan salah satu faktor utamanya adalah disebabkan
bahan mentah import yang mahal, khususnya tepung ikan atau ‘fishmeal’ dan bahan bahan
mentah import lain seperti tepung soya, jagung dan sebagainya. Selain itu, faktor kos operasi
pengeluaran secara komersial yang tinggi serta melibatkan kos mesin dan tanggungan
(overhead) yang tinggi juga telah menyebabkan berlakunya tekanan terhadap harga
pengeluarannya. Bagi menyokong sasaran pengeluaran benih tersebut, salah satu faktor
yang perlu diambil kira ialah pengurusan induk ikan yang bebas penyakit, termasuk aspek
pemakanan dan nutrisi bagi induk ikan mestilah berkualiti (Jabatan Perikanan Malaysia, 2009).

Penghasilan pelet sangat penting dalam industri akuakultur kerana kegiatan ini
mempunyai permintaan yang tinggi dalam kalangan masyarakat dan menjadi tulang
belakang kepada industri makanan akuakultur yang mengandungi protein yang tinggi dan
sangat berguna kepada benih yang diternak. Penggunaan makanan pelet dapat
mempercepatkan kadar tumbesaran ikan dan mudah dikendalikan atau disimpan
(Yulistiana YG., 2016). Tujuan kajian ini dijalankan untuk mengkaji keberkesanan pelet LITINI dari
daun keladi (Colocasia esculenta L), dan mengkudu (Morinda citrifolia) terhadap tumbesaran
Clarias gariepinus.

Dalam sektor akuakultur air tawar pula, sehingga kini tiada makanan khas bagi induk
ikan air tawar yang boleh didapati di pasaran. Oleh itu, amalan pengurusan induk ikan air
tawar tempatan hanya bergantung kepada makanan peringkat ternakan tidak dapat
memenuhi keperluan zat pemakanan induk ikan secara optimum untuk perkembangan benih
yang berkualiti. Oleh yang demikian, pengeluaran telur atau benih ikan di Malaysia terjejas
dari segi kuantiti dan kualiti bagi keperluan industri akuakultur (Jabatan Perikanan Malaysia,
2009).

2. SOROTAN KAJIAN DAN PEMBINAAN HIPOTESIS

2.1 Nutrisi Mengkudu (Morinda citrifolia)
Buah mengkudu atau dalam bahasa saintifik (Morinda citrifolia) memiliki bau yang sangat
kuat. Meskipun bukan buah yang sangat baik untuk kalangan masyarakat, tetapi tanaman ini
boleh tumbuh di dataran rendah mahupun tinggi. Mempunyai banyak manfaat untuk
kesihatan ikan dan industri akuakultur. Pencegahan penyakit ikan merupakan salah satu faktor
yang sangat penting dalam industri akuakultur. Penggunaan buah mengkudu untuk
mencegah penyakit ikan diharapkan dapat membantu para petani untuk mencapai
kejayaan dalam usaha dalam bidang perikanan (Nastiti RA., 2014).

Khasiat mengkudu bagi penternakan ikan, khususnya ikan keli, adalah merawat
penyakit atau gangguan yang disebabkan oleh bakteria Aeromonas hydrophila. Selain itu,
mengkudu juga dapat mengurangi kanibalisme pada ikan keli. Buah mengkudu mempunyai
kandungan yang cukup lengkap, termasuk nutrien, terpenoid, bahan antibakteria, scolopetin,
dan xeronine dan proxeronine. Selain itu, pokok mengkudu juga mengandungi protein 3.9 g,
lemak 0.6 g, air 89.1 g, karbohidrat 2.2g dan kalsium 114 mg
(Wardini TM, 2012).

2.2 Nutrisi Daun Keladi (Colocasia esculenta L)
Kebaikan daun keladi kepada ikan ialah mempercepatkan pertumbuhan ikan. Daun ini akan
dicampurkan ke dalam pelet. Dengan menggunakan daun keladi ini akan mengurangkan
lagi kos pembelian pelet. Kualiti air juga terjaga dan bersih. Daun keladi kaya dengan protein.
Ia mengandungi kira-kira 23% protein pada asas berat kering (Setyowati, 2007). Protein

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merupakan bahan nutrisi untuk ikan. Selain itu, kandungan vitamin yang terkandung dalam
daun tanaman keladi mampu melengkapkan persediaan bahan untuk ketahanan ikan
daripada serangan penyakit serta menambahkan nafsu makan ikan sehingga mampu
membesar dengan baik dan memiliki kadar hidup yang tinggi. Daun Colocasia esculenta
kaya dengan nutrient, termasuk mineral seperti kalsium, fosforus, besi, dan vitamin seperti
vitamin C, tiamin, riboflavin dan niasin (Hartati, 2003).

Kandungan nutrisi daun keladi (Colocasia esculenta L) ialah 71 kkal, protein 4.1 gram,
lemak 2.1 gram, karbohidrat 12.3 gram, kalsium 302 gram, Fosforus 47 mg, zat besi 8 mg, Vitamin
10395 IU, Vitamin B1 0.11 mg Vitamin C 163 mg (Kementerian Kesihatan Republik
Indonesia,1990).

2.3 Kajian terdahulu
Menerusi kajian oleh D. Cholifah, M. Febriani, A.W. Ekawati dan Y.Risjani pada tahun 2012 yang
bertajuk pengaruh penggunaan tepung silase daun mengkudu (Morinda citrifolia) dalam
formula pelet terhadap tumbesaran ikan sidat/belut (Anguilla bicolor) stadia elver. Digunakan
pada ikan linang atau belut yang berukuran panjang 15 cm dan berat 5.5 gram/ ekor. Ikan di
letakan dalam akuarium yang berukuran 40 x 80 x 40 cm³ dan isipadu 96 liter. 21 ekor Ikan
linang atau belut di masukan ke dalam akuarium. Menurut Degani dan Levanon (1983) ikan
dipuasakan dan ditimbang sebelum memberi pelet. Ikan diberi makan sebanyak 3% dari berat
biomass ikan. 40% pada waktu pagi dan 60% pada waktu malam. Pelet akan disimpan di
tempat yang sesuai. Air perlu diganti sebanyak 20 hingga 30 % untuk memastikan kualiti air
terjaga. Ikan perlu ditimbang setiap 15 hari dengan menggunakan timbang digital. Nilai kadar
kelangsungan hidup ikan linang atau belut (A. bicolor) diantara 8412 hingga 9365% hasil ini
cukup tinggi (Sasongko et al 2007).

Hasil penelitian Christianah dan Badirat (2013), bahawa penambahan 20% tepung
daun betik pada pelet ikan tilapia dapat meningkatkan kesan pertumbuhan kepada ikan
dengan protein, pertumbuhan, kadar hidup, vitamin dan zat lainnya. Penggunaan daun betik
dapat meningkatkan sistem pecernaan dan baik untuk ikan maka dilakukan penambahan
yang berasal dari unsur haiwan dan sayur-sayuran.

Tujuan yang hendak dicapai dalam kajian ini adalah untuk menentukan pengaruh
penambahan ekstrak daun betik kepada pertumbuhan makanan, kelangsungan hidup dan
penukaran tilapia. Walaupun kegunaannya adalah sebagai bahan maklumat mengenai
aplikasi pengaruh penambahan ekstrak daun betik kepada makanan, untuk memperbaiki
kecekapan makanan dan pengeluaran tilapia. Data yang diperolehi dalam kajian ini
dipaparkan dalam bentuk grafik dan jadual seterusnya untuk melihat kesannya. Rawatan
tersebut dianalisis menggunakan varians (ANOVA) dan ujian tukey guna tahu perbezaan
pengaruh antara rawatan sementara alat yang digunakan SPSS versi 16.0 untuk Windows.
Manakala untuk persembahan grafik dan tabulasi data menggunakan Microsoft Excel 2007.
Memberi ekstrak daun betik pada pellet menjejaskan pertumbuhan ikan, kadar hidup dan
penukaran makanan dan memberi Ekstrak daun betik paling baik diperolehi pada dos 300 ml
/ Kg makanan.

Antara kajian terdahulu yang dirujuk bagi menjalankan kajian ini telah membuktikan
bahawa sumber protein dari bahan-bahan alternatif selain tepung ikan dapat digunakan
dalam penghasilan pellet makanan ikan. Hal ini kerana bahan lain yang mempunyai
kandungan nutrien penting dapat menggantikan sumber mentah nutrient penting yang lebih
mahal dalam penghasilan pellet makanan ikan.

3. METODOLOGI KAJIAN

Kajian yang dijalankan ini menggunakan empat bekas yang dapat memuatkan sebanyak 50
ekor ikan Keli Afrika bersaiz 3 inci didalam setiap bekas ternakan. Dua bekas ternakan iaitu
sampel A diberi makan pelet LITINI dari daun keladi (Colocasia esculenta L) dan mengkudu

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(Morinda citrifolia) manakala dua bekas ternakan sampel B diberi makan pelet yang tidak
dicampurkan daun keladi dan mengkudu. Kaedah analisis data yang digunakan ialah
dengan menggunakan graf garis dimana data yang diambil seminggu sekali untuk melihat
tumbesaran ikan dari segi berat dan panjang ikan. Ikan diukur dengan menggunakan
pembaris dan penimbang digital. Sampel ikan akan diambil sebayak 10 ekor setiap bekas
ternakan bagi permulaan dan akan ditambah sebanyak sepuluh ekor bagi melihat
tumbesaran ikan.

Kaedah penghasilan pelet ini pertama sekali ialah penyediaan bahan untuk
menghasilkan pelet. Bahan – bahannya ialah tepung gandum, tepung ubi, tepung ikan,
tepung soya, kepala ikan bilis, daun keladi, daun dan buah mengkudu, minyak dan juga
sedikit air. Bahan - bahan ini penting dalam pembuatan pelet LITINI yang dihasilkan untuk
tumbesaran ikan.

Kaedah yang pertama dilakukan ialah bersihkan semua daun keladi dan daun buah
mengkudu yang telah diambil dan dikumpulkan untuk mengelakkan daripada jangkitan
bakteria. Seterusnya, jemur daun keladi mengkudu dibawah cahaya matahari sehingga
kering untuk memastikan daun tersebut mudah untuk dikisar. Selain itu, kisar semua bahan
menggunakan mesin pengisar sehingga halus. Hal ini kerana untuk memudahkan apabila
digaul. Pastikan semua bahan yang telah dikisar diletakkan dalam bekas yang berbeza untuk
memastikan bahan bahan tidak tertukar dan tercampur dengan bahan lain.

Masukkan semua bahan ke dalam bekas pengadun seperti tepung gandum, tepung
ubi, dan bahan yang berprotein tinggi seperti tepung ikan, tepung soya, kepala ikan bilis dan
gaul sehingga sebati. Masukkan air sedikit demi sedikit sehingga menjadi bentuk doh.
Masukkan daun keladi dan daun mengkudu yang telah dikisar kedalam adunan pelet.
Disamping itu, masukkan buah mengkudu kedalam adunan. Hal ini kerana dengan
menambahkan buah mengkudu ia dapat menarik perhatian ikan kerana buah mengkudu ini
mempunyai daya tarikan bau yang kuat.

Seterusnya, Kadar peratusan yang diperlukan untuk penghasilan pelet terapung yang
diberikan kepada ikan ialah bahan seperti tepung ubi dan tepung gandum yang diperlukan
sekitar 10% - 20% sahaja. Bahan sumber protein utama seperti tepung soya, tepung ikan,
kepala ikan bilis, daun mengkudu dan daun keladi perlu dimasukkan dalam sekitar 40%. Selain
itu, air yang digunakan untuk membuat pelet memerlukan 30 – 40%. Ini kerana untuk
memastikan pelet tersebut dapat digaul sehingga sebati. Air juga dikenali sebagai ejen
pelekat untuk memastikan pelet tersebut dibentuk menjadi doh. Kadar peratusan yang
diperlukan untuk minyak masak ialah 10% sahaja. Hal ini kerana minyak masak ini digunakan
untuk membentuk doh. Seterusnya, minyak ini juga berfungsi untuk menghalang daripada
jangkitan kulat pada pelet. (Nasution EZ., 2006).

Dalam tempoh ternakan kualiti air perlu diperiksa dari masa ke semasa. Ini bertujuan
untuk memastikan kualiti air sentiasa berada pada tahap yang baik dan sesuai untuk ikan
hidup dan membesar. Walau bagaimanapun, pada keadaan tertentu air perlu ditukar atau
diganti. Kebiasaannya air didalam tangki sedia ada dibuang sebanyak 50 peratus dan
kemudian dimasukkan air baru. Keadaan ini akan meningkatkan kualiti air. Ikan yang diternak
di dalam dua bekas ternakan akan diberi makan pelet yang dihasilkan sendiri tanpa
campuran daun keladi dan mengkudu dan bekas ternakan selebihnya diberi makan pelet
yang dihasilkan dari daun, buah mengkudu dan juga daun keladi.

Berat badan ikan dan panjang badan ikan akan dibandingkan untuk melihat kadar
perubahan yang berlaku terhadap ikan melalui jadual yang telah dibina untuk kedua dua
pelet iaitu pelet yang dihasilkan tanpa campuran dan pelet LITINI untuk melihat
perbezaannya. Data akan dianalisis dengan melihat perubahan yang berlaku terhadap
tumbesaran ikan keli Afrika dalam pemberian pelet yang berbeza. Data yang diambil setiap
minggu akan dipecahkan kepada bilangan setiap hari. Jadual akan dibina bagi melihat
perbezaan yang berlaku terhadap pembesaran ikan setiap minggu. Microsoft Excel

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digunakan dalam kaedah analisis data. Data-data yang diambil dimasukkan kedalam Excel
untuk ditukarkan kedalam bentuk graf. Analisis terakhir data akan diterjemahkan kepada
bentuk graf garis untuk mendapatkan keputusan dimana pelet LITINI dari daun keladi
(Colocasia esculenta L) dan mengkudu (Morinda citrifolia) adalah yang terbaik bagi kadar
tumbesaran Clarias gariepinus.

4. ANALISIS DAN KEPUTUSAN

Sampel benih ikan yang mempunyai pertumbuhan berat dan panjang yang lebih tinggi
menunjukkan ikan tersebut mempunyai pertumbuhan yang baik. Pertumbuhan bagi benih
ikan keli yang diternak diberi makanan yang berbeza iaitu sampel A diberi pelet LITINI
manakala sampel B diberi makan pelet yang tidak dicampurkan daun keladi (Colocasia
esculenta L) dan mengkudu (Morinda citrifolia) terhadap tumbesaran Clarias gariepinus yang
ditunjukkan dalam rajah 1.

Rajah 1: Kadar tumbesaran berat Clarias gariepinus
Rajah 1 menunjukkan benih ikan yang diberi makan dengan makanan sampel A iaitu
pelet LITINI (pelet yang dicampurkan daun keladi (Colocasia esculenta L) dan mengkudu
(Morinda citrifolia) mempunyai berat badan yang lebih tinggi berbanding benih ikan dalam
sampel B yang diberi pelet yang tidak dicampurkan dengan daun keladi (Colocasia esculenta
L) dan mengkudu (Morinda citrifolia).
Rajah 2 menunjukkan tumbesaran bagi panjang benih ikan yang telah diukur
sepanjang kajian yang dilakukan. Kajian ini dijalankan dengan mengambil sampel sebayak
10 ekor bagi setiap tangki A, B, C dan D untuk minggu pertama manakala pada minggu ke-2
hingga minggu ke-5 mengambil sampel sebanyak 10 ekor bagi setiap tangki untuk sampel A
dan sampel B.

Rajah 2: Kadar tumbesaran panjang Clarias gariepinus

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Berdasarkan graf 4.2, panjang awal untuk sampel A dan B adalah sama iaitu 8.3cm
selepas ternakan selama dua minggu, panjang purata bagi sampel A meningkat kepada
9.45cm berbanding sampel B hanya 8.5cm.

Didapati bahawa sampel A mengalami pertambahan panjang sebanyak 1.15 cm
berbanding sampel B hanya 0.2cm sahaja. Hal ini juga menunjukkan pelet LITINI yang
dicampur daun keladi (Colocasia esculenta L) dan mengkudu (Morinda citrifolia))
menunjukkan pertumbuhan yang lebih ketara berbanding pelet yang tidak dicampur daun
keladi (Colocasia esculenta L) dan mengkudu (Morinda citrifolia).

4. KESIMPULAN

Hasil daripada analisis kajian ini mendapati bahawa pellet LITINI menunjukkan kesan yang
lebih baik berbanding pellet biasa dengan mengukur tumbesaran ikan keli. Ikan keli yang
diberi makan pellet LITINI menunjukkan pertambahan berat dan panjang yang lebih banyak
berbanding ikan keli yang diberi makan pellet biasa. Kesimpulan dapat dibuat bahawa
campuran daun keladi (Colocasia esculenta L) dan mengkudu (Morinda citrifolia) mampu
menjadi pembekal nutrient penting kepada tumbesaran ikan keli apabila dicampurkan dalam
makanan formula. Walaubagaimanapun, tumbesaran ikan keli boleh dipengaruhi oleh
beberapa faktor lain seperti faktor persekitaran. Oleh itu beberapa perkara boleh ditambah
baik dalam kajian seterusnya yang akan dilakukan supaya hasil kajian yang didapati lebih
objektif. Harapnya, kajian ini dapat dipelbagaikan lagi supaya masalah kos penghasilan dan
pemberian makanan ikan dalam akuakultur dapat diselesaikan.

RUJUKAN

Chriatianah.O., S. Badirat (2013). The Effect of Pawpaw (Carica Papaya) Leave Meal On The
Growth Performance And Blood Profile Of African Cat Fish. Transnational Journal of
Science and Technology July 2013 edition vol.3, No.7

D. Cholifah, M. Febriani, A. W Ekawati dan Y. Risjani (2012), Pengaruh Penggunaan tepung
silase daun mengkudu (Morinda citrifolia) dalam formula pakan terhadap pertumbuhan
ikan sidat (Anguilla bicolor) stadia elver. Bidang kelautan.

Hartati, N. S. dan T. K. Prana (2003). Analisis Kadar Pati dan Serat Kasar Tepung beberapa
Kultivar Talas (Colocasia esculenta L. Schott). Jurnal Natur Indonesia 6(1): 29-33.

Jabatan Perikanan Malaysia (2009). penyelidikan makanan rumusan dan makanan induk
ikan/udang marin dan air tawar. Laporan pencapaian projek rmk-10: 2008 – 2009

Nastiti RA, Hermana W & Mutia R. (2014). Penggunaan dedak gandum kasar (wheat bran)
sebagai pengganti jagung dengan kombinasi tepung daun mengkudu (Morinda
citrifolia) untuk menghasilkan telur puyuh sehat rendah kolesterol dan kaya vitamin A.
Buletin Makanan Ternak 101(1):1-12.

Nasution EZ. 2006. Studi Pembuatan Pakan Ikan dari Campuran Ampas Tahu, Ampas Ikan,
Darah Sapi Potong, dan Daun Keladi yang Disesuaikan dengan Standar Mutu Pakan
Ikan. Jurnal Sains Kimia. 10 (1): 40 - 55.

Setyowati, M., I. Hanarida dan Sutoro. 2007. Karakteristik Umbi Plasma Nutfah Tanaman Talas
(Colocasia esculenta). Buletin Plasma Nutfah 13 (2): 49-56.

Syahruddin E, Abbas A, Purwati E & Heryandi Y. 2011. Pengaruh pemberian daun mengkudu
(Morinda citrifolia L.) fermentasi terhadap kandungan kolesterol karkas ayam broiler.
Jurnal Ilmu Ternak dan Veteriner. 16(4):266-271.

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Wardiny TM, Retnani Y & Taryati. 2012. Pengaruh ekstrak daun mengkudu terhadap profil darah

puyuh starter. Jurnal Ilmu Teknologi Peternakan. 2(2):110-120.
Yulistiana YG, Rahayu I, Jannah RN, Setiyowati N, Maladina MR. 2016. Analisis Marketing Mix

Pembuatan Pelet Ikan Alternatif dari Ampas Tahu di Desa Wakah Kecamatan Ngrambe
Kapupaten Ngawi. Dalam: Seminar Nasional Program Studi Peternakan, UNS Surakarta
(ID).

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KADAR PENETASAN TELUR IKAN KELI DALAM KEMASINAN AIR YANG
BERBEZA

Amirul Farhan Bin Othman @ Mustaffa*, Hisyam Bin Che Utama, Mohd Muslim Bin Mustafa
Jabatan Agroteknologi & Bio-Industri, Politeknik Jeli Kelantan
Jalan Raya Timur-Barat,
17600 Jeli Kelantan, Malaysia
*[email protected]

Abstrak. Kajian ini bertujuan mengkaji kadar penetasan telur ikan keli dalam kemasinan yang
berbeza. Kajian ini memerlukan sepasang induk keli afrika jantan dan betina yang matang.
Induk keli jantan disuntik hormon ovaprim dengan kadar 0.25ml/kg dan induk betina disuntik
hormon ovaprim dengan kadar 0.5ml/kg. Selepas 12 jam, perut induk ikan keli betina di urut
bagi mengeluarkan telur. Spermaa diambil dari induk jantan dengan kaedah pembedahan.
Telur di campur dengan spermaa ikan jantan bagi proses persenyawaan. Setiap tangki
penetasan diletakkan sebanyak 1 gram telur ikan keli yang telah disenyawakan. Tahap
kemasinan yang disediakan dalam setiap tangki adalah berbeza 0ppt, 5ppt, 10ppt dan 15ppt.
1gram telur ditimbang dan bilangan telur dikira, jumlah purata bagi telur ikan keli dalam 1
gram berat adalah sebanyak 780 biji telur. Telur dibiarkan dalam tangki penetasan selama 24
jam dan bilangan telur yang menetas dikira. Data yang diambil adalah kadar penetasan
setiap tangki. Keputusan menunjukkan kemasinan yang paling optimum adalah pada tahap
kemasinan 5 ppt dengan kadar 87%. Tiada penetasan berlaku pada tahap kemasinan 15ppt.
Kesimpulan bagi kajian ini adalah, kadar penetasan yang tinggi dicapai pada tahap
kemasinan 5ppt dan telur ikan keli masih boleh menetas dibawah kemasinan 10ppt.

Kata kunci : induk, kemasinan air, kadar penetasan

1. PENGENALAN

Ikan Keli Afrika merupakan haiwan ternakan akuakultur yang biasa diternak. Ia boleh diternak
dalam kepadatan yang tinggi dan mampu bertahan di kawasan yang mutu kualiti air yang
rendah berbanding ikan yang lain. Nama saintifik bagi ikan keli yang dipilih adalah Clarias
gariepinus. Kajian ini dijalankan bagi mengetahui kadar penetasan telur ikan keli dalam tahap
kemasinan yang berbeza. Telur ikan air tawar jika dimasukkan ke dalam air masin akan
mengakibatkan pengembungan dimana cecair luaran telur yang hiperoosmotik akan
menyerap ke dalam telur yang hipoosmotik dan boleh menyebabkan kerosakan pada telur
(Sukendi, 2003). Kajian ini bertujuan mengenalpasti kadar kemasinan yang boleh diterima oleh
telur ikan keli bagi mengekalkan bentuk asal telur. Kehadiran garam dalam air boleh
mengurangkan jangkitan bakteria dan juga parasit yang hidup di air tawar.

2. SOROTAN KAJIAN DAN PEMBINAAN HIPOTESIS

Ikan keli Afrika mampu untuk hidup dalam pelbagai persekitaran air tawar, termasuk perairan
seperti tasik, sungai dan kolam. Ia dapat menyesuaikan diri dengan keadaan persekitaran
yang melampau dan boleh hidup dalam julat pH 6.5 – 8. Ikan keli juga mampu untuk bertahan
dalam air yang keruh dan julat suhu air 8°C hingga 35°C (Jesse M, 2017). Suhu optimum bagi
pertumbuhan ikan keli adalah 28-30°C.

Mengikut kajian yang telah dijalankan oleh Sukendi pada tahun 2003 beliau
menyatakan dua faktor yang mempengaruhi kadar penetasan telur ikan iaitu faktor dalaman
dan faktor luaran. Faktor dalaman adalahseperti hormon dan jumlah kuning telur yang
terkandung dalam telur. Hormon yang dihasilkan oleh hipofisa dan tiroid berperanan dalam
proses metaforfosis disamping mempengaruhi kadar penetasan telur. Faktor luaran yang

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mempengaruhi kadar penetasan telur seperti parameter air, suhu, saliniti, pH, ammonia dan
lain-lain (Hadid et al, 2008).

Penetasan telur ikan keli lebih tinggi dalam air masin berbanding air tawar. Dengan
suhu yang optimum dan kehadiran kemasinan dalam air disyorkan untuk meningkatkan kadar
penetasan benih keli (Burchell et a,l 1822). Kemasinan adalah satu faktor yang mempengaruhi
kadar penetasan telur ikan dan sainiti yang optimum untuk pembesaran ikan adalah pada
2ppt (Tang 2000). Proses penetasan akan dipengaruhi oleh saliniti melalui osmoregulasi telur
ikan (Sukendi 2003). Telur ikan air tawar akan mengembung sekiranya terdedah kepada saliniti
yang tinggi manakala telur ikan air masin akan mengecut apabila dimasukkan ke dalam air

tawar (Hadid et al, 2008).
3. METODOLOGI KAJIAN

Sepasang induk ikan keli matang dipilih bagi mendapatkan telur dan sperma. Teknik yang
digunakan adalah secara aruhan dimana induk betina akan disuntik hormon ovaprim
sebanyak 0.5ml/kg dan induk jantan disuntik sebanyak 0.25ml/kg. Hormon ovaprim perlu
disuntik pada kedua-dua induk bagi mematangkan telur dan sperma sebelum proses leretan
dilakukan. Induk yang telah disuntik di asingkan dan diletakkan dalam 2 tangki yang berlainan
selama 10–12 jam bagi tempoh telur untuk matang. Proses leretan dilakukan bagi
mengeluarkan telur dari induk betina. Perut induk jantan dibelah bagi mendapatkan sperma.
Telur dan sperma dicampurkan dan digaul dengan bulu ayam. Air suling dimasukkan sedikit
bagi meratakan sperma ke semua telur untuk tujuan persenyawaan.

Jadual 1: Susun atur tangki kajian

Empat biji tangki gentian kaca berkapasiti 200L digunakan bagi kajian ini. Kesemua
tangki disusun di tempat yang sama bagi mendapatkan persekitaran yang sama. Oksigen
diberikan dan saliniti setiap tangki diselaraskan mengikut jadual 1.

Ikan keli yang telah matang dipilih bagi mendapatkan telur yang bermutu. Bagi
kajian ini sepasang induk ikan keli berumur 6 bulan keatas digunakan.
3.1. Proses pembenihan ikan keli secara aruhan

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1. Induk matang dipilih berdasarkan 2. Induk disuntik dengan hormon
beberapa faktor, antaranya berat ovaprim
>500g, berumur >6 bulan dan 0.5ml/kg – betina
mempuanyai alat kelamin yang 0.25ml/kg - jantan
kemerahan.

3. Selepas 10-12 jam, perut induk betina 4. Telur dan sperma dicampur dan
dileret bagi mendapatkan telur. Induk diratakan. Air ditambah sedikit
jantan dibedah bagi mendapatkan bagi mempercepatkan proses
sperma. persenyawaan.

Setelah telur ikan keli disenyawakan, sampel telur sebanyak 1g ditimbang dan bilangan
telur dikira sebanyak 3 kali bagi mendapat purata bilangan telur dalam berat 1g. Telur ikan keli
sebanyak 1g dimasukkan ke dalam jaring penetasan yang berada dalam setiap tangki yang
telah diubah kemasinan 0ppt, 5ppt, 10ppt dan 15ppt. Pengudaraan diberikan sepanjang masa
bagi membekalkan oksigen yang mencukupi kepada telur ikan. Pengiraan penetasan dilakukan
setelah 24 jam, bilangan larva yang berjaya menetas dikirabagi mendapatkan kadar peratus
penetasan. Kualiti air diambil sepanjang kajian dijalankan.

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