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A Research Proposal
by
Marionne G. Oliveros
Generose Christy M. Mateo
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February 2011
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One of the main problems concerning our consumer society today is food spoilage.
Many consumers prefer to stock up, while most companies want to preserve their products ±
however, they are unable to do so due to multiple factors which may lead to the shortening of
their shelf life. Food preservatives have offered some assistance, but most are chemical-
based, and, though effective, are not helpful towards the environment as a whole. Persimmon
leaves will be tested for their effectiveness in preventing food spoilage. They will serve as
Four types of canned goods will be tested (cocktail, corned beef, sardines, and meat
loaf). There will be three replicates of each sample. Persimmon leaves were crushed and
filtered thoroughly through a fine nylon mesh and then Rotavapped. Each replicate will have
have extracts 3.33% w/v. The samples would then be stored in an area of constant
temperature, and results will be checked after four days. Statistical data would be tested
through ANOVA, and significance in shelf life lengthening would determine the research's
success.
When proven that persimmon leaves may have a hand in food preservation, this will
encourage businesses to use natural alternatives rather than using chemicals which may harm
the surroundings. Persimmon is common in flora all around the world, including the
preservative
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Persimmon
Mabolo (ñ )
Flavonoids
Extraction of Flavonoids
Food Preservation Techniques
Shelf Life
Food-borne Bacteria
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Procurement and Preparation
of Persimmon Leaves (ñ )
Procurement of Canned Goods
Extraction of Flavonoids from Persimmon Leaves
Addition of Persimmon Extract to Canned Goods
and Storage of Setups
Preparation of Canned Good Samples
after Four Days of Treatment
Gram Staining of Samples
Recording of Observation and Results
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Antibacterial Effects of Persimmon Leaves (ñ ) on the Shelf Life of
Canned Goods
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Since prices of food had been high for the past years, consumers tend to by cheap and
long-lasting goods. Unfortunately, some preservatives utilized during the canning process are
harmful chemicals and inorganic materials that might lead to bodily dysfunctions and might
These were utilized as an anti-allergenic dietary supplement. Chinese and Indians also use
persimmon fruit as an ingredient for ³Holy teas´ and a traditional remedy for hiccups.
Canning has been an extremely popular method for preserving foods. However, there
have been many incidents wherein improper canning has caused food spoilage (and in effect,
a dilemma for consumers worldwide). Plus, several canning techniques involve the addition
of chemicals and inorganic substances which, aside from being inefficient in terms of
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prolonging the shelf life of canned goods since food spoilage has become a huge problem to
both contributors and consumers. This organic preservative may also be a cheaper and
healthier alternative since other preservatives nowadays contain factors that may ail the body,
The extracts from ñ should be able to prevent the multiplication or
existence of non-airborne bacteria that may cause food spoilage, such as species of
and
. The canned goods with ñ extract
should surpass the quality of the control after four days of experimentation in terms of
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Canned goods have always been one of the most affordable products in the market,
appealing to all classes of society. Canning has helped in lengthening the time it took until
the foods inside them would expire. This preservation of foods is helpful for people who
want to stock up on food and for those who have limited budgets. Prolonging of the shelf life
of canned goods would be advantageous not only to the consumers, but also to the
manufacturers. There would be less wasted material and resources. A preservative offering
chemicals which may affect the consumer's diet and health, ñ extracts
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The use of one species of Persimmon, ñ , which is native in the
Philippines, will be the focus of the study. Four types of canned goods were considered,
specifically meat loaf, corned beef, sardines, and cocktail. Replicates of these subjects will
involve the same brand, same amount, and same date of expiration prior to the addition of
ñ The subject of the research is only the preservation of canned goods
using the extracts from ñ The concentration of each extract will be
measured and maintained uniform all throughout the experiment. The only part of the tree
that will be utilized are the leaves, and no extracts from any other part will be taken. For
uniformity, leaves from only one specific tree will be gathered. One brand of each of the
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originated in China, then spread to Korea, Japan, and other nearby countries. Its species also
cultivated in the United States of America when a United States Commander returned with a Japanese
variety to Washington, D.C. (usa.gov., 2006). Persimmons thrive in mild summers and moderate
winters, and due to this range, leaves do not flourish when temperatures reach 26ºF. High
temperatures may sunburn the bark as well (California Rare Fruit Growers, Inc., 1996).
The fruit of persimmons have been used as an ingredient for Chinese and Indian holy teas. It
has also been known as solution for hiccups. Persimmon cakes are known as a traditional remedy for
diarrhea, hemorrhoids, lung infections, and asthma (Kotani, et. al., 2000). Persimmon exhibits rapid
antibacterial effects against food-borne infections, such as and
Persimmon leaves grow up to seven inches long and four inches wide, alternating with each
other in growth on branches. Their colors range from a yellowish green, a darker hue when maturing,
and reddish purple in the autumn season (California Rare Fruit Growers, Inc., 1996). Persimmon
leaves have been known to be a good source of important dietary antioxidants, such as vitamin A, C,
and flavonoids. They are also known to contain bioactive components, like tannins, phenolic
compounds, coumarins, organic acid, carotent, amino acids, and the aforementioned antioxidants (Li-
Lian J. et. al.) They are anti-allergenic and anti-puritic in composition, which makes them an allergy
persimmon leaves improve lipid profiles and suppresses body weight gain in animals that were fed
high-fat diets.
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continuous food resource. Similar to its fruit, leaves (see Fig. 1) also thrive all year round. Mabolo's
main season, wherein the fruits are ³... of a variety, rarer than the red « sweeter, and less astringent
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Flavonoids are widely found in plants, which are non-nitrogenous biological pigments (also
known as biochromes). These are responsible for the red coloring of flower buds and purple-reds of
autumn leaves. Flavonoids are considered as an important part in contribution to the spread of plant
pollen due to the vivid colors it may give the plant, which in turn, attracts insects and other seed-
dispersing agents. (Norton, P., 1991). Apart from giving pigmentation to a plant, flavonoids have
bioactive properties that remain stable against heat, acid, and storage temperature. Research has
shown that hydroxylation of these bioactive properties may have effects on antibacterial activity.
Some flavonoids found in persimmon leaves are astraglin, isquercitrin, kaempferol, quercetin, rutin,
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at Ë5ºC, with a 1:05 material ratio and 75% ethanol as solvent, and four times of extraction within
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two hours (Sathishkumar, T., et. al., 200Ë). The leaves should be shredded and blended until it
becomes fine powder, and added a small volume of ethanol. The solution should be vacuum filtered
thoroughly and then washed with more ethanol (kay, 200Ë). Ethanol is because it, too, is polar,
therefore glycones in the flavonoids are soluble in it. Extracts should be stored in a cool dry place.
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Food prepared using a certain procedure such as cooking, freezing, canning ,dehydrating,
milling, culturing with bacteria, or adding vitamins and minerals, has a lengthened shelf life than
other foods unprepared (Burdon, 196Ë). Foods gradually spoil or rot after hours or minutes after
harvesting, slaughtering or manufacturing. Some foods spoil rapidly, others are kept longer but for a
limited time only. The deterioration in some foods include biotic factors, such as bacteria (like yeast
and molds), and as well as abiotic factors, like temperature, moisture, oxygen, light and time, and
The fundamental principle in food storage is to keep it cool, clean, and covered. High
temperature kills bacteria, while freezing preserves them in suspended animation (Singh, 2007).
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Canned goods that are unopened will store for 12 months or longer from the date of
processing. Canned fruit juices that are unopened will store for nine months
or sealed from the surrounding environment may last up to three or four days. However, its shelf life
may deteriorate depending on the surroundings, both abiotic and biotic factors, such as dampness,
which may corrode cans or metal lids and cause leakage so the food will spoil (QueryCAT, 2007).
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Baby Food 1
Tomato Sauce 1
Fruit Juice 1
Spaghetti Sauce 1
Soup 1
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Putrefactive types of bacteria are the main causes of food spoilage. Multiplication of bacteria
must be prevented to ensure the preservation of food. Once food is dried, or canned, the bacterial
growth will be inhibited due to lack of moisture (Burdon, et. al., 196Ë).
organisms. Most monerans do not make their own food, which means they have to rely on other
organisms to obtain it. These organisms have to break down or decompose other living things to
microorganisms because of their heat-resistant nature. In addition, there are other microorganisms
which are mesophilic, but through the leakage of the container during cooling, may spoil the food.
This would classify food-borne bacteria into two categories: biological spoilage by thermophilic
Most foods have water in them, and that may cause the growth of bacteria. Gram-
negative bacteria are known to be pathogenic especially when found in food (Masschalk, B.,
et. al., 2003). Pathogenic bacteria are usually colorless or odorless (such as or
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In the preparing the persimmon extracts, persimmon (ñ leaves will
be obtained, washed and then shredded into pieces. The shredded leaves will be made into
In this set up, there will be four experimental setups from each of the canned goods,
thus, 12 experimental canned goods to be tested, with the other four canned goods set as
control. The canned goods considered in this experiment are corned beef, cocktail, meat loaf,
and sardines, the samples in each group having the same expiration date as indicated on their
labels.
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In the resulting leaf powder, 96% ethanol was added as a solvent. In order to obtain the
flavonoids, the solution will filtered four times through a fine nylon wire mesh. To remove the
solvent, the solution will go to RotaVaporation for ten minutes. The resulting solution will then
Each of the three replicates of canned good will have persimmon extracts 3.33% of
their weight. Upon adding the extract, the canned good should be covered with parafilm or
foil. The canned goods, including the control, will be placed on a shelf at room temperature
Finally, after four days after the expiration of the canned goods, a loopful of sample
will be obtained from each of the canned goods for observation. Each sample will be placed
In gram staining, each of the sample slides will be heat fixed by quickly passing the
slide over the fire of the alcohol lamp. Crystal violet stain will then be added to the heat fixed
sample for one minute. Next will be washing with distilled water. Iodine will then be added
for three minutes. Then will be washed with distilled water. Afterwards, it will be
decolorized with 95% ethanol for 20 seconds and then will be washed again with water.
Lastly, the sample will be counter-stained with safranin for one to two minutes and then
washed for the last time and blot dried with tissue.
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The sample slides will then be observed under the microscope and will be classified
as gram positive or gram negative. Gram negative bacteria will be regarded as pathogenic
food-borne bacteria. Results and observations will then be recorded, and statistical data will
be determined by ANOVA.
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Baumann, L.S. (2006).
Retrieved from:
http://www.skinandallergynews.com/views/cosmeceutical-critique-by-leslie-s-
baumann/fadf7d9135.html
Burdon, K.L., Williams, R.P. (196Ë).
. Toronto, Ontario, Canada: The Macmillan
Company.
Canned Food Tips and Hints. . (2010). Retrieved from:
http://www.homecooking.about.com/od/specificfood/a/cannedfoodtips.htm
Drummond, K.E. (1996). ñ '( ñ United States of Amercia:
International Thompson Publishing, Inc.
Microbial Spoilage. ' ' Retrieved from:
http://home.pacific.net/hk/~ppleung/Chem/spoilage.htm
Mika, H., Ryuji, S., Takeshi, W., Tokuya, W., Hiroshi, K., Atsushi, N. (2006). *
ñ , '''
-
. Japanese Journal of Conservative Dentistry (Vol. 4Ë
No.2). Japan Science and Technology Agency.
Norton, P., et. al., (1991). ! . Encyclopedia Britannica, Fifteenth Edition. Encyclopaedia
Britannica Inc., USA.
Pirie, N.W. (197Ë). '
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* ( . Oakleigh,
Melbourne, Australia: Cambridge University Press.
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Satishkumar, T., et. al. (200Ë). 3 , ' ! + ' ! '
/ 013 ñ Nature and Science, 6(3),
200Ë, ISSN: 1545-0740
Singh, M. (2007). 2 . New Delhi, India: Anmol Publications, PVT.
LTD.
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Legend:
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Materials Description and Source Estimated Cost
Quantity
4 - Corned Beef Robinsons Market
4 ± Cocktail Robinsons Market
4 ± Meat Loaf Robinsons Market
4 ± Sardines Robinsons Market
Total Budget for Materials
$
Not Applicable
3
Not Applicable
TOTAL BUDGET:
20