research capsule edited edition (scope)

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Phytochemical Screening and Anti-bacterial Potential of Watermelon Seed Extract

(Citrullus lanatus) onP. acnes

Juwan Howard Barlomento1, Joey Lopena1, Joshua Marcial1, Alexis Ane Pa-a1, Kristi Patrisha Sapungan1,

Patricia Anne Yray1and Dean Theresita V. Atienza2

1Biology Student, Department of Biology, College of Science, Polytechnic University of the Philippines2Dean, College of Science, Polytechnic University of the Philippines

OVERVIEW

Watermelon (Citrullus lanatus) is a scrambler and trailer flowering plant which originated fromsouthern Africa and is also cultivated in the Philippines. The fruit it bears is called a watermelon,

classified by botanists as a pepoa berry with thick rind and fleshy center. Though the leaves, roots,flowers, whole plants and stems were examined for useful phytochemicals in many research projects, fewresearches refer to seeds from the fruit as sources of biological properties, specifically anti-bacterial

properties. Also, we found a number of claims in the internet, as well as in Filipino folklore, about the

anti-acne effects of watermelon seeds.

The researchers came up with the following questions with regards to the properties of

watermelon seed extract:

Which phytochemical constituents are present in the watermelon seed extract? How will these phytochemicals affect the growth ofPropionibacterium acnes? Will the 4 extracts (HWE, CWE, ethanol, methanol) yield the same intensity of antibacterial

property?

Thus, the researchershypotheses are:

The seeds contain secondary metabolites such as Saponins, Alkaloids, Tannins, etc. The phytochemical constituents in the seeds could be helpful to inhibit the growth of P. acnes

bacteria.

The four extracts will have different levels of efficacy againstPropionibacterium acnes.

OBJECTIVES

The objective of this study is to determine the presence of phytochemicals and secondarymetabolites on the 4 extracts of Citrullus lanatus, and to test the extracts have antibacterial properties

againstP. acnes. We also aim to find out if which of the HWE, CWE, ethanol, and methanol extracts are

effective in inhibiting the growth of P. acnes

SIGNIFICANCE OF THE STUDY

We are living in this modernized society where commercially available anti-acne products sold in

the market were considered expensive and the worst case brought some adverse effects to the consumers.


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This study would like to unveil whether the phytochemicals in watermelon (Citrullus lanatus) seed

extract have the anti-bacterial potential. Instead of wasting the seeds, it could be offered as one of the

many possible sources of a cheap and accessible supply of readily available anti-acne remedy.

This study may also be of significance to those who have acnes due to some disadvantageous

effects of commercial products (e.g. those who suffered from acnes due to incompatibility or adverse

effects of chemical-based product to their skin), since this extract is natural and organic.

Additionally, considering the types phytochemicals we can screen, this study can be used for

further researches on the possible uses of watermelon seed extract.

RELATED LITERATURE

Watermelon leaves, roots, flowers, whole plants and stems were examined for useful

phytochemicals in many research projects. The fruit has chemical compounds such as: phytonutrient

lycopene (Collins, 2003) an amino acid citrulline (Collins,2007), -carotene (Edwards,2003), and alpha-tocopherol (Charoensiri, 2009).

The seeds are a source of protein (Akpapunam, 1981), B vitamins,minerals (such as magnesium,

potassium, phosphorous,sodium, iron, zinc, manganese and copper) and fat (Vandermark, 2011; Collinset al., 2007). The oil extracted from the seeds are rich in Omega 6 and 9 EFAs, as reported by Natural

Sourcing, LLC.*

Propionibacterium acnesis a gram-positive human skin commensal that prefers anaerobic growthconditions and is involved in the pathogenesis of acne (Kirschbaum and Kligman, 1963). Acne is one of

the most common skin diseases, affecting more than 45 million individuals in the United States.

Also, the researchers found a number of claims in the internet, about the anti-acne effects ofwatermelon seeds. [1]Watermelon seed oils and extracts are being sold as facial serum, [2][3], facial tonic[4], and facial oil [5][6][7], though there are only few studies about the anti-bacterial potential/activity of

watermelon seeds, (Braide et al., 2012), and the researchers was unable to find researches about it havinganti-bacterial activity againstP.acnes.

The results that we will have from this study will give us knowledge on the phytochemical

constituents of watermelon seeds and its anti-bacterial properties, specifically againstPropionibacteriumacnes. The efficacy of each of extract will be compared to each other, to see which extract is moreeffective.

METHODOLOGY

Coll ection and preparation of the seeds of watermelon
http://www.nutritionjrnl.com/article/S0899-9007%2807%2900012-3/abstracthttp://www.nutritionjrnl.com/article/S0899-9007%2807%2900012-3/abstracthttp://www.nutritionjrnl.com/article/S0899-9007%2807%2900012-3/abstract


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Watermelons that will be used for this study will be bought at Divisoria market. The seeds that

will be obtained from the fruits will be washed and dried in laboratory oven maintained at 40C

overnight, and will be stored in a dry container to avoid fungal growth. The seeds will be ground using

laboratory blender, all methods as done by Braide (2012). The ground seeds will be subjected to

extracting methods adopted by Cheesbrough (2000). Filtrate of the extract was obtained by separation the

suspension in a glass funnel and filter paper. Hot and cold water extracts were neatly separated and alsostored. Ethanol and methanol were allowed to evaporate and stored in an airtight conical flask (Braide

2012).

Phytochemical Screening

The analyses for phytochemical constituents will be performed using generally adapting the

methodologies described by Martinez, Miranda, and Doughari (Martinez, et.al, 2003 and Miranda, 2000;

Doughari 2012). The methods were accepted as qualitative methods in determining the presence of

phytochemicals and secondary metabolites.

Preparati on of P. acnes

Pure culture ofPropionibacterium acnes will be ordered from Biotech Philippine National

Collection Of Microorganisms, National Institute of Molecular Biology and Biotechnology, UPLB,

College, Laguna.

Preparati on of paper discs

We will prepare small circular high potency discs with diameter 6.25mm made from Whatman

No.1 grade filter paper with the aid of a mechanical perforator, if available. (Braide et al.,2012) The

circular discs will be sterilized in a glass Petri dish using the hot air oven at a temperature maintained at

160C for 1h (Cheesbrough, 2000; Harrigan and McCance, 1990).

SCOPE AND LIMITATION

The researchers will only use fresh C. lanatusseeds. The C. lanatusspecimens will be acquired

straight from a farm to ensure freshness and availability. Furthermore, the research will only require the

use of the specimen C. lanatus. The specimens will be sent to the Bureau of Plant Industry for

identification and authentication. For the extraction, only polar solvents will be used, thus only the

ethanol, ethyl acetate and maceration methods are to be performed. For the phytochemical analysis, the

presence of only the following phytochemicals will be tested: flavonoids, saponins, terpenes, sterols,

anthocyanins, alkaloids and tannins. Furthermore, the researchers will only use qualitative tests in

determining the presence of the said phytochemicals. For the bacterial specimens, both gram-positive and

negative bacteria are to be used. During bacterial testing, the zones of inhibition will be the determinants

of the antibacterial potential. Additionally, only the antibacterial potential is to be analyzed.


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BUDGET ALLOCATION

Things needed AllocationWatermelon Php 150.00

Laboratory Expenses Php 1500.00

Transportation Php 250.00

Other expenses Php 100.00

TOTAL Php 2000.00

SCHEDULE OF ACTIVITIES

REFERENCES

Nov 20 Nov 27 Dec 4 Dec 11 Dec 18 Dec 25 Jan 1 Jan 8 Jan 15 Jan 22 Jan 29

Research and AdditionalInfo

Gathering Watermelon

Preparation of seeds

Storage

Extraction

Place Order

PhytochemicalScreening

Antibacterial test

Finalizing Paper

Analyzation of Results

Final Writing


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*Natural Sourcing, LLC is and FDA registered specialist in cosmeceutical ingredients thatpublish reports on ingredients (here, watermelon seeds) for evaluation purposes

Akpapunam, M., Markakis, P. (1981)Protein Supplementation of Cowpeas with Sesame and

Watermelon Seeds, Journal of Food ScienceVolume 46, Issue 3, pages 960961, May 1981

Braide, W, Odiong IJ, and Oranusi S (2012)Phytochemical and Antibacterial properties of theseed of watermelon (Citrullus lanatus), Journal of Microbiology Research (PJMR) Vol. 2(3), pp.

99-104, March 2012

Collins, JK, Ph.D., Wu, G, Ph.D., Perkins-Veazie, P, Ph.D., Spears, K, Ph. D., Claypool, LP,

Ph.D., Baker, RA, Ph.D., Clevidence, BA, Ph.D. (2007) Watermelon Consumption IncreasesPlasma Arginine Concentrations In Adults, 11 January 2007.

Collins, JK,Ph.D., Perkins-Veazie, P, Ph.D. (2003) Flesh quality and lycopene stability of fresh-cut watermelon

Charoensiri R, Kongkachuichai R, Suknicom S et al. (2009)Beta-carotene, lycopene, and alpha-

tocopherol contents of selected Thai fruits.Food Chemistry, Volume 113, Issue 1, 1 March 2009,Pages 202-207.

Cheesbrough M (2000).District Laboratory Practice in Tropical Countries Part 2. CambridgeUniversity Press, UK.

Harrigan WF, McCance M (1990).Laboratory Methods in Food and Diary Microbiology.Academic Press Inc., London, pp. 25-28.

Doughari, J. H.

Phytochemicals: Extraction Methods, Basic Structures and Mode of Action as PotentialChemotherapeutic Agents , Phytochemicals -A Global Perspective of Their Role in Nutrition and

Health, 21 March, 2012, pages 1-32

Edwards AJ, Vinyard BT, Wiley ER, Brown ED, Collins JK, Perkins-Veazie P, Baker RA, and

Clevidence BA Consumption of Watermelon Juice Increases Plasma Concentrations of Lycopeneand -Carotene in Humans

Vandermark T (2011). The Health Benefits of Watermelon Seeds. Retrieved Dec. 09, 2011

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