FEU ALABANG

Lot 1, Corporate Woods Avenue corner South Corporate Avenue, Woods District,
Filinvest City, Alabang, Muntinlupa City
SENIOR HIGH SCHOOL PROGRAM
1st Semester School Year 2019-2020
RESEARCH PROPOSAL FORM

Title of the Study

In Vivo Assay for Angiotensin Converting Enzyme (ACE) Ihibitory Activity of Helianthus annuus (sunflower)
Proponent/s Student Number/s Section
Agda, John Felix Magyaya 201810615 12 – SE125
Bolisay, Aaron Elizer Buenaventura 201811088
Contact Number/s Email Address
09773906863 201810615@feualabang.edu.ph
09359912044 201811088@feualabang.edu.ph
Study Duration (in months)
5 months
Study Location
American Bio-Clinical Laboratories, International, Muntinlupa City
Itemized Total Budgetary Requirement (in Php)
Sunflower -₱3,000.00 Experiment fees -₱15,000.00
Personal laboratory equipment -₱500.00 Processing of samples -₱7,500.00
Food -₱2,000.00 Consultation with professionals -₱10,000.00
Transportation -₱1,500.00
Miscellaneous -₱1,500.00
TOTAL: ₱41,000.00
Education
Educational Institution Level Month/Year
Jose Rizal University March 2013
Elementary
St. Joseph’s Academy April 2013
Newville Heights Academy April 2017
Junior High School
Las Piñas City National Science High School March 2017

Agda, John Felix M. Bolisay, Aaron Elizer B.

December 10, 2019 December 10 2019

Signature over Printed Name (Surname, Given Name, M.I.) and Date
Endorsed by (Subject Group Head)

Alegre, Hadji C., Ph. D

December 10, 2019
Signature over Printed Name (Surname, Given Name, M.I.) and Date
Recommending Approval

Eva D. Arcena, EdD

Principal, SHS
Date Submitted Received by

Hadji Alegre, Ph. D

December 10, 2019 Signature over Printed Name of Research Faculty
 CONCEPT PAPER FORMAT

Main Argument/Rationale

Matchar et al. (2008) stated that angiotensin converting enzyme (ACE) inhibitors have long-term effects on blood pressure.
ACE inhibitors are medications that inhibit the activity of the enzyme ACE, which decreases the production of angiotensin II –
a chemical produced in the body causing narrowing of blood vessels. Thus, blood vessels enlarge or dilate, and blood pressure
is reduced. This lower blood pressure makes it easier for the heart to pump blood and improves the function of a failing heart.

According to Daskaya-Dikmen et al. (2017), most researches on ACE inhibitors solely depend on derivatives from animals.
Hence, studies on the characterization of ACE inhibitors from novel food sources, such as plants, should be recognized further.
More specifically, several studies have shown ACE inhibitor peptide derivatives from Helianthus annuus (sunflower) which may
prove beneficial for curing hypertension and, possibly, other cardiovascular diseases. In lined with this, Guo et al. (2017) some
bioactive peptides present in common sunflower seeds and sprouts show different levels of ACE inhibitory effectiveness. Thus,
further research is needed to determine the plant’s concentration of ACE inhibitory peptides.

Theories or Concepts Supporting the Main Argument/Rationale

Flavonoids are the perceived to be most important phenolic compound (Gonçalves, 2017). They are responsible for the vivid
colors of fruits and vegetables. These flavonoids are also present in plants and are used as attractants and UV screens (Lin et
al. 2016) and are commonly extracted for human consumption as they possess various biological and pharmacological activities
including antioxidant, cytotoxic, anticancer, antiviral, and antibacterial, just to name a few (Justino, 2007)

One explanation for these effects might be the flavonoid’s constituent. According to Balasuriya et al. (2011) Flavonoids are
natural ACE-Inhibitors. Competitive ACE-inhibitors were found on the plants’ flavonoids, with the most effective being flavan-3-
ols and anthocyanins. Guo et al. (2017) stated that ACE-inhibitors, along with its parent group, phenolic compounds, were
natural antioxidants, and a study from Karamać et al. (2012) showed that sunflower seeds showed good correlation among the
test results of the antioxidant activity of its phenolic compounds.

The existence of ACE-inhibitors in plant flavonoids opens up the possibility of the development of nature-based medication for
the treatment of hypertension and other cardiovascular diseases. Sowers et al. (2001) Stated that ACE-inhibitors are first line
antihypertensive therapy. Furthermore, Mohler et al. (2006) showed that ACE-inhibitors slow down the prevalence and the
progression of various renal diseases and related cardiovascular complications. One such plant which contains these flavonoids
is the Helianthus annuus or the common sunflower. Studies conducted both by Javed (2011) and Verma (2017) showed that
flavonoids were highly present in the sunflower leaves, followed by the roots and stems, with allelochemical production varying
due to stress. The presence of these flavonoids on the sunflower plant may prove the existence of competitive ACE-inhibitors
which may be sufficient in showing the same effects as synthetic-based ACE-inhibitors.

Based off of the information, it can be concluded that sunflowers having antihypertensive properties is possible. However,
further study is required to prove the validity and the safety of using it as an alternative form of medication to hypertension
and other cardiovascular related diseases.

Statement of the Purpose or Problem

The research aims to determine the concentration of ACE inhibitor peptides in Helianthus annuus (sunflowers) through in vivo
assay. Thus, it also aims to answer the following questions:
1. Does ACE inhibitor peptides in sunflowers exhibit same concentrations to that of commercially used and recognized
ACE inhibitor medications?
2. Can these concentrations prove the utilization of ACE inhibitor peptides derived from sunflower for pharmaceutical
use?
3. What part of the sunflower expresses high concentration or characterization of ACE inhibitor peptides?
Methods: Design and Sampling

The design to be used in this study is case analysis because it will involve reports of past cases and interaction with current
cases, documents and the like which will allow further exploration and understanding of the topic. The sampling method to be
used in this study will be nonprobability purposive sampling for the researchers will identify and select specific cases and
participants which are informative and germane to the study’s interest. This will involve healthcare providers, professionals,
chemists and the like who are experts in the field of pharmacological activities of the aforementioned drug.

Methods: Instrument and Procedures

The instruments that the researchers will use in this study are their electronic devices such as mobile phones and personal
computers in order to record data and look for cases, documents, and such containing information related to the presence of
ACE inhibitory peptides in plant sources, particularly in Helianthus annus (sunflower), hence document analysis. Further, the
researchers will use personal contacts and interactions through survey questionnaires, face-to-face interviews, and focus
groups with experts on the field of interest. Generally, the questions to be used are constructed in a way that will provide
answers for queries regarding the characterization and pharmacological properties of bioactive compounds present in
sunflowers. Once the researchers have gathered sufficient information, obtained data will be compared and analyzed in order
to generate new and novel knowledge that will contribute substantially to the area of interest. To suffice, the researchers will
also gather data from the internet, such as scientific journals and research papers, in order to obtain any lacking information
not attained from previous methods of data collection. As regards to the identification and concentration determination of ACE
inhibitor peptides present in the plant, plant parts will undergo phytochemical analysis to determine medicinally active
substances, particularly bioactive substances capable of characterizing ACE inhibitor peptides. After the identification peptides
present for each part, concentration of such will be determined through the use of a spectrophotometer. This is to determine
the amount of a known substance (concentration) through measurements of detected light intensity.

Data Analysis and Potential Ethical Issues

The data analysis for this study will be non-textual analysis through examination of the data gathered from the sample and
comparing it to other predetermined data obtained from cases, documents, and the like presented in tables and graphs in order
to generate or contribute new body of knowledge. Also, information from the related literatures will also be considered, such as
those indicating the presence of ACE inhibitor peptides in sunflowers.

Potential ethical issues that could arise in the research are the respect for intellectual property, anonymity and confidentiality.
With regard to these, gathering of other data will be from scientific journals and cases, and through the instruments to be used
by the researcher in the sampling method. In carrying out the research, the researchers will make sure that their work will be
conducted in a critical but careful manner. Also, they will make sure that the sources considered will be credible and knowledge-
laden, as well as keeping records of such sources.

References (APA 6th Edition Format)

• Ardehali, R., Perez, M., & Wang, P. J. (Eds.). (2013). A practical approach to cardiovascular medicine.
John Wiley & Sons. Retrieved from:
https://books.google.com.ph/books?hl=en&lr=&id=PQdqaeZCU78C&oi=fnd&pg=PT15&dq=).+A+practical
+approach+to+cardiovascular+medicine&ots=oxTwmOaGal&sig=Nj3i7f8Bnb_qmULVq2e1KjfUspo&redir_
esc=y#v=onepage&q=).%20A%20practical%20approach%20to%20cardiovascular%20medicine&f=false.

• Balasuriya, B. N., & Rupasinghe, H. V. (2011). Plant flavonoids as angiotensin converting enzyme
inhibitors in regulation of hypertension. Functional Foods in Health and Disease, 1(5), 172-188.

• Barnes, J., Anderson, L. A., Phillipson, J. D., & Newall, C. A. (2007). Herbal medicines. London:
pharmaceutical press. Retrieved from:
https://books.google.com.ph/books?id=cTySmwEACAAJ&dq=Herbal+medicines+barnes&hl=en&sa=X&v
ed=0ahUKEwj2wPyxoLflAhVCeXAKHQZfAzUQ6AEIKTAA.

• Daskaya-Dikmen, C., Yucetepe, A., Karbancioglu-Guler, F., Daskaya, H., & Ozcelik, B. (2017).
 Angiotensin-I-Converting Enzyme (ACE)-Inhibitory Peptides from Plants. Nutrients, 9(4), 316.
doi:10.3390/nu9040316.

• Goldstein, L. B. (Ed.). (2009). A primer on stroke prevention and treatment: an overview based on
AHA/ASA guidelines. Hobonken NJ: Wiley-Blackwell. Retrieved from:
https://books.google.com.ph/books?id=3W3MT9vryPYC&printsec=frontcover&dq=A+primer+on+stroke+p
revention+and+treatment:+an+overview+based+on++AHA/ASA+guidelines&hl=en&sa=X&ved=0ahUKEw
jl4K66oLflAhVHA4gKHZvNAY8Q6AEIKTAA#v=onepage&q=A%20primer%20on%20stroke%20prevention
%20and%20treatment%3A%20an%20overview%20based%20on%20%20AHA%2FASA%20guidelines&f
=false.

• Gonçalves, S., & Romano, A. (2017). Inhibitory Properties of Phenolic Compounds Against Enzymes
Linked with Human Diseases. Phenolic Compounds - Biological Activity. doi: 10.5772/66844.

• Grotewold, E. (Ed.). (2006). The science of flavonoids (pp. 239-267). New York: Springer. Retrieved from:
https://books.google.com.ph/books?id=B3yb7TNwsa0C&printsec=frontcover&dq=The+science+of+flavon
oids&hl=en&sa=X&ved=0ahUKEwiokbjRoLflAhWZZt4KHUKaAZkQ6AEIKTAA#v=onepage&q=The%20sci
ence%20of%20flavonoids&f=false.

• Guo, S., Ge, Y., & Na Jom, K. (2017). A review of phytochemistry, metabolite changes, and medicinal
uses of the common sunflower seed and sprouts (Helianthus annuus L.). Chemistry Central journal, 11(1),
95. doi:10.1186/s13065-017-0328-7.

• Iwaniak, A. , Minkiewicz, P. and Darewicz, M. (2014). Food‐Originating ACE Inhibitors, Including

Antihypertensive Peptides, as Preventive Food Components in Blood Pressure Reduction. Comprehensive
Reviews in Food Science and Food Safety, 13: 114-134. doi:10.1111/1541-4337.12051.

• Javed, K. (2011). Quantification of alkaloids, phenols and flavonoids in sunflower (Helianthus annuus L.).
African Journal of Biotechnology, 10(16), 3149–3151. doi: 10.5897/ajb09.1270.

• Justino, G. C. (Ed.). (2017). Flavonoids - From Biosynthesis to Human Health. Rijeka: InTech. Retrieved
from:https://books.google.com.ph/books?hl=en&lr=&id=1dCPDwAAQBAJ&oi=fnd&pg=PR11&dq=Justino,
+G.+C.+(Ed.).+(2017).+Flavonoids+-
+From+Biosynthesis+to+Human+Health.+Rijeka:+InTech.&ots=xGlQnCQKc9&sig=XBPHDr6gjVsyMc3x
Wf4qyEoa26o&redir_esc=y#v=onepage&q&f=false.

• Karamać, M., Kosińska, A., Estrella, I., Hernández, T., & Duenas, M. (2012). Antioxidant activity of
phenolic compounds identified in sunflower seeds. European Food Research and Technology, 235(2), 221-
230.

• Lin, D., Xiao, M., Zhao, J., Li, Z., Xing, B., Li, X., ... & Chen, H. (2016). An overview of plant phenolic
compounds and their importance in human nutrition and management of type 2 diabetes. Molecules, 21(10),
1374.

• Matchar, D. B., McCrory, D. C., Orlando, L. A., Patel, M. R., Patel, U. D., Patwardhan, M. B., ... & Gray,
R. N. (2008). Systematic review: comparative effectiveness of angiotensin-converting enzyme inhibitors
and angiotensin II receptor blockers for treating essential hypertension. Annals of internal medicine, 148(1),
16-29.

• Mohler, E. R., & Townsend, R. R. (Eds.). (2006). Advanced therapy in hypertension and vascular
disease. PMPH-USA. Retrieved from: https://books.google.com.ph/books?id=sCgURxhCJ-
8C&printsec=frontcover&dq=Advanced+therapy+in+hypertension+and+vascular++disease&hl=en&sa=X&
ved=0ahUKEwjp--LMm7flAhVJeXAKHdZFAaAQ6AEIKTAA#v=onepage&q&f=false.
• Packer, L., Wachtel-Galor, S., Ong, C. N., & Halliwell, B. (Eds.). (2004). Herbal and traditional medicine:
biomolecular and clinical aspects. CRC Press. Retrieved from:
https://books.google.com.ph/books?id=7WDgesSflScC&printsec=frontcover&dq=Herbal+and+traditional+
medicine:++biomolecular+and+clinical+aspects&hl=en&sa=X&ved=0ahUKEwictZ3goLflAhUC7WEKHe_t
CeMQ6AEIKTAA#v=onepage&q=Herbal%20and%20traditional%20medicine%3A%20%20biomolecular%
20and%20clinical%20aspects&f=false.

• Verma, Devshree & Sahu, Meena & Haris, Kk. (2017). Phytochemical analysis of helianthus annus lin.,
(Angiosperms:Asteraceae). World journal of pharmacy and pharmaceutical sciences. 6. 825-846. doi:
10.20959/wjpps20173-8725.

• Vermerris, W., & Nicholson, R. (2007). Phenolic compound biochemistry. Springer Science & Business
Media. Retrieved from:
https://books.google.com.ph/books?id=uLzdv8fsRxYC&printsec=frontcover&dq=Phenolic+compound+bio
chemistry&hl=en&sa=X&ved=0ahUKEwiQ9u7poLflAhWHBIgKHeTpDwUQ6AEIKTAA#v=onepage&q=Phe
nolic%20compound%20biochemistry&f=false.
• Sowers, J. R., Epstein, M., & Frohlich, E. D. (2001). Diabetes, hypertension, and cardiovascular disease:
an update. Hypertension, 37(4), 1053-1059.

• Stam, J. (2016). "Flavonoids, catching up with ACE-inhibitors and angiotensin receptor blockers to
treat Diabetic Nephropathy?". Retrieved from
http://fse.studenttheses.ub.rug.nl/14323/1/LST_BC_2016_JStam.pdf.