Republic of the Philippines

Department of Education
REGION III – CENTRAL LUZON
SCHOOLS DIVISION OF BULACAN
SAN MIGUEL NATIONAL HIGH SCHOOL

PRELIMINARY SCREENING OF MUNG BEAN PODS GROWN

A Research Plan Presented to the Faculty of Junior High School Department

San Miguel National High School
San Miguel, Bulacan

NICA ROSE V. ISIDRO

MA. CAZANDRA AYEZA R. LALO

Researchers

MRS. FARRAH M. RAMOS

Research Teacher

July 2023

San Miguel National High School

Scuala St., San Juan, San Miguel, Bulacan
Telephone Number: (044) 327-1123 / (044) 327-1104
 Preliminary Screening of Mung Bean Pods Grown

Mung bean (Vigna radiata L.) Wilczek) is an important grain legume grown in the

tropical and subtropical regions of the world. It is one of the important sources of protein for

both man and domestic animals. Another important feature of mung bean is its ability to fix

atmospheric nitrogen in symbiosis with nodule forming rhizobium bacteria. Nutrient stress

soil was defined soil contains nutrient below the critical level (FRG, 2005). 1

Mung bean productivity is constrained by biotic and abiotic factors. Mung beans are

high in nutrients and antioxidants, which may provide health benefits. In fact, they may

protect against heat stroke, aid digestive health, promote weight loss and lower “bad” LDL

cholesterol, blood pressure and blood sugar levels. 2

Legume, also called pod, fruit of plants in the pea family. Most legumes are dehiscent

fruits that release their seeds by splitting open along two seams, though some do not naturally

open. 3 The fruits come in a variety of sizes and shapes; many, however, are long and narrow

and bear their seeds in a single line. The largest legumes are borne by the monkey ladder

(Entada gigas) and can reach up to 2 meters in length. At maturity, legume fruits are usually

dry and papery or hard and woody; the legumes of certain food crops, such as snow peas

(variety of Pisum sativum), edamame (Glycine max), and green beans (Phaseolus vulgaris),

are harvested while still green and fleshy. 4

1
Ziska, Lewis H., Robert Palowsky, and Danielle R Reed. “A Quantitative and Qualitative Assessment of Mung
Bean (Vigna Mungo (L.) Wilczek) Seed in Response to Elevated Atmospheric Carbon Dioxide: Potential
Changes in Fatty Acid Composition.” Journal of the Science of Food and Agriculture 87, no. 5 (April 15, 2007):
920–23. https://doi.org/10.1002/jsfa.2818.

2
Talib WH, Mahmod AI, Awajan D, Hamed RA, Al-Yasari IH. Immunomodulatory, Anticancer, and
Antimicrobial Effects of Rice Bran Grown in Iraq: An In Vitro and In Vivo Study. Pharmaceuticals (Basel).
2022 Dec 1;15(12):1502. doi: 10.3390/ph15121502. PMID: 36558953; PMCID: PMC9782048.

3
Wikipedia contributors. “Mung Bean.” Wikipedia, March 11, 2023.
https://en.m.wikipedia.org/wiki/Mung_bean.

4
Singh, B., Singh, N., Thakur, S. et al. Ultrasound assisted extraction of polyphenols and their distribution in
whole mung bean, hull and cotyledon. J Food Sci Technol 54, 921–932 (2017). https://doi.org/10.1007/s13197-
 Consequently, mung bean requires hot and dry climate. Cloudy weather, continuous

and heavy rains adversely affect the flowering and podding in mung bean, causing low yields.

Mung bean can be grown on well-drained loamy sand to sandy loam soils. The crop is

sensitive to alkaline, saline or waterlogged soil. Being a short duration crop, mung bean is

cultivated in all three seasons in different parts of country as a pure crop as well as an

associate crop in various cropping systems. 5 However, mung bean has a well-developed root

system. The lateral roots are many and slender, with root nodules grown. Stems are much

branched, sometimes twining at the tips. Young stems are purple or green, and mature stems

are grayish yellow or brown. Mung bean is one of the a few legumes that is composed of

unique macro and micronutrients as well as numerous phytochemicals, including phenolic

acids, flavonoids, sterols, terpenes, alkaloids, and organic acids. 6 The majority of these

products are water-soluble or biodegradable phyto-films, mainly composed of biopolymers

(proteins, polysaccharides, and lipids) and hydrocolloids (carrageenan) gums). Furthermore,

utilization of plant extracts combined with the phyto-film has shown good effects, as they

could increase the stability of the stored product and could provide a barrier against spoilage

and pathogenic microorganisms. They could also control the biochemical degradation in food

induced by enzymes. 7

016-2356-z

5
Antioxidant and Antimicrobial Properties of Mung Bean Phyto-Film Combined with Longkong Pericarp
Extract and Sonication - ProQuest,” n.d.
https://search.proquest.com/openview/93705cfeadeac179057c8dc3c8413d7b/1?pq-
origsite=gscholar&cbl=2032363.

6
Das, A., Parida, S.K. Advances in biotechnological applications in three important food legumes. Plant
Biotechnol Rep 8, 83–99 (2014). https://doi.org/10.1007/s11816-013-0299-7

7
Diao, Jingjing, Chi Zhiping, Zengwang Guo, and Liping Zhang. “Mung Bean Protein Hydrolysate Modulates
the Immune Response Through NF‐κB Pathway in Lipopolysaccharide‐Stimulated RAW 264.7 Macrophages.”
Journal of Food Science 84, no. 9 (September 1, 2019): 2652–57. https://doi.org/10.1111/1750-3841.14691.
 A cytotoxic compound can cause cell damage or death, either through necrosis or

apoptosis. Some substances are more cytotoxic than others and researchers aim to measure a

chemical's cytotoxicity levels to ensure that it is not harmful and/or fatal to patients. Treating

cells with the cytotoxic compound can result in a variety of cell fates. Cytotoxicity studies are

a useful initial step in determining the potential toxicity of a test substance, including plant

extracts or biologically active compounds isolated from plants. 8 The purpose of this research

is to evaluate the cytotoxicity and the Preliminary screening of mung bean pods grown. 9 A

preliminary screening is conducted to determine if a proposed project might be cause for

public concern or might have a significant adverse impact on the environment. The

preliminary screening will determine whether the project should proceed without

environmental assessment or if it will be referred to environmental assessment. 10

8
Brahmer, Julie R., Karen L. Reckamp, Paul Baas, Lucio Crinò, Wilfried Eberhardt, Elena Poddubskaya, Scott
J. Antonia, et al. “Nivolumab versus Docetaxel in Advanced Squamous-Cell Non–Small-Cell Lung Cancer.”
The New England Journal of Medicine 373, no. 2 (July 8, 2015): 123–35.
https://doi.org/10.1056/nejmoa1504627.

9
Chacon, E, Daniel Acosta, and John J. Lemasters. “Primary Cultures of Cardiac Myocytes as In Vitro Models
for Pharmacological and Toxicological Assessments.” Elsevier EBooks, January 1, 1996, 209–23.
https://doi.org/10.1016/b978-012163390-5.50010-7.

10
Hobley L, Ostrowski A, Rao FV, Bromley KM, Porter M, Prescott AR, MacPhee CE, van Aalten DM,
Stanley-Wall NR. BslA is a self-assembling bacterial hydrophobin that coats the Bacillus subtilis biofilm. Proc
Natl Acad Sci U S A. 2013 Aug 13;110(33):13600-5. doi:
Statement of the Problem

1. How does mung bean pods extracts in cytotoxic?

2. How does mung bean pods extracts in phytochemical screening?

3. How does mung bean pods extracts in antioxidant assay?

4. How does mung bean pods extracts in antibacterial assay?

5. How does mung bean pods extracts in brine shrimp lethality assay?

Hypothesis

The mung bean pods has no significant effect.

Significance of the Study

Cancer patients. This study is will help cancer patients to fight cancer by multiple

mechanisms including apoptosis induction, angiogenesis inhibition, and immune system

activation and preventing cancer and stimulating the immune system.

Community. It will benefit the community to reduce cases of cancer. It will also

prevent bacteria may infect humans and animals.

Mung bean cultivators. This study will benefit the mung bean cultivators because

they can sell the Mung Bean pods instead of throwing them away.

Scopes and Delimitations

This project is limited to use of mung bean pods as a cytotoxic. The researchers will

focus on evaluating the potential of mung bean as a cytotoxic by identifying its

phytochemical screening, antioxidant assay, antibacterial assay, and brine shrimp lethality

assay. The collection of mung bean will take place at Salangan, San Miguel, Bulacan.
Meanwhile, the phytochemical screening, antioxidant assay, and antibacterial assay, brine

shrimp lethality assay will take place at the Science Laboratory in Saint Mary University

located at Ponce, Bayombong, Nueva Vizcaya.

Definition of Terms

Preliminary screenings. The gathering of initial information to be used in

determining a person's need for assessment or for referral.

Mung bean pods. The mung bean (Vigna radiata L.) is one of the most important

edible legume crops, grown on more than 6 million worldwide and consumed by most

households in Asia.

Phytochemical screening. It is another name for this process. These extracts are

made from plant samples rich in secondary metabolites such as the leaves and stems, roots,

and bark of the plants studied. The secondary metabolites, such as alkaloids, terpenes, and

flavonoids, are then examined in the plant extracts.

Antimicrobial assays. They are important tools to test and screen the inhibitory

effects of myriad compounds against microorganisms before establishing their inhibitory

spectra (broad vs. narrow).

Brine shrimp lethality bioassay. It is a simple high throughput cytotoxicity test of

bioactive chemicals. It is based on the killing ability of test compounds on a simple

zoological organism-brine shrimp (Artemia salina).

Methodology

General Process:

Mung Beans Phytochemical Antioxidant

Pods Extract Screening Assay

Brine Shrimp Antibacterial Assay

Waste Disposal
Lethality Assay

Figure 1. Flow Chart of Activities

Presented in Figure 1 is the complete procedure of the study. The researchers will

start by gathering materials needed for the study followed by the extracting mung bean pods.

Then, phytochemical screening and antioxidant assay will be next. After that, antibacterial

assay detects and quantifies the cellular events associated with programmed cell death. The

last assay is brine shrimp lethality bioassay that have different concentrations. After the

experimentation, the materials will be disposed properly and the data will be gathered and

results will be interpreted.

Mung Bean Pods Extract

Based on previous studies, they are protein-rich and contain many essential amino

acids we need to maintain our health. We can easily digest mung bean protein and isolates,

too. Mung bean pods was provided by local mills in (location). Mung bean will be

compressed place beaker, add 80% ethanol or water, solid-liquid ratio 1:10,50 ℃ are stirred
and extracted 120 minutes, after filtration, sucking filtration, revolve the inspissation

contracting, get the mung bean extracting solution. 11

Phytochemicals and Antioxidant Activities

Films were extracted prior to phytochemical and antioxidant activities, 1 g of film

samples was homogenized with 15 mL of 80% ethanol, and after that, the homogenate was

centrifuged at 6000× g for 15 min. After centrifugation, a clear supernatant was collected and

used for the following analysis. For total phenolic content [15], 100 µL of WACV was added

into the test tube and followed by the addition of 8.4 mL of distilled water and 500 µL of

Folin–Ciocalteu reagent. After that, 1 mL of sodium carbonate (20%) was added and mixed

well. Then, the reaction mixture was incubated at room temperatures for 30 min, and then it

was measured under a spectrophotometer at 720 nm. The absorbance value of the sample was

compared with the calibration curve made of gallic acid [10–100 µg/ mL; R2 value = 0.9970;

p < 0.0001]. The results were expressed as µg gallic acid equivalence (GAE)/mL. For total

flavonoid content [15], 250 µL of WACV was added into the test tube, and then 2.72 mL of

30% ethanol and 120 µL of 0.5 mol/L sodium nitrite were added and mixed well and kept for

5 min. Then, 120 µL of 0.3 mol/L aluminum chloride was added to the mixture, followed by

adding 800 µL of 1 mol/L sodium hydroxide, and mixed well. After that, the reaction mixture

was measured at 510 nm using a spectrophotometer. The absorbance value of the sample was

compared with the calibration curve made of catechin [10–100 µg/ mL; R2 value = 0.9960; p

11
Chacon, E, Daniel Acosta, and John J. Lemasters. “Primary Cultures of Cardiac Myocytes as In Vitro Models
for Pharmacological and Toxicological Assessments.” Elsevier EBooks, January 1, 1996, 209–23.
https://doi.org/10.1016/b978-012163390-5.50010-7..
Antibacterial Assay

The antimicrobial activity of the films was tested against the two groups of microorganisms,

which are Gram-positive and Gram-negative. After that, 1 mL of Gram, bacteria was added

and mixed. Then, the tubes with a medium that contained L. monocytogens, S. aureus, and B.

subtilis were transferred to the shaker and shaken at 200 rpm at room temperature, whereas

the tubes that contained C. jejuni, E. coli, and P. aeruginosa were incubated. After incubation,

1 mL of microbial aliquots from each tube was collected and tested for its optical density

(OD) at 600 nm. Then, Gram-positive bacteria was plated on the Mueller-Hinton agar plates

and incubated for hours and degree celsius. Whereas Gram-negative bacteria was plated on

Campylobacter agar plates and other bacteria in this group was plated on MacConkey agar

and incubated for hours and degree celsius. After incubation, the plates for the count for

colonies and the results were expressed as log CFU/mL.

BSLA Assay

³⁰ Chacon, E, Daniel Acosta, and John J. Lemasters. “Primary Cultures of Cardiac Myocytes
as In Vitro Models for Pharmacological and Toxicological Assessments.” Elsevier EBooks,
January 1, 1996, 209–23. https://doi.org/10.1016/b978-012163390-5.50010-7..

Brine shrimp lethality bioassay is a simple, high throughput cytotoxicity test of bioactive

chemicals. In this procedure, One important aspect of this assay is that the solvent used in

this assay may give false positive signals due to the toxicity of the solvent itself. This is a

very significant study. That dimethyl sulfoxide (DMSO) is a safer solvent in brine shrimp

lethality bioassay and the maximum working concentration of solvents such as DMSO,

methanol and Tween 20 was suggested. Tween 20 showed the most cytotoxicity in brine

shrimp lethality bioassay. Tween 20 is a detergent, which can help to dissolve natural
products contains oil. Detergents have long alkyl chains and water-soluble functional groups.

Hence they are water-soluble and are able to dissolve hydrophobic compounds with their

alkyl chains.

Waste Disposal
After conducting this study, the chemicals, reagents, and solvents used will be

labelled and disposed into their organic and inorganic containers. Solid wastes will be

collected in a waste container and will be disposed properly 36.

³⁴ Takahashi, Hidenori, Yoko Nomura, Junko Nishida, Yujiro Fujino, Yasuo Yanagi, and
Hidetoshi
Kawashima. “Vascular Endothelial Growth Factor (VEGF) Concentration Is Underestimated
by EnzymeLinked Immunosorbent Assay in the Presence of Anti-VEGF Drugs.”
Investigative Ophthalmology & Visual Science 57, no. 2 (February 1, 2016): 462.
https://doi.org/10.1167/iovs.15-18245.
, Steven A., Michael M. Halford, Sally Roufail, Carol Caesar, and Marc G. Achen. “A
Simple
Bioassay for the Evaluation of Vascular Endothelial Growth Factors.” Journal of Visualized
Experiments, no. 109 (March 15, 2016). https://doi.org/10.3791/53867.
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