Insecticidal Activity of Mustasa (Brassica juncea) and Sampaguita Flower

(Jasminum sambac) Extracts Against Cockroaches (Periplaneta americana)

National Capital Region

Malabon National High School

M. Naval St., Hulong Duhat, Malabon City

Bautista, Joruss Martin R.

Bello, Andrew Q.

Uy, Paolo G.

Ms. Mylene J. Aguinaldo

Project Adviser

Life Science – Team Category

Insecticidal Activity of Mustasa (Brassica juncea) and Sampaguita Flower
(Jasminum sambac) Extracts Against Cockroach (Periplaneta americana)

Bautista, Joruss Martin R., Bello, Andrew Q., Uy, Paolo G.

Mrs. Mylene J. Aguinaldo
Malabon National High School

ABSTRACT
Insects threaten the safety of humans, plants, and animals. However, the toxic
substance which can be found in insecticides poses a problem to health as well. The
goal of this study is to test whether Mustasa(Brassica juncea) leaves and
Sampaguita(Jasminum sambac) flowers can become a safe, affordable, and effective
natural alternative to commercial insecticide against cockroaches found in the market.
The extract was obtained from leaves and flowers of the plant samples. The extracts
were then mixed into several mixtures with different concentrations each and tested
against cockroaches. These concentrations are (A) 50%, 50%, (B) 75%, 25%, (C)
100%, 0% and the control group with a commercial insecticide. Each concentration has
three samples groups. There were two tests carried out: Survival Test, and Time to
Eliminate. Replicate C had the highest percentage in the Survival Test. The F- value is
4.61 which is higher than tabular value of 4.06 with a 0.05 level of confidence. Control
group was the fastest in eliminating cockroaches for the Time to Eliminate test. Both
plants proved to be capable of eliminating cockroaches due to properties which they
contain. Study concludes that both can be used as a natural alternative to insecticide.

Keywords: cockroach, insecticide, survival rate, time to eliminate

INTRODUCTION

Periplaneta americana or American cockroach can measure up to 1.5 inches with

a color of reddish brown. These are the most common cockroaches in the Philippines.

Pests which destroy food supply in restaurants and other facilities must be eliminated.

Spreading diseases is just one of the most dangerous aspects by having an infestation in

homes. By secreting bacteria which emit diseases, communities are in danger of sickness

outbreaks. This happens when the cockroaches get in contact with food or skin and

infects those in the vicinity. This can cause food to be unsafe for consumption. Being safe

from these pests is a way to stay safe from the diseases which are carried by them.

Insecticide is used as a chemical agent to kill off insects. It is used in public health

that can fight off vector of disease, such as cockroaches that carries different kinds of

diseases such as thypoid, polio, and dysentery which can be dangerous for human health.

Cockroach control are widely used around the world including the use of insecticides,

however, insecticides contain chemicals which makes it toxic and bad for the

environment. Baygon is one of these examples, this commercially available insecticide

contains dangerous chemical such as propoxur, organophosphorus chlorpyrifos and

pyrethroids.

The plant Brassica juncea, locally known as Mustasa or Mustard, is a pantropic

vegetable which is introduced from Asia and is known for agricultural and horticultural

crops, specifically here in the Philippines. Mustasa is used mainly for its immature, but

fully expanded tender leaves and is used as a main ingredient for soup and stir-fried

dishes. The Brassica family is known to contain rubefacient. The flower of Sampaguita
plant (Jasminum sambac) ìs a sweetly scented tropical flower belonging to the wide

genus of Jasmines which are native, to tropical and warm temperate regions of Eurasia,

Australia and Oceania. It is normally used for teas, religious offerings, and symbolizing

divine hope and purity. Though commonly used for ornamental purposes, sampaguita’s

active components such as benzyl acetate, benzyl alcohol, linalool, and methyl

anthranilate can be assessed as for an insecticidal purpose that is safe and non-toxic to

humans and environment. This study focuses on utilizing the potential insecticidal

activity of Brassica juncea leaves and Jasminum sambac flower extracts against

Periplaneta americana.

This study focuses on Mustasa (Brassica juncea) leaves and Sampaguita

(Jasminum sambac) flower extracts as an insecticide. The insecticidal activity was

determined through testing several concentrations on Periplaneta americana. The

Mustasa leaves and Sampaguita flower are bought from Hulong Duhat Public Market,

Malabon City. The active chemical composition of the leaves and flowers extracts were

determined and included in the study The Mustasa leaves and Sampaguita flower were

extracted with 95% technical grade ethyl alcohol.

Insecticides have been used by the people and sold in the industry for a very long

time to terminate the threat of cockroaches, but those kinds of insecticides poses a

problem of causing lethal damage to humans and also the environment. This new study

could help the community by producing a new insecticide that is non-toxic for humans

and has the ability to reduce the problems caused by cockroaches.

 MATERIALS AND METHODS

Research Design

Table 1 Proportion of Components in Each Concentration

Concentration Amount of Amount of
Solute (g) Solvent(mL)
Mustasa Sampaguita Ethanol
Flower
A.(50%;50%) 100 100 200
B.(75%;25%) 150 50 200
C.(100%;0) 200 0 200

Table 1 shows the percentage and mass per concentration

from the initial distribution of Mustasa (400g) and Sampaguita

Flower (400g) extracts of the components used in this insecticide.

Concentration A contained both 50% of Mustasa (100g) and 50%

Sampaguita Flower (400g). In Concentration B, 75% of Mustasa

(150g) and 25% of Sampaguita Flower (50g) were used. Finally in

Concentration C, 100% of Mustasa (200g) were used. All solutes

were infused with 200 mL of 95% technical grade ethyl alcohol.

Table 2 Distribution of Cockroaches into three trials and three treatments

Number of cockroaches per Trial
Total Number of
Treatment Trial 1 Trial 2 Trial 3 Cockroaches
A.(50%;50%) 3 3 3 9

B.(75%;25%) 3 3 3 9

B.(75%;25%) 3 3 3 9

Control 3 3 3 9

Total 36
Collection and Preparation of the Extracts

Four (4) kilograms of Mustasa leaves were bought from Hulong Duhat Public

Market, Malabon, Philippines while two (2) kilograms of Sampaguita were bought from a

local market in Maysilo, Malabon, Philippines. The species of the plant samples were

verified in the Bureau of Plant Industry in San Andres street, Sta.Cruz, Manila,

Philippines. One (1) liter of 95% technical grade ethyl alcohol was needed for the

extraction. The Mustasa leaves and Sampaguita Flowers were washed with running water

to remove dirt and other impurities. Separately, the plant samples were put in the dry

oven with a temperature of 100 degrees Celsius over the course of 45 minutes. The dried

Mustasa leaves were cut into thin pieces. Both plant samples were kept safe in a ziplock

bag.

Phytochemical Analysis

The two test tubes were labeled with respective plant extracts after the

procedures. 10mL of Mustasa extract and Sampaguita flower extracts were placed

separately on the proposed test tubes. 12 M hydrochloric acid was added to the extracts.

The mixture was warmed in a water bath for 15-60 minutes. Changes in color were

observed and recorded. The gradual development of a strong red or violet color was

indicated with the presence of flavonoids in both extracts. Bate-smith and Metcalf

Method were used for the analysis. The procedure was conducted at the Institute of

Pharmaceutical Sciences at the University of the Philippines.

Preparation of Concentrations

Four bottles with the measurement of four liters were bought and prepared to be

the container for the cockroaches. These were cut and the top portion was used.

Cheesecloth was soaked in in different concentrations of Mustasa-Sampaguita extract.

Then, air drying those in shaded area is the next step. After three hours, these were taped

under three bottles. Replicate A has 50% Mustasa and 50% Sampaguita concentration.

Replicate B has 75% Mustasa and 25% Sampaguita concentration. Replicate C has 100%

Mustasa concentration. The process is called feedant method.

Rearing of Cockroaches

The American cockroaches (Periplaneta americana) were bought at Arranque

Market, Sta. Cruz, Manila and were placed in the prepared habitat comprising of a

tupperware case, soil for the base and logs to reduce the stress of cockroaches. The

process was carefully done by hand to avoid injuring them. Gloves were used for safety

and hygiene purposes. Test subjects were fed twice a day with sweets, meats, sugar

solution and leftover foods. This routine is to be followed until the experiment is

conducted.

Application

A total of four (4) habitats were used to house the cockroaches. Three of these

constructed habitats were used for the experimental group, and one is allocated for the

control group. Cockroaches were placed in containers where the treatment will be done.

Each container will contain ten (10) cockroaches. Feedant method was used for the

application of extracts.
Testing

The effectiveness of the experimental insecticide was determined by conducting

two tests: Survival Test and Time to Eliminate. The test to be conducted will involve

the observation of alive and knocked down cockroaches in Survival Test and the time it

takes to knock down cockroaches will be recorded in the Time to Eliminate.

Survival Test

The survival test was conducted to the four (4) containers, one which contained

the commercial insecticide. The effect of the insecticide was assessed by counting the

number of cockroaches alive at the end of the testing. The average of the set of three

replicates reported the percentage of survival rate.

The survival rate of the cockroaches of each concentration will be computed by the

following concentration:

Survival Rate (%) = A/P x100

Where; A = alive cockroaches

P = total population of cockroaches used

Time to Eliminate

In determining the speed of which concentration eliminates the test subjects the

fastest, the number of knockdown cockroaches was counted in a logarithmic pattern; 2, 4,

8, 16 hours. This was done for all the trials in each concentration.
 Statistical Treatment
The mean was obtained through adding of the results per concentration and

dividing it to the number of trials. This is used to determine the average knockdown and

survival rate of the cockroaches.

The percentage of survival was obtained by dividing the number of alive

cockroaches by the total population of cockroaches used in the experiment. The answer

was multiplied by 100.

One way ANOVA (Analysis of Variance) was used to determine the effectiveness

of both components based on varying concentration. The researchers used Software

Package for Statistical Analysis (SPSS) to compute the analysis of variance and to

construct the numeric data. The use of bar graphs determined the morphological changes

found in each set of cockroaches within the interval. A 1 significance was used to

evaluate if the hypothesis is effective or not. The data from all replicates were combined.

The overall survival rate and time to eliminate was observed and analyzed.

RESULTS

This study determines the effectiveness of Mustasa and Sampaguita Flower

extracts against cockroaches as prepared with the following tables, graphs and figures.

Survival Test

The researchers conducted the survival test over the set of three trials per

concentration. Each trial contained three (3) cockroaches.

 Table 3 Results of Survival Test
No. of Alive Cockroaches
Concentration Mean Percentage Rank
Trial 1 Trial 2 Trial 3
A.(50%;50%) 0 0 1 0.33 11.11% 3.5

B.(75%;25%) 2 1 2 1.67 55.56% 2

C.(100%;0) 2 3 1 2 66.67% 1

Control 0 0 1 0.33 11.11% 3.5

80
Average Percentage of Survival Rate
70
Percentage of Survival

60

50

40

30

20

10

0
A B Concentrations C Control

Graph 1 Percentage of Survival Rate

F-Test For Survival Test

Table 4 Summary of Results of Survival Test
 Table 5 One-way ANOVA for Survival Test

There is a significant difference on the number of alive cockroaches observed for the

survival test, made by the four different concentrations of submitted samples.

Time to Eliminate

The Time to Eliminate test was observed and recorded in a logarithmic pattern

(1, 2, 4, 8, 16 hours).

Table 9 Time to Eliminate (1 Hour)

Trials
Treatments Mean
1 2 3
A.(50%;50%) 1 0 0 0.33
B.(75%;25%) 0 0 0 0
C.(100%;0) 1 1 0 0.67
Control 0 0 0 0

Table 10 Time to Eliminate (2 Hours)

Trials
Treatments Mean
1 2 3
A.(50%;50%) 1 0 0 0.33
B.(75%;25%) 0 0 0 0
C.(100%;0) 0 0 0 0
Control 0 1 1 0.67
 Table 11 Time to Eliminate (4 Hours)

Trials
Treatments Mean
1 2 3
A.(50%;50%) 0 1 1 0.67
B.(75%;25%) 0 1 0 0.33
C.(100%;0) 0 0 1 0.33
Control 0 1 1 0.67

Table 12 Time to Eliminate (8 Hours)

Trials
Treatments Mean
1 2 3
A.(50%;50%) 2 1 1 1.33
B.(75%;25%) 0 1 0 0.33
C.(100%;0) 1 0 1 0.67
Control 2 1 1 1.33
Table 13 Time to Eliminate (16 Hours)

Trials
Treatments Mean
1 2 3
A.(50%;50%) 0 1 1 0.67
B.(75%;25%) 0 1 0 0.33
C.(100%;0) 0 0 1 0.33
Control 1 0 0 0.33

DISCUSSION

Phytochemical Analysis

In a preliminary study conducted by Duy N., et.al (2017) in a phytochemical screening of

Mustasa (Brassica juncea), the results yield the presence of alkaloids, flavonoids, tannins,

saponins, terpenoids and other phenolic compounds. The same goes for the previous study of

Kunkachan P., (2012) in a phytochemical screening of Sampaguita (Jasminum sambac), the

results showed the presence of carbohydrates, flavonoids, tannins and phenolic compounds,

proteins and amino acids, steroids, glycosides and saponins.

Survival Test

The amount of surviving cockroaches was computed. A higher survival rate

means that more cockroaches survived in a certain concentration. Concentration C had

the highest percentage of 66.67% This means that there is a high chance that cockroaches

will survive in this. Concentration A and control group has the lowest percentage of

Survival Rate with 11.12%. This means that cockroaches are less likely to survive when

exposed to these.

According to a study by (Santos, et.al), the survival rate of the cockroaches in

each concentration was computed. The higher the percentage of survival rate, the higher

the number of cockroaches that survived. Meaning, concentration B with the percentage

of survival rate of 26.6% had the highest chance of the cockroach survival.

Concentrations D and E, both having a percentage of survival rate of 6.6% indicated a

low chance of cockroach survival. To sum it up, the experimental group yielded positive

results in contrast to the control which had the survival percentage of 13.3%

Statistical Analysis for Survival Rate

The difference of Survival Rate between concentrations is presented. The F- value

is 4.61, which is higher than the tabular value of 4.06 with a level of confidence of

0.05. There is significant difference between the surviving cockroaches made by the four

concentrations. The highest Survival Rate belongs to Concentration C with a percentage

of 66.67, while Concentration A and Control Group has the lowest percentage of 11.12%.
Time to Eliminate

The speed of the elimination of cockroaches varies between each concentration.

The fastest in knocking out the most cockroaches for one hour is Concentration C. The

fastest in knocking out the most cockroaches for two hours is the control group. The

fastest in knocking out the most cockroaches on four and eight hours are Concentration A

and the control group. The fastest in knocking out the most cockroaches within 16 hours

is Concentration A.

According to a study by (Santos, et.al), the test indicates the time the extracts took

place to eliminate cockroaches for five treatments. Each concentration involved five

cockroaches. Concentration D and E ranked first with the fastest average time of 15

minutes. Control group ranked second that took 20 minutes to eradicate the cockroaches.

Concentration C ranked third with the average time of 23 minutes in time to eradicate

cockroaches. Meanwhile, Concentration A and B had the longest time. Moreover, the

experimental group eliminated most of the cockroaches faster than the control.

CONCLUSION

The researchers concluded that the Mustasa and Sampaguita Flower extracts can

be used as a biological insecticide against cockroaches.

RECOMMENDATIONS

The proponents would like to recommend to other future researchers to figure out

different components of locally available plants that can be utilized as a biological

insecticide and test it on cockroaches and other insects that causes problems to the

community.
ACKNOWLEDGEMENT

The study cannot be done with only the capability of three students. The

proponents received help and support from others. With all gratefulness and appreciation,

we would like to thank:

God, for His never-ending blessings and guidance throughout the project and for

also giving us this wonderful opportunity.

Dr. Ma. Victoria De Gulan, Principal of Malabon National High School, for the

wisdom and knowledge she shared to us about the study.

Mrs. Mylene J. Aguinaldo, our research adviser, for all the support, morals and

advices she gave to us along the accomplishment of the project

. Family, teachers, classmates, friends who never left our side and continued to

support us physically, mentally, emotionally, and financially.

And lastly, the schools and universities who allowed us to use their facilities and

laboratories in conducting the experiment. This includes but is not limited to; Malabon

National High School (MNHS) and University of Santo Thomas (UST)

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APPENDICES

Appendix A

Collection and Preparation for Extraction

Gathering of Mustasa Plant and Sampaguita Flower

- Oven Drying of Mustasa Plant and

Samapaguita Flower
Appendix B

Preparation of Different Concentrations

Appendix C
Appendix D

Plant Identification from Bureau of Plant Industry

Appendix E

Plant Identification from Bureau of Plant Industry

Appendix F

Plant Identification from Bureau of Plant Industry

Appendix G

Plant Identification from Bureau of Plant Industry

Appendix H

Materials that were borrowed from the laboratory that will be used for the Maceration
Appendix I

An email from the University of Santo Thomas (UST) about the Rotary Evaporation

The crude extracts from the UST

Appendix J

The board for the school based Science Investigatory Project Compitition
Appendix K

The method used for application and testing which is called the Feedant Method
Appendix L
Appendix M