The Efficacy of Pansit-pansitan (Peperomia pellucida Linn.

) Wound

Solutions on BALB/c Mice1

Researchers

Griño, Sophia Liza S.

Montilla, Maria Antonette S.

Portento, Christine Joy C.

Research Adviser

Mr. Fermin A. Clorado Jr.

Consultant

Maria Dalisay G. Maligalig, Ph. D

Animal Physiologist and Endocrinologist

San Pablo City National High School

A Science Investigatory Project Submitted for the Fulfilment of Requirements in Research II-A for the
1

School Year 2015-2016

 TABLE OF CONTENTS

TABLE OF CONTENTS i

ABSTRACT ii

ACKNOWLEDGEMENT iii

INTRODUCTION

Background of the Study 1

Statement of the Objectives 2

Hypotheses 2

Significance of the Study 2

Scope and Limitations 3

Definition of Terms 4

REVIEW OF RELATED LITERATURE 5-8

METHODOLOGY 9-11

RESULTS AND DISCUSSIONS 12-15

SUMMARY AND CONCLUSIONS 16

RECOMMENDATIONS 17

BIBLIOGRAPHY 18

APPENDICES

Appendix A 19

Appendix B 20-22

Appendix C 23-27
 ABSTRACT

Pansit-pansitan (Peperomia pellucida Linn.) is a plant that has been familiar since it is widely
spread in all tropical countries. Pansit-pansitan is considered as a medicinal herb and is traditionally
used to cure fever, cough, common cold, headache, and arthritis. It is proven that it possesses analgesic,
anti-arthritic, diuretic, and anti-bacterial property. This research study was conducted to test the efficacy
of pansit-pansitan extract as wound solution on BALB/c mice.Two different concentrations of pansit-
pansitan extract were tested as wound solution: 50% concentration and the other one is the pure pansit-
pansitan extract. Distilled water served as the negative control while Betadine was considered as the
positive control. The extract was applied to the wounds of the BALB/c mice that were about 2
centimeters long. After the analysis of the results, it was found that pure pansit-pansitan extract can heal
wound but the 50% concentration of extract can heal the wound faster.Based on the results, both
concentrations of pansit-pansitan extract were considered effective.
 ACKNOWLEDGEMENT

We would like to extend our deepest gratitude to GOD for guiding and protecting us every time

we go to UPLB (University of the Philippines Los Baños) and for giving us clear mind to think for all

the papers in our investigatory project.

To our loving parents, for their financial and moral support and for being there for us

throughout our SIP.

To our consultant, Dr. Maria Dalisay G. Maligalig, for the advice that helped us to come up

with a better study.

To our research adviser, Mr Fermin A. Clorado Jr. for his help and guidance in making our

research paper. For entertaining all our questions regarding our study and for helping us in finding our

consultant.

To Mr. Wilson Ray G. Anzures, for helping us in computing our Statistical Analysis and in

making our Post-hoc Test.

Lastly, to our classmates (IOTA) who supported us in our Science Investigatory Project. For

making us happy even we’re tired in preparing our research study and for staying by our side since from

the start.

Thank you!

Team: Tsuchikage

-Sophia, Anette, Christine

 INTRODUCTION

Background of the Study

Up until now, many plants are still ignored without knowing that some of them can be used as

wound solution. One example of this is the pansit-pansitan plant. Compared to other plants, gardeners

consider pansit-pansitan plant as weeds. It was proven that it possesses antibacterial property. Bacteria,

when it infects the wound, tend to prolong its healing time. The purpose of this study was to prove that

plants like pansit-pansitan has its own uses and should not be disregarded. It is also aimed to find out if

it can be natural wound solution with beneficial purposes that could fight against infectious bacteria, so

it has the ability to heal wounds.

Other than that, some studies show that pansit-pansitan can cure coughs, common colds, and

fever.

The study was conducted in search of effective wound solution that is readily available.
Statement of the Objectives

This research study was conducted to produce a wound solution from pansit-pansitan (Peperomia

pellucida Linn) extract. Specifically, this study aims to:

1. Determine the efficacy of pansit-pansitan extract as a wound solution

2. Prove that the pansit-pansitan extract is comparable to commercial wound solution.

3. Determine which concentration of pansit-pansitan will help the wound to heal faster.

Hypotheses

1. If Pansit-pansitan extract is applied to the wound, then the wound will heal faster.

2. If Pansit-pansitan extract is applied to the wound, then the wound will not heal.

Significance of the Study

Taking care of pets is considered as one of the hobbies of people. They will do everything just to

make sure that their pet is in a good condition. And whenever it is wounded, they will search for

different types of wound solutions, and it’s a waste of time.

This study was conducted to test an alternative, natural and readily available wound solution as

compared to other commercial brands and to prove the efficacy of pansit-pansitan

(Peperomia pellucida Linn.) extract to produce an alternative wound solution since it has been proven

to possess antibacterial property.

The success of this study would be a big help to the pet lovers because they don’t need to waste

their time and money to buy different commercial brands of wound solution. Instead, they will just

search for pansit-pansitan that can be found on their backyards.

Scope and Limitation

The main purpose of this study is to test the effect of various concentrations of pansit-pansitan

(Peperomia pellucida Linn.) extract on the wounds of BALB/c Mice each wound measures 2cm in

length and were induce using blade and spring balance to assure that there is the same force applied in

every wound. In this research study, different concentrations of pansit-pansitan extract was used such as

50% and 100% with distilled water as negative control and Betadine as positive control. All parts of the

plant except the roots were used in the experiment. The extract lasts for about 6 days. It was stored at

temperature not exceeding to 30˚ and placed in a plastic bottle. It was applied to the wounds by using

medicine cap. The experiment was observed for about thirty days. This study had undergone 5

replicates to come up with reliable and nonfictional results.

Pansit-pansitan was gathered from Brgy. San Roque, San Pablo City while the experiment was

conducted at San Pablo City National High School and Barangay San Rafael San Pablo City.
Definition of Terms

Betadine- a commercial wound solution; one of the independent variable

BALB/c Mice- animal species that was induced in the experiment

Concentration of pansit-pansitan- different amount of pansit-pansitan extract used in each wound

solution

Healing Time - the moment after inducing the wound until the scab appears from the cuts; it is the

dependent variable

Pansit-pansitan- plant organism that was extracted to gather wound solution; main variable in the

experiment.

Replicates- number of BALB/c mice in the experiment

 Review of Related Literature

Pansit-pansitan

Pansit-pansitan (Peperomia pellucida Linn.) is an annual, shallow-rooted herb that grows to

about 15 to 45 cm in height in damp and lightly shaded areas. This plant is characterized by its shiny

heart-shaped leaves about 4 cm in length with its green, fleshy, branched and translucent stem. Pansit-

pansitan has tiny dot-like flowers that grow from erect and slender green spikes that turn brown when

matured. (Unknown author, 2015)

Pansit-pansitan has been used as food item as well as medicinal herb for its analgesic, anti-

arthritic, diuretic, anti-bacterial activity. The entire plant is edible either cooked or raw. It has been

traditionally used to treat fever, cough, common cold, headache, and arthritis. In study of aerial parts of

peperomia extract in mice indicated that it exhibited anti-inflammatory and analgesic activities.

(Unknown author, 2011)

This plant is native to tropical America. Naturalized in the Philippines and has become a weed in

rice fields and home yards. It is occasionally planted as an ornamental and propagated by seeds.

(Madulid, 2000)

BALB/c Mice

BALB/c mice are a popular strain and are used in many different research disciplines, such as

cardiovascular, aging and pharmacological studies. It is also classified as an inbred from the production

of 20 or more successive brother-sister mating, the BALB/c mice is albino and small in size. (Unknown

author, 2015)

White (albino) mice are the most common type found in pet stores, fancy mice can be twice the

average size, and come in a wide variety of coat colours and types, from curly and shiny to silver and

cinnamon. They originate in Asia. Their size was ranging from five to seven centimeter long weighing
15 grams. They are very sociable creatures and like to have company. They usually live for only 12

months or one year but if well cared for, Albino mice typically live one to three years. Albino Mice are

curious, charming pets, and will be active at various times throughout the day. However, they are fragile

and should be treated gently. (Unknown author, 2015)

Povidone-Iodine

Povidone-Iodine is developing products for major advancement in wound healing. Treating

minor wounds and infections, as well as killing bacteria. Povidone-iodine solution is an antiseptic

combination. It works by killing sensitive bacteria. Povidone-iodine is a broad spectrum antiseptic for

topical application in the treatment and prevention of infection in wounds. May be used in first aid for

minor cuts, grazes, burns, abrasions and blisters. It is equally effective and safe. (drugs.com, 2015)

Wound Solutions

Wound solutions pressure prevention products assist in minimizing pressure, reducing moisture

and eliminating friction, sheer and cleansing power against microorganisms. It is also used to prevent

and treat skin and tissue infections that could result from cuts, scrapes and pressure sores. It is also used

before and after surgery to prevent surgical wound infections. Wound solutions are commonly used to

reduce healing time. (woundsolutions.com, 2015)

Lethal Activity of Pansit-pansitan

This research screens pansit-pansitan (Peperomiapellucida Linn.), a herb accepted by the

populace as an effective cure for a variety of ailments which, unfortunately was proven to exhibit

gene toxicity in the Alliums Test. As part of the school’s herbal project, this research assessed

the plant’s lethal activity using the Brine Shrimp Lethality Bioassay. The testing involved the

exposure of ArtemiaSalina ‘nauplii to different aqueous extract concentrations (10%, 30%, and

50%) of the plant in three serial dilutions. Mortality among nauplii samples was observed for 48

hours in experimental and control groups in three replications, and LC50 value was computed

using the Spearman-Karber Method. Significant differences among sample observations were

determined by employing One-Way Analysis of Variance. Lethal activity of the test plant on the

invertebrate model was proven with 9.18 LC50 value and 50 percent extract concentration as the

most lethal treatment. This established bioactivity of pansit-pansitan

(investigatoryprojectexample.com, 2015)

An Investigative Study about the Effectiveness of Pansit-pansitan (Peperomia pellucida

Linn.) As Organic Facial Cleanser

The researchers conducted a study about the effectiveness of pansit-pansitan (Peperomia

pellucida Linn.) as an organic facial cleanser. The study was conducted as an attempt to discover

readily available, easy to prepare, alternatives to costly anti-parasitic soap, shampoos, and sprays,

sold in pet shops and veterinary clinics. Specifically, the study determines the anti-parasitic

effect of the crude leaf extract of the carabao grass, makahiya, and Pansit-pansitan against adult

red brown dog ticks. Photochemical assays were done to determine the presence of bio-active

chemical compounds in the leaves of the test plants. (studymode.com, 2015)

 METHODOLOGY

Preparation of Pansit-pansitan

` Two kilograms of Pansit-pansitan plants were manually gathered from Barangay San

Roque and Barangay San Francisco San Pablo City, Laguna. The roots were removed from the

Pansit-pansitan plants and were washed under running water and air dried for three minutes.

Preparation of Extract

Using mortar and pestle, the Pansit-pansitan plants were crushed. The crushed plants

were squeezed using cheesecloth to separate the extract from its pulp. The extract was sterilized

by boiling for five minutes. Four beakers were prepared. The first beaker contained pure Pansit-

pansitan extract (100%), second beaker 50% Pansit-pansitan extract and 50% Distilled Water,

third beaker 10% Betadine used as positive control and fourth beaker 100% Distilled Water as

negative control. Every beaker contained 10ml amount of solutions.

Preparation of BALB/c Mice

Twenty BALB/c mice were bought from the University of the Philippines, Biological

technology. They were divided into four groups of five mice each named Group 1, 2, 3, and 4.

All mice were given the same amount of food and water every day. All mice were labelled by

marking their tails with different number of lines to indicate their groups.

Wound Induction Procedure

At their backs, the furs of the BALB/c Mice were shaved using razor. Two centimeter

lateral cut positioned parallel to the groove of the mouse. To assure that the application of force

was the same in inducing the wound of the BALB/c Mice use spring balance. This procedure

was allowed by the consultant (Vertebrate Animal Form).

Application of Treatments

After the wound was induced, four beakers with different solutions were prepared. First

beaker (with 100% Pansit-pansitan extract) was applied on Group 1 by pouring the concentration

in a medicine cap over the wounded part of the mice once a day. Same procedures were done on

the other groups of mice.

Statistical Analysis of Gathered Data

The data were statistically analyzed by One-way analysis of variance (ANOVA). Post-

Hoc Test was used to analyze and interpret the given data.

Raw Data

Wound Solution Results in Days

Betadine
Mice A 11 days
Mice B 13 days
Mice C 14 days
Mice D 15 days
Mice E 17 days
Average: 14 days
50% Pansit-pansitan
Mice A 13 days
Mice B 15 days
Mice C 17 days
Mice D 17days
Mice E 18 days
Average: 16 days
Distilled water
Mice A 18 days
Mice B 19 days
Mice C 21 days
Mice D 23 days
Mice E 24 days
Average: 21 days
100% Pansit-pansitan
Mice A 20 days
Mice B 21 days
Mice C 24 days
Mice D 24 days
Mice E 26 days
Average: 23 days
 RESULTS AND DISCUSSION

Mice of four groups having five replicates each were used to test whether the pansit-

pansitan (Peperomia pellucida Linn.) extract can heal wound. The first group of replicates was

treated with Betadine (positive control). The second group was treated with distilled water

(negative control). The third and fourth groups were treated with pure and 50% Pansit-pansitan

extracts (experimental variable).One-way ANOVA and descriptive was shown on the table

below.

Table 1: Analysis of Variances (ANOVA)

Sum of df Mean Square F-ratio Significance

Squares

Between

Groups 104 15 6.9333333 5.2

Within

Groups 25.333333 19 1.3333333

Total 3258

Critical
F-value: 2.23406292
reject Ho, accept Ha
Table 2: Descriptive Statistics

Dependent Variable: Healing Rate

Wound Solution Mean Std. Deviation N

Betadine 14.0000 2.23607 5

50% Pansit-pansitan 16.0000 2.00000 5

100% Pansit-pansitan 23.0000 2.44949 5

Distilled Water 21.0000 2.54951 5

Total 18.5000 4.29810 20

Table 2 shows that 100% Pansit-pansitan has the highest mean with 23.000 while

Betadine has the lowest mean with 14.0000.

 Table 3: Multiple Comparisons

Dependent Variable: Healing Rate

Scheffe
(I) Wound Solution (J) Wound Solution Mean Std. Sig. 95% Confidence
Difference Error Interval
(I-J) Lower Upper
Bound Bound
50% Pansit-
-2.0000 1.46629 .612 -6.5706 2.5706
pansitan
Betadine 100% Pansit-
-9.0000* 1.46629 .000 -13.5706 -4.4294
pansitan
Distilled Water -7.0000* 1.46629 .002 -11.5706 -2.4294
Betadine 2.0000 1.46629 .612 -2.5706 6.5706
50% Pansit- 100% Pansit-
-7.0000* 1.46629 .002 -11.5706 -2.4294
pansitan pansitan
Distilled Water -5.0000* 1.46629 .029 -9.5706 -.4294
Betadine 9.0000* 1.46629 .000 4.4294 13.5706
100% Pansit- 50% Pansit-
7.0000* 1.46629 .002 2.4294 11.5706
pansitan pansitan
Distilled Water 2.0000 1.46629 .612 -2.5706 6.5706
Betadine 7.0000* 1.46629 .002 2.4294 11.5706
50% Pansit-
5.0000* 1.46629 .029 .4294 9.5706
Distilled Water pansitan
100% Pansit-
-2.0000 1.46629 .612 -6.5706 2.5706
pansitan
Based on observed means.
The error term is Mean Square (Error) = 5.375.
*. The mean difference is significant at the .05 level.

Based on the Table 3, the comparisons between the treatments (Betadine and 50% pansit-

pansitan) shows.612 values as well as the treatments (100% pansit-pansitan and Distilled water).

It means that there are no significant differences between them.

 Based on the observed means, the significant level is at .05 and the comparisons between

the treatments (Betadine and 100% Pansit-pansitan extract); (Betadine and Distilled water);

(100% and 50% Pansit-pansitan extract); (50% Pansit-pansitan extract and Distilled water)

shows values less than .05 which means that there are significant differences between them.
 SUMMARY AND CONCLUSIONS

Summary of Findings

This research study was conducted to test if Pansit-pansitan extract can be an alternative

source for wound solutions. It was also determined whether there were significant differences on

the concentration of the Pansit-pansitan extract.

 Analysis of data showed that among the four concentrations, only the Pansit-

pansitan extract with 100% concentration has no significant difference compared

to Betadine. This was fully supported by the results of One-way ANOVA.

 After careful analysis of the results, it was found that 100% concentration of

Pansit-pansitan was comparable to the positive control. This was also supported

by the results of One-way ANOVA and Post-Hoc Test.

Conclusion

Based on the results, the researchers concluded that 50% and 100% concentration of

Pansit-pansitan extract was acceptable as wound solution because it can heal wounds in animals.

The stated concentrations of Pansit-pansitan extract can prove that it has an ability to heal

wounds and can also be an alternative agent in some degree.

Between 50% and 100% concentrations, 50% pansit-pansitan extract is more applicable

than 100% pansit-pansitan because based on the statistical analysis; the comparisons of 50%

pansit-pansitan from the other extracts have all their significant differences. Whereas, the

concentration of 100% pansit-pansitan have no significant difference compared to distilled water.

This was supported by the results of multiple comparisons on One-way ANOVA.

 RECOMMENDATIONS

Pansit-pansitan (Peperomia pellucida Linn.) extract was proven effective as an

alternative for wound solution. It was also proven that Pansit-pansitan (Peperomia pellucida

Linn.) extract when mixed with distilled water helps fasten the healing time/period of a wounded

mouse. Further study and research should be made to improve the quality of the anti-bacterial

property of Pansit-pansitan extract. It is recommended to:

 More trials could also be done to have much stable results

 Try to search for another plant that can exhibit anti-bacterial property then compare it

with the anti-bacterial property of Pansit-pansitan extract

 Search for other purposes of Pansit-pansitan extract since this can be found throughout

the Philippines
 BIBLIOGRAPHY

BOOKS

L.S. Padua, Medicinal Plants, 1997 Museum of Natural History University of the

Philippines

Dr. D. Madulid, A Pictorial cyclopedia of the Philippines Ornamental Plants, 1995;2000

by Domingo Madulid and Bookmark Inc.

INTERNET

http://www.filipinoherbshealingwonders.filipinovegetarianrecipe.com/pansit-pansitan.htm

http://www.sageresearch.com/research-models/inbred-mice/balbc

http://www.scarletmacawpetshop.com/index.php?option=com_content&view=article&id=357:w

hite-mouse&catid=6:rodents-profiles&Itemid=13

http://www.drugs.com/cdi/povidone-iodine-cream.html

http://www.woundsolutions.com.html

http://www.woundcareinc.com/

http://www.investigatoryprojectexample.com/science/lethal-activity-of-pansit-pansitan.html

http://www.studymode.com/essays/Pansit-Pansitan-As-Organic-Facial-Cleanser-1877128.html
 Appendix A

Experimental Design Diagram

TITLE: The Efficacy of Pansit-pansitan (Peperomia pellucida Linn.) Wound solutions

on BALB/c Mice

HYPOTHESIS: If Pansit-pansitan extract is applied to the wound, then the wound

will heal faster.

IV: Concentration of Pansit-pansitan extract

10% Betadine 50% Pansit-pansitan 100% Pansit-pansitan

Distilled
(positive control) extract extract
water(negative control)

5 replicates 5 replicates 5 replicates 5 replicates

DV:

Healing Time

C:

Number of BALB/c Mice

Size of wounds

Location of the wound

Deepness of the wound

Amount of Solutions
 Appendix B

Figure 1: Preparation of Pansit-pansitan

Figure 2: Preparation of extract

Figure 3: Preparation of BALB/c Mice

Figure 4: Wound Induction Procedure

Figure 5: Wound Healing Process

Wound Solution Results in Days

Betadine
Mice A 11 days
Mice B 13 days
Mice C 14 days
Mice D 15 days
Mice E 17 days
Average: 14 days
50% Pansit-pansitan
Mice A 13 days
Mice B 15 days
Mice C 17 days
Mice D 17days
Mice E 18 days
Average: 16 days
Distilled water
Mice A 18 days
Mice B 19 days
Mice C 21 days
Mice D 23 days
Mice E 24 days
Average: 21 days
100% Pansit-pansitan
Mice A 20 days
Mice B 21 days
Mice C 24 days
Mice D 24 days
Mice E 26 days
Appendi Average: 23 days

xC
 Descriptive Statistics
Dependent Variable: HealingRate

Wound Solution Mean Std. Deviation N

Betadine 14.0000 2.23607 5

50% Pansit-pansitan 16.0000 2.00000 5

100% Pansit-pansitan 23.0000 2.44949 5

Distilled Water 21.0000 2.54951 5

Total 18.5000 4.29810 20

ANOVA

Sum of Df Mean Square F-ratio Significance

Squares

Between

Groups

104 15 6.9333333 5.2

Within Groups
25.333333 19 1.3333333

Total
3258

Critical F-
value: 2.23406292
reject Ho, accept Ha

Post-Hoc
 Levene's Test of Equality of Error Variancesa

Dependent Variable: HealingRate

F df1 df2 Sig.

.209 3 16 .889

Tests the null hypothesis that the error variance of the dependent variable is equal across groups.

a. Design: Intercept + WoundSolution

 Tests of Between-Subjects Effects

Dependent Variable: HealingRate

Source Type III Sum of Df Mean Square F Sig.
Squares

Corrected Model 265.000a 3 88.333 16.434 .000

Intercept 6845.000 1 6845.000 1273.488 .000

WoundSolution 265.000 3 88.333 16.434 .000

Error 86.000 16 5.375

Total 7196.000 20

Corrected Total 351.000 19

a. R Squared = .755 (Adjusted R Squared = .709)

Multiple Comparisons
Dependent Variable: HealingRate
Scheffe
(I) (J) Mean Std. Sig. 95% Confidence Interval
WoundSolution WoundSolution Difference Error
Lower Upper
(I-J) Bound Bound
Betadine 50% Pansit- -2.0000 1.4662 .612 -6.5706 2.5706
pansitan 9
100% Pansit- -9.0000* 1.4662 .000 -13.5706 -4.4294
pansitan 9
Distilled Water -7.0000* 1.4662 .002 -11.5706 -2.4294
9
50% Pansit- Betadine 2.0000 1.4662 .612 -2.5706 6.5706
pansitan 9
100% Pansit- -7.0000* 1.4662 .002 -11.5706 -2.4294
pansitan 9
Distilled Water -5.0000* 1.4662 .029 -9.5706 -.4294
9
100% Pansit- Betadine 9.0000* 1.4662 .000 4.4294 13.5706
pansitan 9
50% Pansit- 7.0000* 1.4662 .002 2.4294 11.5706
pansitan 9
Distilled Water 2.0000 1.4662 .612 -2.5706 6.5706
9
Distilled Water Betadine 7.0000* 1.4662 .002 2.4294 11.5706
9
50% Pansit- 5.0000* 1.4662 .029 .4294 9.5706
pansitan 9
100% Pansit- -2.0000 1.4662 .612 -6.5706 2.5706
pansitan 9
Based on observed means.
The error term is Mean Square(Error) = 5.375.
*. The mean difference is significant at the .05 level.