APPENDICES

APPENDIX A

GANTT CHART

Title: The Diuretic Activity of Euphorbia hirta (Tawa-Tawa) Ethanolic Leaf Extract on Rattus

norvegicus (Sprague Dawley Rats)

Proponents: Mico D. Castillejos, Jan Glicelle E. Malones, John Rouvic V. Sanchez

Research Consultant: Dra. Cynthia N. Ochona, DVM, MSc

2019
ACT

SEPTEMBER

NOVEMBER
FEBRUARY

OCTOBER
AUGUST
MARCH

APRIL

JULY
JUNE
MAY

A
B
C
D
E
F
G
H
I
J
K
L
M
N
O
P
Q
R
Activities:

A. Formulation of Research Problem K. Writing of Chapter 1 (Introduction)

B. Gathering of Research Data L. Extraction of Plant Sample

C. Formulation of Research Title M. Diuretic Test

D. Writing of Research Proposal N. Writing of Chapters 2,3, and 4

E. Writing of Research Plan O. Analyzation of Diuretic Test Result

F. Gathering of Plant Sample P. Revision of Manuscript for Regional

G. Certification of Plant Sample Science and Technology Fair

H. Searching for Research Consultant Q. Preparation for Regional Science and

I. Phytochemical Analysis of the Plant Technology Fair

Sample R. Cliniquing for National Science and

J. Preparation for Research Technology Fair

Experimentation

Prepared by:

MICO D. CASTILLEJOS CELESTE D. CASTILLEJOS

Student Researcher Parent

JAN GLICELLE E. MALONES IGLECERIO S. MALONES

Student Researcher Parent

JOHN ROUVIC V. SANCHEZ AMIE V. SANCHEZ

Student Researcher Parent

Noted: Approved:

ELAIN E. MALONES BELINDA C. LOYOLA

Research Adviser Principal III
 APPENDIX B

CALENDAR OF ACTIVITIES

Title: The Diuretic Activity of Euphorbia hirta (Tawa-Tawa) Ethanolic Leaf Extract on Rattus

norvegicus (Sprague Dawley Rats)

Proponents: Mico D. Castillejos, Jan Glicelle E. Malones, John Rouvic V. Sanchez

Research Consultant: Dra. Cynthia N. Ochona, DVM, MSc

Month Activities People involved Remarks

February  Formulation of Research  Researchers
Problem
2019
 Research II Accomplished
 Gathering of Research Adviser
data
March
 Gathering of Research
 Researchers Accomplished
2019 data

 Gathering of Research
data
April  Researchers
 Formulation of Research
Accomplished
2019 Title  Research II
Adviser
 Writing of Research
Proposal
 Gathering of Research
data  Researchers

May  Gathering of Plant  Research II

Sample Adviser Accomplished
2019

 Certification of Plant  Research

Sample Consultant
  Searching for Research
Consultant

 Phytochemical
Analyzation of Plant
Sample

 Preparation of Research
Experimentation
 Preparation of Research
 Researchers
Experimentation

June  Research II
 Writing of Chapter 1
Adviser
(Introduction) Accomplished
2019

 Research
 Extraction of Plant
Consultant
Sample
 Researchers

July  Extraction of Plant

 Research II
Sample
Adviser Accomplished
2019
 Diuretic Test
 Research
Consultant
 Diuretic Test  Researchers

 Writing of Chapter 2, 3,  Research II

August 2019
and 4 Adviser Accomplished

 Analyzation of Diuretic  Research

Test Result Consultant

 Writing of Chapter 2, 3,
September and 4  Researchers

Accomplished
2019  Analyzation of Diuretic  Research II
Test Result Adviser
 October  Researchers
 Revision of Manuscript
for Regional Science and Accomplished
2019  Research II
Technology Fair
Adviser

 Preparation for Regional  Researchers

November Science and Technology
Fair  Research II
Accomplished
2019  Cliniquing for National Adviser
Science and Technology
Fair  Consultants

Prepared:

MICO D. CASTILLEJOS CELESTE D. CASTILLEJOS

Student Researcher Parent

JAN GLICELLE E. MALONES IGLECERIO S. MALONES

Student Researcher Parent

JOHN ROUVIC V. SANCHEZ AMIE V. SANCHEZ

Student Researcher Parent

Noted: Approved:

ELAIN E. MALONES BELINDA C. LOYOLA

Research Adviser Principal III
APPENDIX C
LETTERS
 Appendix D
Certificate of Analysis and Raw Data
 APPENDIX E

STATISTICAL COMPUTATION

A. 250 mg/kg, 500 mg/kg, and 1000 mg/kg of Euphorbia hirta (Tawa-tawa) Ethanolic Leaf

Extract Induced to Rattus norvegicus (Sprague Dawley Rats)

1. Hypothesis (H0): None of the different dosage (250 mg/kg, 500 mg/kg, 1000 mg/kg) of

Euphorbia hirta (Tawa-tawa) ethanolic leaf extract is the most effective in increasing

the urine output of Sprague Dawley rats.

2. Level of Significance: α = 0.05

3. Type of Test: ANOVA Single Factor

4. Critical Value: 0.05

5. Decision Rule: Reject H0 if p ≤ 0.05

6. Computation:
 7. Result: Since the value of p which is 0.6376 is greater than the level of significance (α)

which is 0.05, therefore we accept H0.

8. Findings showed that none of the different dosage (250 mg/kg, 500 mg/kg, 1000 mg/kg)

of Euphorbia hirta (Tawa-tawa) ethanolic leaf extract is the most effective in increasing

the urine output of Sprague Dawley rats. The result of the ANOVA Single Factor on the

raw data of different dosages of Euphorbia hirta (Tawa-tawa) Ethanolic Leaf Extract

Induced to Rattus norvegicus (Sprague Dawley Rats) showed that the computed p value

of sample which is 0.637 is greater than the level of significance (α) which is 0.05, so it

is within the region of acceptance. Therefore, we accept H0 and reject H1. In conclusion,

Euphorbia hirta (Tawa-tawa) Ethanolic Leaf Extract, regardless of dosage, has no

difference in increasing the urine output of Rattus norvegicus (Sprague Dawley Rats).

B. 250 mg/kg, 500 mg/kg, 1000 mg/kg of Euphorbia hirta (Tawa-tawa) Ethanolic Leaf Extract

and Furosemide Lasix (Positive Control) Induced to Rattus norvegicus (Sprague Dawley Rats)

1. Hypothesis (H0): There is no significant difference between the diuretic properties of

Euphorbia hirta (Tawa-tawa) ethanolic leaf extract and Furosemide Lasix (Positive

Control) on the urine output of Sprague Dawley rats.

2. Level of Significance: α = 0.05

3. Type of Test: ANOVA Single Factor

4. Critical Value: 0.05

5. Decision Rule: Reject H0 if p ≤ 0.05

6. Computation:

7. Result: Since the value of p which is 0.1050 is greater than the level of significance (α)

which is 0.05, therefore we accept H0.

8. Findings showed that there is no significant difference between the diuretic properties

of Euphorbia hirta (Tawa-tawa) ethanolic leaf extract and Furosemide Lasix (Positive

Control) on the urine output of Sprague Dawley rats.The result of the ANOVA Single

Factor on the raw data of different dosages of Euphorbia hirta (Tawa-tawa) Ethanolic

Leaf Extract and the positive control induced to Rattus norvegicus (Sprague Dawley

Rats) showed that the computed p value of sample which is 0.1050 is greater than the

level of significance (α) which is 0.05, so it is within the region of acceptance. Therefore,

we accept H0 and reject H1. In conclusion, Euphorbia hirta (Tawa-tawa) Ethanolic Leaf
 Extract and Furosemide Lasix (Positive Control) has no difference in increasing the

urine output of Rattus norvegicus (Sprague Dawley Rats).

C. 250 mg/kg, 500 mg/kg, 1000 mg/kg of Euphorbia hirta (Tawa-tawa) Ethanolic Leaf Extract

and Normal Saline Solution (Negative Control) Induced to Rattus norvegicus (Sprague Dawley

Rats)

1. Hypothesis (H0): There is no significant difference between the diuretic properties of

Euphorbia hirta (Tawa-tawa) ethanolic leaf extract and Normal Saline Solution

(Negative Control) on the urine output of Sprague Dawley rats.

2. Level of Significance: α = 0.05

3. Type of Test: ANOVA Single Factor

4. Critical Value: 0.05

5. Decision Rule: Reject H0 if p ≤ 0.05

6. Computation:
 7. Result: Since the value of p which is 0.5313 is greater than the level of significance (α)

which is 0.05, therefore we accept H0.

8. Findings showed that There is no significant difference between the diuretic properties

of Euphorbia hirta (Tawa-tawa) ethanolic leaf extract and Normal Saline Solution

(Negative Control) on the urine output of Sprague Dawley rats. The result of the

ANOVA Single Factor on the raw data of different dosages of Euphorbia hirta (Tawa-

tawa) Ethanolic Leaf Extract and the negative control induced to Rattus norvegicus

(Sprague Dawley Rats) showed that the computed p value of sample which is 0.5313 is

greater than the level of significance (α) which is 0.05, so it is within the region of

acceptance. Therefore, we accept H0 and reject H1. In conclusion, Euphorbia hirta

(Tawa-tawa) Ethanolic Leaf Extract and Normal Saline Solution (Negative Control) has

no difference in increasing the urine output of Rattus norvegicus (Sprague Dawley

Rats).

D. Different dosages of Euphorbia hirta (Tawa-tawa) Ethanolic Leaf Extract (250 mg/kg, 500

mg/kg, and 1000 mg/kg) Induced to Rattus norvegicus (Sprague Dawley Rats)

1. Hypothesis (H0): There is no significant difference on the urine output of Sprague

Dawley rats when treated with different dosage (250 mg/kg, 500 mg/kg, 1000 mg/kg)

of ethanolic leaf extract of Euphorbia hirta (Tawa-tawa).

2. Level of Significance: α = 0.05

3. Type of Test: ANOVA Single Factor

4. Critical Value: 0.05

5. Decision Rule: Reject H0 if p ≤ 0.05

6. Computation:

7. Result: Since the value of p which is 0.6376 is greater than the level of significance (α)

which is 0.05, therefore we accept H0.

8. Findings showed that none of the different dosage (250 mg/kg, 500 mg/kg, 1000 mg/kg)

of Euphorbia hirta (Tawa-tawa) ethanolic leaf extract is the most effective in increasing

the urine output of Sprague Dawley rats. The result of the ANOVA Single Factor on the

raw data of different dosages of Euphorbia hirta (Tawa-tawa) Ethanolic Leaf Extract

Induced to Rattus norvegicus (Sprague Dawley Rats) showed that the computed p value

of sample which is 0.637 is greater than the level of significance (α) which is 0.05, so it

is within the region of acceptance. Therefore, we accept H0 and reject H1. In conclusion,

different dosages of Euphorbia hirta (Tawa-tawa) Ethanolic Leaf Extract has no

difference in increasing the urine output of Rattus norvegicus (Sprague Dawley Rats).
 APPENDIX F

EXPENDITURES AND OFFICIAL RECEIPTS

Title: The Diuretic Activity of Euphorbia hirta (Tawa-Tawa) Ethanolic Leaf Extract on Rattus

norvegicus (Sprague Dawley Rats)

Proponents: Mico D. Castillejos, Jan Glicelle E. Malones, John Rouvic V. Sanchez

Research Consultant: Dra. Cynthia N. Ochona, DVM, MSc

Expenditures Amount (Php)

Experimental Fee/Test Fee 3,040.00

Sprague Dawley Rats 3,000.00

IACUC 3,000.00

Sprague Dawley Rats’ Feeds 1,600.00

Phytochemical Analysis 640.00

Plant Extraction 1,010.00

Two Liters Ethanol 550.00

Sample Amber Bottle 20.00

Transportation 9,000.00

Printing 1,000.00

TOTAL 22,860.00
 APPENDIX G

PHOTOGRAPHS

Photograph 1. Collecting Euphorbia hirta (Tawa-tawa) Leaf Samples

Photograph 2. Fixing the Payment for the Crude Extraction

Photograph 3. Wiley-Mill Grinding of Dried Euphorbia hirta (Tawa-tawa) Leaf Samples

Photograph 4. Soaking of Grinded Euphorbia hirta (Tawa-tawa) Leaf Samples

Photograph 5. Filtering of Euphorbia hirta (Tawa-tawa) Ethanolic Extract

Photograph 6. Rotary Evaporating of Euphorbia hirta (Tawa-tawa) Ethanolic Extract

Photograph 7. Water Bathing of Euphorbia hirta (Tawa-tawa) Ethanolic Extract

Photograph 8. Fixing the Payment for Phytochemical Analysis

Photograph 9. Fixing the Payment for Lipschitz Diuretic Test

Photograph 10. Fixing the Payment for Lipschitz Diuretic Test

Photograph 11. Setting of Metabolic Cages of Sprague Dawley Rats

Photograph 12. Fasting of Sprague Dawley Rats

Photograph 13. Oral Dosing of Tawa-tawa Extract on Sprague Dawley Rats

Photograph 14. Baseline Urine Testing of Sprague Dawley Rats

Photograph 15. Recording of Voided Urine Output of Sprague Dawley Rats

Photograph 16. Receiving of Raw Data (Results) from DOST - ITDI

Photograph 17. Computing for the Statistical Test of the Raw Data in Microsoft Excel

Photograph 18. Meeting with the Research Consultant (Dra. Cynthia Nalo-Ochona)
 APPENDIX H

PERSONAL INFORMATION OF THE CONSULTANT

CYNTHIA M. NALO-OCHONA, DVM, MSc

Supervisor at Industrial Technology Development Institute
Department- Department of Science and Technology
(+63)917-7948-787

doc_cyrene@yahoo.com

Specialty: Veterinarian

Years in Practice: 22 years

Areas of Specialization: Dog, Cat, Game fowl, Chicken, Pig, Cow & Buffalo, Goat & Sheep,

Horse, Turkeys, Geese, Ducks, Quails, Pigeons, Rabbits & Fur animals, Fish & Shrimps

Area of Interest: Clinical/Surgical

Clinic Information:

Blk 7, Lot 18 Northbridge Sta. Monica, Novaliches, Metro Manila

Department of Science and Technology

Industrial Technology Development Institute Department (ITDI)

Taguig City, Metro Manila

 APPENDIX I

PERSONAL INFORMATION OF RESEARCHERS

Background Information

1.1 Name Mico De Leon Castillejos

1.2 Birth date March 12, 2002

1.3 Age 17

1.4 Birth place Naic, Cavite

1.5 Gender Male

1.6 Home address Blk 11 Lot 20, Ternate Residential Estate, Ternate,

Cavite

1.7 Contact Number (+63) 966-705-4765

1.8 Email address mico.castillejos12@gmail.com

1.9 Civil Status Single

1.10 Religion Roman Catholic

1.11 Parents Celeste De Leon Castillejos

Marvin Tibayan Castillejos

1.12 Contact Number (+63)927-6076-575

N/A

Educational Background

2.1 Day Care Poblacion 1 Day Care Center (2005-2006)

Brgy. Poblacion 1, Ternate, Cavite 4111

2.2 Nursery Ternate Elementary School (2006-2007)

Brgy. Poblacion 1-A, Ternate, Cavite 4111

 2.3 Kinder Ternate Elementary School (2007-2008)

Brgy. Poblacion 1-A, Ternate, Cavite 4111

2.4 Elementary Dei Gracia Academy (2008-2014)

Brgy. Cabuco, Trece Martires City, Cavite 4109

2.5 Junior High School Cavite National Science High School (2014-18)

Brgy. Garita-B, Maragondon, Cavite 4112

2.6 Senior High School Cavite National Science High School (2018-20)

Brgy. Garita-B, Maragondon, Cavite 4112

Honors / Award Received
3.1 Day Care Best in Science, Mathematics, and Drawing

3.2 Nursery N/A

3.3 Kinder Best in English Writing and Reading

3.4 Grade 1 10th Honors

3.5 Grade 2 N/A

3.6 Grade 3 4th Honors

3.7 Grade 4 4th Honors

3.8 Grade 5 4th Honors, 1st Place Science Quiz Bee

3.9 Grade 6 4th Honors

3.10 Grade 7 N/A

3.11 Grade 8 N/A

3.12 Grade 9 Perfect Attendance (1st Quarter)

With Honors, Perfect Attendance (2nd Quarter)

With Honors, Perfect Attendance (3rd Quarter)

With Honors, Perfect Attendance (4th Quarter)

 3.13 Grade 10 With Honors, Perfect Attendance (1st Quarter)

With Honors, Perfect Attendance (2nd Quarter)

With Honors, Perfect Attendance (3rd Quarter)

With Honors, Perfect Attendance (4th Quarter)

3.14 Grade 11 With Honors, Perfect Attendance (1st Quarter)

With Honors, Perfect Attendance (2nd Quarter)

With Honors, Perfect Attendance (3rd Quarter)

With Honors, Perfect Attendance (4th Quarter)

3.15 Grade 12 With Honors (1st Quarter)

Organizations

4.1 Grade 4 Mathematics Club

4.2 Grade 5 Science Club

4.3 Grade 6 Science Club

4.1 Grade 7 Information, Communications, and Technology Club

Boy Scout of the Philippines

4.2 Grade 8 Calgeotrists’ Club

Boy Scout of the Philippines

4.3 Grade 9 Calgeotrists’ Society

Boy Scout of the Philippines

4.4 Grade 10 Calgeotrists’ Society

Boy Scout of the Philippines

4.5 Grade 11 Calgeotrists’ Society

4.6 Grade 12 Calgeotrists’ Society - Grade 12 Representative

Background Information

1.1 Name Jan Glicelle E. Malones

1.2 Birth date September 07, 2001

1.3 Age 18

1.4 Birth place Naic Medicare Hospital

1.5 Gender Female

1.6 Home address Blk. 7, Lt. 2, Ph. 2, Villa Apolonia, Naic, Cavite

1.7 Contact Number 0915-235-8214

1.8 Email address glicellieeemaloneso@gmail.com

1.9 Civil Status Single

1.10 Religion Iglesia Ni Cristo

1.11 Parents Iglecerio S. Malones

Elain E. Malones

1.12 Contact Number 0930-690-0815

0917-701-0669

Educational Background

2.1 Daycare N/A

2.2 Nursery Naic Star Learning Center (2006-2007)

Villa Apolonia Subdivision, Naic, Cavite 4110

2.3 Kinder Naic Star Learning Center (2007-2008)

Villa Apolonia Subdivision, Naic, Cavite 4110

2.4 Elementary Naic Elemenary School (2008-2014)

01 Captain C. Nazareno, Naic, Cavite 4100

 2.5 Junior High School Cavite National Science High School (2014-18)

Brgy. Garita B, Maragondon, Cavite 4112

2.6 Senior High School Cavite National Science High School (2018-20)

Brgy. Garita B, Maragondon, Cavite 4112

Honors / Award Received

3.1 Day Care N/A

3.2 Nursery 6th Honors, Most Self Reliant

3.3 Kinder 4th Honorable Mention, Most Generous

Best in Writing

3.4 Grade 1 N/A

3.5 Grade 2 N/A

3.6 Grade 3 N/A

3.7 Grade 4 N/A

3.8 Grade 5 10th Honors

3.9 Grade 6 6th Honors

3.10 Grade 7 N/A

3.11 Grade 8 6th Honors

3.12 Grade 9 With Honors, Perfect Attendance (2nd Quarter)

With Honors (3rd Quarter)

With Honors (4th Quarter)

3.13 Grade 10 With Honors, Perfect Attendance (2nd Quarter)

With Honors, Perfect Attendance (3rd Quarter)

With Honors, Perfect Attendance (4th Quarter)

 3.14 Grade 11 1st Semester

With Honors, Perfect Attendance (1st Quarter)

With Honors, Perfect Attendance (2nd Quarter)

2nd Semester

With Honors, Perfect Attendance (1st Quarter)

With Honors (2nd Quarter)

3.15 Grade 12 With Honors, Perfect Attendance (1st Quarter)

IV. Organizations

4.1 Grade 7 Science Club

Girl Scout of the Philippines

4.2 Grade 8 Kamfilan Club

Girl Scout of the Philippines

4.3 Grade 9 Calgeotrists’ Society

Girl Scout of the Philippines

4.4 Grade 10 Sci-Res Club – Assistant Treasurer

Girl Scout of the Philippines

4.5 Grade 11 Calgeotrists’ Society

Yes-O Organization

4.6 Grade 12 Catalyst Club

Yes-O Organization
Background Information

1.1 Name John Rouvic V. Sanchez

1.2 Birth date December 25, 2001

1.3 Age 17

1.4 Birth place Salcedo II, Noveleta, Cavite

1.5 Gender Male

1.6 Home address Mulawin, Tanza, Cavite

1.7 Contact Number 0921-883-9315

1.8 Email address johnrouvic.sanchez@gmail.com

1.9 Civil Status Single

1.10 Religion Roman Catholic

1.11 Parents Amie V. Sanchez

Jose Rouel L. Sanchez

1.12 Contact Number 0925-551-2907

0921-549-3716
Educational Background
2.1 Day Care Mulawin Day Care Center (2005-2006)

Brgy. Mulawin, Tanza, Cavite 4108

2.2 Nursery Holy Nazarene Christian School (2006-2007)

Brgy. Mulawin, Tanza, Cavite 4108

2.3 Kinder Holy Nazarene Christian School (2007-2008)

Brgy. Mulawin, Tanza, Cavite 4108

2.4 Elementary Holy Nazarene Christian School (2008-2014)

Brgy. Mulawin, Tanza, Cavite 4108

 2.5 Junior High School Cavite National Science High School (2014-18)

Brgy. Garita B, Maragondon, Cavite 4112

2.6 Senior High School Cavite National Science High School (2018-20)

Brgy. Garita B, Maragondon, Cavite 4112

Honors / Award Received

3.1 Day Care N/A

3.2 Nursery 4th Honors, Best in Deportment

3.3 Kinder 5th Honorable Mention, Best in Deportment,

Best in Spelling, Best in Reading

3.4 Grade 1 5th Honors

3.5 Grade 2 5th Honors

3.6 Grade 3 Best in Deportment

3.7 Grade 4 6th Honorable Mention

3.8 Grade 5 4th Honors, Perfect Attendance

3.9 Grade 6 1st Honorable Mention, Loyalty Awardee,

Best in Computer, Best in Deportment

3.10 Grade 7 N/A

3.11 Grade 8 N/A

3.12 Grade 9 With Honors, Perfect Attendance (1st Quarter)

With Honors, Perfect Attendance (2nd Quarter)

With Honors, Perfect Attendance (3rd Quarter)

With Honors, Perfect Attendance (4th Quarter)

3.13 Grade 10 With Honors, Perfect Attendance (1st Quarter)

 With Honors, Perfect Attendance (2nd Quarter)

With High Honors, Perfect Attendance (3rd Quarter)

With High Honors, Perfect Attendance (4th Quarter)

Conduct Awardee

3.14 Grade 11 With Honors, Perfect Attendance (1st Quarter)

With Honors, Perfect Attendance (2nd Quarter)

With Honors, Perfect Attendance (3rd Quarter)

With Honors, Perfect Attendance (4th Quarter)

3.15 Grade 12 With High Honors, Perfect Attendance (1st Quarter)

Organizations
4.1 Grade 4 English Club

4.2 Grade 5 Science Club

4.3 Grade 6 Journalism Club

4.1 Grade 7 Information, Communications, and Technology Club

Communication Arts Club

Boy Scout of the Philippines

4.2 Grade 8 SciRes Club

Boy Scout of the Philippines

4.3 Grade 9 Calgeotrists’ Society

Boy Scout of the Philippines

4.4 Grade 10 Calgeotrists’ Society

Boy Scout of the Philippines

4.5 Grade 11 Calgeotrists’ Society – Business Manager

4.6 Grade 12 Calgeotrists’ Society