53

APPENDICES
 54

APPENDIX TABLES
 55

Appendix Table 1. Colony count of before and after application of (T1) 10 mg/mL ethyl
acetate extract of Paragis (Eleusine indica)
SERIAL BEFORE AFTER
DIFFERENCE
NUMBER APPLICATION APPLICATION
1 55 49 6
2 72 64 8
3 42 30 12
4 38 20 18
5 44 40 4
6 104 23 81

Appendix Table 2. Colony count of before and after application of (T2) 20 mg/mL ethyl
acetate extract of Paragis (Eleusine indica)
SERIAL BEFORE AFTER
DIFFERENCE
NUMBER APPLICATION APPLICATION
7 25 14 11
8 41 31 10
9 51 13 38
10 73 48 25
11 71 45 26
12 68 44 24

Appendix Table 3. Colony count of before and after application of (T3) 10 mg/mL
ethanol extract of Paragis (Eleusine indica)
SERIAL BEFORE AFTER
DIFFERENCE
NUMBER APPLICATION APPLICATION
13 101 70 31
14 64 49 15
15 67 51 16
16 4 4 0
17 94 42 52
18 91 67 24
Appendix Table 4. Colony count of before and after application of (T4) 20 mg/mL
ethanol extract of Paragis (Eleusine indica)
SERIAL BEFORE AFTER
DIFFERENCE
NUMBER APPLICATION APPLICATION
19 94 43 51
20 106 71 35
21 127 94 33
22 52 24 28
23 61 40 21
 56

24 113 89 24

Appendix Table 5. Colony count of before and after application of (T5) commercialized
hand sanitizer (positive control)
SERIAL BEFORE AFTER
DIFFERENCE
NUMBER APPLICATION APPLICATION
25 42 8 34
26 53 5 48
27 63 14 49
28 98 3 95

Appendix Table 6. Colony count of (T0) negative control

SERIAL BEFORE AFTER
DIFFERENCE
NUMBER APPLICATION APPLICATION
29 112 112 0
30 10 10 0
31 87 87 0
32 53 53 0

Appendix Table 7. Frequency of Gram-positive and Gram-negative bacteria in samples

obtained before application of treatments
FREQUENCY
SERIALNUMBER Gram + Gram - Both
Cocci Bacilli Cocci Bacilli Coccobacilli Cocci Bacilli Coccobacilli
1 - - - - - - - +
2 - - - - - + - -
3 - - - - - + - -
4 - - - - - + - -
5 - - - - - + - -
6 + - - - - - - -
7 + - - - - - - -
8 + - - - - - - -
9 - - - - - + - -
10 - - - - - + - -
11 - - - - - + - -
12 - - - - + - - -
13 + - - - - - - -
14 + - - - - - - -
15 - - + - - - - -
16 + - - - - - - -
 57

17 - - - - - + - -
18 - - - - - + - -
19 - - + - - - - -
20 + - - - - - - -
21 - - + - - - - -
22 - - - - - + - -
23 - - - - - + - -
24 + - - - - - - -
TOTAL 8 0 3 0 1 11 0 1 24
% 33.33 0 12.5 0 4.17 45.83 0 4.17 100

Appendix Table 8. Frequency of Gram-positive and Gram-negative bacteria in samples

obtained after application of treatments
SERIALNUMBE FREQUENCY
Gram + Gram - Both
R
Cocci Bacilli Cocci Bacilli Coccobacilli Cocci Bacilli Coccobacilli
1 - - - - - + - -
2 - - - - - + - -
3 - - - - - + - -
4 - - + - - - - -
5 - - - - - + - -
6 - - + - - - - -
7 + - - - - - - -
8 + - - - - - - -
9 - - - - - - - +
10 + - - - - - - -
11 - - - - - + - -
12 - - + - - - - -
13 - - - - - + - -
14 + - - - - - - -
15 - - - - - + - -
16 - - - - - + - -
17 + - - - - - - -
18 - - - - - + - -
19 - - + - - - - -
20 - - - - - + - -
21 + - - - - - - -
22 + - - - - - - -
23 - - - - - + - -
24 - - - + - - - -
TOTAL 7 0 4 1 0 11 0 1 24
% 29.17 0 16.67 4.17 0 45.83 0 4.17 100
 58

Appendix Table 9. Phytochemical constituents of Paragis (Eleusineindica)(Morah, F. N.

I., Otuk, M. E. (2015). Antimicrobial and antihelminthic activity of
Eleusine indica.Acta Scientiae etIntellectus, 1(4), p.31)
PHYTOCHEMICALS ETHANOL EXTRACT
Flavonoids ++
Tannins -
Saponins -
Alkaloids ++
Cardiac glycosides +
Anthracene glycosides +
Anthraquinones ++
 59

APPENDIX FIGURES
 60

Appendix Figure 1. Collection of Paragis (Eleusine indica)

 61

Appendix Figure 2. Washing of Paragis (Eleusine indica)

Appendix Figure 3. Drying of Paragis (Eleusine indica)

 62

Appendix Figure 4. Sterilization of glassware

Appendix Figure 5.Weighing of dried Paragis (Eleusine indica)

 63

Appendix Figure 6. Powdering of dried Paragis (Eleusine indica)

Appendix Figure 7. Weighing of powdered Paragis (Eleusine indica)

 64

Appendix Figure 8. Maceration using ethyl acetate and 95% ethanol (1g of powdered
Paragis: 10 mLof solvent)

Appendix Figure 9. Covering of the containers with aluminum foil to protect it from light
 65

Appendix Figure 10. Filtration using gauze pad and whatmann filter paper
 66

Appendix Figure 11. Evaporation of solvents using rotary evaporator

Appendix Figure 12. Ethanolic and ethyl acetate extract

 67

(a) (b)

(b) (d)

(e) (f)

Appendix Figure 13. Formulation of hand sanitizer: a. measure 0.25 grams of carbopol b.
measure 24 mL of distilled water c. in a hotAppendix Figure, mix and allow the carbopol
to dissolve in the water. d. add Paragis extract (0.25 mL in10 mg/mL concentration while
500 uL in 20 mg/mL concentration) e. add 0.75 mL of glycerol f. add 1 drop of
triethanolamine
 68

Appendix Figure 14. Hand sanitizer: 10 mg/mL ethyl acetate, 20 mg/mL ethyl acetate, 20
mg/mL ethanol, 10 mg/mL ethanol, and commercialized hand sanitizer (from left to
right)

Appendix Figure 15. Measuring and adjusting the pH of hand sanitizers by dropping
triethanolamine
 69

(a) (b)

Appendix B-17.

(c) (d)

(e)

Appendix Figure 16. Preparation of nutrient agar: (a) Measure 28 grams of Nutrient Agar
powder, (b) Measure 1000 mL distilled water, (c) Mixtures were subjected to heat, (d)
Mixtures will be autoclaved, (e) Agar after autoclaving
 70

Appendix Figure 17. Selection of participants using random sampling

Appendix Figure 18. Signing of informed consent form (ICF) and providing
questionnaires to be answered after application of hand sanitizer.
 71

Appendix Figure 19. Swabbing before and after application of hand sanitizer

Appendix Figure 20. Streaking on nutrient agar then incubation

 72

(a) Before (b) After

Appendix Figure 21. Colony count of the negative control

(a) Before (b) After

Appendix Figure 22. Colony count of the positive control (Green Cross Hand Sanitizer)
a. before application b. after application
 73

(a) Before (b) After

(b) Before (b) After

Appendix Figure 23. Colony count of 10mg/mL ethyl acetate extract of

Paragis (Eleusine indica): a. before application of hand sanitizer;
b. after application of hand sanitizer

(a) Before
(b) After

Appendix Figure 24. Colony count of 20mg/mL ethyl acetate extract of

Paragis (Eleusine indica): a. before application of hand sanitizer;
b. after application of hand sanitizer
 74

(a) Before
(b)
After

Appendix Figure 25. Colony count of 10mg/mL ethanolic extract of

Paragis (Eleusine indica): a. before application of hand sanitizer;
b. after application of hand sanitizer

(a) Before (b) After

Appendix Figure 26. Colony count of 20mg/mL ethanolic extract of

Paragis (Eleusine indica): a. before application of hand sanitizer;
b. after application of hand sanitizer.
 75

(a) (b) (c)

Appendix Figure 27. Slide preparation: a. obtaining a sample of bacterial colony;

b. mixing of the bacteria with a drop of normal saline solution; c. gently passing
to the flame in order to heat-kill and fixed the bacteria onto the slide.

Appendix Figure 28. Gram staining: Applying of crystal violet to heat-fixed smear
for 1 minute; then gram’s iodine for 1 minute; Decolorization with acetone; and
counterstaining with safranin for 1 minute
 76

(a) (b)

Appendix Figure 29.Microscopic result from negative control: a. before application-

gram negativecocci; b. after application- gram positive cocci

(a) (b)

(c)

Appendix Figure 30. Microscopic result from positive control (commercialized hand
sanitizer): a.before application- gram positive cocci, b. before application- gram negative
cocci; c. after application- gram positive cocci
 77

(a) (b)

(c) (d)

(e)

Appendix Figure 31. Microscopic result : a. before application of 10mg/mL ethyl acetate
extract of Paragis (Eleusine indica) - gram positive cocci, b.before application- gram
negative cocci; c. before application- gram negative bacilli; d. after application- gram
positive cocci; e. after application- gram negative cocci
 78

(a)
(b)

Appendix Figure 32. Microscopic result: a. before application of 20mg/mL ethyl acetate
extract of Paragis (Eleusine indica) - gram positive cocci; b. after application- gram
positive cocci

(a)
(b)

(c)
 79

Appendix Figure 33. Microscopic result: a. before application of 10mg/mL ethanolic

extract of Paragis (Eleusine indica) - gram positive cocci; b. after application- gram
negativecocci; c. after application- gram negative cocci

(a) (b)

Appendix Figure 34. Microscopic result: a. before application of 20mg/mL ethanolic

extract of Paragis (Eleusine indica) - gram negative cocci; b. after application- gram
negative cocci

Gram positive only; 8;

33%
Gram positive only
Both Gram-positive and Gram-negative only
Gram-negative; 12;
50% Both Gram-positive and Gram-
negative

Gram-negative only; 4;
17%

Appendix Figure 35. Percentage of bacterial colonies before application according to

their gram staining characteristics
 80

Gram-positive only; 3;
19%

Gram-positive only
Gram-negative only; 1;
6%
Gram-negative only

Both Gram-positive and

Both Gram-positive Gram-negative
and Gram-negative ;
12; 75%

Appendix Figure 36. Percentage of bacterial colonies after application according to their
gram staining characteristics
 81

Appendix 1. Letter of request for the authentication of the plant

 82

Appendix 2. Letter of request for the use of laboratory

 83

Appendix 3. Letter of request for the use of laboratory

 84

Appendix 4. Letter of request for the use of laboratory

 85

Appendix 5. Letter of request for the use of laboratory

 86

Appendix 6. Letter of request for the use of laboratory

 87

Appendix 7. Letter of request for extension of research

 88

Appendix 8. Letter for the statistician

 89

Appendix 9. Letter of request for the approval of adviser and technical critic
 90

Appendix 10. Letter of request for the approval of the title

 91

Appendix 11. Letter of request for outline proposal defense

 92

Appendix 12. Proposal manuscript routing form

 93

Appendix 13. Letter of request for final oral defense

 94

Appendix 14. Final manuscript routing form

 95

Appendix 15. Sample of an informed consent form for participants

Republic of the Philippines

CAVITE STATE UNIVERSITY
Don Severino de las Alas Campus
Indang, Cavite
 (046) 4150-010 / (046) 4150-013 loc 232
www.cvsu.edu.ph

COLLEGE OF NURSING

Department of Medical Technology

INFORMED CONSENT FORM

This informed consent form is for the respondents who are 4 th year students of

Bachelor of Science in Medical Technology in Cavite State University – Indang,

Campus, whom the researchers are inviting to participate in the study, entitled

“PARAGIS (Eleusine indica) AS ANTIBACTERIAL AGENT USED IN MAKING

HAND SANITIZER”. This study is being conducted by 4th year BSMT students namely

Lairene Mhae L. Feranil, Maria Cristina Gae A. Firme, and Marie Keicelyn R. Libiran

under the supervision of Flordeliza A. de Guzman, RMT, MAT-Phys.

This Informed Consent Form has two parts:

 Information Sheet (to share information about the study with you)

 Certificate of Consent (for signatures if you choose to participate)

 96

Information Sheet

Introduction

We, the researchers are doing an experimental study entitled “PARAGIS

(Eleusine indica) AS ANTIBACTERIAL AGENTUSED IN MAKING HAND

SANITIZER”. We are going to give you information and invite you to be part of this

research. Before you decide, we will welcome your questions for clarifications. This

consent form may also contain words that you do not understand. Please ask us to stop as

we go through the information and we will try to explain. And please do not hesitate to

ask us if you happen to have further questions later.

Purpose of the Research

According to Chassin, Mayer, & Nether (2010), “Hand hygiene remains the most

effective means of breaking transmission of most infectious diseases in and out of

hospital settings.” There are many ways to maintain proper hand hygiene and among

these, hand sanitizers have been recognized for giving convenience to the consumers. In

connection to this, we would like to use Paragis (Eleusine indica) which is known for its

antibacterial property as an antibacterial agent in formulating a hand sanitizer that can

compete on the standards already present in the market because it can minimize possible

harmful effects caused by the chemicals found in commercial sanitizers. To be able to do

this, may we ask for your permission to participate in our study so that we can test the

effectiveness of our product.

 97

Type of Research Intervention

This research will involve your participation in the application of the hand

sanitizer containing Paragis extract, collection of swab sample under the fingernails

before and after application of the hand sanitizer, and answering of the questions based

on your personal experience after application of the product.

Participant Selection

From thirty-five 4th year students of Bachelor of Science in Medical Technology,

the researchers will select thirty-two participants grouped based on the hand sanitizer that

they will use which will differ on the amount and type of Paragis (Eleusine indica)

extract. People with severe skin irritation or eczema, open cuts on their hands, or those

taking antibiotics will be excluded and will not be obliged to participate in the study.

Procedure

The following are the steps with

which the participants will be involved.

The whole process will take around 10

minutes.

1. Collection of swab sample under the

fingernails of the participant’s

dominant hand before application of

 98

the hand sanitizer. Prior to this, they will be asked to kindly remove their rings, if

present, in their fingers.

2. Participants will be provided with specific instructions on how to rub their hands

using a diagram of hand rubbing motions published by the World Health

Organization (please refer to picture on the right side).

3. Collection of swab sample under the fingernails of the participant’s dominant hand

after application of the hand sanitizer. Participants will be asked to allow their hands

to fully air dried after application and shall not rinse nor wipe off the hand sanitizer

before it dries.

4. Answering of the questions based on the participant’s personal experience after

application of the hand sanitizer.

Benefits

Your participation will likely help us test the effect of Paragis (Eleusine indica) as

antibacterial agent in making hand sanitizer which, if found effective, will certainly be

beneficial for the community considering not only the availability and wide distribution

of Paragis (Eleusine indica) throughout the Philippines, but also its potential to minimize

possible harmful effects caused by the chemicals found in commercial sanitizers.

 99

Date and Time

Certificate of Consent

I have been invited to participate in the research about “PARAGIS (Eleusine

indica) AS ANTIBACTERIAL AGENT USED IN MAKING HAND SANITIZER”.
I have read the foregoing information, or it has been read to me. I have had the
opportunity to ask questions about it and any questions I have been asked have been
answered to my satisfaction. I consent voluntarily to be a participant in this study.

Participant’s signature over printed name

Statement by the Researchers

We have accurately read out the information sheet to the potential participant, and
to the best of our ability made sure that the participant understands that the following will
be done:

1. Application of the hand sanitizer containing Paragis (Eleusine indica) extract

2. Collection of swab sample under the fingernails before and after application of the
hand sanitizer
3. Answering of the questions based on personal experience after application of the
hand sanitizer.
We confirm that the participant was given an opportunity to ask questions about
the study, and all the questions asked by the participant have been answered correctly and
to the best of our ability. We confirm that the individual has not been coerced into giving
consent, and the consent has been given freely and voluntarily.

A copy of this ICF has been provided to the participant.

FERANIL, LAIRENE MHAE L. FIRME, MARIA CRISTINA GAE A.

LIBIRAN, MARIE KEICELYN R.

Approved:

FLORDELIZA DE GUZMAN, RMT

 100

Adviser

Appendix 16. Sample of a survey questionnaire

Republic of the Philippines

CAVITE STATE UNIVERSITY
Don Severino de las Alas Campus
Indang, Cavite
 (046) 4150-010 / (046) 4150-013 loc 232
www.cvsu.edu.ph

COLLEGE OF NURSING

Department of Medical Technology

QUESTIONNAIRE

Assessment of Participants’ Response on the Quality of the Formulated Hand

Sanitizers

Please rate how strongly you agree or disagree on the following questions:

5- Strongly Agree
4- Agree
3- Neutral (Neither Agree nor Disagree
2- Disagree
1- Strongly Disagree
QUESTIONS RATINGS
5 4 3 2 1
1. Are you aware of the proper use of hand sanitizer?
2. Do you find the hand sanitizer as a quick and easy
product to use?
3. Is hand rubbing more rapid for hand cleansing than hand
washing?
4. Did you not experience stinging skin irritation?
5. Did it not cause cracking of the skin?
6. Did it not affect the color of the nail?
7. Is it moisturizing to the skin?
8. Didyou not experience excessive sweating of the palm?
9. Are you satisfied with the hand hygiene product?
10. Will you consider this hand sanitizer as effective for
hand hygiene?
 101

Appendix 17. Ethics Review Board Certification

 102

Appendix 18. Plant identification/certification from Bureau of Plant Identification (BPI)

103
 104

Appendix 19. Certification from adviser

 105

Appendix 20. Certification from technical critic

 106

Appendix 21. Certification from statistician

 107

Appendix 22. Certification from english critic

 108

Appendix 23. Timetable of activities

Month April May June July August September October November

Week 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4
Problem
Identification
Gathering
basic and
preliminary
data
Formulating
Research
Problem/
Title
Formulating
the
Research
Design
Proposal
Defense
Preparation
Procurement
of Materials
Making the
Hand
Sanitizer
Evaluation
of the Hand
Sanitizer
Preparation
of Final
Draft for
Defense
First
revision
Second
revision
English
critic: First
reading
English
critic:
Second
reading
Completion
form
Binding of
the
manuscript
 109

Appendix 24. Budgetary requirements of the study

The following are the estimates of the researcher for the budgetary requirements
of the study:

Purpose Budget Proposal

Rental fee for equipment (Rotary Php 1500.00
Evaporator)
Materials to be used Php 10500.00
Commercialized hand gel sanitizer Php 50.00
Transportation Php 1000.00
Adviser Fee Php 1500.00
Technical Critic Fee Php 800.00
Statistician Fee Php 1500.00
TOTAL Php 16 850.00

Appendix 25. Curriculum Vitae of the reseachers

 110

LAIRENE MHAE L. FERANIL

288 Calumpang Cerca,
Indang, Cavite
[email protected]
09651491718

OBJECTIVES:

To be part of a progress and prestigious organization to enhance my knowledge,

characteristics and skills as a Medical Technologist.

PERSONAL INFORMATION

Date of birth: February 13, 1999

Age: 19 years old
Civil Status: Single
Religion: Roman Catholic
Father: Marion B. Feranil
Occupation: Maintenance Personnel
Mother: Lina L. Feranil
Occupation: Housewife

EDUCATIONAL BACKGROUND

2015 – Present Cavite State University – Indang Campus

Tertiary
Bancod, Indang, Cavite
Bachelor of Science in Medical Technology

2015 – 2011 Lumampong National High School- Indang Annex

Secondary
J. Dimabiling Street, Poblacion 4

2011- 2005 Dr. Alfredo Pio de Roda Elementary School

Primary
Calumpang Cerca, Indang, Cavite

TRAININGS AND SEMINARS ATTENDED

 111

November 2016 “Pre-Employment Seminar”

University Gymnasium, Cavite State University-Indang
Campus

January 2017 “Stand Against Vices”

SM Rolle Hall, Cavite State University – Indang Campus

SPECIAL SKILLS

 Computer Literate Can find desired information in the internet within

reasonable time.
Able to use programs such as Office Suites (MS Word,
Power Point, etc.)

 Language Proficiency Can speak English and Filipino

CHARACTER REFERENCES

Flordeliza A. de Guzman, Clinical Instructor

Cavite State University – Indang Campus
0916-230-6569

Rolando P. Antonio, Research Coordinator

Cavite State University – Indang Campus
0927-113-9647

I hereby certify that all information above is true and correct.

LAIRENE MHAE L. FERANIL

MARIA CRISTINA GAE A. FIRME

36 Sangley Street
 112

Greenheights Village
Parañaque City
[email protected]
0906-727-7722/ 0928-128-5034

OBJECTIVES:

To apply and further enhance my skills and knowledge as a Medical Technologist

PERSONAL INFORMATION

Date of birth: November 15, 1997

Age: 21 years old
Civil Status: Single
Religion: Catholic
Father: James Hodgen B. Firme
Occupation: None
Mother: Gay A. Firme
Occupation: Accountant

EDUCATIONAL BACKGROUND

2015 – Present Cavite State University – Indang Campus

Tertiary
Bancod, Indang, Cavite
Bachelor of Science in Medical Technology

2014-2015 San Juan De Dios Educational Foundation Inc.

Tertiary

2014 – 2010 Lycee D’ Regis Marie

Secondary
Parañaque City

2010- 2004 Lycee D’ Regis Marie

Primary
Parañaque City

TRAININGS AND SEMINARS ATTENDED

 113

November 2016 “Pre-Employment Seminar”

University Gymnasium, Cavite State University-Indang
Campus

January 2017 “Stand Against Vices”

SM Rolle Hall, Cavite State University – Indang Campus

SPECIAL SKILLS

 Knowledge in Technology Can use modern technology comfortably and able to

learn
the operations of complex equipment

 Language Proficiency Can speak English and Filipino

CHARACTER REFERENCES

Flordeliza A. de Guzman, Clinical Instructor

Cavite State University – Indang Campus
0916-230-6569

Rolando P. Antonio, Research Coordinator

Cavite State University – Indang Campus
0927-113-9647

I hereby certify that all information above is true and correct.

MARIA CRISTINA GAE A. FIRME

MARIE KEICELYN R. LIBIRAN

 114

Block 90 Lot 1 Phase 1-A

Carissa Homes, Bagtas,
Tanza, Cavite
[email protected]
0936-337-3054/ 0977-850-8475

OBJECTIVES:

To seek opportunities from which I can learn and experience new things in the field
of medical laboratory science that will ultimately enhance my professional skills.

PERSONAL INFORMATION

Date of birth: March 7, 1999

Age: 19 years old
Civil Status: Single
Religion: Born Again Christian
Father: Gilberto S. Libiran
Occupation: Barangay Human Rights Officer
Mother: Alicia R. Libiran
Occupation: Housewife

EDUCATIONAL BACKGROUND

2015 – Present Cavite State University – Indang Campus

Tertiary
Bancod, Indang, Cavite
Bachelor of Science in Medical Technology

2015 – 2011 Dei Gratia School

Secondary
Carissa, Homes Bagtas, Tanza, Cavite

2011- 2005 Bagtas Elementary School

Primary
Pabahay Bagtas, Tanza, Cavite

TRAININGS AND SEMINARS ATTENDED

 115

November 2016 “Pre-Employment Seminar”

University Gymnasium, Cavite State University-Indang
Campus

January 2017 “Stand Against Vices”

SM Rolle Hall, Cavite State University – Indang Campus

SPECIAL SKILLS

 Computer Literate Can find desired information in the internet within

reasonable time.
Able to use programs such as Office Suites (MS Word,
Power Point, etc.)

 Language Proficiency Can speak English and Filipino

CHARACTER REFERENCES

Flordeliza A. de Guzman, Clinical Instructor

Cavite State University – Indang Campus
0916-230-6569

Rolando P. Antonio, Research Coordinator

Cavite State University – Indang Campus
0927-113-9647

I hereby certify that all information above is true and correct.

MARIE KEICELYN R. LIBIRAN