izzati abd malik

36 Pages·2016·0.42 MB·English

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HAZARD CLASSIFICATION OF GAMAT- N,N-DIETHYL-M-
TOLUAMIDE (DEET) MOSQUITO REPELLENT RUBBING OIL USING
THE UNITED NATIONS GLOBALLY HARMONIZED SYSTEM (UN-GHS)
BASED ON RATS ACUTE DERMAL TOXICITY

IZZATI ABD MALIK

UNIVERSITI SAINS MALAYSIA
2015

1
HAZARD CLASSIFICATION OF GAMAT- N,N-DIETHYL-M-
TOLUAMIDE (DEET) MOSQUITO REPELLENT RUBBING OIL USING
THE UNITED NATIONS GLOBALLY HARMONIZED SYSTEM (UN-GHS)
BASED ON RATS ACUTE DERMAL TOXICITY

by

IZZATI ABD MALIK

Thesis submitted in partial fulfilment of the requirements
for the degree of
Master of Science

AUGUST 2015
2
DECLARATION

I hereby declare that I am the sole author of this thesis in title “Hazard
Classification Of Gamat- N,N-Diethyl-M-Toluamide (DEET) Mosquito Repellent
Rubbing Oil Using The United Nations Globally Harmonized System (UN-GHS)
Based On Rats Acute Dermal Toxicity“. I declare that the thesis is being submitted to
Universiti Sains Malaysia (USM) for the purpose of the award of Master of Science
in Health Toxicology. This thesis is the result of my own research under supervision
of Dr. Jahangir bin Kamaldin except as cited in the references. The thesis has being
accepted for the respective study and is not concurrently submitted in candidature of
any other degree.
I authorize Universiti Sains Malaysia (USM) to use this thesis for the purpose
of scholarly publication. Universiti Sains Malaysia (USM) may reproduce this thesis
by photocopying or other means, in total or in part at the request of other institution
or individual for academic reference.

IZZATI ABD MALIK
P-IPM0079/14
i
ACKNOWLEDGEMENT

In the name of Allah, The Most Beneficent, The Most Merciful. Alhamdulillah,
with His blessings, finally, I manage to complete my dissertation after the hard work,
blood, sweat and tears spent during the whole study period. First and foremost, I would
like to express my deep and sincere gratitude to my husband, Mohd Redzuan who is the
main sponsor for this study, spiritual and financial. Without his sacrifice, I will not be
able to complete this study.
Secondly, I would like to present my humble appreciation to my supervisor, Dr.
Jahangir b. Kamaldin for his great effort to guide me and sharing their wonderful
knowledge trough out the research. I also would like to thank my Co-supervisor, Dr.
Noraini for her advises and assistance in making this research a success.
I wish to thank Dr. Sawibah, Dr. Azlina, Mr. Norhisham, Mrs. Rodiah and all
ARC staffs in assisting me during the laboratory works. Also, I would like to thank my
colleague, Norhidayah Ramli who always share a bundle of information, assists me
during the experiments as well as motivate me to the end of this work. May Allah bless
you and your family. Not to forget, my wonderful friends, all my postgraduate friends
who always be my bread and butter, be together with me with laughter and tears. Allah‟s
bless will always be with you guys and in sha allah you will succeed here and hereafter.
Last, but not least, thank you to my wonderful family for their encouragement and moral
support during the production of this dissertation. Thank you Allah for Your wonderful
gift. Wassalam
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TABLE OF CONTENT
DECLARATION..................................................................................................................... i
ACKNOWLEDGEMENT ....................................................................................................... ii
TABLE OF CONTENT ......................................................................................................... iii
LIST OF TABLES ..................................................................................................................v
LIST OF FIGURES ............................................................................................................. vii
LIST OF SYMBOL AND ABBREVIATION ......................................................................... viii
ABSTRACT ...........................................................................................................................ix
ABSTRACT ............................................................................................................................x
Chapter 1 INTRODUCTION ................................................................................................. 1
Chapter 2 LITERATURE REVIEW ...................................................................................... 5
2.1 United Nation of Globally Harmonized System (UN-GHS) ...................................... 5
2.1.1 Hazard Classification ....................................................................................... 6
2.1.1 Health Hazard .................................................................................................. 7
2.1.1 Acute Toxicity ............................................................................................... 14
2.2 Acute Dermal Toxicity Study ................................................................................. 16
2.2.1 Skin Anatomy and Physiology ....................................................................... 16
2.2.2 Mechanism of Absoprtion .............................................................................. 19
2.2.3 Toxicity Effect Upon Skin Exposure .............................................................. 21
2.3 Insect Repellent ...................................................................................................... 23
2.3.1 Formulation ................................................................................................... 23
2.3.2 Mode of Action .............................................................................................. 25
2.3.2 Mammalian Toxicity ...................................................................................... 26
2.4 N,N-Diethyl-m-toluamide (DEET) .......................................................................... 27
2.4.1 Chemical structure and physicochemical of DEET ......................................... 29
2.4.2 Types of DEET formulation ........................................................................... 29
2.4.3 Mode of action ............................................................................................... 29
2.5 Gamat oil ........................................................................................................... 31
2.5.1 Gamat Oil and its health benefits .................................................................... 31
2.5.2 Inert Ingredients in GD MRRO ...................................................................... 31
Chapter 3 MATERIALS AND METHOD ........................................................................... 34
3.1` Materials ............................................................................................................ 34
3.2 Rat as Test System ............................................................................................. 36
3.3 Test Method ....................................................................................................... 36
3.3.1 Acclimatisation & Marking of Rats ................................................................ 36
3.3.2 Dose Preparation ............................................................................................ 43
3.3.3 Administration of Dose – Dosing ................................................................... 43
3.3.4 Observations .................................................................................................. 48
3.3.5 Necropsy gross examination ........................................................................... 54
3.4 Analysis of Data ................................................................................................. 36
3.4.1 Localized Dermal Toxicity Effects ................................................................. 59
3.4.2 Systemic Dermal Toxicity Effects .................................................................. 59
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Chapter 4 RESULT............................................................................................................... 68
4.1 Localized Dermal Toxicity Effect ........................................................................... 68
4.2 Systemic Dermal Toxicity Effect ............................................................................ 68
4.2.1 Body weight change, Feed and Water Consumption ....................................... 70
4.2.2 Clinical Signs of Toxicity............................................................................... 73
4.2.3 Mortality, Moribund and Severe Pain/Distress ................................................ 76
4.2.2 Necropsy Gross Examination ......................................................................... 76
Chapter 5 DISCUSSION ...................................................................................................... 79
5.1 Localized Dermal Toxicity Effect ........................................................................... 79
5.2 Systemic Dermal Toxicity Effect ............................................................................ 80
5.2.1 Body Weight Change ..................................................................................... 80
5.2.2 Feed Consumption ......................................................................................... 81
5.2.3 Water Consumption ....................................................................................... 81
5.2.4 Clinical Signs of Toxicity............................................................................... 81
5.2.5 Mortality, Moribund and Severe Pain/Distress ................................................ 83
5.2.2 Necropsy Gross Examination ......................................................................... 84
5.3 Study Limitation..................................................................................................... 85
Chapter 6 CONCLUSION AND RECOMMENDATION ................................................... 85
REFERENCES...................................................................................................................... 86
APPENDIX ........................................................................................................................... 94
Appendix 1 ..................................................................................................................... 94

iv
LIST OF TABLES

No Title Page

Table 1.1 Hazard class of an insecticide product is based on WHO Recommended 3
Classification of Pesticides by Hazard (WHO, 2010) that has been aligned with
the Acute Toxicity Hazard Categories under the United Nations Globally
Harmonised System for Classification of Chemical Substances and Mixtures
(UN-GHS)

Table 2.1 GHS Criteria, single dose toxicity 9

Table 2.2 GHS criteria, skin corrosion/irritation and serious eye damage/eye irritation 10

Table 2.3 GHS criteria, respiratory or skin sensitization and single or repeated target 11
organ systemic toxicity (TOST)

Table 2.4 GHS criteria, germ cell mutagenicity and reproductive and developmental 12
effects

Table 2.5 GHS criteria, carcinogenicity 13

Table 2.6 DEET Toxicity Test 28

Table 3.1 Environmental control sheet 38

Table 3.2 Acclimatisation sheet 40

Table 3.3 Removal of more than 10% of the rats fur based on Meeh‟s formula 41

Table 3.4 Animal Identification of the test item, DEET MRRO and the placebo item, 42
GRO

Table 3.5 Dose preparation of the test item, DEET MRRO and the placebo item, GRO 44

Table 3.6 Dose preparation sheet 45

Table 3.7 Dosing Sheet 47

Table 3.8 Table observation log 49

Table 3.9 List of criteria for determination of moribund and severe pain/distress of rat 53

Table 3.10 Necropsy Sheet 55

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Table 4.1 Localized dermal toxicity effects exhibited by female Rattus norvegicus of 69
Sprague Dawley rats treated with the test item (DEET MRRO) and the placebo
item (GRO) at the dose of above 5000 mg/ kg body weight via dermal route.

Table 4.2 Mean and standard deviation of body weight of female Rattus norvegicus of 71
Sprague Dawley rats treated with the test item (DEET MRRO) and the placebo
item (GRO) at the dose of above 5000 mg/ kg body weight via dermal route.

Table 4.3 Mean and standard deviation of 14 days feed consumption by female Rattus 72
norvegicus of Sprague Dawley rats treated with the test item (DEET MRRO)
and the placebo item (GRO) at the dose of above 5000 mg/ kg body weight via
dermal route.

Table 4.4 Mean and standard deviation of 14 days water consumption by female Rattus 74
norvegicus of Sprague Dawley rats treated with the test item (DEET MRRO)
and the placebo item (GRO) at the dose of above 5000 mg/ kg body weight via
dermal route.

Table 4.5 Clinical signs shown by female Rattus norvegicus of Sprague Dawley rats 75
treated with the test item (DEET MRRO) and the placebo item (GRO) at the
dose of above 5000 mg/ kg body weight via dermal route.

Table 4.6 UN-GHS Hazard Classification of the test item (DEET MRRO) and the 77
placebo item (GRO) based on acute dermal toxicity response (mortality,
moribund and severe pain/severe distress) of female Rattus norvegicus of
Sprague Dawley rats treated at the dose of above 5000 mg/ kg body weight
using OECD TG402 Test Method

Table 4.7 Necropsy gross examination findings of female Rattus norvegicus-Sprague 78
Dawley rats upon dermal dosing of the test item (GD MRRO) and the placebo
item (GRO) using OECD TG402 test method at the dose of above 5000 mg/ kg
body weight.

vi
LIST OF FIGURES

No Title Page
Figure 2.1 Skin structure and its function 17
Figure 2.2 DEET molecule structure 30
Figure 3.1 Product information of the test item, GD MRRO 35
Figure 3.2 Product information of the placebo item, GRO 37
Figure 3.3 Decision Logic to Acute Toxicity 66

vii
LIST OF SYMBOLS AND ABBREVIATIONS

DEET N, N-diethyl-toluamide
GD MMRO Gamat DEET Mosquito Repellent Rubbing Oil
GRO Gamat Rubbing Oil
OECD Organization for Economic Cooperation and Development
CPL Classification on Packaging and Labelling
GHS Globally Harmonized System
UN-GHS United Nation of Globally Harmonized System
UN United Nations
MW Molecular Weight
EPA Environmental Protection Agency
REACH Registration, Evaluation, Authorisation and Restriction of
Chemicals
WHO World Health Organization
GLP Good Lab Practice
OATP Organic Anion Transporting Polypeptides
K Permeability coefficient
p
LOEL Lowest Observable Effect Level
NOEL No Observable Effect Level
RTBSA Rats Total Body Surface Area
g gram
ml mililiter
mg/kg bw milligram per kilogram body weight

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