THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY NGUYEN THI MAI CHI TOPIC TITLE: THE EFFECT OF BETEL QUID CHEWING ON THE ABSORPTION OF CIGARETTE SMOKE CONTAINING ACROLEIN BARCHELOR THESIS Study Mode: Full-time Major: Environmental Science and Management Faculty: International Programs Office Batch: 2013 - 2017 Thai Nguyen, 20/11/2017 n DOCUMENTATION PAGE WITH ABSTRACT Thai Nguyen University of Agriculture and Forestry Degree Program Bachelor of Environmental Science and Management Student name Nguyen Thi Mai Chi Student ID DTN1353070121 Thesis Title THE EFFECT OF BETEL QUID CHEWING ON THE ABSORPTION OF CIGARETTE SMOKE CONTAINING ACROLEIN Supervisors Prof. Tsung - Yun Liu Dr. Do Thi Ngoc Oanh Abstract: Acrolein, a highly reactive, unsaturated aldehyde and respiratory irritant, is formed during combustion (e., cigarette smoke), high-temperature cooking of foods, and in vivo as a product of oxidative stress and polyamine metabolism. 3-Hydroxypropyl mercapturic acid (3-HPMA) is the main metabolite of acrolein in urine.
Previous studies demonstrated that betel quid can enhance the alkalinity of oral cavity, and might enhance the additional absorption of acrolein(pKa is about 9. However, this has not been shown in literature. This study is designed to describe some characteristics of cigarette smokers in Kaohsiung and test the hypothesis that chewing betel quid will facilitate the absorption of acrolein while smoking cigarette by comparing the level of urinary 3-HPMA in cigarette-smokers with and without betel quid chewing. Urine samples were obtained from healthy volunteers in Kaohsiung, which is located in the southern part of Taiwan.
Urinary 3-HPMA and creatinine were analyzed by LC-MS/MS and HPLC with UV detector, respectively. The results showed that the level of 3-HPMA in cigarette-smokers who chewing betel quid (1.97 ng/ g creatinine) was significant higher (p<0.01) than i n cigarette-smokers without chewing betel quid (1. People in Kaohsiung smoking cigarette and chewing betel quid concentrate in males from 30 to 59-year-old and females from 20 to 49-year-old. This study concluded that the higher level of urinary 3-HPMA in cigarette-smoker who chewing betel quid might be related to the high alkalinity in oral saliva, which facilitate the absorption of cigarette smoke containing acrolein.
Keywords: Acrolein; 3-Hydroxypropyl mercapturic acid (3-HPMA); LC- MS/MS; Toxicity; Cigarette smoke; Betel quid. Number of pages: Date of 20/09/2017 Submission: n ACKNOWLEDGEMENT First and foremost, I would like to express my deepest gratitude and special thanks to School Leadership and teachers at International Training and Development Center - Thai Nguyen University of Agriculture and Forestry those imparted for me the knowledge as well as facilitating learning and help me during the learning process in university. I would like to express my deepest appreciation to my supervisor: Professor. Tsung - Yun Liu of National Yang Ming University Institude of Environtmental and Occupational Health Science - who offered me a warm welcome, guided me wholeheartedly and gave me the best suggestions, explanations, conditions, for complete this study.
In addition, I would like to thank Dr. Do Thi Ngoc Oanh of Department of Agronomy- Thai Nguyen University of Agriculture and Forestry, for her supervision, encouragement, advice, and guidance in writing this thesis. I gratefully acknowledge Mr. Chih - Hung Hu, Mr.
Han - Xing Zou for his help and provided me with the knowledge to use the equipment and guide me during the practice. I wish to thank all the professor National Yang Ming University Institude of Environtmental and Occupational Health Science for their warm welcome and their interesting seminars. My sincere thanks also go to all my classmates – K45 AEP for along with me and helping me finish the study. n Special thanks to Mr Ngo Tuan Hung, Dao Thanh Huyen, Ngan Thi Thu Uyen, Do Van Binh, Kieu Manh Tuan and Taiwanese brothers and sisters in Institute of Environmental & Occupational Health Sciences, all the people who helped me when I stayed in National Yang Ming Univesity, Taiwan.
It was a beautiful time and I really appreciate it. Finally, I would like to thank my family, to my grandparents for their love and supporting me. Many thanks! Nguyễn Thị Mai Chi n TABLE OF CONTENTS LIST OF TABLES. ix LIST OF ABBREVIATIONS.
Background and rationale. Research question and hypotheses. 4 PART II - LITERATURE REVIEW. Physical and Chemical Properties of Acrolein.
Source of Acrolein. Toxicity of Acolein. Environmental standard of Acrolein. Acrolein effects on human beings.
Acrolein effects on animals. Acrolein effects on organisms in the environment. The trends and health risks of chewing betel quid and smoking cigarette in Taiwan. Liquid chromatography / Tandem mass spectrometry (LC / MS / MS) and operational mechanism.
17 PART III - MATERIALS AND METHODS. Time and Location. Adjust the dilution of the urine (Creatinine). Preparing and loading samples to measure levels 3-HPMA.
Listed of chemical compounds used in this study. Listed of Equipment used in this study. 26 PART IV - RESULT AND DISCUSSION. Some characteristics of cigarette smokers in Kaohsiung.
The identifying of sample chromatogram obtained with standard substance upon LC-MS / MS analysis. Effects of smoking cigarette and chewing betel quid to levels of 3-HPMA. 49 n LIST OF FIGURES Figure 3.1: Standard curve of 3-HPMA .2: The chromatograms of 3-HPMA levels in urine .1: The characteristics of cigarette smokers with-and without chewing betel quid of Kaohsiung population .2: The characteristics of using cigarette by gender of Kaohsiung population .3: Compare the percentage of gender between cigarette smokers with- and without chewing betel quid. Chromatograms of standard substance [d3]3-HPMA and 3-HPMA obtained in urine .5: Percentile of 3-HPMA levels in urine of cigarette smokers with - and without chewing betel quid.
37 viii n LIST OF TABLES Table 2. Physical and Chemical Properties of Acrolein .2: Thresholds for the acute effects of acrolein in human beings.1: The characteristics of using cigarette and betel quid by age of Kaohsiung population. 3-HPMA levels of cigarette smokers with –and without chewing betel quid .Effects of smoking cigarette with chewing betel quid on the risk of oral cancer. 35 ix n LIST OF ABBREVIATIONS 1.
ATSDR - Agency for Toxic Substances and Disease Registry 2. EPA - Environmental Protection Agency 3. DHS - Department of Homeland Security 4. 3-HPMA - 3-Hydroxypropyl mercapturic acid 5.
HPLC - High-performance liquid chromatography 6. LC/MS/MS - Liquid chromatography/mass spectrometry tandem mass spectrometer 7. GSH - Glutathione-associated cells 8. (d3)-3-HPMA - N-Acetyl-d3-S-(3-hydroxypropyl) cysteine, standard substance x n PART I - INTRODUCTION 1.
Background and rationale According to Toxicological Profile for Acrolein given by Agency for Toxic Substances and Disease Registry (ATSDR), Department of Homeland Security (DHS) and Environmental Protection Agency (EPA), Acrolein (CAS 107-02-8) was listed as a high toxicity chemical with recommendations on levels of exposure in the environment, food, and water (Toxicological Profile for Acrolein, 2007). In 1839, Berzelius named acrolein, the thermal degradation product of glycerin was characterized as an aldehyde, acrolein is a contraction of ‘acrid’ (referring to its pungent smell) and ‘oleum’ (oil or oil-like consistency) (Berzelius, 1839). In recent years for the industries, acrolein is used as an initial material for acrylate polymers and in the production of acrylic acid, plastics, and technical chemicals. In agriculture, it is known as the herbicide (Stevens & Maier, 2008; Toxicological Profile for Acrolein, 2007).
Environmentally, acrolein exists naturally in food and is formed during the burning of organic matter. Therefore, acrolein is found in all types of smoke including cigarette smoke, in exhaust gases from internal combustion engines, and in superheated cooking oil where there is serious human toxicity (Beauchamp et al. In vivo, acrolein is a metabolite of the anti - cancer drug cyclophosphamide and has been found to be formed from threonine by myeloperoxidase neutrophils at inflammatory sites (Beauchamp et al. Acrolein is a small molecule that is extremely reactive and volatile.
Normally, acrolein is not significant quantity until reaching the smoke phase (Alwis et al. It is estimated to be from 0.08 ppm in ambient air, however, detected at a maximum of 90 ppm in cigarette smoke (Beauchamp et al. Acrolein is a component of cigarette smoke which is known to accumulate in the respiratory system after acute nasal exposure (Tully et al. Long - term cigarette smoking results in significant systemic accumulation of acrolein in humans (Carmella et al.
After the cigarette market opened in Taiwan in 1987, the per capita consumption of betel quid, which has increased by fivefold in 15 years, from 1.4 kg in 1981 to 7. For those 15 years, oral cancer mortality increased fourfold in this country (Wen et al. The doubts began to pose about the relationship of betel quid and cigarette in Taiwan where public health professionals and health professionals have long considered betel to chew and smoke are two separate problems. Betel quid chewing will increase the alkalinity of the saliva, the pH can reach 9 to 11, while healthy saliva is slightly alkaline at a pH 7.
This alkalinity will facilitates the absorption of acrolein with pKa approximately 9.6 (Kaye, 1973); however, this has not been documented in literature. Previous studies have confirmed analysis a potential surrogate marker for acrolein quantification is 3-hydroxypropyl mercapturic acid (3-HPMA) which is a specific and stable metabolite of acrolein in urine (Zheng et al. In contrast to acrolein, 3-HPMA is notably stable in urine, it remains stable for months if stored in −80°C freezer (Moghe et al. Therefore, in this experiment, 3-HPMA, a 2 n typical metabolite of acrolein, was chosen as the analytical object.
3-HPMA data has become increasingly popular as a biomarker to maintain human health because the noninvasive nature of measuring urinary 3-HPMA allows long-term monitoring of cigarette smoke containing acrolein. Investigation on the absorption of acrolein in urine has been performed by liquid chromatography/ Tandem mass spectrometry (LC/MS/MS) of reverse - phase high-performance liquid chromatography (HPLC). This is the basic techique in conducing “The Effect of Betel Quid Chewing on The Absorption of Cigarette Smoke Containing Acrolein” 1. Research’s Objectives The aim of this study was to compare the level of urinary 3-HPMA in cigarette smokers with – and without chewing betel quid and describe some characteristics of cigarette smokers in Kaohsiung.
Research question and hypotheses 1. Research question: - Does the quantitation of acrolein, in term of urinary 3-HPMA, different between cigarette smokers with – and without chewing betel quid? 2. Hypothesis: - HO (Null Hypothesis): The quantitation of acrolein,in term of urinary 3- HPMA, does not different between cigarette smokers with – and without chewing betel quid. - HA (Alternative Hypothesis): The quantitation of acrolein,in term of urinary 3 n 3-HPMA, different between cigarette smokers with – and without chewing betel quid.
Limitations In this survey, due to the low number of samples, the reliability was not high. 4 n PART II - LITERATURE REVIEW 2. Physical and Chemical Properties of Acrolein Table 2. Physical and Chemical Properties of Acrolein Appearance Colorless to yellow liquid.
Colorless gas in smoke. Boiling point 53 °C (127 °F; 326 K) CAS Registry Number 107-02-8 Chemical Name Acrolein Chemical Formula CH2=CHCHO or C3H4O Acraldehyde, Acrylaldehyde, Acrylic aldehyde, Allyl Common Synonyms aldehyde, Ethylene aldehyde, 2-Propenal Density 0.839 g/mL Molecular Weight 56.064 g/mol Melting point −88 °C (−126 °F; 185 K) pH 6.0 Solubility in water Appreciable (> 10%) Structural Formula Specific Gravity 0. Trade names include aqualin, aqualine, biocide, magnacide, magnacide B, and Slimicide (Ghilarducci & Tjeerdema, 1995). At room temperature, Acrolein is a clear, colorless to straw - colored liquid with a pungent, suffocating odor (ACGIH, 1991).
Most people began to smell acrolein in the air at a concentration of 0.25 parts acrolein per million parts air (ppm). It burns and burns easily in the air and highly flammable and burns to produce toxic gases (peroxides, and oxides of carbon) (ATSDR, 2007). Acrolein is volatile, producing toxic concentrations at room temperature. It changes into steam faster than water at normal temperature.
The change of acrolein from liquid to steam becomes faster as the temperature rises.