characterization of chemical compounds in
TRANSCRIPT
CHARACTERIZATION OF CHEMICAL COMPOUNDS IN CIGARETTE
FILTERS LEACHATES
_______________
A Thesis
Presented to the
Faculty of
San Diego State University
_______________
In Partial Fulfillment
of the Requirements for the Degree
Master of Public Health
with a Concentration in
Environmental Health
_______________
by
Veronika Shevchenko
Spring 2012
iii
Copyright © 2012
by
Veronika Shevchenko
All Rights Reserved
iv
DEDICATION
I would like to dedicate this work to my family: my parent for being there for me in
every way throughout my life. And to Eli Grey, the most important person in my life, for
being my inspiration and the driving force behind everything I do.
v
ABSTRACT OF THE THESIS
Characterization of Chemical Compounds in Cigarette Filters Leachates
by Veronika Shevchenko
Master of Public Health with a Concentration in Environmental Health
San Diego State University, 2012
There are more than 4000 identified chemicals in tobacco and more than 7000 identified chemicals in a complex cigarette smoke mix. A lot has been written regarding chemicals in cigarette smoke and the dangers of those chemicals to human lives. But even though a lot has been written on this subject, very little research has been done into identifying the compounds that leachate and can subsequently cause acute toxicity to aquatic environments and life. Still, several studies have been published over the years that do provide evidence that cigarette butt leachates can potentially cause acute toxicity in several different marine and fresh water species including small fish, plankton, bacteria and invertebrates like water fleas. This study is an extension of previously conducted experiments and focuses specifically on leachate compound identification, trying to identify which compounds can possibly be responsible for the acute toxicity to aquatic life. For experiment we utilized two extraction methods: liquid/liquid and solid phase extractions as well as GC×GC/TOF-MS instrument for peak identification. Many chemical compounds were identified with the majority of compounds being identified using the solid phase extraction methods. Liquid/liquid method produced considerably less compounds, but it still yielded several compounds that were unique only to this method. Some of the chemicals identified from smoked cigarette butt leachates could be traced to their original unsmoked cigarette part (i.e. filter or tobacco parts). Chemicals that were retained in the smoked leachate samples, chemicals that disappeared and chemicals that were created could also all be traced and quantified. Some of the chemicals had similar to nicotine structures, an important detail since some of those compounds could be nicotine derivatives and could cause different levels of toxicity. We concluded that further studies into this subject are necessary. Studies mimicking environmental conditions with different condition variations will provide beneficial information. Also, studies using different type and cigarette brands could show variety in chemical composition. We also concluded that it would be best to utilize both extraction methods for future studies and use standards for chemical identification corroboration.
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TABLE OF CONTENTS
PAGE
ABSTRACT ...............................................................................................................................v
LIST OF TABLES ................................................................................................................. viii
LIST OF FIGURES ................................................................................................................. ix
ACKNOWLEDGEMENTS .......................................................................................................x
CHAPTER
1 INTRODUCTION .........................................................................................................1
1.1 Background ........................................................................................................1
1.2 Objective of the Study .......................................................................................2
1.3 Research Plan .....................................................................................................2
1.4 Effects of Cigarette Butt Leachates on Aquatic Life .........................................4
1.5 Composition of Cigarettes .................................................................................5
1.5.1 Filters ........................................................................................................5
1.5.2 Tobacco and Nicotine ...............................................................................6
1.5.3 Cigarette Wrapper and Glue .....................................................................6
1.5.4 Additives ...................................................................................................7
2 LITERATURE REVIEW ..............................................................................................8
2.1 Bioassays............................................................................................................8
2.2 GC×GC/TOF-MS ............................................................................................11
3 METHODOLOGY ......................................................................................................14
3.1 Sample Acquisition and Preparation ................................................................14
3.2 Liquid/Liquid Extraction .................................................................................16
3.3 Solid Phase Extraction .....................................................................................17
3.4 Instrument Operation Procedures ....................................................................17
3.5 Data Analysis ...................................................................................................18
4 RESULTS AND DISCUSSION ..................................................................................19
4.1 Extraction Methods Comparison .....................................................................19
4.2 Chemical Compounds Origins .........................................................................21
vii
4.3 Eco and Human Toxicity .................................................................................22
4.4 Nicotine Similarity ...........................................................................................24
4.5 Study Limitations .............................................................................................26
4.6 Future Studies ..................................................................................................26
5 CONCLUSION ............................................................................................................28
REFERENCES ........................................................................................................................29
APPENDIX
OVERSIZED TABLES ...............................................................................................32
viii
LIST OF TABLES
PAGE
Table 3.1. GC×GC/TOF-MS Conditions of the Analysis .......................................................17
Table 4.1. Number of Chemical Compounds Identified by LLE and SPE Methods ...............19
Table 4.2. Compounds with Structures Similar to Nicotine ....................................................25
Table A.1. Chemicals Identified in the Unsmoked Filter only Leachate Sample ....................33
Table A.2. Chemicals Identified in the Unsmoked Cigarette only Leachate Sample ..............35
Table A.3. Chemicals Identified in the Smoked Cigarette Butts with Remnant Tobacco Leachate Sample ...........................................................................................36
Table A.4. Chemicals Identified in the Smoked Cigarette without Remnant Tobacco Leachate Sample ..........................................................................................................39
Table A.5. Origin of Smoked Cigarette Butt Leachate with Remnant Tobacco .....................43
Table A.6. Origin of Smoked Cigarette Butt Leachate without Remnant Tobacco ................47
Table A.7. Ecotoxicity and Toxicity in Human for a Number of Compounds ........................51
ix
LIST OF FIGURES
PAGE
Figure 3.1. Shows how cigarettes were cut during samples preparation. ................................14
Figure 3.2. Foam container containing all prepared samples. .................................................15
Figure 3.3. Five types of prepared samples. ............................................................................16
x
ACKNOWLEDGEMENTS
Foremost, I would like to express my sincere gratitude to my advisor Dr. Eunha Hoh.
Above all, I am grateful for her continuous support, strong encouragement and motivation.
Her patient guidance and positive approach benefited me enormously during the preparation
of this thesis. I could not have imagined having a better advisor and a mentor. Finally, I
would also like to acknowledge the support and advice of the members of my thesis
committee, Dr. Richard Gersberg and Dr. Victoria Matey as well as Kayo Watanabe for her
greatly appreciated laboratory support.
1
CHAPTER 1
INTRODUCTION
It is a well known fact that thousands of chemicals are found in cigarette smoke with
dozens of these chemicals being identified as human and animal carcinogens (United States
Department of Health and Human Services [USDHHS], 2010). However, it is not known if
cigarette butts, which have become a huge litter problem in recent decades, can be a toxic
risk and become a health threat to marine as well as freshwater habitats. This study focuses
on detecting specific chemicals that can contribute to the toxicity of the cigarette butt litter in
the aquatic environments.
1.1 BACKGROUND
In 2009, cigarette and cigarette butts were the most common type of debris found in
natural water sources (Ocean Conservancy, 2010). According to the 2010 International
Coastal Cleanup Report, an annual report describing the results of a worldwide oceans and
beaches cleanup efforts, “on the list of top ten items found worldwide, cigarettes and
cigarette filters were the most prevalent debris items found” with more than 2.2 million
cigarettes and cigarette filters removed from oceans and inland waterways. 1.3 million of
these were collected from United States alone (Ocean Conservancy, 2010, web page).
Over 7,000 chemicals have been found in particulate matter and mainstream smoke
and over 60 of these are known carcinogens (USDHHS, 2010). Moriwaki, Kitajima, and
Katahira (2009) concluded through their study that heavy metals like arsenic and cadmium,
nicotine and polyaromatic hydrocarbons (PAHs) seep up into the environment from the
inland discarded cigarette butts. There is concern that these same chemicals could potentially
contaminate aquatic ecosystems through littered cigarettes and cigarette butts. These littered
cigarette butts are swept by rain from gutters and streets into storm drains which connect to
open waterways and can negatively impact aquatic life (Ocean Conservancy, 2010).
Multiple studies have discovered that chemicals often found in cigarettes have toxic
effects on marine life. Konar (1977) found that nicotine causes liver damage in fish and upon
2
extended exposure leads to death. Another study, by Micevska, Warne, Pablo, and Patra
(2006) established that cigarette butt leachates are toxic to freshwater invertebrates,
specifically the freshwater flea Ceriodaphnia cf. dubia and marine bacterium Vibrio fischeri.
Most recently, a study was conducted looking specifically at leachate toxicity from
unsmoked cigarette filters, smoked cigarette filters, and smoked cigarette filters with remnant
tobacco in both marine and freshwater fish. Results from this study suggested that all three
were toxic to both marine and fresh water fish, but cigarette filters with remnant tobacco
were the most toxic (Slaughter, Gersberg, Watanabe, Rudolph, & Novotny, 2011).
It is a well known fact that cigarette smoke and specifically the chemicals contained
within cigarette smoke have a negative impact on human health. However, what about
marine and freshwater life? What are the chemicals that leachate into the environment from
littered cigarette butts and do those chemicals have any harmful effects on the aquatic life?
The novelty of the above studies is in the fact that they are the only few that explore this
subject and explore the dangers posed by cigarette butt chemicals to aquatic life.
1.2 OBJECTIVE OF THE STUDY
The objectives of this study are to investigate the risk posed by cigarette butt waste,
specifically by focusing on the characterization of the chemicals in cigarette butts leachates
using a comprehensive two-dimensional gas chromatography with time-of-flight mass
spectrometry (GC×GC /TOF-MS) instrument. This study will determine the specific
chemicals that leach into water and can be the source of toxicity to life in marine and
freshwater environments. Another important goal of the study is to determine which
extraction method was the most practical for these kinds of analyses.
1.3 RESEARCH PLAN
Although we know a great deal about chemicals in cigarette smoke having adverse
effects on human health, we do not really know which specific chemicals cause the toxicity
in fish and other aquatic life in previously conducted experiments. A review of the literature
on toxic chemicals identified in cigarettes, sidestream and mainstream smokes follows.
Stabbert, Schäfer, Biefel, and Rustemeier (2003) found four aromatic amines in the
mainstream smoke. The four aromatic amines were identified as smoke constituents by IARC
3
and were also classified as known human or animal carcinogens. They found that there are
different conditions determining the aromatic amine amounts in smoke, the most important
of which was the combustion temperature. Forehand, Dooly, and Moldoveanu (2000)
identified polycyclic aromatic hydrocarbons (PAHs), phenols and aromatic amines in
particulate phase mainstream smoke using different preparation techniques. In comparing the
different techniques good agreement between the results was established and different types
of PAHs, phenols and aromatic amines were found present in cigarette smoke.
Using an electron monochromator-mass spectrometry (EM-MS) Dane, Crystal, and
Kent (2006) analyzed and showed the presence of three different dinitroaniline pesticides
(pesticides containing nitro group), flumetralin, pendimethalin, and trifluralin in both
mainstream and sidestream tobacco smokes. Their analysis showed the presence of all three
pesticides in smoke with trifluralin levels reaching up to 47 (± 17) nanograms/cigarette
(ng/cig), flumetralin levels of up to 37 (± 9) ng/cig and pendimethalin levels of up to 10.4 (±
0.6) ng/cig. Dane and colleagues (2006) also presented acute toxicity information and stated
that all three pesticides are endocrine disruptors, and trifluralin and pendimethalin are
possible human carcinogens.
Liu, Feng, van Heemst, and McAdam (2000) monitored a group of volatile smoke
analytes from mainstream cigarette smoke and discovered that for some volatile species a
significant fraction in the cigarette mainstream smoke had been generated during the
smoldering period. The tested analytes included nitric oxide, acetaldehyde, acetone, benzene,
toluene, 1, 3 butadiene, isoprene and carbon dioxide.
Moriwaki and colleagues (2009) actually obtained the cigarette butts from collected
waste from the roadside in suburb Japan. The cigarette butts were soaked creating a leachate
in order to measure heavy metals (arsenic, cadmium, copper, lead and chromium), nicotine
and PAH’s in that solution as well as assessing the amounts of these compounds that can
potentially enter the environment. In this study the researchers concluded that heavy metals,
nicotine and PAH’s were discharged into the environment from cigarette butts in waste. They
also speculated that nicotine which was released into the environment from the cigarette butts
could have an adverse effect on living organisms.
4
1.4 EFFECTS OF CIGARETTE BUTT LEACHATES ON
AQUATIC LIFE
The subject of the toxicity of cigarette butts leachates to the aquatic environments is
rather new, although the subject of cigarettes and cigarette smoke toxicity to humans has
been tackled for decades. Some ground breaking studies have been conducted and presented
a picture of an acute toxicity of cigarette butt leachates to marine and freshwater organisms.
These studies include a study conducted in 1977 by Konar showing nicotine, a known major
component in cigarettes to be toxic to fish, aquatic insects and plankton. Upon exposure to 40
percent nicotine solution for 10-20 minutes, the fish surfaced and some jumped up. After 30-
40 minute exposure fish became paralyzed and sunk. Death followed 5-50 hours after the
initial exposure. Aquatic insects became lethargic 10-15 minutes after the exposure and the
most tolerant insects died within 2-7 days. Konar (1977) concluded that the non-air breathing
organisms were more susceptible to nicotine toxicity than the air breathing ones.
In 2006, Micevska and colleagues determined the acute toxicity to Ceriodaphnia cf.
dubia (freshwater flea) and Vibrio fischeri (marine bacterium) using leachates from
artificially smoked cigarette butts. The study concluded that cigarette butts leachates were
acutely toxic to both species, with Ceriodaphnia cf. dubia being more sensitive each time
than Vibrio fischeri. The study also recognized nicotine and ethylphenol to be the key causes
of the toxicity to C. cf. dubia.
Slaughter and colleagues (2011) were the first in literature to investigate the toxicity
of cigarette butt leachates to fish as well as assessing the potential ecological risks of
cigarette butts to the aquatic environments. Slaughter and colleagues (2011) also compared
the results of their study with the study conducted by Micevska and colleagues (2006) and
found that leachates from smoked cigarette butts with tobacco were more toxic to the water
flea Ceriodaphnia cf. dubia then to the two species of fish they studied. For their study
Slaughter and colleagues (2011) tested three conditions: smoked cigarette filters with 1-2cm
of remnant tobacco, smoked cigarettes with no remnant tobacco and unsmoked cigarette
filters with no tobacco. As expected, of the three conditions tested, leachate from smoked
cigarette butts with remnant tobacco showed to be the most toxic to both the saltwater Pacific
topsmelt (Atherinops affinis) and the freshwater minnow (Pimephales promelas) tested.
Smoked cigarette with no tobacco leachate also showed toxicity, but not as great as the
5
toxicity in previous condition. The surprise of the study was that even unsmoked filters with
all tobacco removed showed toxicity to both species, which could mean that even unsmoked
filters contain chemicals acutely toxic to aquatic ecosystems.
1.5 COMPOSITION OF CIGARETTES
In general, a standard cigarette will consist of four parts: a filter, tobacco, cigarette
wrapper and glue, and additives. Cigarette butts are discarded and end up in the environment
as litter. Presumably these cigarette butts will still contain same four parts when they end up
in the environment and each of the residual components can potentially pose different
problems and cause harmful effects in the environment.
1.5.1 Filters
Cigarette filters are intended to have several main tasks: one is to absorb vapors.
Others are to prevent tobacco from entering a smoker’s mouth by collecting particulate
smoke components as well as to provide a form to the cigarette so it does not collapse when
smoked (Register, 2000). Until 1950, only about 1 percent of cigarettes had filters present.
Today, more than 90 percent of the cigarettes sold around the world and 95 percent sold in
the United States have filters (Pauly, Mapani, Lesses, Cummings, & Streck, 2002).
According to Register (2000), 95 percent of cigarette filters are made of a natural plastic
called cellulose acetate. Cellulose acetate is usually derived from cotton and wood pulp. It is
photodegradable, but even under ultimate degradation conditions the base materials will be
diluted in water or soil rather than disappear (Novotny, Lum, Smith, Wang, & Barnes, 2009).
The addition of cellulose acetate filter to the cigarette significantly reduces many of the toxic
chemicals in the smoke. Although the filter seems to barely change tar and nicotine
concentrations in smoke it is still efficient in removing up to 80 percent of semi volatile
phenols, up to 75 percent of carcinogenic and volatile N-nitrosamines, alkylphenols and
volatile pyridines (Hoffmann, Hoffmann, & El-Bayoumy, 2001; USDHHS, 2010). The
fibers of cellulose acetate are thin, white in color and are tightly packed to form a filter.
These fibers are bond with a plasticizer, triacetin (glycerol triacetate), which softens and
welds them together into a cross-linked structure to provide firmness to the filter (Pauly et
al., 2002). Triacetin is a liquid soluble in water, biodegradable in activated sludge and is
6
capable of forming a homogeneous mixture with alcohols, aromatic hydrocarbons and diethyl
ether (United Nations Environment Programme, 2002).
1.5.2 Tobacco and Nicotine
Tobacco is a broad-leafed plant; however, the word tobacco often refers only to the
dry leafs of the tobacco plants. It is a member of the nightshade family (Solonaceae), which
includes potatoes and tomatoes among other members. There are many known species of
tobacco, one of which - Nicotiana tabacum - is cultivated tobacco containing a powerful
alkaloid drug nicotine. The latter is produced by the roots of tobacco plants and stored in
their leaves. Nicotine is a powerful neurotoxin, and its likely purpose is to protect tobacco
plants from insects. Leafs are dried and cured by tobacco producers, and nicotine is inhaled
when tobacco is smoked although it can also be consumed in other ways. Different types of
tobacco and various source regions produce leafs with different tastes, colors, burning and
smoking properties and nicotine content (Hoffmann & Hoffmann, 1997; Register, 2000;
Rodgman & Perfetti, 2008).
Nicotine is an alkaloid drug found in tobacco plants, which is particularly harmful to
insects. In its pure form, it is a colorless liquid. As an antiherbivore chemical, it has been
used as a powerful insecticide although in recent years its popularity has diminished and
various nicotine analogs are used instead. In cigarettes, nicotine is present in very small
doses. In commercial tobacco products nicotine normally constitutes up to 1.8 percent of the
tobacco weight, so that an average tobacco product will contain up to 18 milligram per gram
of nicotine. When consumed by humans (or other large mammals), nicotine acts as a
stimulant, which is the main cause of addiction to tobacco (Register, 2000; USDHHS, 2010).
1.5.3 Cigarette Wrapper and Glue
An essential element of any cigarette is the paper used to wrap the tobacco. Such
paper is usually made from linen fiber or flax. To make it whiter, calcium carbonate is added
during the production process. The addition of calcium carbonate also produces soft ash that
enhances smoking experience when a cigarette burns. Other chemicals are often added to the
paper to control the burning rate, which affects different aspects of smoking, including a
potential number of puffs and the quality of smoke. Such chemicals may include sodium and
7
potassium citrates, monoammonium phosphate and various salts. Another important property
of the paper is its porosity. It is important because the higher the porosity, the greater is the
gas diffusion in and out of the cigarette. Gas diffusion, specifically the diffusion of oxygen
gas into the cigarette facilitates greater airflow and burn rate which dilutes the smoke and in
turn reduces the inhaled tar and nicotine by having the smoker make fewer puffs. When the
tobacco is wrapped, seems of the paper are glued with modified starch. Natural gum is also
frequently used (Hoffmann et al., 2001; Longwood University, 2011).
1.5.4 Additives
There are 599 additives in U.S. manufactured cigarettes that have been regarded as
safe for use in food by the U.S. Food and Drug Administration (USDHHS, 2010). Some are
added for pH adjustment purposes, as well as rate of burning control. Others include
flavorings like cocoa, licorice, menthol, honey, nutmeg and fruit extracts as well as
humectants to keep the tobacco moist (USDHHS, 2010). Unfortunately, the act of burning a
cigarette changes the properties of those chemicals and additives to create more than 7,000
chemical compounds, some of which are toxic and carcinogenic like lead, cyanide and
formaldehyde (USDHHS, 2010).
8
CHAPTER 2
LITERATURE REVIEW
This chapter explores previously investigated bioassays spanning back to 1977
analyzing cigarette butt leachates and their toxicity to a variety of aquatic life ranging from
plankton and invertebrates to small vertebrate freshwater and marine fish species. Also
included in this chapter is information on GC×GC/TOF-MS instrument, its function, and its
benefits over one-dimensional GC and applications.
2.1 BIOASSAYS
This current study does not examine the toxicity of the cigarette butt leachates in a
specific aquatic environment or to any specific aquatic life forms. Rather, this study looked at
specific chemicals that leachate from cigarette butts into water. Previously conducted
experiments detailed in this chapter almost exclusively examine the toxicity of non specific
compounds that leachate from cigarette filters and cigarette butts. Therefore, the current
study is very novel in that it investigates particular chemical compounds rather than general
toxicity of the cigarette butts leachates.
In 1977, Konar researched nicotine as a potential fish poison as well as investigated
nicotine toxicity to freshwater insects and plankton. Different from the rest of the studies
detailed in this chapter, no cigarette butt leachates were used but rather water soluble nicotine
sulfate that contained 40 percent pure nicotine. Another difference is that Konar (1977) also
investigated sodium carbonate and lime as potential compounds that increase nicotine
toxicity. 10 fish species, 8 species of aquatic insects and 7 species of plankton were used in
this experiment. Plankton population was treated with 1, 2, 5, 10, 20, 40 and 50 parts per
million (ppm) of nicotine. Fish and aquatic insect population were treated with mixtures of
nicotine and 100 ppm of sodium carbonate or nicotine and 100 ppm of lime. The results of
Konar (1977) revealed that the toxicity of nicotine varied between species of all three tested
subjects. Fishes surfaced within 20 minutes of exposure, became paralyzed within 40
minutes and dies between 5-50 hours of exposure. Some of the aquatic insects became
9
lethargic and sunk to the bottom within 15 minutes of exposure. Other species like bugs and
beetles became inactive 24 hours before death and died within 2-7 days of exposure. Some of
the zooplankton species were killed at 2 ppm in 168 hours, but others tolerated up to 50 ppm
for 168 hours and all phytoplankton survived 5 ppm for 168 hours. One of the study’s
conclusions was that lime and sodium carbonate do increase the toxicity of nicotine. The
study also demonstrated that air breathing organisms were more tolerant to the exposure than
the non air breathing ones.
In 2000, Register conducted several range finding experiments in which smoked
filters, residue tobacco and new unused filters were used to develop a dose-response curve
and to determine the concentration that would result in LC50 or a lethal concentration 50,
where 50 percent of the test population would result in death during continuous exposure
over a specific period of time. The first experiment used two filters from two smoked
cigarette butts soaked in 500 mL of distilled water for one hour at room temperature (RT)
and exposed 20 daphnids, water fleas, to concentration levels of 4, 2, 3, 0.5, 0.25, 0.125 butts
per liter. The second experiment involved residue tobacco from two smoked cigarette butts
being soaked in 500 mL of distilled water for one hour at RT and again exposed 20 daphnids
to concentration levels of 4, 2, 3, 0.5, 0.25, 0.125 butts per liter. Third experiment did not use
any tobacco, but rather exposed 20 daphnids to leachate from new, unused filters to establish
whether there were chemicals in unsmoked filters that could be toxic to them. New filters
were soaked in 500 mL of distilled water for one hour at RT and daphnids were exposed to
concentration levels of 16, 8, 4, 2, 1 and 0.5 filters per liter. For the first experiment, Register
(2000) found that LC50 was between one and two used cigarette filters per liter. LC50 for
experiment two was between 0.25 and 0.125 residue tobacco cigarette butts per liter. LC50
for the third experiment was not verified because it was established that the concentrations
were not sufficient enough to cause lethality in 50 percent of the test subjects. The third
experiment demonstrated that even at concentrations equivalent to 64 unsmoked filters per
gallon, the filters’ leached chemicals killed less than 50 percent of the water fleas. Thus,
Register (2000) concluded that the chemicals that leached from the residue tobacco (per
second experiment) were the most toxic to daphnids, followed by chemicals leached from
smoked filters (per first experiment) with unsmoked filters being the least toxic of all three.
10
Micevska and colleagues (2006) investigated the variation in the acute toxicity of
cigarette butt leachates to Ceriodaphnia cf. dubia and Vibrio fischeri. Micevska and
colleague’s (2006) study detailed here had three objectives: inspect which factors contribute
to the variation in leachate toxicity of both species, what is the relative sensitivity of C. cf.
dubia and V. fischeri to the leachates and whether any toxicity relationship could be
developed. Micevska and colleagues (2006) used 19 filtered cigarette types from 6 brands of
cigarettes that varied in the amounts of tar and nicotine ranging from 1 to 16 milligrams (mg)
of tar and 0.1 to 1.5 mg of nicotine per cigarette. The cigarettes were artificially smoked to
imitate the smokers’ action of puffing, placed in glass bottles containing 1 liter (L) of
cladoceran water, shaken for 24 hours to create the leachates and then filtered using
1micrometer pore diameter using plankton net. 1993 US EPA protocols were used to test for
cladoceran toxicity, modified Microtox methods were used to test for cigarette butt leachates
toxicity to V. fischeri and GC-MS analysis was used for toxicity identification and
evaluation. The study showed that there was an increase in toxicity to both species with an
increase in tar and nicotine contents. But the tar/nicotine contents still could not explain the
majority of the toxicity variation, which was attributed by researchers to other parameters
like cigarette brand. The EC50 values for two species ranged between 8.9 and 25.9 mg
butts/L (a threefold variation in toxicity) to C. cf. dubia and between 104 and 832 mg butts/L
(eightfold variation in toxicity) to V. fischeri. Micevska and colleagues (2006) concluded
that cigarette butts leachates are acutely toxic to both tested species with C. cf. dubia
consistently demonstrating higher sensitivity to the leachates then V. fischeri. Other analyses
also showed that nicotine and ethylphenol were the main compounds of the toxicity to C. cf.
dubia.
Up until recently, most bioassays on the subject of cigarette butt leachates toxicity
were conducted using non-vertebrate species like water fleas (daphnids) and marine bacteria.
Slaughter and colleagues (2011) studied the effects of cigarette butt leachates on marine and
freshwater fish. For their project, Slaughter and colleagues (2011) used marine fish, topsmelt
Atherinops affinis and fresh water fathead minnow Pimephales promelas. The objectives of
this project were to determine whether cigarette butt leachates were acutely toxic to the two
subjects, if any specific part of the cigarette butt was more toxic than the other components,
compare the sensitivity of the two subjects to organisms tested in previous works and
11
determine whether smoked cigarettes increased the toxicity of the cigarette filters. For the
study, three different leachates were prepared: smoked cigarette butts (SCB) with 1-2
centimeters (cm) of remnant tobacco, smoked filters (SF) with no tobacco and unsmoked
cigarette filters (USF). SCB leachate was prepared by soaking 8 cigarette butts in 2 liters of
dilution water and acute toxicity was measured at 4, 2, 1, 0.5, 0.25, 0.125 cigarette butts/liter
concentrations using 20 fish per concentration. SF leachate was prepared by soaking 16
cigarette butts in 2 liters of dilution water and acute toxicity to 20 was measured at 8, 4, 2, 1,
0.5, 0.25, 0.125 cigarette butts/liter concentrations using 20 fish per concentration. USF
leachate was prepared by soaking 32 cigarette butts in 2 liters of dilution water and acute
toxicity was measured at 16, 8, 4, 2, 1, 0.5 cigarette butts/liter concentrations using 20 fish
per concentration. SCB leachates were found to be acutely toxic to both species of fish and
an LC50 of approximately 1 cigarette butt/L was recorded for both species. SF leachates
were found to be acutely toxic to both species of fish but at different concentrations: 1.8
cigarette butts/L for topsmelt and 4.3 cigarette butts/L for fathead minnows. USF leachates
were found to be acutely toxic to both species of fish with an LC50 value of 5.1 cigarette
butts/L for topsmelt and of 13.5 cigarette butts/L for fathead minnows. Even thought all three
cigarette components showed to be acutely toxic to both fish species, Slaughter and
colleagues (2011) observed that smoked filters with remnant tobacco were the most toxic out
of three leachates and unsmoked filters were the least toxic. They also concluded that fish
species in their experiment showed similar sensitivity to cigarette butt leachates as did
marine bacteria from Micevska and colleague’s study conducted in 2006, but at the same
time showed less sensitivity to the leachates in comparison to water fleas tested by Micevska
and colleagues (2006).
2.2 GC×GC/TOF-MS
Overcrowding of compounds and broad peaks were the norms when complex samples
were used in one dimensional chromatogram (Bertsch, 1999). In turn, the need for a
multidimensional GC system was necessitated by scientific interest to increase resolution of
peak capacity of congested areas of one dimensional chromatogram (Dimandja, 2004). This
leads us to a discussion of a vastly improved multidimensional GC×GC system used in
currently presented study.
12
The methodologies of the multidimensional GC system enhances sensitivity to give it
a superior resolution in peak capacity in comparison to the one dimensional GC system (Ong
& Marriott, 2002). Chromatographic resolution may be enhanced and improved by either
employing selective detectors which are capable of deconvoluting merged peaks or
increasing peak capacity (Bertsch, 2000). The terminology of a peak capacity is the
maximum number of components that can be placed, side by side, into available separation
space at a given resolution (Bertsch, 1999). In principal, when the multidimensional GC×GC
system is coupled to mass spectroscopy with a hard ionization methods, individual
compounds can be identified according to their deconvoluted mass spectrometric fragment
patterns (Gröger, Welthagen, Mitschke, Schäffer, & Zimmermann, 2008).
The GC×GC separation method uses an orthogonal chromatography alignment. The
enhanced resolution arises by serially connecting two columns such that all samples
emerging from the first column enter a second column of different selectivity and samples
are analyzed sequentially (Phillips & Beens, 1999). The second dimension offers additional
extra available spaces to help disperse and spread complex compounds (Bertsch, 2000). In
principal, samples exiting the first column and entering the second column are considered
sub-samples. These sub-samples are composed of simple substances with similar volatility.
Since the two column separation are independent of one another, the multidimensional GC
system has a much more powerful separation and analysis; yielding more accurate and
relevant scientific results (Phillips & Beens, 1999).
The detection method of TOF-MS considers the time of the flight mass spectrometry
analyzer. TOF-MS is a powerful tool used for GC detection due to its high acquisition rates.
In order to obtain enhanced peak resolution, high acquisition rates provide researchers with a
great number of data points per peak. Thus, the GC×GC combined with TOF-MS technique
provides more readily identified and quantified separation of complex compounds (Ong &
Marriott, 2002).
The role of GC×GC separation method in analyzing complex matrices such as
cigarette particulate matter is important due to the fact that there are great amounts of
different compounds in cigarette smoke (Gröger et al., 2008). The enormous number of peaks
in as complex a matrix such as cigarette particulate matter requires a detection technique
powerful enough to collect data in a timely manner (Adahchour, Beens, Vreuls, & Brinkman,
13
2006). The TOF-MS detection technique combined with the GC×GC separation method
helps lead to the detection of a series of peaks in cigarette particulate matter which would be
difficult to evaluate manually (Gröger et al., 2008). In addition, the GC×GC/TOF-MS is a
powerful detection tool which yields the acquisition analysis peak of complex matrix of
compounds at a higher rapidity rate in comparison to one-dimensional GC system
(Adahchour et al., 2006). Where one-dimensional GC would yield merges peaks in complex
matrix of cigarette particulate matters, two-dimensional GC×GC separation method analog
with TOF-MS detection technique offers improved collection analysis of peak capacity by
deconvoluting merged peaks (Bertsch, 2000).
14
CHAPTER 3
METHODOLOGY
Two different extraction methods, liquid/liquid extraction and solid phase extraction
(SPE) were used in parallel to maximize extraction of a wide range of chemicals. Each
extracted solution was analyzed by a non-targeted method using a comprehensive two-
dimensional gas chromatography with time-of-flight mass spectrometry (GC×GC /TOF-MS)
instrument.
3.1 SAMPLE ACQUISITION AND PREPARATION
The unsmoked cigarettes used for this experiment were regular strength type
Marlboro cigarettes by Phillip Morris brand. The smoked cigarette butts were collected from
public ash trays within 24 hours of being smoked and were of the same type and brand as the
unsmoked cigarettes. The cigarettes were cut into two parts (see Figure 3.1): cigarette parts
(1) were used with tobacco and wrapping paper only (no filter). The filter part (2) was cut 0.5
centimeters from the tobacco part using scissors and was also used with the filter wrapping
paper.
Figure 3.1. Shows how cigarettes were cut during samples preparation.
15
All glassware was washed with Fisher Brand Versa Clean liquid concentrate, rinsed
with deionized water and then rinsed with purified Millipore lab water. And then, the
glassware was baked at 450˚C for 6 hours prior to use. Tweezers and scissors used during
preparation were rinsed with HPLC grade methanol to avoid cross contamination. Purified
Millipore water was transferred into test glass bottles using a glass cylinder and was also the
water used in leachate preparation to avoid any contamination from non purified water. Four
grams of cigarette samples (i.e. cigarettes, filters, butts) were soaked in 100 mL of purified
water for seven days. Each container was sealed and all five placed in a styrene foam
container covered with aluminum foil to block the light (see Figure 3.2). The five samples
were prepared (see Figure. 3.3):
1. Blank leachate: purified water 100 mL
2. 4 g of unsmoked filter part in 100 mL of purified water
3. 4 g of unsmoked cigarette part in 100 mL of purified water
4. 4 g of smoked butts without remnant tobacco in 100 mL of purified water
5. 4 g of smoked butts with remnant tobacco in 100 mL of purified water
Figure 3.2. Foam container containing all prepared samples.
After 7 days, each sample was filtered using a Buchner funnel and a P2 qualitative
grade filter paper made of cellulose fiber. The filter paper was 9.0 cm in diameter, with
particle retention of 1-5 micrometer, fine porosity and a slow flow rate of 5 milliliter per
16
Figure 3.3. Five types of prepared samples.
minute. There was approximately 75-100 mL of liquid present in each bottle after the 7 day
period soaking. The volume of each leachate sample depended on the sample itself. Blank
had almost a full amount of liquid initially put in, but the rest of the samples had less volume
due to loss from absorption to cigarettes. The amount of sample that was filtered was evenly
divided and stored in a glass vials. Two different extraction methods, liquid/liquid extraction
and solid phase extraction, were used and description of the procedures of each method is
followed.
3.2 LIQUID/LIQUID EXTRACTION
One half of the filtered sample was transferred into three glass test tubes (size =
32mL) equally. 5 mL of 1:4 ratio of dichloromethane (DCM)/hexane mixture was prepared,
added into each test tubes and vortexed for 1 minute. Carefully, the organic phase (the upper
division) in each test tube was transferred and combined into a separate, 32 mL glass tube.
This procedure was repeated again using 3 mL of the DCM/hexane and the second extract
was combined with the first extract. In order to dry the samples, sodium sulfate (Na2SO4)
was added to the extract and centrifuged for 15 minutes. Applying a Zymark TurboVap
(Caliper Life Sciences, Hopkinton, MA, USA), at 40˚C using nitrogen, the extract was
reduced to 2 mL. 5 mL of hexane was added and reduced to 2 mL by evaporation again. The
17
final extract was then transferred into a 4 mL amber glass vial and stored in -10°C freezer
until GC×GC/TOF-MS analysis. The liquid/liquid extraction was applied to all five samples.
3.3 SOLID PHASE EXTRACTION
Waters Sep-Pak Vac 12cc C18 pre-packaged solid phase extraction (SPE) cartridges
were used to extract the other half of the filtered samples. Prior to use, each C18 cartridge
SPE was conditioned with 2 mL of dichloromethane (DCM), 2 mL of acetone and then 2 mL
of HPLC grade water. After conditioning, the samples were loaded onto the SPEs and eluted
in vacuum, vacuuming continued for another 5 minutes to elute water completely. 5 mL of
acetone and 3 mL of DCM were used as extraction solvents. To eliminate water in the
extract, Na2SO4 was added to the extract. Applying a Zymark TurboVap (Caliper Life
Sciences, Hopkinton, MA, USA), at 40˚C using nitrogen, the extract was reduced to 2 mL. 5
mL of hexane was added and reduced to 2 mL by evaporation again. The final extract was
then transferred into a 4 mL amber glass vial and stored in -10°C freezer until GC×GC/TOF-
MS analysis. The SPE extraction was applied to all five samples. .
3.4 INSTRUMENT OPERATION PROCEDURES
Once both extractions were completed, the samples were injected into GC×GC/TOF-
MS equipped with splitless mode. Table 3.1 reflects the conditions used during this analysis.
Table 3.1. GC×GC/TOF-MS Conditions of the Analysis
Inlet SplitlessInjection volume 1 µLInlet temperature 300˚C for an entire run
Column combination 1st Dimension Rtx-5 35m × 0.25mm × 0.25um2nd Dimension Rtx-17 1m × 0.10mm × 0.10um
Carrier gas HeliumCarrier gas flow Constant flow 1mL/min
First dimension oven programIsothermal at 60˚C for 1 min. with 6˚C/min. to 300˚C, isothermal at 300˚C
for 3 min. with 20˚C/min. to 320˚C, isothermal at 320˚C for 15 min.Second dimension oven
programIsothermal at 80˚C for 1 min. with 6˚C/min. to 320˚C, isothermal at 320˚C
for 3 min. with 20˚C/min. to 340˚C, isothermal at 340˚C for 15 min.Modulator temperature offset
relative to 1st GC oven 35˚CModulation period 5 sec
Modulation timing Hot pulse time 1 secCool time between stages 1.5 sec
Transfer mass line temperature 285˚CTOF-MS temperatures Ion Source 250˚C
Solvent delay 8 minScan rate 151 spectra/second
18
3.5 DATA ANALYSIS
After the instrument ran all 10 samples, instrument software ChromaTOF (version
4.34) was used to perform data analysis. Mass spectra of all peaks collected by the
instrument were run through the National Institute of Standards and Technology (NIST)
Mass Spectral Library for identification. Once we had all instrument identified peaks, a
procedure for data analysis peak selection and data cleanup was devised. This procedure was
developed in order to manually review each peak selected by the software. The processing
method used for this data analysis was 2DKayo. All results were copied by choosing to
“copy selected DP results” within the software before starting the cleanup procedures.
This was done in order to retain the original results of the run. Then, the copy of the
original data was picked to make any changes and the original data was preserved as is. For
data cleaning, three rules were established that had to be followed during the analysis:
1. Mass spectral similarity with the library hit: 700 and above (70% and above)
2. Minimum of three ions to be present for comparison between the peak and the library hit
3. Similarity by human eye between the mass spectra of the peak to the mass spectra of the library hit
First step was to delete all peaks (compounds) that contained silicon (Si). The reason
is that the silica based columns bled during the run and those compounds were picked up by
the software. Next, all peaks that resulted in a lower than 700 library similarities were also
manually erased because the lower the similarity, the lower are the chances that the
compound selected by the software is the actual compound and 700 was a threshold we
decided to use. The maximum similarity achievable by the software is 999. Next step was to
compare each and every sample to its corresponding blank sample (4 LLE samples to LLE
blank and other 4 SPE samples to SPE blank) and erase those compounds that showed up in
both the blank and the sample and had the same or very close retention times (R.T.). The last
step in the data cleanup was to compare each compound from each sample to its compound
in the library hit (chosen by the software) and erase those compounds that did not follow the
three previously developed rules (i.e. compounds that had less than 3 ions in common and
compounds that had a different mass spectra).
19
CHAPTER 4
RESULTS AND DISCUSSION
Presented in this section are mass spectral characterizations of the chemicals
identified in the cigarette butt leachate samples. This section compares two extraction
methods used in this study, identifies the origins of several of the compounds, and looks at
available ecotoxicological risk data of selected compounds.
4.1 EXTRACTION METHODS COMPARISON
Both extraction methods proved to be beneficial in identifying chemical compounds
in different samples of cigarette butt leachates. Table 4.1 shows the number of compounds
that were unique to each extraction method (only SPE/only LLE) and each sample as well as
the number of compounds that were not unique and were found in both LLE and SPE
methods (common compounds).
Table 4.1. Number of Chemical Compounds Identified by LLE and SPE Methods
Test/Sample Filters Cigarettes
Smoked Butts With
Tobacco
Smoked Butts Without Tobacco
Common Compounds (#) 27 20 88 76Total SPE Compounds (#) 41 30 106 99Total LLE Compounds (#) 28 20 90 77
Unique to SPE Compounds (#) 14 10 18 23Unique to LLE Compounds (#) 1 0 2 1
Total # of Compounds (SPE+LLE) 42 30 108 100
The two rows in Table 4.1 named “total SPE compounds” and “total LLE
compounds” demonstrate the number of compounds that were identified in each of the
methods for each of the four samples.
Tables A.1 - A.4 in the Appendix present the actual chemical compounds that were
identified by the GC×GC/TOF-MS instrument. All four Tables identify the chemicals, show
the CAS numbers, both retention times (one per each column) and a similarity number that is
20
derived from how close the identified compound is to the actual compound in the library.
LLE and SPE columns have check marks for each compound that was identified during any
specific method. The grayed areas mark the compounds that were unique to that particulate
extraction method. In all four tables one or more compounds like benzamide, N-propyl- are
numbered and could be observed as different peak numbers with same names. These
compounds were identified by the software to be the same compounds, but because the
retention times for those compounds are absolutely different we suspect that these
compounds are definitely not the same compounds. Also, during the analysis, several
compounds had the same or close retention times but were identified by the software to be
different compounds. We manually corrected them to be the same, placing them in the
category of common compounds but using the compound with higher similarity as the main
and principal compound. Table A.1 in the Appendix presents chemicals identified in the
unsmoked filter only (no tobacco) leachate sample. Table A.2 in the Appendix presents
chemicals identified in the unsmoked cigarette only (no filter) leachate sample. Table A.3 in
the Appendix presents chemicals identified in the smoked cigarette with remnant tobacco
leachate sample and Table A.4 in the Appendix presents chemicals identified in the smoked
cigarette with no remnant tobacco leachate sample.
As it was mentioned previously in this section, unique compounds were identified
using both extraction methods. Table 4.1 refers specifically to compound quantity. In
unsmoked filter leachate samples, 41 compounds were identified from SPE extraction and 28
compounds from LLE extraction. 27 of total compounds were common to both SPE and
LLE extraction. Only one compound, 4-ehtyl octane, was unique to LLE and 14 compounds
were found to be unique to SPE. These included compounds like p-xylene, 2-methyl-3-
hexanone, acetic acid, methoxy-, methyl ester and 2,6-dimethyl-pyrazine. Cigarette leachate
samples yielded a total of 30 compounds for SPE method and 20 compounds for LLE with
all 20 compounds from LLE being common to compounds extracted using SPE method.
None of the compounds in this sample were unique to LLE and 10 were unique to SPE.
Some of the SPE unique compounds for this leachate were 3-peridinol butanoic acid, and 3-
methyl-pentanoic acid. In the leachate sample of smoked cigarette butts with remnant
tobacco, 88 common compounds were identified from both methods. Out of 106 compounds
identified from SPE extraction, 18 compounds were unique to SPE (these included
21
compounds like 1H-pyrrole-2-carboxaldehyde, 2,3-dimethyl-pyrazine, and 3-methyl-1,2-
cyclopentanedione) and out of 90 compounds identified from LLE extraction two
compounds, specifically 2,2,4-trimethyl-1,3-pentanediol diisobutyrate and 2-acetyl-5-
methylfuran were unique to the method. For the smoked cigarette butts without remnant
tobacco sample 76 compounds were common to both methods, 99 compounds were
identified in SPE extraction with 23 compounds being unique to SPE. The SPE unique
compounds for this leachate sample included cotinine, 2(1H)-pyridinone, 1-(4-pyridinyl)-3-
methyl-ethanone, and 2-benzoyloxyacetophenone among others. 77 compounds were
identified in LLE extraction with 3,5-dimethyl-cyclohexene, being the only compound
unique to that specific method of extraction.
Because SPE extraction yielded a higher count of unique compounds in each of four
samples, it is evident that SPE extraction is definitely superior to LLE extraction. It is also a
fact that a few compounds that were detected in LLE method were not detected in SPE.
Since it is still largely unclear which particular compounds cause the toxicity in previously
detailed bioassays, we cannot exclude any evidence and thus it is essential to use both
extraction methods when researching cigarette butts leachate samples.
4.2 CHEMICAL COMPOUNDS ORIGINS
In Tables A.5 and A.6 in the Appendix, only smoked cigarette butt samples (with and
without tobacco) are included. This is because most of the cigarette litter that we are
concerned about are the already smoked cigarette butts. We look at the filter as a sort of
storage that accumulates and retains all the chemicals that were initially present as well as
chemicals that were created during the process of smoking. Tables A.5 and A.6 in the
Appendix present chemicals detected in four tested samples (LLE and SPE compounds
combined) and mark out (check marked) which chemicals were initially in unsmoked filters,
which were initially in unsmoked cigarette part (in tobacco) and which chemicals appeared in
smoked samples in both filters with and without remnant tobacco leachate samples. For
Table A.5 in the Appendix chemicals from both extraction methods and three samples
(smoked cigarette butts with tobacco, filters and cigarettes) were combined. The same thing
was done with Table A. 6 in the Appendix with three samples used being smoked cigarette
butts without tobacco, filters and cigarettes). Copies of the same compounds were erased to
22
avoid recurrence. Both Tables clearly show which of the compounds were found in filters,
cigarettes, or the two smoked samples. We can also calculate how many compounds were
retained or disappeared from smoked samples. For example, in Table A.5 in the Appendix 18
compounds out of initial 42 filter sample compounds were also found in the smoked cigarette
with tobacco sample but the other 24 compounds disappeared and were no longer found in
the smoked sample. If we look at the cigarette sample, 17 compounds out of initial 30 were
retained and 13 compounds disappeared. Looking at Table A.5 in the Appendix we can also
see several compounds like benzyl alcohol, phenylethyl alcohol, diethyl phthalate, and
triacetin that were found in all three samples. Table A.6 in the Appendix has the same
principal, but here we are looking at a sample of smoked cigarette butts without tobacco,
filters and cigarettes. Here, out of 42 initial filter compounds, 17 compounds were retained,
and 25 disappear. Out of 30 initial cigarette compounds, 11 compounds are preserved and 19
disappear. So, for the smoked cigarette butts with tobacco sample, out of 108 compounds
(from both SPE and LLE), 35 compounds were retained from both unsmoked samples of
filters and cigarettes. This means that 73 compounds were most likely created during the
smoking process. For the smoked cigarette butts without tobacco sample, out of 100
compounds, a total of 28 compounds were retained from filters and cigarettes and 72
compounds were presumably created during the smoking process.
4.3 ECO AND HUMAN TOXICITY
To further this study, we looked more closely at some of the compounds identified. In
recent years several previously discussed studies have measured the acute toxicity of nicotine
and cigarette butt leachates of several different species of freshwater flea, marine bacterium
and several species of marine and freshwater fish. However, except for Micevska and
colleagues (2006), who concluded that nicotine and ethylphenol were the likely toxicants in
their study, none of the other studies identify and specify which chemicals caused the
toxicity. Using a TOXNET toxicology database (http://toxnet.nlm.nih.gov/cgi-
bin/sis/htmlgen?HSDB) and identified in the study chemicals’ CAS numbers we searched for
available data on ecotoxicity values as well as some existing human toxicity information.
Table A.7 in the Appendix presents some selected information found in the TOXNET
database on some of the identified compounds. The following compounds are considered to
23
be ecotoxic (i.e. toxic to aquatic organisms): hexanoic acid, 3,5-dimethyl-phenol, quinoline,
phenol, benzonitrile, benzyl alcohol, 2-methyl-phenol (aka o-cresol), 4-methyl-phenol (aka
p-cresol), acetophenone, 2-methoxy-phenol (aka o-methoxyphenol), 2,3-dimethyl-phenol
(aka 2,3-dimethylphenol), 2,4-dimethyl-phenol (aka 2,4-dimethylphenol), 3,4-dimethyl-
phenol, triacetin, nicotine, and diethyl phthalate. Database ecotoxicity was offered only for a
small amount of identified compounds. In view of the fact that the most recently conducted
bioassays focused on water flea organism (i.e. daphnids) and small fresh water fish
Pimephales promelas, most of the ecotoxicity used for this table mainly focuses on Daphnia
magna (a water flea) and Pimephales promelas (a fresh water fathead minnow). The majority
of ecotoxicity information consists of LC50 (median lethal dose) values, an exposure dose
that is lethal to 50 percent of the test subjects and TLm (median tolerance limit), a chemical
concentration at which 50 percent of the test subjects survive after a specific period of time.
Human toxicity data was presented in this table to supplement the limited ecotoxicity
provided in the toxicology database. Most human toxicity data obtained from the database
does not contain specific toxicity values, but rather refers to whether the chemicals in
question are toxic and/or carcinogenic to humans, to what extent are they toxic, possible
routes of exposures and clinical signs of toxicity for some. Only a small number of
compounds contain human toxicity values. These include nicotine, with 60 mg being the fatal
dose for an adult; phenol, with a probable oral lethal dose to humans of 50-500 mg/kg;
phenylethyl alcohol with a probable oral lethal dose for humans between 0.5-5 g/kg; and 2-
chloro-acetophenone, to which a 10 minute exposure to 0.85 mg/L is estimated to be lethal in
man. Most phenol compounds presented in the table considered to be toxic to humans by
ingestions and skin absorption. A few phenol compounds presented in the table are also
classified as possible human carcinogens. These are 4-methyl-phenol (aka p-cresol) and 2-
methyl-phenol, (aka o-cresol). Compounds like 3-methyl-benzaldehyde (aka 3-
methylbenzaldehyde), 1,2,3-Propanetriol, monoacetate (aka Glyceryl monoacetate) and
triacetin are considered to be practically nontoxic to humans.
We found toxicity data of several individual compounds in the leachates to be
ecotoxic, human toxic, and animal toxic. However, when we discuss chemical toxicity,
especially in a complex mixture such as cigarette smoke or cigarette butt leachates (where
multiple compounds are present) we also have to consider the fact of multiple compound
24
interactions. Are the compounds identified in the leachate samples additive? Could they be
synergistic? We have no data on possible accumulation or concentrations of these chemicals
in nature. Maybe an individual compound is safe even at high concentration, but in
combination with other chemicals could result in producing a toxic effect on the environment
the compounds leach into. This is a concern even with compounds that are not considered
toxic and dangerous in the toxicology database. To extend on this idea, there are studies that
evaluate the negative effect of wastewater treatment plants (possibly containing the
chemicals in cigarettes and cigarette butts) on fish and aquatic life. For example, Liney,
Jobling, Shears, Simpson, and Tyler (2005) found that fish exposure to alkylphenolic
chemicals from wastewater plants at early stages of development has a negative effect on
testicular growth. Another study conducted by Woodling, Lopez, Maldonado, Norris, and
Vajda in 2006 also reported female biased sex ratio intersex fish, delayed gonadal
development and other gonadal abnormalities in fish downstream of wastewater plants.
These and similar to these studies confirm that chemical compounds released into the aquatic
systems may have a negative effect on the health of the aquatic environments.
4.4 NICOTINE SIMILARITY
Many of the chemical compounds identified in this study were not found in the
unsmoked samples but rather appeared only in smoked samples (both with and without
tobacco). This fact supports previously conducted studies on tobacco smoke mixtures that
recognized that majority of the harmful chemicals are created during the act of smoking at
which time the temperature within a cigarette can be quite high. The high temperatures can
change the already existing compounds and create many more compounds that could also be
the derivatives of compounds previously found in unsmoked samples of filters and cigarettes.
From previously conducted studies we know that nicotine (and possibly its derivatives) is
one of the most potent poisons and could alone pose an increasing threat to aquatic life. This
fact is supported by Konar’s (1977) research, which showed nicotine to be toxic to both fish
species and aquatic insects. Micevska and colleagues (2006) also identified nicotine as one of
the toxicants in their study. This is why for Table 4.2 we decided to focus on compounds
similar in their structure to nicotine since these could be nicotine derivatives and as such
could also
25
Table 4.2. Compounds with Structures Similar to Nicotine
Compound Nicotine
Similarity Compound
Nicotine Similarity
Compound Nicotine
Similarity
(1's,2's)-Nicotine-N'-oxide
Ethanone, 1-(3-
pyridinyl)-
Pyridine, 3-(1-methyl-1H-pyrrol-
2-yl)-
2,3'-Dipyridyl
Ethanone, 1-(4-
pyridinyl)-
Pyridine, 3-(3,4-
dihydro-2H-pyrrol-5-yl)-
3-Pyridinami
ne, 2,6-dimethyl-
Isoquinoline
Pyridine, 3,5-
dimethyl-
3-Pyridinecar
bonitrile
Nicotine Pyridine, 3-
ethenyl-
5H-1-Pyrindine
Pyrazine,
2,3- dimethyl
Pyridine, 4-ethenyl-
Cotinine
Pyridine, 2,4-
dimethyl-
Quinoline
N
N
O
O
N
N
N
N
N
O
N
N
N
NNH2
N
N
N
NN
NN
N
N
N
N
N
O
N N
N
26
pose a threat if allowed to leachate from littered cigarette butts. Table 4.2 shows compounds
with structural similarity to nicotine. 17 compounds were found to be structurally similar to
nicotine. These compounds consist mostly of pyridines, pyrazines, ethanones and cotinine, a
nicotine metabolite. In addition, nicotine is a compound that is known to be present in
tobacco and cigarette butt waste. Nicotine in cigarettes can be transformed into different
chemicals during smoking, creating many different but still toxic derivatives. Furthermore,
these compounds can be used as markers for cigarette butt leachates in the field water
analysis. Future study should confirm the identities of identified compounds by comparing
them with their authentic standards.
4.5 STUDY LIMITATIONS
There are more than 7000 estimated chemical compounds in sidestream, mainstream
and environmental tobacco smokes (USDHHS, 2010). It is unknown which chemical
compounds and at what quantities enter the aquatic environments from the littered cigarettes
and cigarette butts. Their health effects on aquatic life are also largely unknown.
In this study, the chemicals identified from the leachate of cigarette butts are dependent on our analytical method.
In this study we investigated only one specific brand of cigarettes, the most common brand in the market. Most likely there is a lot of variability in the chemical composition between the different types of cigarettes and brands.
The leachates were prepared by soaking the cigarette butts for 7 days in filtered pure water in a laboratory. Therefore, these were not real environmental conditions.
4.6 FUTURE STUDIES
Chemical composition will vary among different types and brands of cigarettes. Since in our study we only used regular type of cigarettes a future comparison between different types (i.e. regular, lights and ultralights) and different brands of cigarettes could provide additional information of chemical residues in leachates of cigarette butts.
Current study is based on leachates produced by soaking the cigarette butts for only one, 7 day period. A future study involving several different soaking periods with different conditions such as pH, temperature, or salinity could show additional information in chemicals’ behavior in environment.
Another interesting question to answer is whether we can find compounds identified in the lab in actual aquatic environments. Can they be found at all? What is the concentration they are found in nature? If there is a significant concentration of a
27
specific compound or compounds and what are the potential dangers to the overall health of aquatic systems.
28
CHAPTER 5
CONCLUSION
For decades now, thousands of studies have been conducted researching and
identifying chemicals found in tobacco smoke mixture and the effects of those chemicals on
human and animal health. Up to date, only a handful of studies have been conducted
identifying and researching the effects of cigarette butt leachates on marine and freshwater
aquatic environments. This is at the time when cigarette butt litter is reaching massive
proportions as fewer places allow inside smoking and people are forced to smoke outside.
The discarded cigarette butts eventually end up in our oceans, lakes and smaller bodies of
water where they leachate the chemicals retained within the filters and remnant tobacco and
potentially reach concentration that could negatively affect aquatic life. In recent years
several studies have been published that touch on this previously almost unexplored subject.
Most of those studies centered on toxicity of cigarette butt leachates to different aquatic
organisms. This study is an evolution of previous studies but its’ main focus was on
identifying the compounds in the cigarette butts leachate that could be responsible for the
toxicity recorded in previous studies. This study succeeded in identifying many compounds,
several of which we expected to find (i.e. nicotine and triacetin, known to be present in
tobacco and cigarette filters respectively). We also realized that although solid phase
extraction method revealed more of the unique compounds, liquid/liquid extraction method
also produced compounds unique to it and in order to acquire the best possible spectra of all
of the compounds in cigarette butt leachates, both extraction methods should be utilized for
future studies.
But the real question is, how concerned should we really be? Could these chemicals
really reach high enough concentrations to cause real harm? By confirming the true identities
of the chemicals we can use them as markers and get a little closer to the much needed
answers.
29
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32
APPENDIX
OVERSIZED TABLES
33
Table A.1. Chemicals Identified in the Unsmoked Filter only Leachate Sample
Name CAS Similarity R.T. (s) LLE SPE2-Propanone, 1-(acetyloxy)- 592-20-1 878 484.996 , 1.030
p-Xylene 106-42-3 844 489.992 , 0.865 Acetic acid, methoxy-, methyl ester 6290-49-9 841 489.992 , 1.003
1,2-Propanediol, 2-acetate 1-3-6214 898 504.981 , 1.010 1-Propanamine, N,N-diethyl- 4458-31-5 837 509.977 , 1.016
Nonane 111-84-2 907 519.97 , 0.785 1,3,5,7-Cyclooctatetraene 629-20-9 859 519.97 , 0.944
1,2-Epoxy-3-propyl acetate 6387-89-9 923 519.97 , 1.043 Pyrazine, 2,6-dimethyl- 108-50-9 920 539.954 , 1.023
Octane, 4-ethyl- 15869-86-0 921 584.92 , 0.818 3-Penten-1-ol, 2,2,4-trimethyl- 5842-53-5 800 584.92 , 0.838
Benzaldehyde 100-52-7 893 609.901 , 1.129 Carbamic acid, phenyl ester 622-46-8 847 619.894 , 1.109 1,2-Propanediol, diacetate 623-84-7 887 679.848 , 1.063
1-Hexanol, 2-ethyl- 104-76-7 900 684.844 , 0.964 Benzyl Alcohol 100-51-6 923 704.829 , 1.181
3-Acetyl-1H-pyrroline 1072-82-8 869 739.802 , 1.234 Phenylethyl Alcohol 60-12-8 938 819.742 , 1.214
Cyclohexanol, 5-methyl-2-(1-methylethyl)-, [1R-(1à,2á,5à)]- 2216-51-5 890 899.681 , 1.036
3-Hexanone, 2-methyl- 12-6-7379 988 934.654 , 1.379 1,2,3-Propanetriol, monoacetate 26446-35-5 961 984.616 , 1.287
Triacetin 102-76-1 874 1119.51 , 1.313 Nicotine 54-11-5 941 1139.5 , 1.267
Orthoformic acid, triisobutyl ester 16754-49-7 905 1224.43 , 1.201 Pyridine, 3-(3,4-dihydro-2H-pyrrol-5-yl)- 532-12-7 912 1249.41 , 1.399
Acetophenone, 2-chloro- 532-27-4 859 1259.41 , 1.320 1,3,7,7-Tetramethyl-9-oxo-2-oxabicyclo[4.4.0]dec-5-ene 20194-67-6 786 1309.37 , 1.214
Pyridine, 3-(1-methyl-1H-pyrrol-2-yl)- 487-19-4 886 1314.37 , 1.439 2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-
4,4,7a-trimethyl- 15356-74-8 877 1379.32 , 1.406 2,3'-Dipyridyl 581-50-0 904 1384.31 , 1.485
(table continues)
34
Table A.1. (continued)
Name CAS Similarity R.T. (s) LLE SPE2,2-Bis[4-(benzoyloxy)phenyl]propane 2297-14-5 973 1394.3 , 1.241
2-Dodecanol 10203-28-8 912 1409.29 , 0.944 Benzamide, 4-benzoyl-N-
(immino)(methylthio)methyl- 351417-57-7 844 1409.29 , 1.247 Diethyl Phthalate 84-66-2 903 1429.28 , 1.366
2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-oxo-1-butenyl)- 20194-68-7 867 1504.22 , 1.333
Butanenitrile, 2,3-bis(benzoyloxyimino)- N/A 919 1554.18 , 1.439 1,2(4H)-Oxazine-3-ol, 5,6-dihydro-6-oxo-,
benzoate N/A 842 1574.17 , 1.327 Benzoic acid, 1-methylethyl ester 939-48-0 693 1589.16 , 1.326
Benzamide, N-propyl- (1) 10546-70-0 919 1594.15 , 1.346 l-Alanyl-l-alanyl-l-alanine methyl ester 30802-27-8 860 1784.01 , 1.353
Benzamide, N-propyl- (2) 10546-70-0 925 2238.66 , 1.624 Diethylene glycol dibenzoate 120-55-8 872 2243.66 , 1.756
35
Table A.2. Chemicals Identified in the Unsmoked Cigarette only Leachate Sample
Name CAS Similarity R.T. (s) LLE SPEButanoic acid 107-92-6 891 554.943 , 0.812
Pentanoic acid, 3-methyl- 105-43-1 914 554.943 , 0.970 Hexanoic acid 142-62-1 755 574.928 , 0.944 Benzaldehyde 100-52-7 904 609.901 , 1.129
Phenol 108-95-2 935 619.894 , 1.109 Benzyl Alcohol 100-51-6 938 704.829 , 1.181
Phenol, 2-methoxy- 90-05-1 939 784.768 , 1.195 Phenylethyl Alcohol 60-12-8 941 819.742 , 1.214
Phenol, 4-ethyl- 123-07-9 919 879.696 , 1.181 Phenol, 4-ethyl-2-methoxy- 2785-89-9 884 1044.57 , 1.201
Triacetin 102-76-1 963 1119.51 , 1.201 Nicotine 54-11-5 938 1139.5 , 1.353 4-Carene N/A 850 1149.49 , 1.063
Cyclohexane, 1,1,3,5-tetramethyl-, cis- 50876-32-9 772 1204.45 , 1.274 Pyridine, 3-(3,4-dihydro-2H-pyrrol-5-yl)- 532-12-7 928 1244.42 , 1.412 Pyridine, 3-(1-methyl-1H-pyrrol-2-yl)- 487-19-4 885 1309.37 , 1.452
2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-oxo-1-butenyl)- (1) 20194-68-7 747 1324.36 , 1.234
2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl- 15356-74-8 822 1379.32 , 1.406
2,3'-Dipyridyl 581-50-0 928 1379.32 , 1.492 Diethyl Phthalate 84-66-2 819 1429.28 , 1.366
Propanoic acid, 2-methyl-, 2-phenylethyl ester 103-48-0 756 1459.26 , 1.373
3-Pyridinol 109-00-2 776 1464.25 , 1.313 2-Cyclohexen-1-one, 4-(3-hydroxy-1-
butenyl)-3,5,5-trimethyl-, [R-[R*,R*-(E)]]- 52210-15-8 842 1494.23 , 1.287 2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-
oxo-1-butenyl)- (2) 20194-68-7 912 1504.22 , 1.340 2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-
oxobutyl)- 74233-41-3 819 1529.2 , 1.366 4-Fluorobenzoic acid, undec-2-enyl ester N/A 894 1534.2 , 1.333
Propanoic acid, 2,2-dimethyl-, 2-phenylethyl ester 67662-96-8 749 1554.18 , 1.366
2-Cyclohexen-1-one, 4-(3-hydroxybutyl)-3,5,5-trimethyl- 36151-02-7 812 1559.18 , 1.327
Rishitin 18178-54-6 821 1684.08 , 1.360 4,7-Methano-1H-indene, octahydro-2-(1-
methylethylidene)- 74793-54-7 714 1754.03 , 1.346
36
Table A.3. Chemicals Identified in the Smoked Cigarette Butts with Remnant Tobacco Leachate Sample
Name CAS Similarity R.T. (s) LLE SPE2-Cyclopentene-1,4-dione 930-60-9 851 509.977 , 1.102
1,2-Epoxy-3-propyl acetate 6387-89-9 916 519.97 , 1.043 2-Cyclopenten-1-one, 2-methyl- 1120-73-6 920 534.958 , 1.069
Ethanone, 1-(2-furanyl)- 1192-62-7 930 539.954 , 1.076 Pyrazine, 2,3-dimethyl- 5910-89-4 792 549.947 , 1.043 Pyridine, 2,4-dimethyl- 108-47-4 933 564.935 , 1.036 2-Cyclohexen-1-one 930-68-7 883 569.932 , 1.148
2(5H)-Furanone, 5-methyl- 591-11-7 909 574.928 , 1.228 2-Furanmethanol, 5-methyl- 3857-25-8 873 584.92 , 1.082
1,5-Pentanediamine 462-94-2 922 594.913 , 1.228 2-Cyclopenten-1-one, 3-methyl- 2758-18-1 935 609.901 , 1.175
Phenol 108-95-2 973 619.894 , 1.115 4-Methyl-5H-furan-2-one (1) 6124-79-4 852 624.89 , 1.294
Pyridine, 3-ethenyl- 1121-55-7 895 634.882 , 1.089 Benzonitrile 100-47-0 795 639.878 , 1.175
2-Cyclopenten-1-one, 3,4-dimethyl- 30434-64-1 910 649.871 , 1.129 2,4,6-Cycloheptatrien-1-one, 2-hydroxy- 533-75-5 798 654.867 , 1.082
2-Furanone, 2,5-dihydro-3,5-dimethyl N/A 888 654.867 , 1.214 1H-Pyrrole-2-carboxaldehyde 1003-29-8 870 664.859 , 1.234
3-Pyridinecarbonitrile 100-54-9 952 664.859 , 1.247 4(H)-Pyridine, N-acetyl- 67402-83-9 719 679.848 , 1.214
1,4-Pentadiene, 2,3,3-trimethyl- 756-02-5 752 689.84 , 1.162 1,2-Cyclopentanedione, 3-methyl- 765-70-8 919 689.84 , 1.188
2-Cyclohexene-1,4-dione # 4505-38-8 725 699.833 , 1.241 2-Acetyl-5-methylfuran 1193-79-9 867 704.829 , 1.162
Benzyl Alcohol 100-51-6 897 704.829 , 1.188 2-Cyclopenten-1-one, 2,3-dimethyl- 1121-05-7 784 709.825 , 1.175
4-Methyl-5H-furan-2-one (2) 6124-79-4 922 719.818 , 1.379 Phenol, 2-methyl- 95-48-7 960 724.814 , 1.148
Cyclohexane, 1,1,2-trimethyl- 7094-26-0 763 729.81 , 1.135 2-Cyclohexen-1-one, 3-methyl- 1193-18-6 875 739.802 , 1.214 Ethanone, 1-(1H-pyrrol-2-yl)- 1072-83-9 937 734.806 , 1.247
Ethanone, 1-(1-cyclohexen-1-yl)- 932-66-1 818 744.799 , 1.128 Phenol, 4-methyl- 106-44-5 956 749.795 , 1.162
Acetophenone 98-86-2 943 749.795 , 1.221 2,3-Dimethyl-4-hydroxy-2-butenoic lactone
(1)1575-46-8 801 754.791 , 1.300
2-Cyclopenten-1-one, 3-ethyl- 5682-69-9 874 764.783 , 1.214
(table continues)
37
Table A.3. (continued)
Name CAS Similarity R.T. (s) LLE SPE4-Hexen-2-one, 3-methyl- 72189-24-3 903 769.78 , 1.327
Phenol, 2-methoxy- 90-05-1 938 779.772 , 1.201 2,5-Pyrrolidinedione, 1-methyl- 1121-07-9 950 779.772 , 1.426 4-Hexen-3-one, 4,5-dimethyl- 17325-90-5 710 789.764 , 1.228
3-Ethenyl-3-methylcyclopentanone (1) 49664-66-6 744 799.757 , 1.221 Phenol, 2,3-dimethyl- 526-75-0 794 804.753 , 1.176 Phenylethyl Alcohol 60-12-8 911 814.745 , 1.228
Ethanone, 1-(3-pyridinyl)- 350-03-8 901 814.745 , 1.287 2,3-Dimethyl-4-hydroxy-2-butenoic lactone
(2)1575-46-8 869 814.745 , 1.340
2-Cyclopenten-1-one, 3-ethyl-2-hydroxy- 21835-01-8 820 819.742 , 1.195
Phenol, 2-ethyl- (1) 90-00-6 857 839.726 , 1.175 2-Ethylidenecyclohexanone 1122-24-3 754 844.723 , 1.181 1H-Pyrrole-2-carbonitrile 9513-94-4 813 849.719 , 1.327
Phenol, 2,4-dimethyl- 105-67-9 879 854.715 , 1.175 Benzyl nitrile 140-29-4 924 854.715 , 1.333
3-Ethenyl-3-methylcyclopentanone (2) 49664-66-6 727 859.711 , 1.254 Phenol, 2-ethyl- (2) 90-00-6 929 879.696 , 1.188
1-Propanone, 1-phenyl- 93-55-0 898 889.688 , 1.208 3-Ethenyl-3-methylcyclopentanone (3) 49664-66-6 737 894.685 , 1.294
Cyclohexanol, 5-methyl-2-(1-methylethyl)-, (1à,2á,5à)-(ñ)-
15356-70-4 943 899.681 , 1.043
Phenol, 3,4-dimethyl- 95-65-8 782 899.681 , 1.214 Ethanone, 1-(3-methylphenyl)- 585-74-0 792 904.677 , 1.207 Cyclohexene,3-(2-propenyl)- 15232-95-8 637 909.673 , 1.252 Phenol, 2-methoxy-4-methyl- 93-51-6 791 924.662 , 1.208 Phenol, 4-ethenyl-, acetate 2628-16-2 849 954.639 , 1.241 Phenol, 2-ethyl-6-methyl- 1687-64-5 799 964.631 , 1.182
1H-Pyrrole-2,5-dione, 3-ethyl-4-methyl- 20189-42-8 788 974.624 , 1.261 Phenol, 4-ethyl-3-methyl- 1123-94-0 925 984.616 , 1.195
1,2,3-Propanetriol, monoacetate 26446-35-5 885 984.616 , 1.300 Benzenepropanenitrile 645-59-0 952 994.609 , 1.379
Isoquinoline 119-65-3 723 999.605 , 1.366 1H-Imidazole-4-carboxaldehyde 3034-50-2 867 999.605 , 1.419
1,1'-Bicyclopentyl 1636-39-1 771 1004.6 , 1.267 Acetic acid, 2-phenylethyl ester 103-45-7 742 1009.6 , 1.213
Phenol, 4-ethyl-2-methoxy- 2785-89-9 862 1044.57 , 1.208 1H-Inden-1-one, 2,3-dihydro- 83-33-0 917 1059.56 , 1.393
(table continues)
38
Table A.3. (continued)
Name CAS Similarity R.T. (s) LLE SPE5H-1-Pyrindine 270-91-7 916 1069.55 , 1.452
1H-Inden-1-one, 2,3-dihydro-2-methyl- 17496-14-9 862 1089.54 , 1.333 1-Methylindan-2-one 35587-60-1 760 1104.53 , 1.349
Triacetin 102-76-1 880 1119.51 , 1.333 2,2,4-Trimethyl-1,3-pentanediol diisobutyrate 6846-50-0 869 1139.5 , 1.122
Nicotine 54-11-5 889 1139.5 , 1.333 Phenol, 2,6-dimethoxy- 91-10-1 749 1139.5 , 1.426
4-Carene N/A 845 1149.49 , 1.069 Methylchromone 85-90-5 662 1149.49 , 1.304
1(3H)-Isobenzofuranone 87-41-2 959 1149.49 , 1.538 1H-Indole, 3-methyl- 83-34-1 940 1194.46 , 1.432
Orthoformic acid, triisobutyl ester 16754-49-7 910 1224.43 , 1.201 7-Methylindan-1-one 39627-61-7 750 1229.43 , 1.406
Pyridine, 3-(3,4-dihydro-2H-pyrrol-5-yl)- 532-12-7 959 1244.42 , 1.419 Acetophenone, 2-chloro- 532-27-4 874 1254.41 , 1.327
(1's,2's)-Nicotine-N'-oxide 29419-55-4 778 1304.37 , 1.346 Pyridine, 3-(1-methyl-1H-pyrrol-2-yl)- 487-19-4 897 1309.37 , 1.459
2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl-, (R)-
17092-92-1 813 1379.32 , 1.412
2,3'-Dipyridyl 581-50-0 935 1374.32 , 1.511 2,2-Bis[4-(benzoyloxy)phenyl]propane 2297-14-5 996 1394.3 , 1.241
Diethyl Phthalate 84-66-2 910 1429.28 , 1.373 Diphenyl sulfide 139-66-2 895 1439.27 , 1.432
2-Cyclohexen-1-one, 4-(3-hydroxy-1-butenyl)-3,5,5-trimethyl-, [R-[R*,R*-(E)]]-
52210-15-8 919 1494.23 , 1.294
3-Oxo-à-ionone 79734-43-3 878 1504.22 , 1.346 2,4-Dibenzoylpentanedioic acid, dimethyl
esterN/A 932 1544.19 , 1.459
6áBicyclo[4.3.0]nonane, 5á-iodomethyl-1á-
isopropenyl-4à,5à-dimethyl-,N/A 745 1564.18 , 1.287
1,2(4H)-Oxazine-3-ol, 5,6-dihydro-6-oxo-,
benzoateN/A 843 1574.17 , 1.333
Benzamide, N-propyl- (1) 10546-70-0 913 1594.15 , 1.353
2-Cyclohexen-1-one, 4-hydroxy-3,5,5-trimethyl-4-(3-oxo-1-butenyl)-
7070-24-8 894 1644.11 , 1.472
Rishitin 18178-54-6 845 1684.08 , 1.365 4,7-Methano-1H-indene, octahydro-2-(1-
methylethylidene)-74793-54-7 728 1754.03 , 1.352
Benzamide, N-propyl- (2) 10546-70-0 935 2238.66 , 1.630
Diethylene glycol dibenzoate 120-55-8 865 2243.66 , 1.762
39
Table A.4. Chemicals Identified in the Smoked Cigarette without Remnant Tobacco Leachate Sample
Name CAS Similarity R.T. (s) LLE SPE1,2-Propanediol, 2-acetate 1-3-6214 892 504.981 , 0.997 2-Cyclopentene-1,4-dione 930-60-9 866 509.977 , 1.102
1,2-Epoxy-3-propyl acetate 6387-89-9 913 519.97 , 1.043 2-Cyclopenten-1-one, 2-methyl- 1120-73-6 933 534.958 , 1.069
4,4-Dimethyl-2-cyclopenten-1-one (1) 22748-16-9 794 539.954 , 1.076 2,5-Hexanedione 110-13-4 928 554.943 , 1.109 Hexanoic acid 142-62-1 902 559.939 , 0.957
Pyridine, 3,5-dimethyl- 591-22-0 889 564.935 , 1.036 2(5H)-Furanone, 5-methyl- 591-11-7 927 574.928 , 1.228 2-Furanmethanol, 5-methyl- 3857-25-8 818 584.92 , 1.076
1,5-Pentanediamine 462-94-2 914 594.913 , 1.228 2-Furancarboxaldehyde, 5-methyl- 620-02-0 791 604.905 , 1.142 2-Cyclopenten-1-one, 3-methyl- 2758-18-1 954 609.901 , 1.168
Phenol 108-95-2 971 619.894 , 1.109 2H-Pyran-2-one 504-31-4 870 619.894 , 1.300
4-Methyl-5H-furan-2-one 6124-79-4 849 624.89 , 1.287 Pyridine, 4-ethenyl- 100-43-6 870 634.882 , 1.089
Benzonitrile 100-47-0 724 639.878 , 1.176 4,4-Dimethyl-2-cyclopenten-1-one (2) 22748-16-9 858 649.871 , 1.122
3-Pyridinamine, 2,6-dimethyl- 3430-33-9 724 654.867 , 1.082 2-Furanone, 2,5-dihydro-3,5-dimethyl N/A 900 654.867 , 1.214
3-Pyridinecarbonitrile (1) 100-54-9 898 664.859 , 1.247 4(H)-Pyridine, N-acetyl- 67402-83-9 705 679.848 , 1.214
1,2-Cyclopentanedione, 3-methyl- 765-70-8 934 689.84 , 1.188 2-Cyclohexene-1,4-dione # 4505-38-8 730 699.833 , 1.234
2-Acetyl-5-methylfuran 1193-79-9 679 704.829 , 1.168 Benzyl Alcohol 100-51-6 943 699.833 , 1.188
Propanoic acid, 2-methyl-, 2-propenyl ester 15727-77-2 918 704.829 , 1.241 2-Cyclopenten-1-one, 2,3-dimethyl- 1121-05-7 765 709.825 , 1.175
Phenol, 2-methyl- 95-48-7 948 724.814 , 1.148 trans-7-Methyl-3-octene N/A 773 729.81 , 1.135
Ethanone, 1-(1H-pyrrol-2-yl)- 1072-83-9 948 734.806 , 1.247 2-Cyclohexen-1-one, 3-methyl- 1193-18-6 656 739.802 , 1.214
Ethanone, 1-(1-cyclohexen-1-yl)- 932-66-1 773 744.799 , 1.122 Phenol, 4-methyl- 106-44-5 954 749.795 , 1.155
3,4,5,6-Tetrahydrophthalic anhydride 2426-02-0 720 749.795 , 1.247 3-Pyridinecarbonitrile (2) 100-54-9 893 749.795 , 1.340
Acetophenone 98-86-2 858 749.795 , 1.214 (table continues)
40
Table A.4. (continued)
Name CAS Similarity R.T. (s) LLE SPE2,3-Dimethyl-4-hydroxy-2-butenoic lactone
(1) 1575-46-8 845 754.791 , 1.300 2-Cyclopenten-1-one, 3-ethyl- 5682-69-9 875 764.783 , 1.208
2,5-Furandicarboxaldehyde 823-82-5 915 764.783 , 1.327 1H-Imidazole-4-carboxaldehyde (1) 3034-50-2 775 779.772 , 1.148
2,5-Pyrrolidinedione, 1-methyl- 1121-07-9 958 779.772 , 1.432 Phenol, 2-methoxy- 90-05-1 937 784.768 , 1.197
4-Ethyl-2-hydroxycyclopent-2-en-1-one 28017-62-1 640 789.764 , 1.218 5-Hexen-2-one, 5-methyl-3-methylene- 51756-18-4 840 799.757 , 1.168
3-Ethenyl-3-methylcyclopentanone 49664-66-6 729 799.757 , 1.214 Phenol, 3,5-dimethyl- 108-68-9 744 804.753 , 1.168
2(1H)-Pyridinone, 3-methyl- 1003-56-1 857 809.749 , 1.247 Phenylethyl Alcohol 60-12-8 961 814.745 , 1.228
Ethanone, 1-(4-pyridinyl)- 1122-54-9 811 814.745 , 1.287 2,3-Dimethyl-4-hydroxy-2-butenoic lactone
(2) 1575-46-8 913 814.745 , 1.340 2-Cyclopenten-1-one, 3-ethyl-2-hydroxy- 21835-01-8 717 819.742 , 1.195
Phenol, 2-ethyl- (1) 90-00-6 727 839.726 , 1.168 Benzene, (2-methylpropyl)- 538-93-2 643 839.726 , 1.227 2-Ethylidenecyclohexanone 1122-24-3 812 844.723 , 1.181 1H-Pyrrole-3-carbonitrile 7126-38-7 856 849.719 , 1.320
Phenol, 3,4-dimethyl- 95-65-8 878 854.715 , 1.175 Benzyl nitrile 140-29-4 877 854.715 , 1.327
Phenol, 2-ethyl- (2) 90-00-6 929 879.696 , 1.181 Benzamide, 4-benzoyl-N-
(immino)(methylthio)methyl- 351417-57-7 954 889.688 , 1.208 Cyclohexanol, 5-methyl-2-(1-methylethyl)-,
[1R-(1à,2á,5à)]- 2216-51-5 953 899.681 , 1.043 Ethanone, 1-(2-methylphenyl)- 577-16-2 859 904.677 , 1.208 4,4-Dimethylcyclohexadienone 1073-14-9 721 909.673 , 1.240
1H-Pyrrole, 1-(2-furanylmethyl)- 1438-94-4 818 909.673 , 1.247 Ethanone, 1-(4-methylphenyl)- 122-00-9 717 919.666 , 1.214
Cyclohexene, 3,5-dimethyl- 823-17-6 790 924.662 , 1.175 Phenol, 2-methoxy-4-methyl- 93-51-6 820 924.662 , 1.201
Benzaldehyde, 3-methyl- 620-23-5 807 954.639 , 1.241 Phenol, 2-ethyl-6-methyl- 1687-64-5 604 964.631 , 1.184
1H-Pyrrole-2,5-dione, 3-ethyl-4-methyl- 20189-42-8 790 974.624 , 1.261 2-Furancarboxaldehyde, 5-(hydroxymethyl)- 67-47-0 776 974.624 , 1.393
Phenol, 4-ethyl-3-methyl- 1123-94-0 848 979.62 , 1.201 (table continues)
41
Table A.4. (continued)
Name CAS Similarity R.T. (s) LLE SPE2,3-Dimethyl-4-hydroxy-2-butenoic lactone
(1) 1575-46-8 845 754.791 , 1.300 2-Cyclopenten-1-one, 3-ethyl- 5682-69-9 875 764.783 , 1.208
2,5-Furandicarboxaldehyde 823-82-5 915 764.783 , 1.327 1H-Imidazole-4-carboxaldehyde (1) 3034-50-2 775 779.772 , 1.148
2,5-Pyrrolidinedione, 1-methyl- 1121-07-9 958 779.772 , 1.432 Phenol, 2-methoxy- 90-05-1 937 784.768 , 1.197
4-Ethyl-2-hydroxycyclopent-2-en-1-one 28017-62-1 640 789.764 , 1.218 5-Hexen-2-one, 5-methyl-3-methylene- 51756-18-4 840 799.757 , 1.168
3-Ethenyl-3-methylcyclopentanone 49664-66-6 729 799.757 , 1.214 Phenol, 3,5-dimethyl- 108-68-9 744 804.753 , 1.168
2(1H)-Pyridinone, 3-methyl- 1003-56-1 857 809.749 , 1.247 Phenylethyl Alcohol 60-12-8 961 814.745 , 1.228
Ethanone, 1-(4-pyridinyl)- 1122-54-9 811 814.745 , 1.287 2,3-Dimethyl-4-hydroxy-2-butenoic lactone
(2) 1575-46-8 913 814.745 , 1.340 2-Cyclopenten-1-one, 3-ethyl-2-hydroxy- 21835-01-8 717 819.742 , 1.195
Phenol, 2-ethyl- (1) 90-00-6 727 839.726 , 1.168 Benzene, (2-methylpropyl)- 538-93-2 643 839.726 , 1.227 2-Ethylidenecyclohexanone 1122-24-3 812 844.723 , 1.181 1H-Pyrrole-3-carbonitrile 7126-38-7 856 849.719 , 1.320
Phenol, 3,4-dimethyl- 95-65-8 878 854.715 , 1.175 Benzyl nitrile 140-29-4 877 854.715 , 1.327
Phenol, 2-ethyl- (2) 90-00-6 929 879.696 , 1.181 Benzamide, 4-benzoyl-N-
(immino)(methylthio)methyl- 351417-57-7 954 889.688 , 1.208 Cyclohexanol, 5-methyl-2-(1-methylethyl)-,
[1R-(1à,2á,5à)]- 2216-51-5 953 899.681 , 1.043 Ethanone, 1-(2-methylphenyl)- 577-16-2 859 904.677 , 1.208 4,4-Dimethylcyclohexadienone 1073-14-9 721 909.673 , 1.240
1H-Pyrrole, 1-(2-furanylmethyl)- 1438-94-4 818 909.673 , 1.247 Ethanone, 1-(4-methylphenyl)- 122-00-9 717 919.666 , 1.214
Cyclohexene, 3,5-dimethyl- 823-17-6 790 924.662 , 1.175 Phenol, 2-methoxy-4-methyl- 93-51-6 820 924.662 , 1.201
Benzaldehyde, 3-methyl- 620-23-5 807 954.639 , 1.241 Phenol, 2-ethyl-6-methyl- 1687-64-5 604 964.631 , 1.184
1H-Pyrrole-2,5-dione, 3-ethyl-4-methyl- 20189-42-8 790 974.624 , 1.261 2-Furancarboxaldehyde, 5-(hydroxymethyl)- 67-47-0 776 974.624 , 1.393
Phenol, 4-ethyl-3-methyl- 1123-94-0 848 979.62 , 1.201 (table continues)
42
Table A.4. (continued)
Name CAS Similarity R.T. (s) LLE SPE1,2,3-Propanetriol, monoacetate 26446-35-5 875 989.612 , 1.247
Benzenepropanenitrile 645-59-0 941 994.609 , 1.373 1H-Imidazole-4-carboxaldehyde (2) 3034-50-2 840 999.605 , 1.412
Quinoline 91-22-5 690 999.605 , 1.366 Naphthalene, decahydro-, trans- 493-02-7 816 1004.6 , 1.260
Phenol, 4-ethyl-2-methoxy- 2785-89-9 650 1044.57 , 1.202 1H-Inden-1-one, 2,3-dihydro- 83-33-0 909 1059.56 , 1.393
5H-1-Pyrindine 270-91-7 908 1069.55 , 1.452 1H-Inden-1-one, 2,3-dihydro-2-methyl- 17496-14-9 729 1089.54 , 1.329
Triacetin 102-76-1 859 1119.51 , 1.426 Nicotine 54-11-5 939 1139.5 , 1.294
1(3H)-Isobenzofuranone 87-41-2 937 1149.49 , 1.538 1H-Indole, 3-methyl- 83-34-1 869 1194.46 , 1.432
Orthoformic acid, triisobutyl ester 16754-49-7 909 1224.43 , 1.201 7-Methylindan-1-one 39627-61-7 515 1229.43 , 1.403
Pyridine, 3-(3,4-dihydro-2H-pyrrol-5-yl)- 532-12-7 954 1244.42 , 1.412 Pyridine, 3-(1-methyl-1H-pyrrol-2-yl)- 487-19-4 874 1309.37 , 1.452
2,3'-Dipyridyl 581-50-0 924 1379.32 , 1.498 Acetamide, 2-benzoylthio-N-(3-
acetylphenyl)- 309282-67-5 987 1394.3 , 1.241 2-Cyclohexen-1-one, 4-(3-hydroxy-1-
butenyl)-3,5,5-trimethyl- 34318-21-3 821 1494.23 , 1.294 3-Oxo-à-ionone 79734-43-3 791 1504.22 , 1.333
2-Benzoyloxyacetophenone 4010-33-7 947 1544.19 , 1.459 Cotinine 486-56-6 896 1569.17 , 1.676
1,2(4H)-Oxazine-3-ol, 5,6-dihydro-6-oxo-, benzoate N/A 838 1574.17 , 1.333
Benzamide, N-propyl- (1) 10546-70-0 901 1594.15 , 1.353 Benzamide, N-propyl- (2) 10546-70-0 937 2238.66 , 1.624
Diethylene glycol dibenzoate 120-55-8 860 2243.66 , 1.762
43
Table A.5. Origin of Smoked Cigarette Butt Leachate with Remnant Tobacco
Compounds
Smoked With
Tobacco Filter Cigarette2-Cyclopentene-1,4-dione
1,2-Epoxy-3-propyl acetate 2-Cyclopenten-1-one, 2-methyl-
Ethanone, 1-(2-furanyl)- Pyrazine, 2,3-dimethyl- Pyridine, 2,4-dimethyl- 2-Cyclohexen-1-one
2(5H)-Furanone, 5-methyl- 2-Furanmethanol, 5-methyl-
1,5-Pentanediamine 2-Cyclopenten-1-one, 3-methyl-
Phenol 4-Methyl-5H-furan-2-one (1)
Pyridine, 3-ethenyl- Benzonitrile
2-Cyclopenten-1-one, 3,4-dimethyl- 2,4,6-Cycloheptatrien-1-one, 2-hydroxy-
2-Furanone, 2,5-dihydro-3,5-dimethyl 1H-Pyrrole-2-carboxaldehyde
3-Pyridinecarbonitrile 4(H)-Pyridine, N-acetyl-
1,4-Pentadiene, 2,3,3-trimethyl- 1,2-Cyclopentanedione, 3-methyl-
2-Cyclohexene-1,4-dione # 2-Acetyl-5-methylfuran
Benzyl Alcohol 2-Cyclopenten-1-one, 2,3-dimethyl-
4-Methyl-5H-furan-2-one (2) Phenol, 2-methyl-
Cyclohexane, 1,1,2-trimethyl- 2-Cyclohexen-1-one, 3-methyl- Ethanone, 1-(1H-pyrrol-2-yl)-
Ethanone, 1-(1-cyclohexen-1-yl)- Phenol, 4-methyl-
Acetophenone 2,3-Dimethyl-4-hydroxy-2-butenoic lactone (1)
2-Cyclopenten-1-one, 3-ethyl- (table continues)
44
Table A.5. (continued)
Compounds
Smoked With
Tobacco Filter Cigarette
4-Hexen-2-one, 3-methyl- Phenol, 2-methoxy-
2,5-Pyrrolidinedione, 1-methyl- 4-Hexen-3-one, 4,5-dimethyl-
3-Ethenyl-3-methylcyclopentanone (1) Phenol, 2,3-dimethyl- Phenylethyl Alcohol
Ethanone, 1-(3-pyridinyl)- 2,3-Dimethyl-4-hydroxy-2-butenoic lactone (2)
2-Cyclopenten-1-one, 3-ethyl-2-hydroxy- Phenol, 2-ethyl- (1)
2-Ethylidenecyclohexanone 1H-Pyrrole-2-carbonitrile
Phenol, 2,4-dimethyl- Benzyl nitrile
3-Ethenyl-3-methylcyclopentanone (2) Phenol, 2-ethyl- (2)
1-Propanone, 1-phenyl- 3-Ethenyl-3-methylcyclopentanone (3)
Cyclohexanol, 5-methyl-2-(1-methylethyl)-, (1à,2á,5à)-(ñ)- Phenol, 3,4-dimethyl-
Ethanone, 1-(3-methylphenyl)- Cyclohexene,3-(2-propenyl)- Phenol, 2-methoxy-4-methyl- Phenol, 4-ethenyl-, acetate Phenol, 2-ethyl-6-methyl-
1H-Pyrrole-2,5-dione, 3-ethyl-4-methyl- Phenol, 4-ethyl-3-methyl-
1,2,3-Propanetriol, monoacetate Benzenepropanenitrile
Isoquinoline 1H-Imidazole-4-carboxaldehyde
1,1'-Bicyclopentyl Acetic acid, 2-phenylethyl ester
Phenol, 4-ethyl-2-methoxy- 1H-Inden-1-one, 2,3-dihydro-
(table continues)
45
Table A.5. (continued)
Compounds
Smoked With
Tobacco Filter Cigarette5H-1-Pyrindine
1H-Inden-1-one, 2,3-dihydro-2-methyl- 1-Methylindan-2-one
Triacetin 2,2,4-Trimethyl-1,3-pentanediol diisobutyrate
Nicotine Phenol, 2,6-dimethoxy-
4-Carene Methylchromone
1(3H)-Isobenzofuranone 1H-Indole, 3-methyl-
Orthoformic acid, triisobutyl ester 7-Methylindan-1-one
Pyridine, 3-(3,4-dihydro-2H-pyrrol-5-yl)- Acetophenone, 2-chloro-
(1's,2's)-Nicotine-N'-oxide Pyridine, 3-(1-methyl-1H-pyrrol-2-yl)-
2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl-, (R)- 2,3'-Dipyridyl
2,2-Bis[4-(benzoyloxy)phenyl]propane Diethyl Phthalate Diphenyl sulfide
2-Cyclohexen-1-one, 4-(3-hydroxy-1-butenyl)-3,5,5-trimethyl-, [R-[R*,R*-(E)]]-
3-Oxo-à-ionone 2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-oxobutyl)-
2,4-Dibenzoylpentanedioic acid, dimethyl ester 6áBicyclo[4.3.0]nonane, 5á-iodomethyl-1á-isopropenyl-4à,5à-
dimethyl-,
1,2(4H)-Oxazine-3-ol, 5,6-dihydro-6-oxo-, benzoate Benzamide, N-propyl- (1)
Benzeneacetic acid, 3,4-dihydroxy- 2-Cyclohexen-1-one, 4-hydroxy-3,5,5-trimethyl-4-(3-oxo-1-
Rishitin 4,7-Methano-1H-indene, octahydro-2-(1-methylethylidene)-
Benzamide, N-propyl- (2) Diethylene glycol dibenzoate
(table continues)
46
Table A.5. (continued)
Compounds
Smoked With
Tobacco Filter Cigarette2-Propanone, 1-(acetyloxy)-
p-Xylene Acetic acid, methoxy-, methyl ester
1,2-Propanediol, 2-acetate 1-Propanamine, N,N-diethyl-
Nonane 1,3,5,7-Cyclooctatetraene
Pyrazine, 2,6-dimethyl- Octane, 4-ethyl-
3-Penten-1-ol, 2,2,4-trimethyl- Benzaldehyde
Carbamic acid, phenyl ester 1,2-Propanediol, diacetate
1-Hexanol, 2-ethyl- 3-Acetyl-1H-pyrroline
Cyclohexanol, 5-methyl-2-(1-methylethyl)-, [1R-(1à,2á,5à)]- 3-Hexanone, 2-methyl-
1,3,7,7-Tetramethyl-9-oxo-2-oxabicyclo[4.4.0]dec-5-ene 2-Dodecanol
Benzamide, 4-benzoyl-N-(immino)(methylthio)methyl- Butanenitrile, 2,3-bis(benzoyloxyimino)-
Benzoic acid, 1-methylethyl ester l-Alanyl-l-alanyl-l-alanine methyl ester
2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-oxo-1-butenyl)- (1) Butanoic acid
Pentanoic acid, 3-methyl- Hexanoic acid
Phenol, 4-ethyl- Cyclohexane, 1,1,3,5-tetramethyl-, cis-
Propanoic acid, 2-methyl-, 2-phenylethyl ester 3-Pyridinol
4-Fluorobenzoic acid, undec-2-enyl ester Propanoic acid, 2,2-dimethyl-, 2-phenylethyl ester
2-Cyclohexen-1-one, 4-(3-hydroxybutyl)-3,5,5-trimethyl- 2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-oxo-1-butenyl)- (2)
47
Table A.6. Origin of Smoked Cigarette Butt Leachate without Remnant Tobacco
Compounds
Smoked Without Tobacco Filter Cigarette
1,2-Propanediol, 2-acetate 2-Cyclopentene-1,4-dione
1,2-Epoxy-3-propyl acetate 2-Cyclopenten-1-one, 2-methyl-
4,4-Dimethyl-2-cyclopenten-1-one (1) 2,5-Hexanedione Hexanoic acid
Pyridine, 3,5-dimethyl- 2(5H)-Furanone, 5-methyl- 2-Furanmethanol, 5-methyl-
1,5-Pentanediamine 2-Furancarboxaldehyde, 5-methyl- 2-Cyclopenten-1-one, 3-methyl-
Phenol 2H-Pyran-2-one
4-Methyl-5H-furan-2-one Pyridine, 4-ethenyl-
Benzonitrile 4,4-Dimethyl-2-cyclopenten-1-one (2)
3-Pyridinamine, 2,6-dimethyl- 2-Furanone, 2,5-dihydro-3,5-dimethyl
3-Pyridinecarbonitrile (1) 4(H)-Pyridine, N-acetyl-
1,2-Cyclopentanedione, 3-methyl- 2-Cyclohexene-1,4-dione #
2-Acetyl-5-methylfuran Benzyl Alcohol
Propanoic acid, 2-methyl-, 2-propenyl ester 2-Cyclopenten-1-one, 2,3-dimethyl-
Phenol, 2-methyl- trans-7-Methyl-3-octene
Ethanone, 1-(1H-pyrrol-2-yl)- 2-Cyclohexen-1-one, 3-methyl-
Ethanone, 1-(1-cyclohexen-1-yl)- Phenol, 4-methyl-
3,4,5,6-Tetrahydrophthalic anhydride 3-Pyridinecarbonitrile (2)
(table continues)
48
Table A.6. (continued)
Compounds
Smoked Without Tobacco Filter Cigarette
Acetophenone 2,3-Dimethyl-4-hydroxy-2-butenoic lactone (1)
2-Cyclopenten-1-one, 3-ethyl- 2,5-Furandicarboxaldehyde
1H-Imidazole-4-carboxaldehyde (1) 2,5-Pyrrolidinedione, 1-methyl-
Phenol, 2-methoxy- 4-Ethyl-2-hydroxycyclopent-2-en-1-one 5-Hexen-2-one, 5-methyl-3-methylene-
3-Ethenyl-3-methylcyclopentanone Phenol, 3,5-dimethyl-
2(1H)-Pyridinone, 3-methyl- Phenylethyl Alcohol
Ethanone, 1-(4-pyridinyl)- 2,3-Dimethyl-4-hydroxy-2-butenoic lactone (2)
2-Cyclopenten-1-one, 3-ethyl-2-hydroxy- Phenol, 2-ethyl- (1)
Benzene, (2-methylpropyl)- 2-Ethylidenecyclohexanone 1H-Pyrrole-3-carbonitrile
Phenol, 3,4-dimethyl- Benzyl nitrile
Phenol, 2-ethyl- (2) Benzamide, 4-benzoyl-N-(immino)(methylthio)methyl-
Cyclohexanol, 5-methyl-2-(1-methylethyl)-, [1R-(1à,2á,5à)]- Ethanone, 1-(2-methylphenyl)- 4,4-Dimethylcyclohexadienone
1H-Pyrrole, 1-(2-furanylmethyl)- Ethanone, 1-(4-methylphenyl)-
Cyclohexene, 3,5-dimethyl- Phenol, 2-methoxy-4-methyl-
Benzaldehyde, 3-methyl- Phenol, 2-ethyl-6-methyl-
1H-Pyrrole-2,5-dione, 3-ethyl-4-methyl- 2-Furancarboxaldehyde, 5-(hydroxymethyl)-
Phenol, 4-ethyl-3-methyl- (table continues)
49
Table A.6. (continued)
Compounds
Smoked Without Tobacco Filter Cigarette
1,2,3-Propanetriol, monoacetate Benzenepropanenitrile
1H-Imidazole-4-carboxaldehyde (2) Quinoline
Naphthalene, decahydro-, trans- Phenol, 4-ethyl-2-methoxy-
1H-Inden-1-one, 2,3-dihydro- 5H-1-Pyrindine
1H-Inden-1-one, 2,3-dihydro-2-methyl- Triacetin Nicotine
1(3H)-Isobenzofuranone 1H-Indole, 3-methyl-
Orthoformic acid, triisobutyl ester 7-Methylindan-1-one
Pyridine, 3-(3,4-dihydro-2H-pyrrol-5-yl)- Pyridine, 3-(1-methyl-1H-pyrrol-2-yl)-
2,3'-Dipyridyl Acetamide, 2-benzoylthio-N-(3-acetylphenyl)-
2-Cyclohexen-1-one, 4-(3-hydroxy-1-butenyl)-3,5,5-trimethyl- 3-Oxo-à-ionone
2-Benzoyloxyacetophenone Cotinine
1,2(4H)-Oxazine-3-ol, 5,6-dihydro-6-oxo-, benzoate Benzamide, N-propyl- (1) Benzamide, N-propyl- (2)
Diethylene glycol dibenzoate 3-Acetyl-1H-pyrroline
2-Propanone, 1-(acetyloxy)- p-Xylene
Acetic acid, methoxy-, methyl ester 1-Propanamine, N,N-diethyl-
Nonane 1,3,5,7-Cyclooctatetraene Pyrazine, 2,6-dimethyl-
Octane, 4-ethyl- (table continues)
50
Table A.6. (continued)
Compounds
Smoked Without Tobacco Filter Cigarette
3-Penten-1-ol, 2,2,4-trimethyl- Benzaldehyde
Carbamic acid, phenyl ester 1,2-Propanediol, diacetate
1-Hexanol, 2-ethyl- 3-Hexanone, 2-methyl-
Acetophenone, 2-chloro- 1,3,7,7-Tetramethyl-9-oxo-2-oxabicyclo[4.4.0]dec-5-ene
2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl- 2,2-Bis[4-(benzoyloxy)phenyl]propane
2-Dodecanol Diethyl Phthalate
2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-oxo-1-butenyl)- Butanenitrile, 2,3-bis(benzoyloxyimino)-
Benzoic acid, 1-methylethyl ester l-Alanyl-l-alanyl-l-alanine methyl ester
Butanoic acid Pentanoic acid, 3-methyl-
Phenol, 4-ethyl- 4-Carene
Cyclohexane, 1,1,3,5-tetramethyl-, cis- 2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-oxo-1-butenyl)- (1)
Propanoic acid, 2-methyl-, 2-phenylethyl ester 3-Pyridinol
2-Cyclohexen-1-one, 4-(3-hydroxy-1-butenyl)-3,5,5-trimethyl-, [R-[R*,R*-(E)]]-
2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-oxo-1-butenyl)- (2) 2-Cyclohexen-1-one, 3,5,5-trimethyl-4-(3-oxobutyl)-
4-Fluorobenzoic acid, undec-2-enyl ester Propanoic acid, 2,2-dimethyl-, 2-phenylethyl ester
2-Cyclohexen-1-one, 4-(3-hydroxybutyl)-3,5,5-trimethyl- Rishitin
4,7-Methano-1H-indene, octahydro-2-(1-methylethylidene)-
51
Table A.7. Ecotoxicity and Toxicity in Human for a Number of Compounds
Compounds Ecotoxicity Human Toxicity
Hexanoic acid
LC50 Daphnia magna (water flea) 22 mg/L/24 hr LC50 Pimephales promelas (Fathead minnow)
140 mg/L/1 hr LC50 Fathead minnow 88 mg/L/24 hr LC50 Fathead minnow 88 mg/L/48 hr LC50 Fathead minnow 88 mg/L/72 hr
N/A
Phenol, 3,5-dimethyl-
LC100 Tetrahymena pyriformis (protozoa ciliate) 2.3 mmole/l/24 hr
TLm Carassius carassius (Crucian carp) 53 mg/l/24 hr
TLm Tinca tinca (tench) 52 mg/l/24 hr TLm Salvelinus (trout embryos) 50 mg/l/24 hr
Toxic by ingestion and skin absorption
Cyclohexanol, 5-methyl-2-(1-methylethyl)-, [1R-(1à,2á,5à)]- (aka L-
Menthol)N/A
Moderate irritant to mucous membranes on inhalation
Benzaldehyde, 3-methyl- (aka 3-Methylbenzaldehyde)
N/ANo evidence of skin sensitization was found in volunteers treated
with dilute solutions
Quinoline
LC50 Xenophus laevis (South African clawed toad) embryo 26.3 mg/l/96 hr
LC100 Tetrahymena pyriformis (ciliate) 6.19 mmole/l/24 hr
Clinical signs of toxicity include lethargy, respiratory distress.
Irritating to skin and toxic to the retina or optic nerve in man
Cotinine N/A
Cotinine is the main stable metabolite of nicotine and has
been shown to have a biological half-life approximately 10 times
longer than nicotine
Phenol
LC50 Daphnia magna (young) 17 mg/l/24-48 hr LC50 Daphnia magna (adult) 21; 61 mg/l
LC50 Fathead minnow 41-36 mg/l/48-96 hr LC50 Fathead minnow >50 mg/l/1 hr, >50 mg/l/24
hr, >50 mg/l/48 hr, 33 mg/l/72 hr, 32 mg/l/96 hr
Phenol is toxic with a probable oral lethal dose to humans of 50-
500 mg/kg
BenzonitrileTLm Fathead minnow 78 mg/l/96 hr
TLm Fathead minnow 135 mg/l/96 hr
Clinical signs of toxicity include reddening & blister formation
upon skin exposure, respiratory distress, tonic convulsions, thirst, pyrexia, anxiety and tachycardia
3-Pyridinecarbonitrile (aka 3-Pyridinecarbonitrile)
N/A
Pyridine and its derivatives cause local irritation on contact with the
skin, mucous membranes and cornea. Occupational exposure
may occur through inhalation and dermal contact (table continues)
52
Table A.7. (continued)
Compounds Ecotoxicity Human Toxicity
Benzyl AlcoholLC50 Fathead minnows 770 mg/l/48 hr LC50 Fathead minnows 480 mg/l/72 hr LC50 Fathead minnows 460 mg/l/96 hr
Benzyl alcohol is slightly irritating to the skin nose and throat.
Paraparesis and flaccid paraplegia after epidural or intrathecal
administration of more than 7.5 mg of benzyl alcohol as a diluent are
Phenol, 2-methyl- (aka o-Cresol)
LC50 Fathead minnow, size 17.9 mm, age 29 days 14 mg/L/96 hr
LC50 Fathead minnow, size 3.8-6.4 cm 12.55 mg/L/96 hr
LC50 Fathead minnow, size 3.8-6.4 cm 13.42 mg/L/96 hr
LC50 Fathead minnow, size 5.0 cm 18.2 mg/L/96 hr
Possible human carcinogen (classification C). Irritating to skin,
eyes, and respiratory system. Serious or even fatal poisoning may result if large areas of the skin are wet with cresol and the cresol is
not removed
Phenol, 4-methyl- (aka p-Cresol)
LC50 Fathead minnow, juvenile 26 mg/L/24 hrLC50 Fathead minnow, juvenile 21 mg/L/48 hrLC50 Fathead minnow, juvenile 21 mg/L/72 hrLC50 Fathead minnow, juvenile 19 mg/L/96 hr
Possible human carcinogen (classification C). Irritating to skin,
eyes, and respiratory system. Serious or even fatal poisoning may result if large areas of the skin are wet with cresol and the cresol is
not removed
Acetophenone
LC50 Fathead minnow >200 mg/l/1 hr; >200 mg/l/24 hr; 103 mg/l/48 hr; 158 mg/l/72 hr; 155
mg/l/96 hr LC50 Fathead minnow 162 mg/l/96 hr
Has a slightly depressant action on pulse and slight but continuous
decrease of hemoglobin. May result in marked irritation and transient
corneal injury
Phenol, 2-methoxy- (aka o-Methoxyphenol)
EC50 Daphnia magna age 12 hr; 25900 ug/L for 48 hr; Effect: intoxication, immobilization
Eye, skin, gastrointestinal, and/or respiratory tract irritation uopn
exposurePhenol, 2,3-dimethyl- (aka 2,3-
Dimethylphenol)LC50 Daphnia magna 16.0 mg/l/48 hr
Toxic by ingestion and skin absorption
Phenylethyl Alcohol (aka 2-Phenylethanol)
N/AProbable oral lethal dose for human 0.5-5 g/kg; between 1 oz and pint
(or 1lb) for 70 kg person
Phenol, 2,4-dimethyl- (aka 2,4-Dimethylphenol)
LC50 Fathead minnow 17 mg/l/96 hr LC50 Fathead minnow 16.6 mg/l/96 hr
LC50 Fathead minnow 13,650 ug/l/192 hr LC50 Daphnia magna 2,120 ug/l/48 hr
Moderately toxic by ingestion and skin contact. 2,4-dimethylphenol
may be a carcinogen, but its role as a primary cancer-producing agent
is uncertainBenzyl nitrile (aka Phenylacetonitrile) N/A N/A
Phenol, 2-ethyl- (aka 2-Ethylphenol) N/AToxicity is similar to, but less
severe than, phenol1-Propanone, 1-phenyl- (aka Phenyl
ethyl ketone)N/A
Vapors are irritating to eyes and respiratory system
Phenol, 3,4-dimethyl-LC50 Fathead minnows 15 mg/l/48 hr LC50 Fathead minnows 14 mg/l/72 hr LC50 Fathead minnows 14 mg/l/96 hr
Toxic by ingestion and skin absorption
(table continues)
53
Table A.7. (continued)
Compounds Ecotoxicity Human Toxicity
1,2,3-Propanetriol, monoacetate (aka Glyceryl monoacetate)
N/APractically nontoxic: probable oral lethal dose is above 15 g/kg, more
than 2.2 lb for 70 kg person
Triacetin LC50 Cyprinus carpio (Carp) 174 mg/L/48 hrLC50 Leuciscus idus (Ide) 170 mg/L/48 hr
Appears to be innocuous when swallowed, inhaled or in contact
with the skin, but may cause slight irritation in sensitive individuals
Nicotine
EC50 Daphnia magna age <24 hr 0.035 mmol/L for 24 hr; Effect: intoxication, immobilization
EC50 Daphnia pulex age <24 hr 0.00857 mM for 24 hr; Effect: intoxication, immobilization
Nicotine is one of the most lethal poisons known. The fatal adult dose
is 60 mg.
1H-Indole, 3-methyl- (aka 3-Methylindole)
N/A
Tested at 2% in petrolatum in produced no irritation after a 48-hour closed-patch test on human
subjects.
Acetophenone, 2-chloro- (aka 2-Chloroacetophenone)
N/AA 10 minute exposure to 0.85 mg/L
is estimated to be lethal in man
Diethyl Phthalate
LC50 Daphnia magna age <24 hr 52000 ug/L for 48 hr
LC50 Daphnia magna age <24 hr 56470 ug/L for 48 hr
LC50 Fathead minnow, juvenile, length 29-40 mm 17000 ug/L for 96 hr
LC50 Fathead minnow, juvenile, length 29-40 mm 16800 ug/L for 96 hr
Regarded as showing little acute or chronic toxic properties. May
cause transient irritation of nose or throat when heated
Diethylene glycol dibenzoate (ala Diethylene glycol, dibenzoate)
N/A N/A