**Ecological Risk Assessment**

**Registration Case 2460**

**NICOTINE** (CAS Number 54-11-5; PC Code 056702)

**NICOTINE SULFATE** (CAS Number 65-30-5; PC Code 056703)

**TOBACCO DUST** (CAS Number 8037-19-2; PC Code 056704)

![](media/image1.png){width="1.7965277777777777in"
height="1.4965277777777777in"}

**IUPAC***: (S)* -3-(1-Methyl-2-pyrrolidinyl) pyridine

**Synonyms**: Pyridine, 3-\[(2S)-1-methyl-2-pyrrolidinyl\]-; Pyridine,
3-(1-methyl-2-pyrrolidinyl)-, (SL-Nicotine; Pyridine,
(S)-3-(1-methyl-2-pyrrolidinyl); 1-Methyl-2-(3-pyridyl)pyrrolidine;
Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S);
(S)-3-(1-Methyl-2-pyrrolidinyl)pyridine; (-)-Nicotine

**End-Use Products**: Shotgun^®^ Rabbit and Dog Repellent (EPA Reg. No.
4-465; 0.35% nicotine; 15% naphthalene; 15% dry blood); Fulex Nicotine
Fumigator (EPA Reg. No. 1327-41; 13.4% nicotine). "Nicotine Dust" (CAS
Reg. No. 8037-19-2) is a source of nicotine in the products.

**Prepared By:** Colleen M. Flaherty, M.S., Biologist

Silvia C. Termes, Ph.D., Chemist

Ecological Risk Branch III

Environmental Fate and Effects Division

**Secondary Reviewers:** Stephanie Syslo, M.S., Environmental Scientist

James Hetrick, Ph.D, Branch Senior Scientist

Ecological Risk Branch III

Environmental Fate and Effects Division

**Branch Chief:** Karen Whitby, Ph.D.

Ecological Risk Branch III

Environmental Fate and Effects Division

**Date:** 30 July 2007

**Table of Contents**

[**I.** **Executive Summary** 4](#__RefHeading___Toc173552872)

[**A. Nature of Chemical Stressor** 4](#__RefHeading___Toc173552873)

[**B. Potential Risks to Non-target Organisms**
4](#__RefHeading___Toc173552874)

[**C.** **Environmental Fate and Exposure**
5](#__RefHeading___Toc173552875)

[**D.** **Ecological Effects** 6](#__RefHeading___Toc173552876)

[**E.** **Uncertainties and Data Gaps** 7](#__RefHeading___Toc173552877)

[**1.** **Environmental Fate and Exposure Assessment**
7](#__RefHeading___Toc173552878)

[**2.** **Ecological Effects** 8](#__RefHeading___Toc173552879)

[**II.** **Problem Formulation** 10](#__RefHeading___Toc173552880)

[**A.** **Stressor Source and Distribution**
10](#__RefHeading___Toc173552881)

[**1.** **Chemical Identification** 11](#__RefHeading___Toc173552882)

[**2.** **Physical/Chemical/Fate and Transport Properties**
12](#__RefHeading___Toc173552883)

[**3.** **Pesticide Type, Class, and Mode of Action**
13](#__RefHeading___Toc173552884)

[**4**. **Overview of Pesticide Usage**
14](#__RefHeading___Toc173552885)

[**B.** **Receptors** 15](#__RefHeading___Toc173552886)

[**C.** **Assessment Endpoints** 15](#__RefHeading___Toc173552887)

[**D.** **Conceptual Model** 15](#__RefHeading___Toc173552888)

[**1.** **Risk Hypotheses** 15](#__RefHeading___Toc173552889)

[**2.** **Diagram** 16](#__RefHeading___Toc173552890)

[**F.** **Data Gaps** 18](#__RefHeading___Toc173552891)

[**1.** **Environmental Fate** 18](#__RefHeading___Toc173552892)

[**2.** **Ecological Effects** 20](#__RefHeading___Toc173552893)

[**III.** **Analysis** 20](#__RefHeading___Toc173552894)

[**A.** **Use Characterization** 20](#__RefHeading___Toc173552895)

[**B.** **Exposure Characterization** 20](#__RefHeading___Toc173552896)

[**1.** **Environmental Fate and Transport Characterization**
20](#__RefHeading___Toc173552897)

[**2.** **Aquatic Exposure Characterization**
24](#__RefHeading___Toc173552898)

[**a.** **Simulation Modeling** 24](#__RefHeading___Toc173552899)

[**b.** **Monitoring Information** 27](#__RefHeading___Toc173552900)

[**3.** **Terrestrial Exposure Characterization**
27](#__RefHeading___Toc173552901)

[**C.** **Ecological Effects Characterization**
28](#__RefHeading___Toc173552902)

[**1.** **Aquatic Effects Characterization**
29](#__RefHeading___Toc173552903)

[**a.** **Aquatic Animals** 29](#__RefHeading___Toc173552904)

[**(1)** **Acute Effects** 29](#__RefHeading___Toc173552905)

[**(2)** **Chronic Effects** 31](#__RefHeading___Toc173552906)

[**b.** **Aquatic Plants** 32](#__RefHeading___Toc173552907)

[**2.** **Terrestrial Effects Characterization**
32](#__RefHeading___Toc173552908)

[**a.** **Terrestrial Animals** 32](#__RefHeading___Toc173552909)

[**(1)** **Acute Effects** 32](#__RefHeading___Toc173552910)

[**(2)** **Chronic Effects** 33](#__RefHeading___Toc173552911)

[**b.** **Terrestrial Plants** 36](#__RefHeading___Toc173552912)

[**IV.** **Risk Characterization** 36](#iv.-risk-characterization)

[**A.** **Risks to Aquatic Organisms** 36](#__RefHeading___Toc173552914)

[**B.** **Risks to Terrestrial Organisms**
37](#__RefHeading___Toc173552915)

[**C.** **Ecological Incident Information**
38](#__RefHeading___Toc173552916)

[**D.** **Federally Threatened and Endangered (Listed) Species
Concerns** 38](#__RefHeading___Toc173552917)

[**E.** **Assumptions, Limitations, Uncertainties, Strengths and Data
Gaps** 38](#__RefHeading___Toc173552918)

[**1.** **Exposure For All Taxa** 38](#__RefHeading___Toc173552919)

[**a.** **Maximum Use Scenario** 38](#__RefHeading___Toc173552920)

[**b.** **Additive and/or Synergistic Effects**
38](#__RefHeading___Toc173552921)

[**2.** **Exposure for Aquatic Species**
39](#__RefHeading___Toc173552922)

[**a.** **Data Gaps and Uncertainties**
39](#__RefHeading___Toc173552923)

[**b.** **Aquatic Exposure Model** 39](#__RefHeading___Toc173552924)

[**a.** **Location of Wildlife Species**
41](#__RefHeading___Toc173552925)

[**b.** **Routes of Exposure** 42](#__RefHeading___Toc173552926)

[**c.** **Dietary Intake** 44](#__RefHeading___Toc173552927)

[**4.** **Ecological Effects Assessment**
44](#__RefHeading___Toc173552928)

[**a.** **Data Gaps and Uncertainties**
44](#__RefHeading___Toc173552929)

[**b.** **Sublethal Effects** 45](#__RefHeading___Toc173552930)

[**APPENDIX A. Environmental Fate and Exposure Assessment**
46](#__RefHeading___Toc173552931)

[**APPENDIX B. Aquatic Exposure Model (GENEEC2) Output**
56](#__RefHeading___Toc173552932)

[**APPENDIX C. ECOTOX Open Literature Study Summaries and Bibliography**
57](#__RefHeading___Toc173552933)

[]{#__RefHeading___Toc173552872 .anchor}**I. Executive Summary**

[]{#__RefHeading___Toc173552873 .anchor}**A. Nature of Chemical
Stressor**

There are three active ingredients in the List B, nicotine and
derivatives case number 2460: nicotine (PC code 056702), nicotine
sulfate (PC code 056703), and tobacco dust (PC code 056704). There are
no active products for either nicotine sulfate or tobacco dust; this
ecological risk assessment focuses solely on nicotine use as a
pesticide.

Nicotine ((*S*) -3-(1-Methyl-2-pyrrolidinyl) pyridine; CAS Reg. No.
54-11-5) is an insecticide used primarily for insects with
piercing-sucking mouthparts, such as aphids, whiteflies, leafhoppers,
thrips, and mites. Nicotine is fast-acting, easily absorbed through the
eyes, skin, and mucous membranes.

There are two active products for nicotine: Fulex Nicotine Fumigator
(EPA Reg. No. 1327-41) and Shotgun^®^ Rabbit and Dog Repellent (EPA
Reg.No. 4-465). Fulex Nicotine Fumigator (13.4% nicotine) is a
RESTRICTED USE pesticide (due to very high acute inhalation, oral,
dermal, and eye toxicity to humans) for use only in greenhouses on
ornamental plants. This product is used to control aphids and thrips.
Since this formulation may only be used in greenhouses, environmental
exposures and ecological risk to non-target species are presumed to be
negligible.

Shotgun^®^ Rabbit and Dog Repellent (Formerly F&B Rabbit and Dog Chaser)
is a dust containing 0.35% nicotine, 15% naphthalene, and 15% dried
blood and is exclusively for residential and/or homeowner use. The
product is applied in the perimeter of plants or areas to be protected
from dogs and rabbits. The mode of action of nicotine when used as an
active ingredient in a dog and rabbit chaser is unclear.

[]{.mark}

[]{#__RefHeading___Toc173552874 .anchor}**B. Potential Risks to
Non-target Organisms**

Aquatic EECs for nicotine when used as an active ingredient in
Shotgun^®^ Rabbit and Dog Repellent were estimated using GENEEC (Version
2.0) and adjusted for the percent area treated assuming a generic
residential setting. The highest peak aquatic EEC was 40 ng/L (parts per
trillion). There were no acceptable toxicity data available to
quantitatively assess the potential risks to of Shotgun^®^ Rabbit and
Dog Repellent to aquatic organisms. Acute and chronic risks to aquatic
organisms cannot be precluded; however, given the extremely low
predicted aquatic exposures, the likelihood of risk is presumed to be
very low. Acute toxicity data for freshwater fish (Guideline 72-1)
freshwater invertebrates (Guideline 72-2), and aquatic plants (Guideline
123-2) would reduce the uncertainty in this risk assessment.

Using the T-REX model (Version 1.3.1), terrestrial dietary exposures
were estimated for nicotine as an active ingredient in Shotgun^®^ Rabbit
and Dog Repellent, a granular (dust) formulation for use in residential
settings. Assuming a generic residential setting, the estimated exposure
for a terrestrial animal is about 67 mg a.i./A. Acute mammalian toxicity
data from the open literature suggest that nicotine is very highly toxic
to mice, with an acute oral LD~50~ of 3 mg/kg,[^1] and based on the
modeled exposure, there is a potential for risk. However, this nicotine
product is a rabbit and dog repellent, and if small mammals (*e.g.*,
field mice) are similarly repelled, terrestrial dietary exposure may be
unlikely.

There are no avian toxicity data available for nicotine, and it is
unclear whether Shotgun^®^ Rabbit and Dog Repellent is capable of
repelling birds as well. Risk to birds cannot be precluded at this time.
Acute and chronic avian toxicity data (Guidelines 71-1, 71-2, and 71-4)
would help reduce this uncertainty regarding the risk of nicotine to
birds.

There are no terrestrial invertebrate data available for consideration
in this risk assessment. Nicotine shares a common mode of action with
neonicotinoid insecticides, implicated in the decline of honey bee
populations. Since Shotgun^®^ Rabbit and Dog Repellent is a dust
(granular) to be applied in a band around the perimeter of gardens,
exposure to beneficial insects (*e.g.*, honey bees) may be unlikely. An
acute contact toxicity test with honey bees (Guideline 141-1) would help
reduce the uncertainty in this risk assessment.

There are no terrestrial plant toxicity data available for consideration
in this risk assessment. At this time, risks to terrestrial plants
cannot be precluded. Tier I seedling emergence and vegetative vigor
studies (Guideline 123-1(a,b)) would help reduce the uncertainty in this
assessment.

[]{#__RefHeading___Toc173552875 .anchor}**C. Environmental Fate and
Exposure**

The description of environmental fate of nicotine is based on model
estimates using EPISuite Version 3.20 and relevant open literature
sources. EPISuite is only a screening level used to assess the
environmental fate and transport of a chemical (i.e., persistence and
transport). The environmental fate profile of nicotine is summarized
below:

a.  Nicotine is stable to abiotic hydrolysis.

b.  Indirect photolysis in water is likely to be an important
    dissipation pathway in water, but how fast indirect photolysis takes
    place is unknown. The rate of indirect photolysis will depend on the
    nature and concentration of photosensitizers is surface water, which
    is temporally and spatially variable. Direct photolysis in water is
    not likely to be an important dissipation pathway for nicotine.

c.  The biotransformation (aerobic) of nicotine in soils is not rapid
    (EPISuite-estimated half-live of 37 days). The biotransformation
    products of nicotine are well known and involve sequential oxidative
    steps brought about by microbial enzymes. Formation of cotinine in
    soils has not been reported, Cotinine, however, is a major human
    metabolite, which used as a marker for nicotine exposure.

d.  In water-sediment systems, oxic conditions appear to favor
    mineralization of cotinine, with complete recovering as carbon
    dioxide (i.e., complete mineralization) observed after 72 days,
    which falls within the "weeks-to-months" time frame for the ultimate
    biodegradation of nicotine estimated by EPISuite. Under anoxic
    conditions, the biotransformation of nicotine and cotinine in stream
    sediments involves microbial demethylation of the N-methyl group of
    both nicotine and cotinine.

e.  In aerobic soils, the enzyme-catalyzed biotransformation of nicotine
    proceeds as a series of sequential oxidative processes.

f.  The mobility of nicotine in soil is moderate, based on a single
    EPISuite Koc of 2376. However, estimates of Koc in EPISuite are
    based on neutral compounds. Nicotine is a weak acid (pKa 8.5), and
    the relative concentrations of the protonated and deprotonated forms
    is dependent on pH. Theoretically, mobility would increase with
    increasing pH. Volatilization of nicotine from soils depends on how
    tight it binds to soils. The half-life and rate volatilization are
    indirectly proportional to the Koc, which means that volatilization
    would be faster from soils on which nicotine does not bind strongly.

g.  Nicotine is unlikely to volatilize from water, based on a Henry's
    Law Constant of 3.0 x 10^-9^ atm-m^3^mole^-1^ ( 25° C) and a Log of
    air-water partition coefficient (Log K~aw~) of -6.91 .

h.  If released to air (e.g., from soils on which it does not bind
    tightly), nicotine will react rapidly with photochemically generated
    hydroxyl radicals (half-life 1.14 days).

i.  Atmospheric transport of nicotine sorbed to colloidal particulates
    and wet deposition may result in off-site exposure.

j.  Nicotine is not likely to bioaccumulate or bioconcentrate in fish.

    []{#__RefHeading___Toc173552876 .anchor}**D. Ecological Effects**

There are no acceptable guideline ecotoxicity studies for nicotine. In
1994[^2], the EFED denied waiver requests for the following studies:
acute avian oral toxicity (71-1(a)), acute avian dietary toxicity
(71-2(a)), and acute fish toxicity (72-1(c); however, none of these
studies were subsequently provided to the Agency. A search of the open
literature (ECOTOX database) identified 177 studies that were acceptable
for ECOTOX and OPP (**Appendix C**). Of these, 10 studies were reviewed
for this risk assessment; however, none of them were deemed acceptable
for quantitative use in this ecological risk assessment.

Several studies from the open literature suggest that nicotine is at
least moderately toxic to freshwater fish and at least highly toxic to
freshwater invertebrates; however, given the high uncertainty in the
actual exposure concentrations in the tests, the reported toxicity
thresholds (*i.e.,* LC50, NOAEC) are unreliable. Exposures were
analytically verified in only one study, and the recovery rates were
extremely poor. The following statement regarding aquatic toxicity is on
the product label: "This product is toxic to fish. Do not apply directly
to water. Do not contaminate water by cleaning of equipment or disposal
of waste."

There are no avian acute or chronic toxicity data available for
nicotine. One study for tobacco dust indicated that the acute oral
LD~50~ was determined to be greater than 2150 mg/kg bw for bobwhite
quail; however, the percent nicotine in the test substance was not
reported.

Based on information provided by the Health Effects Division (HED),
there are no acceptable guideline acute or chronic toxicity studies for
mammals. Data from the open literature suggest that nicotine is very
highly toxic to mice, with an acute oral LD~50~ of 3 mg/kg; however,
these data have not been formally reviewed by the Agency. There are no
chronic reproductive mammalian toxicity data available for nicotine;
however, there are several studies in the open literature that provide
information to qualitatively assess the potential impacts of nicotine on
mammals.

There are no available aquatic or terrestrial plant toxicity data for
nicotine. The product label states, "Do not apply the product directly
to foliage or stems," which suggests that phytotoxic effects are
possible.

[]{#__RefHeading___Toc173552877 .anchor}**E. Uncertainties and Data
Gaps**

[]{#__RefHeading___Toc173552878 .anchor}**1. Environmental Fate and
Exposure Assessment**

No guideline studies (USEPA, FIFRA Subdivision N) studies conducted with
nicotine as the test substance are available. On September 7, 1994
(DPBarcode D206473), the Environmental Fate and Effects Division (EFED)
concluded that the environmental fate data requirements for nicotine
could be waived. However, the registrant did not request any waiver nor
provided the open literature information requested by EFED.

The environmental fate data used in the present ecological risk
assessment comes from estimates using EPISuite (Version 3.20) and from
the open scientific literature. When integrated, the data provide an
overview of how nicotine would behave in the environment when used as a
pesticide.

The source of nicotine in the end-use products being considered is
"tobacco dust" (as defined in the USEPA's "Substance Registry
Inventory") and not pure ("neat") nicotine. The environmental fate
exposure assessment is based on "neat" nicotine. Assumptions had to be
made to estimate nicotine content in tobacco dust (*i.e.*, the source of
nicotine). It is unclear if the "0.35%" nicotine content in Shotgun^®^
Rabbit and Dog Repellent product is based on a nominal or measured
concentration. For this reason, the nicotine content in the product(s)
was assumed to be 0.35% "neat nicotine" (i.e., pure nicotine). How this
assumption over- or underestimates the exposure of nicotine in the
environment, when used as a pesticide, is unknown. It should also be
kept in mind that nicotine content in the tobacco plant depends on the
strain of tobacco, where it is cultivated, and varies in the plant
itself (leaves, stems, etc).

The manner in which Shotgun^®^ Rabbit and Dog Repellent is used can be
envisioned as a localized application, given that the product is to be
used on the perimeter of the area to be protected (ornamental beds;
vegetable gardens). To express the application rate in terms of pounds
of active ingredient (nicotine) per acre (lb a.i,/acre), as required by
the exposure models, several assumptions had to be made to extrapolate
localized applications to lb a.i./acre. Among others these assumptions
include: (a) typical size of the treatment area for ornamental beds and
vegetable gardens; (b) width of bands of product applied; (c) amount of
product per square foot; (d) number of applications and interval between
applications.

The present ecological risk assessment is based only on estimated
exposure concentrations that result exclusively from the use of nicotine
as a pesticide. Other sources of nicotine exposure in the environment
come from tobacco smoking/chewing, use as a non-prescription drug, and
from releases by the tobacco industry. Nicotine is listed in the Toxic
Release Inventory (TRI), and the tobacco industry is required to report
releases of nicotine to the environment on a yearly basis. Because
exposure to nicotine can come from other sources, it becomes difficult
to correlate level of exposure to nicotine due solely from its use as a
pesticide. This ecological risk assessment only considers nicotine used
as a pesticide and does not consider aggregate exposure from all other
sources. Therefore, risk from pesticide use of nicotine may
underestimate the overall ecological risk of nicotine.

Although the nature of enzyme-catalyzed oxidation products in soils is
known, there are no environmental data to estimate their concentrations
in water sources.

The T-REX model is designed to calculate risk indices from pesticide
applications on an entire one-acre agricultural field by broadcast
spray/granular application or by rows/bands. Shotgun^®^ Rabbit and Dog
Repellent is recommended to be applied in 2 or more inch-wide bands
around the perimeter of a flowering bed, house, or garden, and T-REX
assumptions likely overestimate the exposure estimates. Further,
Shotgun^®^ Rabbit and Dog Repellent is a as a dog and rabbit repellent,
and assuming small mammals (*e.g.*, field mice) are similarly repelled,
the terrestrial dietary exposure pathway for mammals is unlikely.

Currently, the T-REX model does not have the capability to estimate
chronic exposure to terrestrial animals from banded granular
applications.

[]{#__RefHeading___Toc173552879 .anchor}**2. Ecological Effects**

As described above (in Section I.D), there are no acceptable guideline
aquatic or terrestrial toxicity data for nicotine. **Table I.2**
summarizes the ecotoxicity data gaps for nicotine.

+------+-------------+------+-----------------------------------------+
| **T  |             |      |                                         |
| able |             |      |                                         |
| I.2  |             |      |                                         |
| Ec   |             |      |                                         |
| olog |             |      |                                         |
| ical |             |      |                                         |
| Eff  |             |      |                                         |
| ects |             |      |                                         |
| Data |             |      |                                         |
| Requ |             |      |                                         |
| irem |             |      |                                         |
| ents |             |      |                                         |
| for  |             |      |                                         |
| Ni   |             |      |                                         |
| coti |             |      |                                         |
| ne** |             |      |                                         |
+======+=============+======+=========================================+
| *    | **Data      | **MR | **Are More Data Needed?**               |
| *Gui | Re          | ID** |                                         |
| deli | quirement** |      |                                         |
| ne** |             |      |                                         |
+------+-------------+------+-----------------------------------------+
| 71-1 | Avian Oral  | No   | **Yes**. The data waiver request for    |
|      | LD~50~      | data | these guideline studies was denied in   |
|      |             |      | 1994, and the data are still needed.    |
|      |             |      | There are no data to quantitatively     |
|      |             |      | assess the potential acute risk to      |
|      |             |      | birds. Nicotine exposure to birds is    |
|      |             |      | possible. The assessed nicotine product |
|      |             |      | is a mammalian repellent, but it is     |
|      |             |      | unclear if it also repels avian         |
|      |             |      | species. Acute mammalian toxicity data  |
|      |             |      | suggest possible high toxicity of       |
|      |             |      | nicotine to terrestrial animals.        |
+------+-------------+------+-----------------------------------------+
| 71-2 | Avian       | No   |                                         |
|      | Dietary     | data |                                         |
|      | LC~50~      |      |                                         |
+------+-------------+------+-----------------------------------------+
| 71-4 | Avian       | No   | **Yes**. There are no data to           |
|      | R           | data | quantitatively assess the potential     |
|      | eproduction |      | chronic risk to birds. Chronic nicotine |
|      |             |      | exposure to birds is possible given     |
|      |             |      | that the product may be applied         |
|      |             |      | multiple times (as needed). The         |
|      |             |      | assessed nicotine product is a          |
|      |             |      | mammalian repellent, but it is unclear  |
|      |             |      | if it also repels avian species.        |
|      |             |      | Mammalian toxicity data suggest         |
|      |             |      | potential chronic toxicity effects of   |
|      |             |      | nicotine to terrestrial animals.        |
+------+-------------+------+-----------------------------------------+
| 72-1 | Freshwater  | No   | **Yes**. The data waiver request for    |
|      | Fish LC~50~ | data | this guideline study was denied in      |
|      |             |      | 1994, and the data are still needed.    |
|      |             |      | Data from the open literature suggest   |
|      |             |      | that nicotine is [at least moderately   |
|      |             |      | toxic]{.underline} to freshwater fish.  |
+------+-------------+------+-----------------------------------------+
| 72-2 | Freshwater  | No   | **Yes**. Data from the open literature  |
|      | I           | data | suggest that nicotine is [at least      |
|      | nvertebrate |      | highly toxic]{.underline} to freshwater |
|      | Acute       |      | invertebrates.                          |
|      | LC~50~      |      |                                         |
+------+-------------+------+-----------------------------------------+
| 72-  | Estua       | No   | Not at this time.                       |
| 3(a) | rine/Marine | data |                                         |
|      | Fish LC~50~ |      |                                         |
+------+-------------+------+-----------------------------------------+
| 72-  | Estua       | No   | Not at this time.                       |
| 3(b) | rine/Marine | data |                                         |
|      | Mollusk     |      |                                         |
|      | EC~50~      |      |                                         |
+------+-------------+------+-----------------------------------------+
| 72-  | Estua       | No   | Not at this time.                       |
| 3(c) | rine/Marine | data |                                         |
|      | Shrimp      |      |                                         |
|      | EC~50~      |      |                                         |
+------+-------------+------+-----------------------------------------+
| 72-  | Freshwater  | No   | Pending results of Guideline 72-1       |
| 4(a) | Fish Early  | data |                                         |
|      | Life-Stage  |      |                                         |
+------+-------------+------+-----------------------------------------+
| 72-  | Aquatic     | No   | Pending results of Guideline 72-2       |
| 4(b) | I           | data |                                         |
|      | nvertebrate |      |                                         |
|      | Life-Cycle  |      |                                         |
+------+-------------+------+-----------------------------------------+
| 123- | Seedling    | No   | **Yes**. There are no data to assess    |
| 1(a) | Emergence   | data | the potential phytotoxic effects of     |
|      |             |      | nicotine. There is a label statement    |
|      | (Tier I)    |      | that suggests potential                 |
|      |             |      | phytotoxicity.^1^                       |
+------+-------------+------+-----------------------------------------+
| 123- | Vegetative  | No   | **Yes**. There are no data to assess    |
| 1(b) | Vigor (Tier | data | the potential phytotoxic effects of     |
|      | I)          |      | nicotine. There is a label statement    |
|      |             |      | that suggests potential                 |
|      |             |      | phytotoxicity.^1^                       |
+------+-------------+------+-----------------------------------------+
| 1    | Aquatic     | No   | **Yes**. There are no data to assess    |
| 23-2 | Plant       | data | the potential phytotoxic effects of     |
|      | Growth      |      | nicotine. There is a label statement    |
|      | (Tier I)    |      | that suggests potential                 |
|      |             |      | phytotoxicity.^1^                       |
+------+-------------+------+-----------------------------------------+
| 1    | Honey Bee   | No   | **Yes**. There are no data to assess    |
| 41-1 | Acute       | data | the potential effects of nicotine to    |
|      |             |      | beneficial insects. Nicotine shares a   |
|      | Contact     |      | common mode of action with a class of   |
|      | LD~50~      |      | pesticides (neonicotinoids) that has    |
|      |             |      | been implicated in honey bee incidents. |
+------+-------------+------+-----------------------------------------+

^1^ The label states, "Do not apply the product directly to foliage or
stems." It is unknown if this refers to nicotine or one of the two other
active ingredients in the formulation.

Number range CHAPTER[]{#__RefHeading___Toc173552880 .anchor}**II.
Problem Formulation**

[]{#__RefHeading___Toc173552881 .anchor}**A. Stressor Source and
Distribution**

There are three active ingredients in the List B, nicotine and
derivatives case number 2460: nicotine (CAS \# 54-11-5; PC code 056702),
nicotine sulfate (CAS \# 65-30-5; PC code 056703), and tobacco dust (CAS
\# 8037-19-2; PC code 056704). There are no active products for either
nicotine sulfate or tobacco dust; this ecological risk assessment
focuses solely on nicotine use as a pesticide.

There are two[^3] end-use products with active registration that contain
nicotine as an active ingredient (**Table II.1**). Shotgun^®^ Rabbit and
Dog Repellent (Formerly F&B Rabbit and Dog Chaser) is exclusively for
residential and/or homeowner use. The product is applied in the
perimeter of plants or areas to be protected from dogs and rabbits. The
other product, Fulex Nicotine Fumigator, is a restricted use insecticide
for greenhouse use only; environmental exposures and ecological risk are
assumed to be negligible for this product. There are no active Special
Local Needs (SLN) products containing nicotine as the active ingredient.

+--------------------+----------------------+--------------------------+
| **Table II.1.**    |                      |                          |
| End-use Products   |                      |                          |
| Containing         |                      |                          |
| Nicotine as the    |                      |                          |
| Active Ingredient  |                      |                          |
+--------------------+----------------------+--------------------------+
| **Name of          | **Percent (%)        | **Use and Target Pests** |
| Product**          | Nicotine Active      |                          |
|                    | Ingredient**         |                          |
| **EPA Reg.         |                      |                          |
| Number**           |                      |                          |
+--------------------+----------------------+--------------------------+
| Shotgun^®^ Dog and | 0.35 (Derived from   | Use around flower        |
| Rabbit Repellent   | tobacco dust)        | gardens, ornamentals,    |
|                    |                      | trees, and shrubs. May   |
| 4-465 (Formerly    | Other Active         | be used around the       |
| 779-29)            | Ingredients:         | perimeter of vegetable   |
|                    |                      | gardens, but not in      |
|                    | Naphthalene 15%      | vegetable gardens or on  |
|                    |                      | food crops               |
|                    | Dried blood 15%      |                          |
+--------------------+----------------------+--------------------------+
| Fulex Nicotine     | 13.4                 | RESTRICTED USE           |
| Fumigator          |                      | PESTICIDE^1^ Greenhouse  |
|                    |                      | use only on ornamental   |
| 1327-41            |                      | plants, except violets   |
|                    |                      |                          |
|                    |                      | Control of aphids and    |
|                    |                      | most thrips              |
+--------------------+----------------------+--------------------------+

^1^ Due to very high acute inhalation, oral, dermal, and eye toxicity to
humans

Although there are no active products for nicotine sulfate or tobacco
dust in this reregistration case, the source of nicotine for the
Shotgun^®^ Rabbit and Dog Repellent product (EPA Reg. No 4-465) is
"tobacco dust". The USEPA's "Substance Registry System" defines tobacco
dust (CAS Reg. No. 8037-19-2) as "extractives and their physically
modified derivatives obtained from Nicotiana (*Solanaceae*) of
unspecified molecular formula". "Tobacco dust" carries many other
synonyms. The Office of Pesticides Programs Information Network (OPPIN)
uses the "tobacco dust" synonym, whereas tobacco oil, tobacco leaf
extract, tobacco leaf oil, tobacco leaf absolute, tobacco resinoid,
tabac oil, tobacco and tobacco absolute area synonyms used in the Toxic
Substance Control Act (TSCA) Inventory[^4].

Nicotine is listed in the Toxic Release Inventory (TRI)[^5], but it is
not in the High Production Volume (HPV) Listing.

[]{#__RefHeading___Toc173552882 .anchor}**1. Chemical Identification**

The insecticide-active chemical species in the nicotine-containing
products is the alkaloid nicotine, but the source of nicotine is tobacco
leaf dust (see USEPA's "Substance Registry System"). That is, nicotine
in a matrix of unspecified molecular weight. However, the present risk
assessment is based on "neat" (pure nicotine). Actual or estimated
exposure to nicotine carries a high degree of uncertainties for the
following reasons:

-   Variability in nicotine content in tobacco plants. Some of the
    factors that determine the content of nicotine found in tobacco
    leaves include: (a) the species of the plant, variety and
    [strain](javascript:void(0);); (b) growing conditions (particularly
    soil and climate) (c) the methods of [culture](javascript:void(0);),
    methods of [curing](javascript:void(0);), and from where on the
    [stalk](javascript:void(0);) the leaves are picked. Although the
    labels of the end-use products state a percentage of nicotine, it is
    unclear if this is a theoretical content normalized to tobacco
    leaves from multiple sources. Or an actual, measured nicotine
    content. That is, nominal versus actual concentrations.

-   Nicotine in the tobacco leaves exists in a complex plant matrix from
    which nicotine and other alkaloids are extracted. Given that
    nicotine content is variable (see above), the "application rate" and
    actual exposure to nicotine would depend on the extraction
    efficiency from leaves as well as the rate and efficiency of release
    from the "tobacco dust/oil". The latter would control the actual
    exposure to nicotine (i.e., its bioavailability and mechanism of
    bioavailability). Since there are no such data, the EFED is basing
    its assessment on the application rate calculated from the 0.35%
    specified in the label for the dog and rabbit chaser product
    (terrestrial outdoor, non-crop use pattern). No ecological risk
    assessment is performed for the greenhouse use pattern since
    environmental exposures are assumed to be negligible for this use
    pattern.

*Nicotine as a pure chemical substance*

Nicotine is an alkaloid synthesized by members of the nightshade family
(*Solanaceae*), which includes potatoes, tomatoes, eggplant, peppers,
and tobacco. The concentration of nicotine in tobacco plants (*Nicotiana
tabacum; Nicotiana* spp*.*) far exceeds the concentration in other
members of the nightshade family. Nicotine is the primary alkaloid in
tobacco.

Nicotine is a bicyclic compound containing a pyridine and a pyrrolidine
ring (**Table II.2**). The molecule possesses an asymmetric (chiral)
carbon in the C-2 position of the pyrrolidine ring. Thus, the molecule
would exist as a pair of enantiomers. However, in nature it only exists
as the *S-* enantiomer (absolute configuration), which is levorotatory
(i.e., the sign of rotation of plane polarized light is negative;
anticlockwise rotation). Therefore, it is more appropriate to identify
nicotine as *(S)* -3-(1-Methyl-2-pyrrolidinyl) pyridine (IUPAC).

+-------------------------+--------------------------------------------+
| **Table II.2**.         |                                            |
| Chemical Identification |                                            |
| of Nicotine             |                                            |
+-------------------------+--------------------------------------------+
| **Empirical Formula**   | C~10~H~14~N~2~                             |
+-------------------------+--------------------------------------------+
| **Molecular Structure** | ![](medi                                   |
|                         | a/image2.png){width="1.1034722222222222in" |
|                         | height="1.229861111111111in"}              |
|                         | ![](medi                                   |
|                         | a/image3.png){width="2.2916666666666665in" |
|                         | height="2.2916666666666665in"}             |
+-------------------------+--------------------------------------------+
| **Chemical Name**       | 3-(1-methylpyrrolidin-2-yl)pyridine;       |
|                         | *S*-enantiomer                             |
|                         |                                            |
|                         | [Synonyms]{.underline}: Pyridine,          |
|                         | 3-\[(2S)-1-methyl-2-pyrrolidinyl\]-;       |
|                         | Pyridine, 3-(1-methyl-2-pyrrolidinyl)-,    |
|                         | (S)-;                                      |
|                         |                                            |
|                         | L-Nicotine; Pyridine,                      |
|                         | (S)-3-(1-methyl-2-pyrrolidinyl);           |
|                         | 1-Methyl-2-(3-pyridyl) pyrrolidine;        |
|                         |                                            |
|                         | Pyridine, 3-(1-methyl-2-pyrrolidinyl)-,    |
|                         | (S);                                       |
|                         | (S)-3-(1-Methyl-2-pyrrolidinyl)pyridine;   |
|                         |                                            |
|                         | (-)-Nicotine                               |
+-------------------------+--------------------------------------------+
| **Chemical Abstracts    | 54-11-5                                    |
| Registry Number**       |                                            |
+-------------------------+--------------------------------------------+
| **Molecular Weight,     | 162.234                                    |
| g.mol^-1^**             |                                            |
+-------------------------+--------------------------------------------+

[]{#__RefHeading___Toc173552883 .anchor}**2. Physical/Chemical/Fate and
Transport Properties**

Nicotine (pure nicotine; "neat" nicotine) is a liquid alkaloid. It is
water soluble (4.2 x 10^5^ -- 1.0 x 10^6^ mg/L at 25° C). Nicotine has a
pK~a~ of 8.5 (*i.e*., it is a weak acid). It is a bitter-tasting liquid
which is strongly alkaline in reaction and forms salts with acids. It is
incompatible with strong oxidizers.

  --------------------------------- -------------------------------------
  **Table II.3.** Physical          
  Properties of Nicotine (Pure      
  nicotine; "Neat" nicotine)        

  **Property**                      **Value**

  Melting point                     -7,9 ° C

  Boiling point                     247 ° C

  Rotatory index^1^, α~D~           α~D~ = -168    at 20° C

  Density                           1.010
  --------------------------------- -------------------------------------

> ^1^Nicotine is the *S*-enantiomer of
> 3-(1-methylpyrrolidin-2-yl)pyridine, which is levorotatory (*S*)

Pure nicotine would remain liquid within in the range of temperature
when the target insects are most active. At temperatures below -7.9 ° C,
nicotine would be found in the solid state.

Physical and chemical properties are often used to identify potential
behavior of a chemical in the environment, mostly transport. The
following physical and chemical properties of nicotine were estimated
using EPISuite Version 3.20[^6].

+----------------------------------------------+-----------------------+
| **Table II.4** Physical and Chemical         |                       |
| Properties of Nicotine Relevant to Its Fate  |                       |
| in the Environment (Estimated)               |                       |
+----------------------------------------------+-----------------------+
| **Property**                                 | **Value**             |
+----------------------------------------------+-----------------------+
| Solubility in Water, mgL^-1^ at 25° C        | 1 x 10^6^             |
+----------------------------------------------+-----------------------+
| Vapor Pressure, mmHg at 25° C                | 3.2 x 10^-2^          |
|                                              |                       |
| Vapor Pressure increases with increasing     |                       |
| temperature                                  |                       |
+----------------------------------------------+-----------------------+
| Henry's Law Constant, atm-m^3^mole^-1^ at    | 3.0 x 10^-9^          |
| 25° C                                        |                       |
+----------------------------------------------+-----------------------+
| Log K~ow~                                    | 0.99                  |
+----------------------------------------------+-----------------------+
| K~oc~                                        | 2376                  |
|                                              |                       |
| Note: Nicotine is a weak acid (pK~a~ 8.5).   |                       |
| Therefore, sorption to soils is expected to  |                       |
| be pH dependent. EPISuite does not have the  |                       |
| capability of estimating sorption            |                       |
| coefficients as function of pH               |                       |
+----------------------------------------------+-----------------------+

The moderate vapor pressure of nicotine suggests that nicotine could
volatilize from soil. However, how fast it may volatilize from soil
depends on the sorption behavior of the chemical on soil (i.e., how
tight it may bind to soil). With an estimated K~oc~ of 2376, nicotine is
moderately mobile, and thus, the rate of volatilization is controlled by
sorption and would vary according to the K~oc~. The low Henry's Law
Constant is indicative of unlikely volatilization from water.
Furthermore, the logarithm of the air/water partitioning coefficient of
-6.911 is also indicative that nicotine partitions mainly to the water
column and not to air.

[]{#__RefHeading___Toc173552884 .anchor}**3. Pesticide Type, Class, and
Mode of Action**

Nicotine is classified as a natural product and a "botanical"
insecticide. It is used primarily for insects with piercing-sucking
mouthparts, such as aphids, whiteflies, leafhoppers, thrips, and mites.
Nicotine is fast-acting, easily absorbed through the eyes, skin, and
mucous membranes. The mode of action of nicotine when used as a dog and
rabbit chaser is unclear.

Nicotine, the primary alkaloid in tobacco binds stereoselectively to
nicotinic-cholinergic receptors. Nicotine mimics acetylcholine (**Figure
II.1**) in the nerve synapse, causing tremors, loss of coordination, and
eventually death. Acetylcholine is a major excitatory neurotransmitter
in the insect central nervous system. After acetylcholine is released by
the pre-synaptic cell, it binds to the postsynaptic nicotinic
acetylcholine receptor and activates an intrinsic cation channel,
resulting in a depolarization of the postsynaptic cell due an influx of
sodium and calcium ions.

The synaptic action of acetylcholine is terminated by the enzyme
acetylcholinesterase, which rapidly hydrolyzes the ester linkage in
acetylcholine. Nicotine also activates the nicotinic acetylcholine
receptor and does so persistently[^7]. It is this persistent activation
what leads to an over stimulation of cholinergic synapses, and results
in hyperexcitation, convulsions, paralysis, and death of the insect.

  -----------------------------------------------------------------------
  ![](media/image4.png){width="1.8833333333333333in"
  height="0.7229166666666667in"} Acetylcholine

  -----------------------------------------------------------------------

**Figure II.1** -- Chemical Structure of acetylchloline

Nicotine shares a mode of action with neonicotinoids, a class of
insecticides that has been implicated in honey bee population declines.
There are no honey bee toxicity data available for nicotine.

Although the role of nicotine in the rabbit and dog repellent product is
not well understood, it is reasonable to assume that effects on dogs and
rabbits may be related to the neurotoxicity of nicotine. It is also
unclear why the product is specific to dogs and rabbits and not for
other mammals. This dog and rabbit repellent product contains two other
active ingredients, naphthalene (15%) and dry blood (15%).

[]{#__RefHeading___Toc173552885 .anchor}**4**. **Overview of Pesticide
Usage**

Shotgun^®^ Rabbit and Dog Repellent (EPA Reg.No. 4-465; Formerly F&B
Rabbit and Dog Chaser) is a dust containing 0.35% nicotine, 15%
naphthalene, and 15% dried blood). It has exclusively residential
homeowner uses. The product is applied in bands around the perimeter of
gardens or areas to be protected from dogs and rabbits. It is not to be
used inside a vegetable garden or on food/feed crops. Shotgun^®^ Rabbit
and Dog Repellent is claimed to discourage domestic dogs (*Canis
familiaris*) from defecating on or near treated lawns or areas around
ornamental plants, trees, shrubs, and pavements. The product may be also
used to repel cottontail rabbits (*Sylvilagus floridanus*).

Fulex Nicotine Fumigator (EPA Reg. No. 1327-41; 13.4% nicotine) is a
RESTRICTED USE pesticide (due to very high acute inhalation, oral,
dermal, and eye toxicity to humans) for use only in greenhouses on
ornamental plants to control aphids and thrips. Since this product may
only be applied in greenhouses, environmental exposures and subsequent
risk to non-target organisms are assumed to be negligible.

[]{#__RefHeading___Toc173552886 .anchor}**B. Receptors**

This ecological risk assessment focuses on the potential risk of
nicotine when used as a pesticide in Shotgun^®^ Rabbit and Dog Repellent
(EPA Reg.No. 4-465). This repellent is used in residential/homeowner
settings and is the only active product for nicotine that has the
potential to result in environmental exposures to non-target organisms.
The other active product, Fulex Nicotine Fumigator, may only be used in
greenhouses, and environmental exposures as a result of this use pattern
are presumed to be negligible.

For Tier I assessment purposes, risk will be assessed to aquatic animals
assumed to occur in small, static ponds receiving runoff from adjacent
treated areas. The terrestrial ecosystems potentially at risk include
the treated area where nicotine is applied in residential settings. For
Tier I assessment purposes, risk will be assessed to terrestrial animals
assumed to exclusively occur in the treated area directly exposed to
nicotine dust.

[]{#__RefHeading___Toc173552887 .anchor}**C. Assessment Endpoints**

There are no acceptable ecotoxicity data available to assess the
potential effects of nicotine (in Shotgun^®^ Rabbit and Dog Repellent)
to non-target aquatic and terrestrial organisms. Risk cannot be
quantitatively estimated, but will be described qualitatively using
available information from the open literature.

[]{#__RefHeading___Toc173552888 .anchor}**D. Conceptual Model**

[]{#__RefHeading___Toc173552889 .anchor}**1. Risk Hypotheses**

For this assessment, the risk is stressor-initiated, where the stressor
is the release of nicotine from the end-use product, Shotgun^®^ Rabbit
and Dog Repellent. The following risk hypothesis is presumed for this
screening-level assessment:

> *Non-target aquatic and terrestrial animals and plants may be exposed
> to nicotine when used according to the label to repel dogs and rabbits
> from vegetation in a residential setting. Release of these active
> ingredients has the potential to compromise survival and/or elicit
> sublethal effects in non-target aquatic and terrestrial animals and
> plants.*

The exposure to nicotine for the ecological risk assessment is
exclusively based on the environmental fate, transport, and
transformation of nicotine when used as a pesticide and accordingly to
the existing labels of the end-use product. The exposure assessment does
not consider the fate of nicotine in tobacco smoking/chewing, as
released from the tobacco industry, or as a non-prescription drug[^8].

[]{#__RefHeading___Toc173552890 .anchor}**2. Diagram**

The conceptual model is used to depict the potential routes of exposure
from nicotine when used around flower gardens, ornamentals, trees, or
shrubs as well as around the perimeter of vegetable gardens to repel
pest mammals. All potential routes of exposure are considered and
presented in the conceptual model (**Figure II.2**). The conceptual
model generically depicts the potential source of nicotine, release
mechanisms, abiotic receiving media, biological receptor types, and
effects endpoints of potential concern.

In order for a chemical to pose an ecological risk, it must reach
ecological receptors in biologically significant concentrations. An
exposure pathway is the means by which a contaminant moves in the
environment from a source to an ecological receptor. For an ecological
exposure pathway to be complete, it must have a source, an environmental
transport medium, a point of exposure for ecological receptors, and a
feasible route of exposure. The assessment of ecological exposure
pathways, therefore, includes an examination of the source and potential
migration pathways for constituents, and the determination of potential
exposure routes (*e.g.*, ingestion, inhalation, dermal contact).

![](media/image1.wmf)

**E. Analysis Plan**

This ecological risk assessment focuses on the potential risk of
nicotine when used as a pesticide in Shotgun^®^ Rabbit and Dog Repellent
(EPA Reg.No. 4-465; formerly known as "F&B Rabbit and Dog Chaser"). This
repellent is used in residential/homeowner settings around flower
gardens, ornamentals, trees, and shrubs and the perimeter of vegetable
gardens and is generally classified as a "terrestrial non-food/non-feed
use pattern". This is the only active product for nicotine that has the
potential to result in environmental exposures to non-target organisms.
The other active product, Fulex Nicotine Fumigator, may only be used in
greenhouses, and environmental exposures (and risk) as a result of this
use pattern are presumed to be negligible.

Aquatic ecosystems potentially at risk as a result of nicotine use as a
repellent include water bodies adjacent to or downstream from the
treated residential site and might include impounded bodies such as
ponds, lakes, and reservoirs, or flowing waterways, such as streams or
rivers. There are no acceptable aquatic toxicity data with which to
quantitatively estimate risk (*i.e.*, calculate risk quotients);
however, risk to fish (surrogate for aquatic-phase amphibians), aquatic
invertebrates, and aquatic plants will be characterized qualitatively.
Aquatic exposure estimates will be generated for nicotine using the Tier
I exposure model, GENEEC2[^9] (GENeric Estimated Exposure Concentration;
Version 2.0; May 1, 2001), which assumes 100% of the area is treated.
Since Shotgun^®^ Rabbit and Dog Repellent is applied in bands around
vegetation, the modeled EECs will be adjusted to account for the uneven
distribution of nicotine in the treated area (see **Section III.B.2**
for specific calculations). It is assumed that the 0.35% nicotine
specified in the label is the actual percentage of nicotine ("neat"
nicotine) in the product regardless of the source and composition of the
"tobacco dust" and that all of the nicotine is available for runoff.

Terrestrial ecosystems potentially at risk as a result of nicotine use
as a repellent in a dust formulation include the treated area and areas
immediately adjacent to the treated residential area. There are no
acceptable toxicity data with which to quantitatively estimate risk to
terrestrial organisms; however, risk to birds (surrogate for reptiles
and terrestrial-phase amphibians), mammals, terrestrial invertebrates,
and terrestrial plants will be characterized qualitatively. For
terrestrial exposure modeling purposes, granules are used as a surrogate
for dust in the Tier 1 model, T-REX (Version 1.3.1, dated December 7,
2006). It is assumed that a banded granular application is 100%
unincorporated in soil. Risk to terrestrial animals from exposure to
granules will be based on LD~50~/ft^2^ values. The LD~50~/ft^2^ values
are calculated using a toxicity value (adjusted LD~50~ of the assessed
animal and its weight classes) and the EEC (mg ai/ft^2^) and are
directly compared with Agency's levels of concern (LOCs). Since nicotine
is used only for granular applications, exposures to animals from
foraging on food items with nicotine residues (short and tall grass,
leaves, seeds) are not estimated.

[]{#__RefHeading___Toc173552891 .anchor}**F. Data Gaps**

[]{#__RefHeading___Toc173552892 .anchor}**1. Environmental Fate**

As per 40CFR §158.290 (Subdivision N), the data requirements for a
"terrestrial, non-food use pattern" are:

1.  \[Abiotic\] Hydrolysis

2.  \[Direct\] Photolysis in Water

```{=html}
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```
1.  Aerobic Soil Metabolism (Conditionally Required)

```{=html}
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```
1.  Mobility in Soil (Conditional Required)

164-1 Terrestrial Field Dissipation

For domestic, outdoor use patterns, the data requirements are 161-1
(Conditionally required) and 162-1.

For greenhouse uses, non-food the data requirements are 161-1 and 162-1.
Volatility from soil (163-2/-3) is conditionally required on a
case-by-case basis as it is triggered by the vapor pressure of the
active ingredient. The vapor pressure of nicotine (3.2 x 10^-2^ mmHg at
25°C) suggests that these data are required.

There are no available environmental fate guideline studies (Subdivision
N) available for nicotine (or tobacco dust). For this reason, the
environmental fate and exposure assessment is predominantly performed
using EPISuite Version 3.20[^10] estimates. The estimates are based on
pure nicotine ("neat" nicotine) and not on the nicotine in tobacco dust.
The exposure assessment is performed only for the outdoor use
(Shotgun^®^ Rabbit and Dog Repellent product).

On September 7, 1994 (DPBarcode D206473), EFED recommended that the
following data requirements could be waived:

1.  \[Abiotic\] Hydrolysis

2.  \[Direct\] Photolysis in Water

3.  Photolysis on Soil

162-1 Aerobic Soil Metabolism

162-2 Anaerobic Soil Metabolism

163-1 Mobility in Soil (Conditional Required

163-2 Volatility from Soil (laboratory scale)

It was recommended that the registrant(s) request waivers and provide
articles from the open scientific literature related to the chemical
transformation of nicotine when ignited. To EFED's knowledge, this
information has not been provided. However, only the product used in
greenhouses is ignited, and since no ecological risk assessment is being
performed for greenhouse uses, the information may no longer needed by
EFED. The EFED has no record on waiver petitions by the registrant(s) or
any additional information submitted after 1994.

Because no open, scientific literature information was provided, the
EFED relied on model estimates (EPISuite) and its own search of the
open, scientific literature.

[]{#__RefHeading___Toc173552893 .anchor}**2. Ecological Effects**

There are no acceptable guideline ecotoxicity studies for nicotine. In
1994[^11], the EFED denied waiver requests for the following studies:
acute avian oral toxicity (71-1(a)), acute avian dietary toxicity
(71-2(a)), and acute fish toxicity (72-1(c); however, none of these
studies were subsequently provided to the Agency. A search of the open
literature (ECOTOX database) identified 177 studies that were acceptable
for ECOTOX and OPP (**Appendix C**). Of these, 10 studies were formally
reviewed for this risk assessment; however, none of them were deemed
acceptable for quantitative use in this ecological risk assessment. A
full list of data gaps can be found in **Table I.2.**

[]{#__RefHeading___Toc173552894 .anchor}**III. Analysis**

[]{#__RefHeading___Toc173552895 .anchor}**A. Use Characterization**

Shotgun^®^ Rabbit and Dog Repellent (EPA Reg. No. 4-465), formerly known
as F&B Dog and Rabbit Chaser, is used in residential settings around the
perimeter of ornamental plants, trees, shrubs, and vegetable gardens.
The product is not to be used within vegetable gardens. The product is
claimed to discourage domestic dogs from defecating on or near the
treated area. It is also claimed that the product discourages cottontail
rabbits from entering, feeding in, or defecating within treated areas.

*Application*

Traces of animal droppings and urine should be removed prior to treating
the areas to be protected. As per the current label, the product is
sprinkled directly \[to soil\] in two or more inch-wide bands in the
perimeter of plants or areas to be protected. The product should not be
applied directly to foliage or stems.

Applications can be repeated as needed. Heavy rains, heavy snowfalls,
hot weather, or high winds would require more frequent applications. It
can be applied year round, typically 4 to 5 times per season or year.

The product is mostly used East of the Rocky Mountains, but use areas
are not specified in the label (May 8, 2003).

*Warnings (Environmental Hazards) in the Label*

"This pesticide is toxic to fish. Do not apply directly to water. Do not
contaminate water by cleaning of equipment or disposal of waste".

[]{#__RefHeading___Toc173552896 .anchor}**B. Exposure Characterization**

[]{#__RefHeading___Toc173552897 .anchor}**1. Environmental Fate and
Transport Characterization**

There are no targeted, environmental fate guideline studies (Subdivision
N) conducted with nicotine as the test substance submitted by the
registrant(s) in support of nicotine as a pesticide-active ingredient,
as required under FIFRA. The environmental fate assessment of nicotine
is based on quantitative structure-activity relationships (QSAR) using
EPISuite (Version 3,20) as the estimation model, supplemented by open
literature data. Therefore, this environmental fate assessment is only
at the screening level. The EPISuite estimates for nicotine appear in
**Appendix A.**

According to the model estimates, the following general behavior of
nicotine in the environment is anticipated:

*Persistence*

[Abiotic Hydrolysis]{.underline} - Nicotine lacks hydrolysable groups.
Thus, abiotic hydrolysis does not constitute a dissipation pathway.
Nicotine is expected to be persistent in water under abiotic conditions.

[Biodegradation]{.underline} - The half-life of nicotine under biotic
(aerobic) conditions is estimated by EPISuite as 37.5 days. This
estimate is based on the survey model for ultimate (i.e., complete
mineralization to carbon dioxide and water) and primary (i.e., initial
step in the biodegradation process that forms a new compound), which
indicate that the biodegradation of nicotine takes place within
"weeks-to-months" time frame. The half-life of 37.5 days is assigned in
the EPISuite biodegradation model to the "weeks-to months" time frame.

[Photodegradation]{.underline}- If released to air, nicotine can undergo
rapid photooxidation reactions with photolytically generated hydroxyl
free radicals (half-life 1.14 hours). In addition, the estimated Log of
the air/water partition coefficient (Log K~aw~) of -6.91 is indicative
that volatilization from water does not release nicotine into air.

The electronic absorption spectrum of nicotine shows an intense
absorption band (chromophore) at 260-262 nm (Log of molar absorptivity,
ε, 3.46- 3.8)[^12], but some absorption occurs above 290 nm (i.e.,
towards the sunlight spectral region. As the necessary condition to
undergo direct photolysis is met, nicotine has the potential to undergo
direct photolysis, provided that the absorbed energy is sufficient to
change the molecular structure of nicotine (e.g., bond breaking,
rearrangement, photooxidation and so forth). Even if nicotine degrades
by direct photolysis, the reaction would only be of importance in clear,
shallow water. It is most likely that nicotine degrades in surface water
via indirect photolysis brought about by naturally occurring
photosensitizers (e.g., photochemically generated excited states of
dissolved organic matter, DOM; photochemically formed hydroxyl radicals,
OH•; singlet oxygen, O~2~(^1^∆~g~)). The importance of indirect
photolysis in reducing the persistence of pharmaceuticals, including
nicotine, has been recognized[^13]. Thus, indirect photolysis is likely
to play an important role in the degradation of nicotine in aqueous
environments.

*Mobility/Transport*

Nicotine is a weak acid (pK~a~ 8.5).The mobility of nicotine in acid and
neutral conditions is moderate (estimated K~oc~ 2376), but mobility
would be much higher in alkaline pH. That is, as the pH increases the
amount of the non-protonated form (i.e., the "anion") increases, which
results in increased mobility. However, the EPISuite Koc model is not
designed to estimate pH-dependent sorption coefficients as it does not
take into account the Henderson-Hasselbach equation[^14]. The K~oc~ of
2376 is only a rough estimate of the mobility of nicotine in soils.
Therefore, K~oc~ values, rate of volatilization (see below), and
environmental exposure concentrations in surface water (EECs) may be
overestimated/ underestimate because they are not soil/site specific.

Although a moderate vapor pressure of 3.2 x 10^-2^ mmHg (25°C) suggests
that nicotine could volatilize from soil, an estimate of the rate of
volatilization from moist soil surfaces using the rough K~oc~ of 2376
indicates that volatilization from soils is not a fast process (5.91 x
10^-4^ day^-1^); Appendix A.2). Because sorption to soil decreases with
pH (i.e., increased mobility), theoretically, the volatilization rate
would increase[^15] as the pH increases. In addition, the vapor pressure
and the volatility of a chemical substance would increase with
temperature. Table III.1 provides an example on how K~oc~ affects the
volatility of nicotine from soil, assuming arbitrary K~oc~ values. Even
at lower K~oc~ values, the volatilization of nicotine from moist soil is
slow.

  -------------------------------------------- --------- --------- ---------
  **Table III.1**- Theoretical Volatilization                      
  of Nicotine from Moist Soil as a Function of                     
  K~oc~ (25° C). These estimates apply only to                     
  25° C. Volatilization is expected to                             
  increase with increasing temperature                             
  (Appendix A)                                                     

  **Kinetics Information**                     **K~oc~   **K~oc~   **K~oc~
                                               500**     2376**    3000**

  Volatilization Half-life, days               247       1.173 x   1.481 x
                                                         10^3^     10^3^

  Volatilization Rate Constant, day^-1^        2.81 x    5.91 x    4.68 x
                                               10^-3^    10^-4^    10^-4^
  -------------------------------------------- --------- --------- ---------

Volatilization from water would not be a significant dissipation route
for nicotine (Henry's Law Constant, 3.0 x 10^-9^ atm-m^3^mole^-1^ at 25°
C; Log air/water partition coefficient -- 6.91). That is, nicotine
predominantly partitions to the water phase. Therefore, the low rate of
volatilization from soil and/or water reduces the amount of nicotine
transported to air. Although the rate of photooxidation in air is rapid
(half-life 1.14 hours), the low rate of volatilization decreases the
availability of nicotine in air.

Long-range atmospheric transport of nicotine sorbed to colloidal size
soil particulates, as well as wet deposition, are feasible. Because the
rabbit and dog repellent product is a dust, off-site exposure may result
from drifting during application or by wind after application.

*Accumulation*

Based on an estimate of the *n*-octanol/water partition coefficient (Log
K~ow~ = 0.99) and the estimated bioconcentration factor (Log BCF= 0.2),
there is low concern for bioconcentration of nicotine in fish.

*Transformation Products*

The bacterial oxidation of nicotine by *Arthrobacter oxydans* (now known
as *Arthrobacter nicotinovarans*) has been recognized *in vitro* for a
long time. The enzyme-catalyzed oxidative products have been identified,
isolated and characterized[^16]. The first oxidative product is
(1)-6-hydroxynicotine, which is subsequently catabolized to
6-hydroxypseudonicotine (oxynicotine), 2,6-dihydroxypseudonicotine,
2,6-dihyroxy-N-methymyosmine, and other oxidation products (either at
the pyridine and/or pyrrolidine rings). In addition, a "crystalline,
purple-blue pigment" (nicotine blue) has been identified as an
"end-product" of nicotine.

The bacterial soil population is known to play an important role in the
biotransformation of nicotine in soils, where *Arthrobacter
nicotinovarans* (previously known as *A. oxydans*) is involved in the
enzyme-catayzed oxidative pathway in the catabolism of nicotine. The
first metabolite of nicotine is 6-hydroxynicotine. Nicotine further
breakdown leads to "nicotine blue"[^17] from the hydroxylated pyridine
ring and γ-N-methylaminobutyrate, where the latter forms from the
pyrrolidine ring. Furthermore, and alternative pathways (deamination
versus demethylation) have been identified in the final steps in the
catabolism of nicotine[^18]

Biotransformation products of nicotine in soil include oxynicotine,
3-pyridinylmethyl ketone, N-methylmyosamine, and a "purple-blue
crystalline pigment", but the relative amounts and conditions of
formation in soils is likely to vary with soil and bacterial population.
The nature of transformation products in the environment is not as well
known as it is in living systems. The major metabolite of nicotine,
however, is cotinine (*S*)-1-Methyl-5-(3-pyridyl)-2-pyrrolidinone),
which is an oxidation product of nicotine. Although cotinine pathway of
formation in the liver is well known, its formation in soils has not
been reported so far.

The biotransformation of nicotine and cotinine has been studied in
water-sediment systems[^19]. Oxic conditions appear to favor
mineralization of cotinine, with complete recovery as carbon dioxide
(i.e., complete mineralization) observed after 72 days, which falls
within the "weeks-to-months" time frame for the ultimate biodegradation
of nicotine estimated by EPISuite. Under anoxic conditions, the
biotransformation of nicotine and cotinine in stream sediments involves
microbial demethylation of the N-methyl group of both nicotine and
cotinine.

Appendix A.3 presents a comparison of the physical, chemical, and
environmental fate of nicotine and cotinine. Cotinine primary
biodegradation (half-life estimated 8.6 days) is faster than that of
nicotine (37.5 days), but the ultimate biodegradation for both is
estimated as 37.5 days. Both are stable towards abiotic hydrolysis. The
vapor pressure and Henry's Law Constant of cotinine are lower than those
of nicotine and, theoretically, the partitioning cotinine formed on soil
(if formed) and/or water would be higher than nicotine. If formed in
soils, cotinine is more mobile than nicotine.

[]{#__RefHeading___Toc173552898 .anchor}**2. Aquatic Exposure
Characterization**

[]{#__RefHeading___Toc173552899 .anchor}**a. Simulation Modeling**

[]{.mark}

Aquatic exposures for nicotine were estimated using the Tier I exposure
model, GENEEC Version 2.0, which assumes 100% of the area is treated.
Since Shotgun^®^ Rabbit and Dog Repellent is applied in bands around
vegetation, the modeled EECs needed to be adjusted to account for the
uneven distribution of nicotine in the treated area. Specifically, the
product is applied in bands 2 inches (0.1667 ft) wide (as specified in
the label) or wider (upper limit not specified in the label, but assumed
to be 6 and 12 inches or 0.5 and 1 ft, respectively). Further, based on
United States 2000 Census data[^20], a typical house footprint is 1000
square feet located on a 0.25 acre plot. Assuming that the house is
square, each side is 31.6 ft, and the perimeter is 126.4 feet. If there
is a 10-foot wide flower bed around the house, the total perimeter
increases to 206.4 feet. Further, if there is a garden within the 0.25
acre plot measuring 20 feet by 100 feet, there is an additional 240 feet
in perimeter. Based on this theoretical residential site, the percent
treatment area as a function of band width of Shotgun^®^ Rabbit and Dog
Repellent can be calculated (**Table III.2**). These percentages are
used to adjust the GENEEC-generated EECs for nicotine.

  ----------- ----------------- ---------------- ----------- ---------------
  **Table                                                    
  III.2.**                                                   
  Percent                                                    
  Treatment                                                  
  Area as a                                                  
  Function of                                                
  Band Width                                                 
  Based on                                                   
  0.25 Acre                                                  
  Plot                                                       

  **Band      **Area Around     **Area Around    **Total     **Percent (%)
  Width,      House With Flower Garden^2^,       Area^3^,    Treatment
  feet**      Bed^1^, sq-ft**   sq-ft**          sq-ft**     Area^4^**

  0.1667      34.4              40               74.4        0.68

  0.5         103.5             120              223.5       2.05

  1.0         206.4             240              446.4       4.1
  ----------- ----------------- ---------------- ----------- ---------------

^1^ Assuming the house (with flower bed) has a perimeter of 206.4 feet
as described above.

^2^ Assuming the garden has perimeter of 240 feet as described above.

^3^ Total Area = Area Around House With Flower Bed + Area Around Garden

^4^ Percent (%) Treatment Area = (Total Area / (0.25 acre or 10,890
sq-ft)) \* 100

According to the product label[^21], three pounds of Shotgun^®^ Rabbit
and Dog Repellent will produce a band of product 1 inch wide and 85 feet
long, or 7.08 feet^2^. Based on the assumed total area treated
(calculated above), the application rate in terms of pound of nicotine
active ingredient per acre (lbs a.i./A) were calculated for each of the
three assumed band width scenarios (**Table III.3**).

+-------------+------------------+------------------+-----------------+
| **Table     |                  |                  |                 |
| III.3.**    |                  |                  |                 |
| Application |                  |                  |                 |
| Rates of    |                  |                  |                 |
| Nicotine as |                  |                  |                 |
| lb          |                  |                  |                 |
| ai(         |                  |                  |                 |
| nicotine)/A |                  |                  |                 |
| for         |                  |                  |                 |
| exposure    |                  |                  |                 |
| modeling    |                  |                  |                 |
| purposes.   |                  |                  |                 |
| It is       |                  |                  |                 |
| assumed     |                  |                  |                 |
| that        |                  |                  |                 |
| nicotine is |                  |                  |                 |
| pure (neat) |                  |                  |                 |
| nicotine    |                  |                  |                 |
| and that    |                  |                  |                 |
| all of the  |                  |                  |                 |
| nicotine in |                  |                  |                 |
| the tobacco |                  |                  |                 |
| dust is     |                  |                  |                 |
| available   |                  |                  |                 |
| for runoff  |                  |                  |                 |
+-------------+------------------+------------------+-----------------+
| **Band      | **Treated Area   | **Application    | **Application   |
| width       | (sq-ft/acre)**   | Rate**           | Rate (lbs       |
| (ft)**      |                  |                  | a.i.(ni         |
|             |                  | **(lbs           | cotine)/acre)** |
|             |                  | product)/acre)** |                 |
+-------------+------------------+------------------+-----------------+
| 0.1667      | 298              | 126              | 0.044           |
+-------------+------------------+------------------+-----------------+
| 0.5         | 894              | 377              | 0.132           |
+-------------+------------------+------------------+-----------------+
| 1.0         | 1786             | 754              | 0.264           |
+-------------+------------------+------------------+-----------------+

Although model-estimated physical and chemical properties and
environmental fate input parameters are not routinely used to estimate
environmental exposures, EPISuite-generated input parameters were used
in this risk assessment (**Table III.4**). This was done because of
insufficient and poorly documented data in the open literature. Most of
the available data on nicotine targets human exposure from tobacco
smoking.

+-----------+-------+--------------------------------------------------+
| **Table   |       |                                                  |
| III.4.**  |       |                                                  |
| Envi      |       |                                                  |
| ronmental |       |                                                  |
| Fate      |       |                                                  |
| Input     |       |                                                  |
| P         |       |                                                  |
| arameters |       |                                                  |
+-----------+-------+--------------------------------------------------+
| **Envi    | **Va  | **Source and Comments**                          |
| ronmental | lue** |                                                  |
| Fate      |       |                                                  |
| Input     |       |                                                  |
| Par       |       |                                                  |
| ameters** |       |                                                  |
+-----------+-------+--------------------------------------------------+
| S         | 1 x   | EPISuite, but corroborated by experimental data  |
| olubility | 10^6^ |                                                  |
| in Water, |       |                                                  |
| mgL^-1^   |       |                                                  |
| at 25° C  |       |                                                  |
+-----------+-------+--------------------------------------------------+
| Abiotic   | 0     | Nicotine is presumed to be stable towards        |
| H         |       | abiotic hydrolysis. It does not contain any      |
| ydrolysis |       | hydrolysable group                               |
| Half-life |       |                                                  |
+-----------+-------+--------------------------------------------------+
| P         | 0     | Nicotine is not likely to undergo direct         |
| hotolysis |       | photolysis in water. There is no indirect        |
| is Water  |       | photolysis quantitative data, although indirect  |
| Half-life |       | photolysis is likely to be an important          |
|           |       | dissipation route in natural water               |
+-----------+-------+--------------------------------------------------+
| Aerobic   |       | 37.5 days (EPISuite)                             |
| Soil      |       |                                                  |
| M         |       | (Refer to "Environmental Fate Characterization") |
| etabolism |       |                                                  |
| Half-life |       |                                                  |
+-----------+-------+--------------------------------------------------+
| Anaerobic |       | 75 days                                          |
| Aquatic   |       |                                                  |
| M         |       | Twice the aerobic soil metabolism as per         |
| etabolism |       | recommended by GENEEC (default value)            |
| Half-life |       |                                                  |
|           |       | (Refer to "Environmental Fate Characterization") |
+-----------+-------+--------------------------------------------------+
| Soil      |       | 2376                                             |
| sorption  |       |                                                  |
| coe       |       | Estimated by EPISuite, which does not estimate   |
| fficient, |       | pH-dependency of sorption coefficients for weak  |
| as Koc    |       | acids, such as nicotine                          |
+-----------+-------+--------------------------------------------------+
| Ap        | 100%  | Default value                                    |
| plication |       |                                                  |
| E         | 0%    | A non-incorporated, granular formulation was     |
| fficiency |       | used to run the model, for which spray drift is  |
|           |       | not likely.                                      |
| Spray     |       |                                                  |
| Drift     |       | Note: A granular formulation was used as a       |
|           |       | surrogate for a dust product that is applied     |
|           |       | directly to the perimeter around plants.         |
|           |       | However, there is a potential for drifting of    |
|           |       | the dust (unknown particle size distribution)    |
|           |       | during application. Therefore, the contribution  |
|           |       | of dust drift to environmental exposures is      |
|           |       | uncertain.                                       |
+-----------+-------+--------------------------------------------------+

The GENEEC estimated environmental concentrations in surface water are
shown in **Table III.4. (**See **Appendix B** for model output). The
maximum number of applications per year and the minimal application
interval were not specified on the product label; it was assumed that
Shotgun^®^ Rabbit and Dog Repellent applications were made 6 times per
year with an application interval of 60 days. The GENEEC-estimated
concentrations in surface water as the result the residential, outdoor
use of Shotgun^®^ Rabbit and Dog Repellent product are low. None of the
adjusted peak EECs exceed 40 ng/L (ppt). Exposure concentrations
increase with increasing band widths.

+----------+-------+-----------+------------+-----------+------------+
| **Table  |       |           |            |           |            |
| III.4.** |       |           |            |           |            |
| GENEEC-g |       |           |            |           |            |
| enerated |       |           |            |           |            |
| Envir    |       |           |            |           |            |
| onmental |       |           |            |           |            |
| Exposure |       |           |            |           |            |
| Concen   |       |           |            |           |            |
| trations |       |           |            |           |            |
| of       |       |           |            |           |            |
| Nicotine |       |           |            |           |            |
| in       |       |           |            |           |            |
| Surface  |       |           |            |           |            |
| Water as |       |           |            |           |            |
| the      |       |           |            |           |            |
| result   |       |           |            |           |            |
| of its   |       |           |            |           |            |
| use as a |       |           |            |           |            |
| p        |       |           |            |           |            |
| esticide |       |           |            |           |            |
| (rabbit  |       |           |            |           |            |
| and dog  |       |           |            |           |            |
| rep      |       |           |            |           |            |
| ellent). |       |           |            |           |            |
| All      |       |           |            |           |            |
| concen   |       |           |            |           |            |
| trations |       |           |            |           |            |
| are in   |       |           |            |           |            |
| ng/L     |       |           |            |           |            |
| (ppt).   |       |           |            |           |            |
| Numbers  |       |           |            |           |            |
| in bold  |       |           |            |           |            |
| are the  |       |           |            |           |            |
| concen   |       |           |            |           |            |
| trations |       |           |            |           |            |
| adjusted |       |           |            |           |            |
| for      |       |           |            |           |            |
| percent  |       |           |            |           |            |
| treated  |       |           |            |           |            |
| area.    |       |           |            |           |            |
+----------+-------+-----------+------------+-----------+------------+
| **Band   | *     | **Maximum | **Maximum  | **Maximum | **Maximum  |
| width**  | *Peak | 4-day     | 21-day     | 60 day    | 90-day     |
|          | GEN   | Average   | Average    | Average   | Average    |
| **(%     | EEC** | GENEEC**  | GENEEC**   | GENEEC**  | GENEEC**   |
| treated  |       |           |            |           |            |
| area)**  |       |           |            |           |            |
+----------+-------+-----------+------------+-----------+------------+
| 2 in     | 163   | 108       | 29         | 10        | 6.9        |
|          |       |           |            |           |            |
| (0.68)   | **    | **0.73**  | **0.2**    | **0.07**  | **0.05**   |
|          | 1.1** |           |            |           |            |
+----------+-------+-----------+------------+-----------+------------+
| 6 in     | 489   | 325       | 87         | 31        | 21         |
|          |       |           |            |           |            |
| (2.05)   | *     | **6.6**   | **1.8**    | **0.64**  | **0.43**   |
|          | *10** |           |            |           |            |
+----------+-------+-----------+------------+-----------+------------+
| 12 in    | 978   | 650       | 174        | 62        | 41         |
|          |       |           |            |           |            |
| (4.1)    | *     | **26.6**  | **7.1**    | **2.5**   | **1.7**    |
|          | *40** |           |            |           |            |
+----------+-------+-----------+------------+-----------+------------+

EECs resulting from the use of nicotine as a pesticide carry a high
degree of uncertainty because both EPISuite and GENEEC2 are only
screening-level models. Further, assumptions were made regarding the
maximum annual rate and minimum application interval since this
information was not specified on the label. To the extent that actual
use practices result in more than 6 applications per year and/or there
is a shorter minimum re-treatment interval than the assumed 60-day
interval, the aquatic EECs would be correspondingly higher.

b.  []{#__RefHeading___Toc173552900 .anchor}**Monitoring Information**

Nicotine and its metabolite cotinine are analytes considered in
monitoring studies of anthropogenic organic compounds in water, in which
both chemicals are classified as "non-prescription drugs".[^22] Most of
the available environmental monitoring data on nicotine and cotinine
target human exposure from tobacco smoking (excreted; human wastes) in
pre- and post water treatment, where cotinine is used as a marker to
nicotine exposure. The U.S. Geological Survey has shown 38% detection
(84 collected samples) at a maximum of 0.9 μg/L (0.9 ppb). Because the
aquatic exposure assessment applies only to the use of nicotine as a
pesticide, the USGS data are not included in the ecological risk
assessment.

As previously noted, this assessment only considers the exposure of
nicotine in water as a result of its use as a pesticide. Nicotine and
its salts are listed in the Toxic Release Inventory (TRI) for the
release of nicotine from the tobacco industry which must be reported to
the Agency annually, including release to surface water. In 2005, 755
pounds of "nicotine"[^23] were released into surface water.

[]{#__RefHeading___Toc173552901 .anchor}**3. Terrestrial Exposure
Characterization**

Terrestrial exposures for nicotine are estimated using the conceptual
approach given in the Tier-1 model, T-REX Version 1.3.1. Potential risk
to terrestrial animals was estimated in terms of LD~50~/ft^2^. The T-REX
model is designed to estimate terrestrial exposures for applications in
agricultural settings (i.e., not residential settings), and some
assumptions had to be made for this nicotine assessment. T-REX model
input parameters are tabulated below (**Table III.5**).

  -----------------------------------------------------------------------
  **Table III.5.** T-REX Input          
  Parameters for Shotgun^®^ Rabbit and  
  Dog Repellent granules                
  ------------------------------------- ---------------------------------
  **Input Parameter**                   **Value**

  Percent active ingredient             0.035

  Application Rate (lbs product per     754
  acre)                                 

  Half-life (days)                      35

  Application Interval (days)           60

  Number of Applications                6

  Row Spacing (inches)                  291.6^1^

  Band width (inches)                   12

  Percent incorporated                  0
  -----------------------------------------------------------------------

^1^ One of the important inputs in the 'banded application' mode for
granular formulations is row spacing, which is the amount of space (in
inches) between crop rows. Based on the assumed generic residential
application setting (Section III.B.2), there are 1786 linear feet per
acre that can be treated with Shotgun^®^ Rabbit and Dog Repellent.
Assuming a square, 1-acre plot, maximum row length would be 208.7 feet
(√43560 square feet per acre). The number of possible rows (bands) per
acre is 8.6 (1786 ft./208.7 ft = 8.6). To calculate the row spacing, the
maximum row length, 208.7 feet, was divided by the number of rows, 8.6,
which equals 24.3 feet (or 291.6 inches).

The T-REX model predicted a terrestrial EEC of 66.78 mg a.i./ ft^2^ for
Shotgun^®^ Rabbit and Dog Repellent (**Table III.6**). This is the
estimated exposure for a bird (surrogate for reptile and
terrestrial-phase amphibian) or mammal if 100% of the animal's diet was
bands of Shotgun^®^ Rabbit and Dog Repellent.

  ----------------------------------- -----------------------------------
  **Table III.6.** T-REX Output for   
  Shotgun^®^ Rabbit and Dog Repellent 
  granules                            

  \# rows acre-1:                     8.59

  row length (ft):                    208.71

  lb ai/1000 ft row:                  420.62

  bandwidth (ft):                     1.00

  mg ai/ft^2^ (EEC):                  66.78

  exposed EEC (mg ai/ft^2^):          66.78
  ----------------------------------- -----------------------------------

[]{#__RefHeading___Toc173552902 .anchor}**C. Ecological Effects
Characterization**

Summaries of the available ecotoxicity studies can be found in
**Appendix C.**

There are no acceptable guideline acute or chronic toxicity data with
which to quantitatively estimate risk to aquatic animals for the use of
nicotine as a dog and rabbit repellent in residential settings. Several
studies from the open literature suggest that nicotine is [at least
moderately toxic]{.underline} to fish and [at least highly
toxic]{.underline} to aquatic invertebrates; however, given the high
uncertainty in the actual exposure concentrations in the tests, the
reported toxicity thresholds (*i.e.,* LC50, NOAEC) are unreliable.
Further, the following statement is on the label: "This product is toxic
to fish. Do not apply directly to water. Do not contaminate water by
cleaning of equipment or disposal of waste." It is unclear whether the
assumed toxicity of the product is attributed to nicotine or the other
active ingredients, naphthalene or dried blood.

Terrestrial risk assessment for mammals relies on mammalian toxicity
information provided by the Health Effects Division (HED) as well as
studies identified through a search of the ECOTOX database. There are no
acceptable guideline mammalian toxicity studies with which to
quantitatively estimate risk to aquatic animals for the use of nicotine.
However, the assessed formulated product is a as a dog and rabbit
repellent; assuming small mammals (*e.g.*, field mice) are similarly
repelled, the terrestrial dietary exposure pathway for mammals may be
unlikely. There is currently no information to suggest that birds are
repelled by Shotgun^®^ Rabbit and Dog Repellent; thus, there is a
potential for avian terrestrial dietary exposure. There are no avian
acute or chronic toxicity data available for consideration in this risk
assessment.

[]{.mark}

No terrestrial or aquatic plant studies are available. The product label
states, "Do not apply the product directly to foliage or stems," which
suggests that there is a possibility of phytotoxicity. However, it is
unclear if this statement refers to nicotine or one of the other two
active ingredients in the formulation.

[]{#__RefHeading___Toc173552903 .anchor}**1. Aquatic Effects
Characterization**

[]{#__RefHeading___Toc173552904 .anchor}**a. Aquatic Animals**

[]{#__RefHeading___Toc173552905 .anchor}**(1) Acute Effects**

***Fish***

Two acute toxicity studies suggest that nicotine is moderately toxic to
freshwater fish (**Table III.6**). However, there is considerable
uncertainty in both of these toxicity estimates since test
concentrations were not analytically verified. The actual exposures in
these studies are unknown and likely considerably lower than the nominal
concentrations suggest. There is additional uncertainty in the bluegill
sunfish study (MRID 00107188) since it appears that there were no
control groups included in the study. These studies cannot be used to
quantitatively estimate risk.

+---------------+-----------+---------+-------+----------+----------+
| **Table III.6 |           |         |       |          |          |
| Acute         |           |         |       |          |          |
| toxicity of   |           |         |       |          |          |
| nicotine to   |           |         |       |          |          |
| freshwater    |           |         |       |          |          |
| fish.**       |           |         |       |          |          |
+---------------+-----------+---------+-------+----------+----------+
| **Test        | **Test    | **End   | **Va  | **Eco    | **MRID   |
| Organism**    | Substance | point** | lue** | toxicity | or       |
|               | (         |         |       | Ca       | ECOTOX.  |
|               | Purity)** |         | **(mg | tegory** | No.**    |
|               |           |         | a.i.  |          |          |
|               |           |         | /L)** |          |          |
+---------------+-----------+---------+-------+----------+----------+
| Bluegill      | Nicotine  | 96-hour | 5.    | Mo       | 00107188 |
| Sunfish       | (98%)     | LC~50~  | 45^1^ | derately |          |
|               |           |         |       | toxic    |          |
| *Lepomis      |           |         |       |          |          |
| macrochirus*  |           |         |       |          |          |
+---------------+-----------+---------+-------+----------+----------+
| Rainbow trout | Nicotine  | 96-hour | 4     | Mo       | Eco. 138 |
|               | (≥ 95%)   | LC~50~  | .0^1^ | derately |          |
| *             |           |         |       | toxic    |          |
| Onchorhynchus |           |         |       |          |          |
| mykiss*       |           |         |       |          |          |
+---------------+-----------+---------+-------+----------+----------+

^1^ Nominal test concentration

In addition, there is a rainbow trout toxicity 96-hour acute toxicity
study available for tobacco dust (0.5% nicotine; MRID 42625503). Test
concentrations were 0 (control), 1, 10, 100, and 1000 mg/L.
Concentrations were not analytically verified. It was reported that test
material was observed floating on the surface of media, coating the
bottom of test vessels, and suspended throughout the test media at all
tested concentrations. The 96-hour LC~50~ was determined to be between
100 and 1000 mg/L tobacco dust; however, this is very uncertain given
that the test material was not in solution. This study cannot be used to
quantitatively estimate risk.

***Amphibians***

The acute toxicity of nicotine to early embryos of *Xenopus laevis* was
assessed over 96 hours in static renewal tests. The average LC~50~ (of
two tests) for nicotine was 136 mg/L. Several malformations were
reported for nicotine-exposed frogs. Nicotine exposure beginning at 0.25
mg/L induced contorted posture (lateral body flexure) and incomplete
development of the underside of the mouth. Gill hyperplasia was also
noted. All embryos were malformed above 0.8 mg/L nicotine. At 1.0 mg/L,
skeletal kinking was observed along with incomplete mouth development.
Above 90 mg/L the head and brain were reduced in size and the mouth was
poorly developed, if present at all. The gut was poorly coiled and the
heart swollen, and generalized pericardial and fin edema were noted. The
eyes were reduced in size and incompletely developed at levels higher
than 110 mg/L nicotine. The average EC~50~ for these malformations was
0.45 mg/L nicotine. This study cannot be used to quantitatively estimate
nicotine risk to amphibians for several reasons: 1) exposure
concentrations were not analytically verified; thus, there is
uncertainty regarding the actual exposures of the test organisms; 2)
nicotine was dissolved in FETAX solution, which contained other
chemicals (*i.e.,* antibiotics); and 3) the test substance purity was
not reported.

***Aquatic Invertebrates***

Nicotine acute toxicity data for freshwater invertebrates is limited to
a single study from the open literature. Perry and Smith (1988; Ecotox
No. 13161) assessed the 48-hour toxicity of nicotine to *Daphnia pulex*
in a static study (**Table III.7**). Concentrations were not
analytically verified in this study; thus, actual exposures are unknown
and may have been considerably lower than the nominal treatments
suggest. This study cannot be used to quantitatively estimate risk of
nicotine to freshwater invertebrates.

+--------------+-----------+-----------+-------+----------+----------+
| **Table      |           |           |       |          |          |
| III.7. Acute |           |           |       |          |          |
| toxicity of  |           |           |       |          |          |
| nicotine to  |           |           |       |          |          |
| freshwater   |           |           |       |          |          |
| inve         |           |           |       |          |          |
| rtebrates.** |           |           |       |          |          |
+--------------+-----------+-----------+-------+----------+----------+
| **Test       | **Test    | **E       | **Va  | **Eco    | *        |
| Organism**   | Substance | ndpoint** | lue** | toxicity | *ECOTOX. |
|              | (         |           |       | Ca       | No.**    |
|              | Purity)** |           | **(mg | tegory** |          |
|              |           |           | a.i.  |          |          |
|              |           |           | /L)** |          |          |
+--------------+-----------+-----------+-------+----------+----------+
| Water flea   | Nicotine  | 48-hour   | 0.2   | Highly   | 13161    |
|              | (≥ 97%)   | EC~50~    | 42^1^ | toxic    |          |
| *Daphnia     |           | (Immobi   |       |          |          |
| pulex*       |           | lization) |       |          |          |
+--------------+-----------+-----------+-------+----------+----------+

^1^ Nominal test concentration

In another study, the acute toxicity of tobacco dust (0.5% nicotine) to
*Daphnia magna* was assessed (MRID 42625502). Test concentrations were 0
(control), 1, 10, 100, and 1000 mg/L. Concentrations were not
analytically verified. It was reported that test material was observed
floating on the surface of media and coating the bottom of test vessels
at all tested concentrations. The 24-hour LC~50~ for tobacco dust was
determined to be greater than 1000 mg/L if the test vessels were aerated
and between 100 to 1000 mg/L if they were not aerated. These toxicity
estimates are very uncertain given that the test material was not in
solution, and aeration affected the test results.

Estuarine/marine invertebrate acute toxicity data is also limited to one
open literature study that assessed the effects of tobacco dust (2.8%
nicotine) to brackishwater pond snails (*Cerithidea cingulata* Gmelin).
The 72-hour LC~50~ for the juveniles, sub-adult, and adult snails were
30, 87, and 166 kg/ha (0.75, 2.17, and 4.15 lbs a.i./A), respectively.
This study is of limited use in this risk assessment since actual
exposures were not determined.

[]{#__RefHeading___Toc173552906 .anchor}**(2) Chronic Effects**

***Fish***

No guideline chronic fish toxicity studies have been submitted for
nicotine; however, there is one study available in the open literature
that provides information to characterize the potential chronic effects
of nicotine to fish. Passino-Reader, *et al.* (1995; EcoReference No.
16362) conducted sixty-day bioassays to test the effects of nicotine on
the survival, growth, and behavior of rainbow trout fry in a
constant-flow, temperature-controlled water system. The standard
procedures of ASTM (1988) and USEPA (1986) were followed. Nicotine
concentrations in a geometric progression from 0.06 to 1.0 mg/L resulted
in no significant effects on survivorship, weight, or length. At
nicotine concentrations of 1.4 to 6.0 mg/L, complete mortality occurred
at 6 mg/L, and the estimated 60-day LC50 was 5 mg/L. The median lethal
time (LT50) was for fry exposed to 6.0 mg/L nicotine were 22.0 and 19.8
days in replicate tanks. Length and weight decreased linearly with
increasing concentration in the range of 1.4 to 4.2 mg/L. The NOAEC and
LOAEC for length and weight were 2.9 and 4.2 mg/L, respectively.

However, this study is of limited value in this risk assessment because
the exposures were not analytically verified and are very uncertain. The
study author described the difficulty of maintaining nominal
concentrations of nicotine in large test systems and referred to Savino
and Tanabe (1989; Ecotox ref 390), a study in which the nicotine
treatments were undetectable 72 hours after dosing.

***Aquatic Invertebrates***

No guideline freshwater invertebrate chronic toxicity studies have been
submitted for nicotine; however, there is one study available in the
open literature that provides some information. A static-renewal, 16-day
chronic toxicity test was conducted to assess the effects of nicotine on
*Daphnia pulex* growth and reproduction. Test nicotine concentrations
were 0 (control), 0.02, 0.07, 0.12, 0.18, and 0.24 mg/L and were renewed
three times a week during the 16-day study. According to the study
author, nicotine significantly reduced growth and fecundity of daphnids
at nominal concentrations from 0.02 -- 0.24 mg/L. The LOAEC for length
was 0.07 mg/L, and the LOAEC for fecundity was 0.18 mg/L. It is unclear
if the NOAEC was 0.02 mg/L or \<0.02 mg/L.

The actual exposure concentrations in this study are uncertain. Test
concentrations were analytically verified at 1 hour, 48 hours, and 72
hours after preparation to simulate exposure in test media at the
beginning and end of the renewal cycle. At 48 hours after preparation,
the nicotine recovery rate in the test system was only 3% of the nominal
treatment, and it was undetectable at 72 hours. Therefore, the actual
NOAEC would be considerably less than 0.02 mg/L.

[]{#__RefHeading___Toc173552907 .anchor}**b. Aquatic Plants**

There are no aquatic plant toxicity data available for consideration in
this risk assessment.

[]{#__RefHeading___Toc173552908 .anchor}**2. Terrestrial Effects
Characterization**

[]{#__RefHeading___Toc173552909 .anchor}**a. Terrestrial Animals**

[]{#__RefHeading___Toc173552910 .anchor}**(1) Acute Effects**

***Birds***

There are no avian acute oral or dietary toxicity studies available for
nicotine. There is one study available for tobacco dust (MRID 42625501);
however, the percent nicotine in the test substance was not reported. In
this 14-day acute oral toxicity test with 25-week old bobwhite quail,
the acute oral LD~50~ was determined to be greater than 2150 mg/kg bw.
There were no mortalities or sublethal effects reported.

***Mammals***

The following summary of mammalian acute toxicity data for nicotine is
based on information provided by the Health Effects Division (HED).
There are no acceptable registrant-submitted acute toxicity data for
mammals[^24]. According to the National Institute for Occupational
Safety and Health[^25], the acute oral LD~50~ for mice, rats, and dogs
are of 3 mg/kg, 50 mg/kg, and 9.2 mg/kg, respectively. It should be
noted that these data have not been reviewed by the Agency.

The HED toxicity summary suggests that nicotine is readily absorbed
through the skin. When free nicotine was applied to the backs of cats
weighing 1.8-4.5 kg (treated area, 5-7 cm, in diameter was clipped with
scissors and not shaven) at 2-10 ml per cat, nicotine was fatal within a
few minutes (Faulkner, 1933). In other tests with cats receiving dermal
doses of 200 mg of nicotine or nicotine sulfate per cat, 81% of the cats
receiving the nicotine base died within 21-195 minutes (Travell, 1960).
Poisoning occurred very rapidly within 1-4 minutes including nausea,
vomiting, salivation, swallowing difficulty, increased rate of
respiration. The remaining nicotine treated cats were moribund after 4
hours of exposure. Symptoms of the nicotine sulfate treated cats were
milder and none of the animals died. In experiments with dogs using
nicotine dermal patches, nicotine was absorbed and produced clinical
signs in 15% of the treated dogs with plasma concentrations reaching 43
ng/ml (Matsushima *et al,* 1995).

***Terrestrial Invertebrates***

There are no terrestrial invertebrate toxicity data available for
nicotine. Nicotine shares a common mode of action with neonicotinoid
insecticides, including imidacloprid, which has been implicated in the
decline of honey bee populations[^26]. A honey bee acute contact study
(Guideline 141-1) would help reduce the uncertainty in this assessment.

[]{#__RefHeading___Toc173552911 .anchor}**(2) Chronic Effects**

***Birds***

There are no avian chronic toxicity data available for nicotine.

***Mammals***

A search of the ECOTOX database discovered many mammalian chronic
(developmental, behavioral) toxicity studies; however, none of them are
acceptable for use to quantitatively estimate chronic risk to mammals in
this ecological risk assessment. Summaries of each of the reviewed
studies are found below. (See **Appendix C** for study details).

-   Ajarem and Ahmad (1998; ECOTOX ref. 84721) investigated the effects
    of prenatal nicotine exposure on development and behavior of mice.
    Pregnant dams were given daily subcutaneous injections of normal
    saline (control) or 0.5 mg/kg bw nicotine dissolved in normal saline
    for 9-10 days. Nicotine treatment significantly reduced postnatal
    body weight gain, and delayed eye opening, the appearance of body
    hairs, and sensory motor reflexes. However, motor activity was
    stimulated in early adulthood of pups prenatally exposed to
    nicotine. Since nicotine was administered via injection, these data
    cannot be used quantitatively to estimate risk in this ecological
    risk assessment.

-   Johns *et al.* (1982; ECOTOX ref. 84597) studied the behavioral
    effects of prenatal nicotine exposure in guinea pigs. The guinea
    pigs exposed to nicotine (3 mg/kg SC injection twice daily)
    prenatally demonstrated severe behavioral impairments. Treatment
    offspring alternated at chance or below chance levels while the
    controls alternated at normal levels. Most of the nicotine-treated
    guinea pigs failed to enter the novel alley while 80% of the control
    animals entered. Treated animals were severely impaired compared to
    the control subjects on the reversal and discrimination problems.
    Since nicotine was administered via injection, these data cannot be
    used quantitatively to estimate risk in this ecological risk
    assessment.

-   Kita *et al.* (1988; ECOTOX ref. 84604) studied the effects of
    nicotine on ambulatory activity in seven-week-old mice were injected
    subcutaneously with nicotine (0.1, 0.5, and 1.0 mg/kg) or a control
    solution (saline) and then placed into an ambulo-cage. Activity
    counts were recorded during a 180-minute period. Mice treated with
    the highest nicotine dose (1.0 mg/kg) demonstrated significantly
    decreased ambulatory activity in a dose-dependent manner from 5 to
    60 minutes after the administration. Mice dosed with 0.5 mg/kg
    nicotine showed depressed activity for 40 minutes after
    administration. On the contrary, the lowest nicotine dose, 0.1
    mg/kg, stimulated activity in the first 20 minutes. Further analysis
    of the data revealed that all nicotine treatments resulted in an
    initial increase in activity, and then the ataxic phase developed.
    Since nicotine was administered via injection, these data cannot be
    used quantitatively to estimate risk in this ecological risk
    assessment.

-   Romero and Chen (2004; ECOTOX ref. 84725) examined the effects of
    developmental nicotine exposure on rat offspring somatic growth and
    behavioral performance in an open-field test. Female rats were
    implanted with nicotine (35 mg for 21-day release) or placebo
    pellets on gestational day 8. There was no significant difference in
    offspring body weight across the treatments. The amount of activity,
    measured by the total number of crossings in the open-field test,
    revealed less activity in male offspring and an increase in female
    offspring activity as a function of testing day. The increase in
    female ambulatory activity was observed in the placebo and normal
    control, but not in the nicotine treatment group. This suggests that
    the control subjects adapted to the testing apparatus and were less
    anxious or afraid whereas the nicotine-exposed subjects failed to
    adapt to the test system, learn the contextual cues, or retrieve the
    learned information. Since nicotine implants were used as the route
    of exposure, these data cannot be used quantitatively to estimate
    risk in this ecological risk assessment.

In addition, the HED provided information regarding mammalian toxicity
of nicotine. The following summary of mammalian toxicity data for
nicotine is based on this information. There are no chronic reproductive
toxicity data available for nicotine; however, there are several studies
in the open literature that provide information to qualitatively assess
the potential impacts of nicotine on mammals.

In a nicotine sulfate 300-day dietary toxicity study (Wilson & DeEds,
1936), nicotine retarded rat growth with a NOAEL of 0.006% nicotine base
(4 mg/kg/day). Retarded growth in rats was reversed upon discontinuing
the nicotine diets and rats resumed normal growth.

Nicotine has a detrimental effect on general growth and development as
well as on palatogenesis in mice (Saad *et al*, 1990). Pregnant CD-1
mice (N=19), exposed to i.p. injection of 0.1% nicotine sulfate at a
dose of 1.67 mg/kg body weight/day on gestational days 6-15 had
significantly depressed maternal weight gain (37% decrease on GD18) and
fetal weight (64% decrease) compared to the controls. Fetal crown-rump
length and fetal head dimensions (width, height and circumference) were
significantly reduced. Histological examination revealed that 9.6% of
fetuses of nicotine injected mothers presented clefts of the palate,
whereas none of the control fetuses had that anomaly. Nicotine treatment
also had teratogenic effects on first molar odontogenesis in the mouse
(Saad *et al*, 1991). It was suggested that nicotine, or its metabolic
byproducts, interfere with normal interaction between the epithelial and
mesenchymal components of the developing tooth.

Mean fetal body fat was significantly increased in fetuses of rats
administered nicotine (2.46 ± 0.18 mg/kg/day in drinking water) during
pregnancy throughout gestation day 20 (Williams & Kanagasabai, 1984).
Rate for maternal lypolysis were higher in the nicotine treated animals.
Maternal body weights gains were significantly lower (77.2% of controls,
p \< 0.001).

Nicotine - delivering transdermal patches applied on the back of
pregnant female rats resulted in 100% pregnancy failure in two animals
treated with 3.5 mg/day during the entire pregnancy (GD 2-19) and 50% in
8 animals exposed to the same amount during the first trimester (GD 2-7)
and 55% in 13 animals exposed to 1.75 mg/day during the entire pregnancy
(Witschi *et al*, 1994). Litter size and pup weights were not affected
by the nicotine treatment. Nicotine and cotinine plasma levels in the
sacrificed animals were not detected in animals that had carried a patch
during the first trimester of pregnancy. In animals exposed the entire
pregnancy at 1.75 mg/day patches, 3 pregnant animals out of six had
measurable nicotine levels (43 ± 22 ng/mL) and all had cotinine levels
(100 ± 48 ng/mL). The non pregnant females of the 1.75 mg/day patches
had 70 ± 57 ng/mL of plasma nicotine and 231 ± 84 ng/mL of plasma
cotinine. In the two animals exposed to 3.5 mg/day patches and were
nonpregnant, nicotine plasma levels were 241 ± 51 ng/mL and cotinine
levels of 302 ± 94 ng/mL.

Offspring of groups of guinea pig dams injected twice daily throughout
gestation with 0, 0.5, 1.5 or 2.5 mg/kg of nicotine-hydrogen tartarate
(15/dose) exhibited performance deficits in both learned and innate
behavioral measures throughout development and adulthood (Johns *et al*,
1992). Offspring birth weight was not affected by treatment nor the 32
day weight gain. Initial weight, weight gain, gestation length, number
of live or dead offspring, or mean food consumption during pregnancy did
not differ significantly in the treated groups versus the controls. Dams
receiving the nicotine injections reacted with aversion.

Nicotine may limit the physical performance of exposed animals. Thus
subcutaneous injection of nicotine (0.125 - 0.375 mg/kg) in Swiss albino
adult male rats (200-400 g weight) decreased the endurance time in
swimming exercise significantly (10 minute after the injection, in a
dose-dependent manner compared to the control group (Temocin *et al*,
1993). At the dose of 0.125 mg/kg nicotine, the endurance time in
swimming exercise remained unchanged, while at the doses of 0.25 and
0.375 mg/kg, it decreased significantly (p \< 0.05 and p \< 0.01,
respectively). This effect was antagonized by pretreatment with
hexamethonium 5 mg/kg s.c., suggesting that nicotine may limit physical
performance.

Male and female offspring of rats exposed to nicotine (1.5 mg/kg/day,
subcutaneous implanting of pregnant rats) during gestation demonstrated
an increase in spontaneous locomotor activity when compared with
saline-exposed controls and the total number of pups born to the treated
group was significantly less than the controls (Fung, 1988).

Pre-natal exposure to nicotine causes significant changes in behavior in
later life (Peters *et al*, 1979). Thus, 60 - 80 days old offspring of
rats treated with nicotine in drinking water (6 mg/kg/day intake) four
weeks before mating and during pregnancy and throughout nursing and 6
weeks after weaning showed an increased spontaneous motor activity in
the light which was not prevented by cross-fostering to control dams at
birth. The paternal rats had a marked reduction in body weight gain (55%
of controls for males and 63% of controls for females after 4 month of
nicotine treatment). The dams were more active during the day and
exhibited a reduced plasma corticosterone response to stress. Male but
not female offspring of nicotine treated rats were significantly lighter
at birth than control males.

Nicotine administered chronically in low doses (0, 1.5, 3 or 6.0 mg/kg
per day; subcutaneously) throughout gestation causes subtle neurological
changes which are manifested as behavioral alterations in the newborn
(the righting reflex, temperature regulation, adherence to the inclined
screen, and in organ/body weight ratios for brain, heart, lung, liver,
and kidney) and adult offspring (prolonged time required and an increase
in number of mistakes made, during food maze testing and an increased
brain protein content) (Peters & Ngan, 1982).

[]{#__RefHeading___Toc173552912 .anchor}**b. Terrestrial Plants**

There are no terrestrial plant toxicity data available for consideration
in this risk assessment. The product label states, "Do not apply the
product directly to foliage or stems," which indicates that there is a
possibility of phytotoxicity. However, it is unclear if this statement
refers to the potential phytotoxicity of nicotine or one of the other
two active ingredients in the formulation.

# **IV. Risk Characterization** 

Since there were no acceptable ecotoxicity data, aquatic and terrestrial
risks cannot be quantitatively estimated. However, the available data
are useful for qualitatively characterizing the potential risks
associated with use of Shotgun^®^ Rabbit and Dog Repellent.

[]{#__RefHeading___Toc173552914 .anchor}**A. Risks to Aquatic
Organisms**

Due to the lack of acceptable toxicity data for nicotine, acute and
chronic risk of Shotgun^®^ Rabbit and Dog Repellent to aquatic organisms
cannot be precluded; however, given the extremely low predicted aquatic
exposures, the likelihood of risk is presumed to be very low. Aquatic
EECs for nicotine when used as Shotgun^®^ Rabbit and Dog Repellent were
estimated using GENEEC2 and adjusted for the percent area treated
assuming a generic residential setting (Section III.B.2.A). The highest
peak aquatic EEC was 40 ng/L (parts per trillion). Based on this
exposure estimate, in order to exceed the Agency's acute listed species
LOC of 0.05 for aquatic animals, the acute toxicity threshold (*e.g.*,
LC~50~) would need to be 800 ng/L or less. A rainbow trout toxicity
study from the open literature (ECOTOX ref. 138) reported a 96-hour
LC~50~ of 4 mg/L; although there is uncertainty in the actual test
concentrations, the reported LC~50~ is about 5000 times greater than the
level that would trigger acute listed species concerns. Similarly, a
freshwater invertebrate toxicity study from the open literature (ECOTOX
ref. 13161) reported a 48-hour EC~50~ of 0.242 mg/L for *Daphnia pulex*,
which is about 300 times greater than 800 ng/L. For aquatic plants,
there are no available toxicity data; an aquatic plant NOAEC would need
to be less than 2.5 ng/L to be below the acute listed species plant LOC
of 1.0. Acute toxicity data for freshwater fish (Guideline 72-1),
freshwater invertebrates (Guideline 72-2), and aquatic plants (Guideline
123-2) would reduce the uncertainty in this risk assessment.

For chronic risk, the highest predicted 60-day EEC was 2.5 ng/L. A
chronic study from the open literature (ECOTOX ref. 16362) reported a
60-day NOAEC of 2.9 mg/L for rainbow trout growth. The study author
admitted difficulty of maintaining nominal concentrations of nicotine in
large test systems and referred to Savino and Tanabe (1989; Ecotox ref
390), a study in which the nicotine treatments were undetectable 72
hours after dosing. In addition, reproductive effects were not measured
in this study. However, based on the predicted 60-day EEC, in order to
exceed the Agency's chronic LOC of 1.0, the chronic NOAEC must be 2.5
ng/L (ppt) or lower. This level is more than 6 orders of magnitude lower
than the reported NOAEC (2.9 mg/L). A 16-day chronic *Daphnia pulex*
toxicity study from the open literature (Ecotox ref 390) determined a
NOAEC of 0.02 mg/L for length. As explained above, this study had
extremely poor recovery of the test compound. This NOAEC is 8000 times
greater than the level that would trigger chronic risk.

[]{#__RefHeading___Toc173552915 .anchor}**B. Risks to Terrestrial
Organisms**

Using the T-REX model, terrestrial dietary exposures were estimated for
Shotgun^®^ Rabbit and Dog Repellent, a granular (dust) formulation for
use in residential settings. Assuming a generic residential setting
(Section III.B.2.A), the estimated exposure for a terrestrial animal is
about 67 mg a.i./A. Since the T-REX model is designed to calculate
exposures for pesticides used in agricultural settings (*i.e.*, complete
treatment of a one-acre plot), this exposure estimate for nicotine is
likely an overestimate given that the product is applied in 2 to 12-inch
bands around the perimeter of gardens in residential settings.

Acute mammalian toxicity data from the open literature suggest that
nicotine is very highly toxic to mice, with an acute oral LD~50~ of 3
mg/kg.[^27] Compared to the predicted exposure level of 67 mg a.i./A,
the LD~50~/ft^2^ values are 677, 359, and 29 for the modeled 15, 35, and
1000 g small mammals, respectively. These risk estimates all exceed the
acute risk LOC of 0.5. However, this is the estimated risk for a mammal
if 100% of the animal's diet consists of Shotgun^®^ Rabbit and Dog
Repellent. Since this product is a rabbit and dog repellent, it is
reasonable to assume that small mammals (*e.g.*, field mice) are
similarly repelled, and terrestrial dietary exposure may be unlikely.

There are no avian toxicity data available for nicotine. It is unclear
whether Shotgun^®^ Rabbit and Dog Repellent is capable of repelling
birds as well; thus, the dietary exposure route is presumed to be
possible. At this time, the potential risk to birds cannot be precluded.
Acute and chronic avian toxicity data (Guidelines 71-1, 71-2, and 71-4)
would help reduce this uncertainty regarding the risk of nicotine to
birds.

There are no terrestrial invertebrate data available for consideration
in this risk assessment. Nicotine shares a common mode of action with
neonicotinoid insecticides, including imidacloprid, which has been
implicated in the decline of honey bee populations. Since Shotgun^®^
Rabbit and Dog Repellent is a dust (granular) to be applied in a band
around the perimeter of gardens, exposure to beneficial insects (*e.g.*,
honey bees) may be unlikely; however, risk cannot be precluded at this
time. An acute contact toxicity test with honey bees (Guideline 141-1)
would help reduce the uncertainty in this risk assessment.

There are no terrestrial plant toxicity data available for consideration
in this risk assessment. At this time, risks to terrestrial plants
cannot be precluded. Tier I seedling emergence and vegetative vigor
terrestrial plant toxicity studies (Guidelines 123-1(a,b)) would help
reduce uncertainty in this risk assessment.

[]{#__RefHeading___Toc173552916 .anchor}**C. Ecological Incident
Information**

There are no reports for nicotine, nicotine sulfate, or tobacco dust in
the Ecological Incident Information System (EIIS) database.

[]{#__RefHeading___Toc173552917 .anchor}**D. Federally Threatened and
Endangered (Listed) Species Concerns**

Due to a lack of toxicity data, risks to aquatic and terrestrial
organisms were not quantitatively estimated. At this time, direct and
indirect effects to freshwater fish (surrogate for aquatic-phase
amphibians), freshwater invertebrates, aquatic plants, mammals, birds
(surrogate for terrestrial-phase amphibians and reptiles), terrestrial
invertebrates, and terrestrial plants as a result of nicotine use (in
Shotgun^®^ Rabbit and Dog Repellent) cannot be precluded. In the event
nicotine toxicity data become available, a more detailed discussion of
the potential direct and indirect effects to listed species would be
possible.

[]{#__RefHeading___Toc173552918 .anchor}**E. Assumptions, Limitations,
Uncertainties, Strengths and Data Gaps**

[]{#__RefHeading___Toc173552919 .anchor}**1. Exposure For All Taxa**

[]{#__RefHeading___Toc173552920 .anchor}**a. Maximum Use Scenario**

Since the maximum application rate of Shotgun^®^ Rabbit and Dog
Repellent was not specified on the label, it was assumed that 6
applications are made annually, 60 days apart. The frequency at which
actual uses approach this maximum use scenario may be dependent on
insecticide resistance, timing of applications, cultural practices, and
market forces.

[]{#__RefHeading___Toc173552921 .anchor}**b. Additive and/or Synergistic
Effects**

It was assumed that aquatic and terrestrial organisms were exposed only
to nicotine in a formulation called Shotgun^®^ Rabbit and Dog Repellent.
Ecological risks associated with exposure to a mixture of nicotine and
its degradates, other pesticides, adjuvants, heavy metals, industrial
chemicals, pharmaceuticals, *etc.* were not considered in this risk
assessment.

[]{#__RefHeading___Toc173552922 .anchor}**2. Exposure for Aquatic
Species**

[]{#__RefHeading___Toc173552923 .anchor}**a. Data Gaps and
Uncertainties**

The assessment of the aquatic exposure relies on the environmental fate
behavior of the chemical (that is, its persistence in water/soil,
transformation products, and movement of the parent chemical and/or in
water/soil/air).

No guideline studies (USEPA, FIFRA Subdivision N) were conducted with
nicotine as the test substance are available. In 1994 (DPBarcode
D206473), the Environmental Fate and Effects Division concluded that the
environmental fate data requirements for nicotine could be waived.
Environmental fate data were estimated using EPISuiteVersion 3.20, and
data from the open literature were considered. When integrated, the data
provide an overview on how nicotine would behave in the environment when
used as a pesticide. EPISuite is a screening level tool used to
delineate the behavior of a chemical in the environment.

[]{#__RefHeading___Toc173552924 .anchor}**b. Aquatic Exposure Model**

No aquatic risk assessment was performed for the greenhouse uses. The
ecological risk assessment was only performed for nicotine in Shotgun^®^
Rabbit and Dog Repellent product used in a residential outdoor setting.

The aquatic exposure for nicotine was performed using GENEEC Version 2.0
Tier I simulation model, following the recommended selection of input
parameters. Uncertainties in the aquatic exposure assessment come from
two sets of major data sources. One set of uncertainties come from the
assumptions that had to be made for selecting the use input parameters
(i.e., application rate(s); frequency and method of application;
estimates of treated area). The other set of uncertainties come from the
model estimated (EPISuite), screening level environmental fate input
parameters.

[Use Input Parameters]{.underline}

*Source of Nicotine*

Assumptions had to be made to estimate exposure concentrations of
nicotine from application of the product in which "tobacco dust" is the
source of nicotine. It is unclear if the "0.35%" nicotine content in the
Shotgun^®^ Rabbit and Dog Repellent product is based on a nominal
concentration or measured concentration. For this reason, the nicotine
content in the product(s) was assumed to be "neat nicotine" (pure
nicotine). How this assumption overestimates/underestimates the exposure
of nicotine in the environment, when used as a pesticide, is not known.
Furthermore, it should kept in mind that nicotine content in tobacco
plant depends on the strain of tobacco, where it is cultivated, and
varies in the plant itself (leaves, stems, etc).

*Nicotine Available for Runoff*

It was assumed that all of the nicotine in the tobacco dust becomes
available for runoff and the available nicotine is pure nicotine ("neat"
nicotine). Coupled with the uncertainty on actual nicotine in tobacco
dust (see above), the overestimation/underestimation of aquatic exposure
concentration is not known.

*Estimation of Application Rate and Application Site*

The manner in which the Dog & Rabbit Chaser is used can be envisioned as
a localized application, given that the product is to be used on the
perimeter of the area to be protected. To express the application rate
in terms of pounds of active ingredient (nicotine) per acre (lb
a.i,/acre), as required by the exposure models, several assumptions had
to be made to extrapolate localized applications to lb a.i./acre. These
assumptions include:

-   The potential surface area to be treated was based on a typical
    house and garden perimeters, as per U.S.A. 2000 Census data, which
    served to estimate the percent of treated area within a typical 0.25
    acre lot.

-   The band widths were 2 (minimum width recommended in label) inches
    and assumed widths of 6 and 12 inches because upper limit widths are
    not specified in the label.

-   The amount of product to be applied was taken from the label and the
    percent of nicotine (0.35%) was taken as the available nicotine
    (pure nicotine; "neat nicotine") for each width. The amount of
    nicotine (pounds of nicotine) was estimated for a square-foot
    treatment area and then converted to the equivalent application rate
    in terms of nicotine per acre. Note that this constitutes an
    extrapolation of a localized treatment to treatment of a larger
    area.

    *Method of Application*, *frequency, and Spray Drift Assumptions*

    The product is a dust, but a granular product was assumed because
    GENEEC does not consider dusts. Because the product is a dust, a 1%
    drift (default value) was assumed to take into account any potential
    drift (wind) of the product to adjacent areas.

    The number of applications and application intervals are not
    specified in the label. Six applications per year, two months apart
    were assumed.

    [Environmental Fate Input Parameters]{.underline}

    Environmental fate/physical-chemical properties were selected from
    screening level model estimates (EPISuite Version 3.20).
    Uncertainties that may underestimate/ overestimate environmental
    exposure concentrations in surface water are linked to the
    assumptions and limitations of the model:

```{=html}
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```
-   That the Koc estimates were developed for neutral compounds.
    Nicotine is a weak acid, for which the concentration of the
    protonated and nonprotonated forms varies with pH and hence the
    mobility. Increase in pH would increase the mobility of the
    chemical.

-   That the half-life of biotransformation is based on assigned days
    according to estimated time frames. For nicotine, both the primary
    and ultimate biotransformation time frames of "weeks-to-months" are
    assigned the same half-life of 37.5 days.

-   That the aerobic aquatic metabolism half-life uses a default value,
    assuming that the biotransformation half-life of 37.5 days applies
    to aerobic soil metabolism.

-   That the photolysis in water half-life is zero for direct
    photolysis.

    *Transformation Products*

    Although bacteria have been shown to oxidize nicotine in soils in a
    sequence of oxidative reactions, the environmental fate of these
    metabolites is not known (i.e., pertistence and transport).

    [Additional Uncertainties]{.underline}

Other sources of nicotine exposure in the environment come from tobacco
smoking/chewing, "non-prescription drug," and from releases by the
tobacco industry. Nicotine (tobacco industry) is listed in the Toxic
Release Inventory (TRI) and the tobacco industry is required to report
releases of nicotine to the environment on a yearly basis. Because
exposure to nicotine in actual surface water can result from other
sources, it would become difficult to distinguish the exclusive
contribution of its use as a pesticide from other sources.

The present ecological risk assessment is based only on estimated
exposure concentrations that result exclusively from the use of nicotine
as a pesticide. It is not an aggregate exposure assessment that takes
into consideration other sources. Therefore, risk from pesticide use of
nicotine alone may underestimate the overall exposure and ecological
risk of nicotine.

The environmental fate and physical/chemical properties needed to run
the model are selected from EPISuite 3.20 estimates, unless otherwise
specified. Both EPISuite and GENEEC2 are screening level models, and,
therefore, the resulting aquatic exposure estimates are likely to be
overestimated.

**3. Exposure For Terrestrial Species**

[]{#__RefHeading___Toc173552925 .anchor}**a. Location of Wildlife
Species**

For this screening-level terrestrial risk assessment, a generic bird or
mammal was assumed to occupy either the treated area or adjacent areas
receiving nicotine at the treatment rate on the field. Actual habitat
requirements of any particular terrestrial species were not considered,
and it was assumed that species occupy, exclusively and permanently, the
modeled treatment area.

[]{#__RefHeading___Toc173552926 .anchor}**b.** **Routes of Exposure**

For screening-level terrestrial risk assessments, a generic bird or
mammal is assumed to occupy either the treated field or adjacent areas
receiving the pesticide at a rate commensurate with the treatment rate
on the field. The actual habitat requirements of any particular
terrestrial species are not considered, and it is assumed that species
occupy, exclusively and permanently, the treated area being modeled.
This assumption leads to a maximum level of exposure in the risk
assessment. Screening‑level risk assessments for pesticides consider
dietary exposure alone. Other routes of exposure, not considered in this
assessment, are discussed below.

***Incidental Soil Ingestion Exposure***

This risk assessment does not consider incidental soil ingestion.
Available data suggest that up to 15% of an animal's diet can consist of
incidentally ingested soil depending on the species and feeding strategy
(Beyer *et al*., 1994). The simple first approximation of soil
concentration of pesticide from spray application shows that ingestion
of soil at an incidental rate of up to 15% of the diet would not
increase dietary exposure.

***Inhalation Exposure***

The screening risk assessment does not consider inhalation exposure of
spray applications. Such exposure may occur through three potential
sources: (1) spray material in droplet form at the time of application
(2) vapor phase pesticide volatilizing from treated surfaces, and (3)
airborne particulate (soil, vegetative material, and pesticide dusts).

Available data suggest that inhalation exposure at the time of
application is not an appreciable route of exposure for birds. According
to research on mallards and bobwhite quail, respirable particle size in
birds (particles reaching the lung) is limited to a maximum diameter of
2 to 5 microns. Theoretically, inhalation of a pesticide's active
ingredient in the vapor phase may be another source of exposure for some
pesticides under some exposure situations. Although there is potential
for volatilization (soil, water, and possibly plant surfaces),
atmospheric reactions with hydroxy radicals (half-lives less than 10
hours) and diffusion will decrease concentration in the vapor phase.

The impact from exposure to dusts contaminated with the pesticide cannot
be assessed generically as partitioning issues related to application
site soils and chemical properties render the exposure potential from
this route highly situation-specific.

***Dermal Exposure***

The screening assessment does not consider dermal exposure, except as it
is indirectly included in calculations of RQs based on lethal doses per
unit of pesticide treated area. Dermal exposure may occur through three
potential sources: (1) direct application of spray or granules to
terrestrial wildlife in the treated area or within the drift footprint,
(2) incidental contact with contaminated vegetation for foliar spray
applications, or (3) contact with contaminated water or soil.

The available measured data related to wildlife dermal contact with
pesticides are extremely limited. The Agency is actively pursuing
modeling techniques to account for dermal exposure via direct
application of spray and by incidental contact with vegetation.

***Drinking Water Exposure***

Drinking water exposure to a pesticide's active ingredient may be the
result of consumption of surface water or consumption of the pesticide
in dew or other water on the surfaces of treated vegetation. For a
pesticide containing an active ingredient with the potential to dissolve
in runoff, puddles on the treated field may contain the chemical.

***Dietary Intake ‑ The Differences Between Laboratory and Field
Conditions***

The acute and chronic characterizations of risk rely on comparisons of
wildlife dietary residues with LC~50~ or NOAEC values expressed in
concentrations of pesticides in laboratory feed. These comparisons
assume that ingestion of food items in the field occurs at rates
commensurate with those in the laboratory. Although the screening
assessment process adjusts dry‑weight estimates of food intake to
reflect the increased mass in fresh‑weight wildlife food intake
estimates, it does not allow for gross energy and assimilative
efficiency differences between wildlife food items and laboratory feed.

On gross energy content alone, direct comparison of a laboratory dietary
concentration‑based effects threshold to a fresh‑weight pesticide
residue estimate would result in an underestimation of field exposure
through food consumption by a factor of 1.25 to 2.5 for most food items.
Only for seeds would the direct comparison of dietary threshold to
residue estimate lead to an overestimate of exposure.

Differences in assimilative efficiency between laboratory and wild diets
suggest that current screening assessment methods do not account for a
potentially important aspect of food requirements. Depending upon
species and dietary matrix, bird assimilation of wild diet energy ranges
from 23 to 80%, and mammal assimilation ranges from 41 to 85% (U.S. EPA,
1993). If it is assumed that laboratory chow is formulated to maximize
assimilative efficiency (e.g., a value of 85%), a potential for
underestimation of exposure may exist by assuming that consumption of
food in the wild is comparable with consumption during laboratory
testing. In the screening process, exposure may be underestimated
because metabolic rates are not related to food consumption.

Finally, the screening procedure does not account for situations where
the feeding rate may be above or below requirements to meet free living
metabolic requirements. Gorging behavior is a possibility under some
specific wildlife scenarios (e.g., bird migration) where the food intake
rate may be greatly increased. Kirkwood (1983) has suggested that an
upper‑bound limit to this behavior might be the typical intake rate
multiplied by a factor of 5.

In contrast is the potential for avoidance, operationally defined as
animals responding to the presence of noxious chemicals in their food by
reducing consumption of treated dietary elements. This response is seen
in nature where herbivores avoid plant secondary compounds.

[]{#__RefHeading___Toc173552927 .anchor}**c. Dietary Intake**

It was assumed that ingestion of food items in the field occurs at rates
commensurate with those in the laboratory. Although the screening
assessment process adjusts dry-weight estimates of food intake to
reflect the increased mass in fresh-weight wildlife food intake
estimates, it does not allow for gross energy differences. Direct
comparison of a laboratory dietary concentration- based effects
threshold to a fresh-weight pesticide residue estimate would result in
an underestimation of field exposure by food consumption by a factor of
1.25 - 2.5 for most food items.

Differences in assimilative efficiency between laboratory and wild diets
suggest that current screening assessment methods do not account for a
potentially important aspect of food requirements. Depending upon
species and dietary matrix, bird assimilation of wild diet energy ranges
from 23 - 80%, and mammal's assimilation ranges from 41 - 85% (U.S.
Environmental Protection Agency, 1993). If it is assumed that laboratory
chow is formulated to maximize assimilative efficiency (e.g., a value of
85%), a potential for underestimation of exposure may exist by assuming
that consumption of food in the wild is comparable with consumption
during laboratory testing. In the screening process, exposure may be
underestimated because metabolic rates are not related to food
consumption.

Finally, the screening procedure does not account for situations where
the feeding rate may be above or below requirements to meet free living
metabolic requirements. Gorging behavior is a possibility under some
specific wildlife scenarios (e.g., bird migration) where the food intake
rate may be greatly increased. Kirkwood (1983) has suggested that an
upper-bound limit to this behavior might be the typical intake rate
multiplied by a factor of 5. In contrast, there may be potential for
avoidance (animals respond to the presence of noxious chemicals in food
by reducing consumption of treated dietary elements).

[]{#__RefHeading___Toc173552928 .anchor}**4. Ecological Effects
Assessment**

[]{#__RefHeading___Toc173552929 .anchor}**a. Data Gaps and
Uncertainties**

There are no acceptable guideline ecotoxicity studies for nicotine. In
1994[^28], the EFED denied waiver requests for the following studies:
acute avian oral toxicity (71-1(a)), acute avian dietary toxicity
(71-2(a)), and acute fish toxicity (72-1(c); however, none of these
studies were subsequently provided to the Agency. In addition, the
following studies would reduce uncertainty in this risk assessment: Tier
I aquatic plant growth (123-2), avian chronic reproduction (71-4), acute
contact toxicity to honey bees (Guideline 141-1), and Tier I seedling
emergence and vegetative vigor terrestrial plant toxicity studies
(Guidelines 123-1(a,b)).

[]{#__RefHeading___Toc173552930 .anchor}**b. Sublethal Effects**

For an acute risk assessment, the screening risk assessment relies on
the acute mortality endpoint as well as a suite of sublethal responses
to the pesticide, as determined by the testing of species response to
chronic exposure conditions and subsequent chronic risk assessment.
Consideration of additional sublethal data in the assessment is
exercised on a case-by-case basis and only after careful consideration
of the nature of the sublethal effect measured and the extent and
quality of available data to support establishing a plausible
relationship between the measure of effect (sublethal endpoint) and the
assessment endpoints.

[]{#__RefHeading___Toc173552931 .anchor}**APPENDIX A. Environmental Fate
and Exposure Assessment**

**1. EPI Suite Summary**

The EPI (Estimation Programs Interface) EPI Suite™ is a Windows^®^ based
suite of physical/chemical property and environmental fate estimation
models developed by the EPA's Office of Pollution Prevention Toxics and
Syracuse Research Corporation (SRC). EPI Suite^™^ uses a single input to
run the following estimation models: KOWWIN™, AOPWIN™, HENRYWIN™,
MPBPWIN™, BIOWIN™, BioHCWIN, PCKOCWIN™, WSKOWWIN™, WATERNT™, BCFWIN™,
HYDROWIN™, KOAWIN and AEROWIN™, and the fate models STPWIN™, WVOLWIN™,
and LEV3EPI™.

^™^

EPI Suite is a screening level tool used to assess the environmental
fate and exposure of a chemical in the environment .

• [KOWWIN]{.underline}™: Estimates the log octanol-water partition
coefficient, log KOW, of chemicals using an atom/fragment contribution
method.\
• [AOPWIN]{.underline}™: Estimates the gas-phase reaction rate for the
reaction between the most prevalent atmospheric oxidant, hydroxyl
radicals, and a chemical. Gas-phase ozone radical reaction rates are
also estimated for olefins and acetylenes. In addition, AOPWIN™ informs
the user if nitrate radical reaction will be important. Atmospheric
half-lives for each chemical are automatically calculated using assumed
average hydroxyl radical and ozone concentrations.\
• [HENRYWIN]{.underline}™: Calculates the Henry's Law constant
(air/water partition coefficient) using both the group contribution and
the bond contribution methods.\
• [MPBPWIN]{.underline}™: Melting point, boiling point, and vapor
pressure of organic chemicals are estimated using a combination of
techniques.  Included is the subcooled liquid vapor presssure, which is
the vapor pressure a solid would have if it were liquid at room
temperature.  It is important in fate modeling.\
• [BIOWIN]{.underline}™: Estimates aerobic and anaerobic
biodegradability of organic chemicals using 7 different models; two of
these are the original Biodegradation Probability Program (BPP™).  The
seventh and newest model estimates anaerobic biodegradation potential.\
• [BioHCWIN]{.underline}: Estimates biodegradation half-life for
compounds containing only carbon and hydrogen (i.e. hydrocarbons).\
• [PCKOCWIN]{.underline}™: The ability of a chemical to sorb to soil and
sediment, its soil adsorption coefficient (Koc), is estimated by this
program. EPI\'s Koc estimations are based on the Sabljic molecular
connectivity method with improved correction factors.\
• [WSKOWWIN]{.underline}™: Estimates an octanol-water partition
coefficient using the algorithms in the KOWWIN™ program and estimates a
chemical's water solubility from this value. This method uses correction
factors to modify the water solubility estimate based on regression
against log Kow.\
• [WATERNT]{.underline}™: Estimates water solubility directly using a
\"fragment constant\" method similar to that used in the KOWWIN™ model.\
• [HYDROWIN]{.underline}™: Acid- and base-catalyzed hydrolysis constants
for specific organic classes are estimated by HYDROWIN™. A chemical's
hydrolytic half-life under typical environmental conditions is also
determined. Neutral hydrolysis rates are currently not estimated.\
• [BCFWIN]{.underline}™: This program calculates the
[B]{.underline}io[C]{.underline}oncentration [F]{.underline}actor and
its logarithm from the log Kow. The methodology is analogous to that for
WSKOWWIN™. Both are based on log Kow and correction factors.\
• [KOAWIN]{.underline}: KOA is the octanol/air partition coefficient and
has multiple uses in chemical assessment.  The model estimates KOA using
the ratio of the octanol/water partition coefficient (KOW) from KOWWIN™,
and the dimensionless Henry\'s Law constant (KAW) from HENRYWIN™.

**EPISuite Calculations for Nicotine (CAS Reg. No. 54-11-5)**

**Photodegradation Air**

SMILES : n(cccc1C(N(CC2)C)C2)c1

CHEM : Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S)-

MOL FOR: C10 H14 N2

MOL WT : 162.24

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-- SUMMARY (AOP v1.92): HYDROXYL
RADICALS \-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

Hydrogen Abstraction = 24.1139 E-12 cm3/molecule-sec

Reaction with N, S and -OH = 66.0000 E-12 cm3/molecule-sec

Addition to Triple Bonds = 0.0000 E-12 cm3/molecule-sec

Addition to Olefinic Bonds = 0.0000 E-12 cm3/molecule-sec

\*\*Addition to Aromatic Rings = 0.8778 E-12 cm3/molecule-sec

Addition to Fused Rings = 0.0000 E-12 cm3/molecule-sec

OVERALL OH Rate Constant = 90.9918 E-12 cm3/molecule-sec

HALF-LIFE = 0.118 Days (12-hr day; 1.5E6 OH/cm3)

HALF-LIFE = 1.411 Hrs

\...\...\...\...\...\...\...\... \*\* Designates Estimation(s) Using
ASSUMED Value(s)

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-- SUMMARY (AOP v1.91): OZONE
REACTION \-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

\*\*\*\*\*\* NO OZONE REACTION ESTIMATION \*\*\*\*\*\*

(ONLY Olefins and Acetylenes are Estimated)

Experimental Database: NO Structure Matches

**Biodegr. Hydrocarbon**

SMILES : n(cccc1C(N(CC2)C)C2)c1

CHEM : Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S)-

MOL FOR: C10 H14 N2

MOL WT : 162.24

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-- BioHCwin v1.01
Results \-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

NO Estimate Possible \... Structure NOT a Hydrocarbon

(Contains atoms other than C, H or S (-S-))

**Biodegradation BIOWIN**

SMILES : n(cccc1C(N(CC2)C)C2)c1

CHEM : Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S)-

MOL FOR: C10 H14 N2

MOL WT : 162.24

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-- BIOWIN v4.10
Results \-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

Biowin1 (Linear Model Prediction) : Does Not Biodegrade Fast

Biowin2 (Non-Linear Model Prediction): Does Not Biodegrade Fast

Biowin3 (Ultimate Biodegradation Timeframe): Weeks-Months

Biowin4 (Primary Biodegradation Timeframe): Days-Weeks

Biowin5 (MITI Linear Model Prediction) : Not Readily Degradable

Biowin6 (MITI Non-Linear Model Prediction): Not Readily Degradable

Biowin7 (Anaerobic Model Prediction): Does Not Biodegrade Fast

Ready Biodegradability Prediction: NO

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

TYPE \| NUM \| Biowin1 FRAGMENT DESCRIPTION \| COEFF \| VALUE

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

Frag \| 1 \| Pyridine ring \| -0.1546 \| -0.1546

Frag \| 1 \| Tertiary amine \| -0.2053 \| -0.2053

MolWt\| \* \| Molecular Weight Parameter \| \| -0.0772

Const\| \* \| Equation Constant \| \| 0.7475

============+============================================+=========+=========

RESULT \| Biowin1 (Linear Biodeg Probability) \| \| 0.3105

============+============================================+=========+=========

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

TYPE \| NUM \| Biowin2 FRAGMENT DESCRIPTION \| COEFF \| VALUE

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

Frag \| 1 \| Pyridine ring \| -1.6381 \| -1.6381

Frag \| 1 \| Tertiary amine \| -2.2229 \| -2.2229

MolWt\| \* \| Molecular Weight Parameter \| \| -2.3038

============+============================================+=========+=========

RESULT \| Biowin2 (Non-Linear Biodeg Probability) \| \| 0.0409

============+============================================+=========+=========

A Probability Greater Than or Equal to 0.5 indicates \--\> Biodegrades
Fast

A Probability Less Than 0.5 indicates \--\> Does NOT Biodegrade Fast

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

TYPE \| NUM \| Biowin3 FRAGMENT DESCRIPTION \| COEFF \| VALUE

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

Frag \| 1 \| Pyridine ring \| -0.2142 \| -0.2142

Frag \| 1 \| Tertiary amine \| -0.2548 \| -0.2548

MolWt\| \* \| Molecular Weight Parameter \| \| -0.3585

Const\| \* \| Equation Constant \| \| 3.1992

============+============================================+=========+=========

RESULT \| Biowin3 (Survey Model - Ultimate Biodeg) \| \| 2.3717

============+============================================+=========+========

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

TYPE \| NUM \| Biowin4 FRAGMENT DESCRIPTION \| COEFF \| VALUE

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

Frag \| 1 \| Pyridine ring \| -0.0187 \| -0.0187

Frag \| 1 \| Tertiary amine \| -0.2880 \| -0.2880

MolWt\| \* \| Molecular Weight Parameter \| \| -0.2341

Const\| \* \| Equation Constant \| \| 3.8477

============+============================================+=========+=========

RESULT \| Biowin4 (Survey Model - Primary Biodeg) \| \| 3.3069

============+============================================+=========+=========

Result Classification: 5.00 -\> hours 4.00 -\> days 3.00 -\> weeks

(Primary & Ultimate) 2.00 -\> months 1.00 -\> longer

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

TYPE \| NUM \| Biowin5 FRAGMENT DESCRIPTION \| COEFF \| VALUE

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

Frag \| 1 \| Pyridine ring \| -0.0335 \| -0.0335

Frag \| 1 \| Tertiary amine \| -0.0848 \| -0.0848

Frag \| 1 \| Aromatic-CH \| -0.0098 \| -0.0098

Frag \| 4 \| Aromatic-H \| 0.0082 \| 0.0329

Frag \| 1 \| Methyl \[-CH3\] \| 0.0004 \| 0.0004

Frag \| 3 \| -CH2- \[cyclic\] \| 0.0197 \| 0.0592

MolWt\| \* \| Molecular Weight Parameter \| \| -0.4827

Const\| \* \| Equation Constant \| \| 0.7121

============+============================================+=========+=========

RESULT \| Biowin5 (MITI Linear Biodeg Probability) \| \| 0.1939

============+============================================+=========+========

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

TYPE \| NUM \| Biowin6 FRAGMENT DESCRIPTION \| COEFF \| VALUE

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

Frag \| 1 \| Pyridine ring \| -0.4599 \| -0.4599

Frag \| 1 \| Tertiary amine \| -0.8396 \| -0.8396

Frag \| 1 \| Aromatic-CH \| 0.2624 \| 0.2624

Frag \| 4 \| Aromatic-H \| 0.1201 \| 0.4806

Frag \| 1 \| Methyl \[-CH3\] \| 0.0194 \| 0.0194

Frag \| 3 \| -CH2- \[cyclic\] \| 0.2365 \| 0.7096

MolWt\| \* \| Molecular Weight Parameter \| \| -4.6836

============+============================================+=========+=========

RESULT \|Biowin6 (MITI Non-Linear Biodeg Probability)\| \| 0.1207

============+============================================+=========+========

A Probability Greater Than or Equal to 0.5 indicates \--\> Readily
Degradable

A Probability Less Than 0.5 indicates \--\> NOT Readily Degradable

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

TYPE \| NUM \| Biowin7 FRAGMENT DESCRIPTION \| COEFF \| VALUE

\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

Frag \| 1 \| Pyridine ring \| 0.6411 \| 0.6411

Frag \| 1 \| Tertiary amine \| -1.0749 \| -1.0749

Frag \| 1 \| Aromatic-CH \| 0.0331 \| 0.0331

Frag \| 4 \| Aromatic-H \| -0.0954 \| -0.3817

Frag \| 1 \| Methyl \[-CH3\] \| -0.0796 \| -0.0796

Frag \| 3 \| -CH2- \[cyclic\] \| -0.1200 \| -0.3600

Const\| \* \| Equation Constant \| \| 0.8361

============+============================================+=========+=========

RESULT \| Biowin7 (Anaerobic Linear Biodeg Prob) \| \| -0.3860

============+============================================+=========+=========

A Probability Greater Than or Equal to 0.5 indicates \--\> Biodegrades
Fast

A Probability Less Than 0.5 indicates \--\> Does NOT Biodegrade Fast

**Octanol/Air Partitioning**

**Log Koa: 8.08**

SMILES : n(cccc1C(N(CC2)C)C2)c1

CHEM : Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S)-

MOL FOR: C10 H14 N2

MOL WT : 162.24

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-- KOAWIN v1.10
Results \-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

Log Koa (octanol/air) estimate: 8.081

Koa (octanol/air) estimate: 1.206e+008

Using:

Log Kow: 1.17 (exp database)

HenryLC: 3e-009 atm-m3/mole (HenryWin est)

**Log Kaw: -6.911 (air/water part.coef.)**

LogKow : 1.17 (exp database)

LogKow : 1.00 (KowWin estimate)

Henry LC: \-\-- atm-m3/mole(exp database)

Henry LC: 3e-009 atm-m3/mole (HenryWin bond estimate)

Log Koa (octanol/air) estimate: 7.911 (from KowWin/HenryWin)

**Boiling Point; Vapor Pressure; Melting Point**

Experimental Database Structure Match:

Name : NICOTINE

CAS Num : 000054-11-5

Exp MP (deg C): -79

Exp BP (deg C): 247

Exp VP (mm Hg): 3.80E-02 (extrapolated)

Exp VP (deg C): 25

Exp VP ref : BOUBLIK,T ET AL. (1984)

SMILES : n(cccc1C(N(CC2)C)C2)c1

CHEM : Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S)-

MOL FOR: C10 H14 N2

MOL WT : 162.24

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-- SUMMARY MPBPWIN v1.42
\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

Boiling Point: 247.51 deg C (Adapted Stein and Brown Method)

Melting Point: 110.31 deg C (Adapted Joback Method)

Melting Point: 30.86 deg C (Gold and Ogle Method)

Mean Melt Pt : 70.59 deg C (Joback; Gold,Ogle Methods)

Selected MP: 57.34 deg C (Weighted Value)

Vapor Pressure Estimations (25 deg C):

(Using BP: 247.00 deg C (exp database))

(MP not used for liquids)

VP: 0.0329 mm Hg (Antoine Method)

VP: 0.031 mm Hg (Modified Grain Method)

VP: 0.0546 mm Hg (Mackay Method)

Selected VP: 0.032 mm Hg (Mean of Antoine & Grain methods)

\-\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

TYPE \| NUM \| BOIL DESCRIPTION \| COEFF \| VALUE

\-\-\-\-\-\--+\-\-\-\--+\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\-\--+\-\-\-\-\-\-\-\--

Group \| 1 \| -CH3 \| 21.98 \| 21.98

Group \| 3 \| -CH2- (ring) \| 26.44 \| 79.32

Group \| 1 \| \>CH- (ring) \| 21.66 \| 21.66

Group \| 4 \| CH (aromatic) \| 28.53 \| 114.12

Group \| 1 \| -C (aromatic) \| 30.76 \| 30.76

Group \| 1 \| \>N- (ring) \| 32.77 \| 32.77

Group \| 1 \| N (aromatic) \| 39.88 \| 39.88

\* \| \| Equation Constant \| \| 198.18

=============+====================+==========+=========

RESULT-uncorr\| BOILING POINT in deg Kelvin \| 538.67

RESULT- corr \| BOILING POINT in deg Kelvin \| 520.67

**Water Sol: 1e+006 mg/L**

Experimental Water Solubility Database Match:

Name : NICOTINE

CAS Num : 000054-11-5

Exp WSol : 1E+006 mg/L ( deg C)

Exp Ref : SEIDELL,A (1941)

**LogKow**

SMILES : n(cccc1C(N(CC2)C)C2)c1

CHEM : Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S)-

MOL FOR: C10 H14 N2

MOL WT : 162.24

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--
WSKOW v1.41 Results \-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

Log Kow (estimated) : 1.00

Log Kow (experimental): 1.17

Cas No: 000054-11-5

Name : Nicotine

Refer : Hansch,C et al. (1995)

Log Kow used by Water solubility estimates: 1.17

Equation Used to Make Water Sol estimate:

Log S (mol/L) = 0.796 - 0.854 log Kow - 0.00728 MW + Correction

(used when Melting Point NOT available)

Correction(s): Value

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-- \-\-\-\--

Amine, aliphatic 1.008

Pyridine, alkyl 1.300

Log Water Solubility (in moles/L) : 0.924

Log Water Solubility (in moles/L) : 0.790 (Applied Upper Limit)

Water Solubility at 25 deg C (mg/L): 1e+006

**Koc (estimated): 2.38e+003**

Koc may be sensitive to pH!

SMILES : n(cccc1C(N(CC2)C)C2)c1

CHEM : Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S)-

MOL FOR: C10 H14 N2

MOL WT : 162.24

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-- PCKOCWIN v1.66
Results \-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

First Order Molecular Connectivity Index \...\...\..... : 5.877

Non-Corrected Log Koc \...\...\...\...\...\...\...\...\...\... : 3.7484

Fragment Correction(s):

3 Nitrogen to Carbon (aliphatic) (-N-C).. : -0.3726

Corrected Log Koc \...\...\...\...\...\...\...\...\...\...\.... : 3.3758

Estimated Koc: 2376

NOTE:

The Koc of this structure may be sensitive to pH! The estimated

Koc represents a best-fit to the majority of experimental values;

however, the Koc may vary significantly with pH.

**2.** **Kinetics of Volatilization of Nicotine from Soil**

> [Rate of Volatilization from Soil- Methodology]{.underline}

The Dow Method, as described in the "Handbook of Chemical Property
Estimation Methods"[^29] was used to estimate the first-order rate of
volatilization and half-life nicotine from the surface of soil. This
method was selected because all of the required information was
available.

> [Information Required]{.underline}:
>
> Koc= Soil adsorption coefficient,
>
> P~vp~ = Vapor Pressure, mmHg
>
> S= Solubility, in mg/L
>
> **Table 1- Estimated; EPISuite v3.20**

  ---------------------------------------------------- ------------------
  **Required Information**                             **Nicotine**

  Koc                                                  2376

  Vapor Pressure, mmHg, 25°C                           3.2 x 10^-2^

  Solubility in Water, mg/L, 25°C                      1 x 10^6^
  ---------------------------------------------------- ------------------

[Half-life Estimation]{.underline}

The following equation was used to estimate half-lives of volatilization
from soil surfaces using the Dow Method

> Half-life (t ~½~)= 1.58 x 10^-8^(K~oc~S)/P~vp~ days
>
> Half-life (t ~½~)= 1.58 x 10^-8^ \[2376 x (1 x 10^6^)\]/ 3.2 x 10^-2^,
> days
>
> (t ~½~)= 1.173 x 10^3^ days

[Rate Constant Estimation]{.underline}

The following equation was used to estimate rates of volatilization from
soil surfaces using the Dow Method

> k~v~= 0.693/ t ~½\ =~ day^-1^= 4.4 x 10^7^ (P~vp~/ K~oc~S)= 5.91 ^-4^
> day^-1^

For a Koc value of 2376, the volatilization t~1/2~ = 1.173 x 10^3^ days
corresponding to a rate constant of 0.693/1.173 x 10^3^ days = 5.91 x
10^-4^ day^-1^

For a Koc value of 3000, the volatilization t~1/2~ = (3000/2376) x
(1.173 x 10^3^ days) = 1.481 x 10^3^ days corresponding to a rate
constant of 0.693/1.481 x 10^3^ days = 4.68 x 10^-4^ day^-1^

For a Koc value of 500, the volatilization t~1/2~ = (500/2376) x (1.173
x 10^3^ days) = 247 days corresponding to a rate constant of 0.693/247
days = 2.81 x 10^-3^ day^-1^

**Table 2- Comparison of the Kinetics of Volatilization of Nicotine at
Different Values of Koc (25°C)**

  --------------------------------- ----------- ------------ ------------
  **Kinetics Information**          **K~oc~     **K~oc~      **K~oc~
                                    500**       2376**       3000**

  Volatilization Half-life, days    247         1.173 x      1.481 x
                                                10^3^        10^3^

  Volatilization Rate Constant,     2.81 x      5.91 x       4.68 x
  day^-1^                           10^-3^      10^-4^       10^-4^
  --------------------------------- ----------- ------------ ------------

**Note**: Nicotine is a weak acid. The Koc is expected to decrease with
increasing pH, as the ratio of the deprotonated to the nonprotonated
increases with pH The Koc estimated by EPISuite does not consider the pH
dependency of sorption. However, the Table shows the sensitivity of Koc
to the dissipation of nicotine by volatilization. In addition, nicotine
is expected to volatilize faster with increasing temperature.

**3.** **Comparison of Physical/Chemical Properties and Environmental
Fate of Nicotine and the Human Metabolite, Cotinine (EPISuite
estimates)**

+--------------+----------------------------+-------------------------+
| Physi        | Nicotine                   | Cotinine                |
| cal/Chemical |                            |                         |
| Properties   | *(S)*                      | (*S*)-1-Methyl-5-(3-pyr |
|              | -3                         | idinyl)-2-pyrrolidinone |
|              | -(1-Methyl-2-pyrrolidinyl) |                         |
|              | pyridine                   | 1-Methyl-5-(3-pyri      |
|              |                            | dinyl)-2-pyrrolidinone\ |
|              | *(S)*(-)                   | *S*-(-)-Cotinine        |
|              | -3                         |                         |
|              | -(1-Methyl-2-pyrrolidinyl) | CAS Reg.No. 486-56-6    |
|              | pyridine                   |                         |
|              |                            |                         |
|              | CAS Reg.No. 54-11-5        |                         |
+--------------+----------------------------+-------------------------+
| Chemical     | ![](media/image2.png){w    | ![](                    |
| Structure    | idth="2.438888888888889in" | media/image3.png){width |
| and          | heig                       | ="2.0819444444444444in" |
| Molecular    | ht="2.0215277777777776in"} | height=                 |
| Weight       |                            | "1.0527777777777778in"} |
|              | 162,2 g/mol                |                         |
|              |                            | 176.22 g/mol            |
+--------------+----------------------------+-------------------------+
| Melting      | -7,9 ° C degree C          | 41 degree C             |
| Point        |                            |                         |
|              | 247 degree C               | 324 degree C            |
| Boiling      |                            |                         |
| Point        | 3.2 x 10^-2^               | 3.8 x 10^-4^            |
|              |                            |                         |
| Vapor        | 3 x 10^-9^                 | 3.3 x 10^-12^           |
| Pressure     |                            |                         |
| mmHg at 25C  |                            |                         |
|              |                            |                         |
| Henry's Law  |                            |                         |
| C            |                            |                         |
| onstant(atm- |                            |                         |
| m3 /mole) at |                            |                         |
| 25C          |                            |                         |
+--------------+----------------------------+-------------------------+
| Solubility   | 1 x 10^6^                  | 5.6 x 10^5^             |
| in water     |                            |                         |
+--------------+----------------------------+-------------------------+
| Log K~ow~    | 0.9                        | 0.34                    |
+--------------+----------------------------+-------------------------+
| Koc          | 2376                       | 808                     |
+--------------+----------------------------+-------------------------+
| Hydrolysis   | \> 1yr                     | \> 1 yr                 |
+--------------+----------------------------+-------------------------+
| Biotr        | Primary and ultimate time  | Primary time frame,     |
| ansformation | frame weeks to months      | Days-to-Weeks           |
|              |                            | (half-life 8.6 days)    |
|              | Half-life 37.5             |                         |
|              |                            | Ultimate time frame     |
|              |                            | weeks to months         |
|              |                            | (half-life 37.5 days)   |
+--------------+----------------------------+-------------------------+
| Air-water    | -6.9                       | -9.8                    |
| partitioning |                            |                         |
| (Log k~aw)~  |                            |                         |
+--------------+----------------------------+-------------------------+
| Photo (atm), | 1.411 hour                 | 4.9 hrs                 |
| half-life    |                            |                         |
+--------------+----------------------------+-------------------------+

**[Environmental Fate and Exposure References]{.underline}**

Baitsch, D., Sandu, Brandsch, R., and Igloi, G.L. 2001. Gene Cluster on
pAO1 Of *Arthobacter nicotinovorans* involved in Degradation of the
Plant Alkaloid Nicotine: Cloning, Purification, and Characterization of
2,6-Dihydroxypyridine 3-Hydroxylase. Journal of Bacteriology. Sept.
2001, pp.5262-5267.

Bradley, P.M., Barber, L.B., Kolpin, D.W., McMahon, P.B., and Chapell,
F.H. 2007. Biotransformation of Caffeine, Cotinine, and nicotine in
stream sediments: Implications for use as wastewater indicators.
Environmental Toxicology and Chemistry, Vol. 26 (6), pp

Carter, J.M., Delzer, G.C., Kingsbury, J.A., and Hopple, J.A. 2007.
Concentration data for anthropogenic organic compounds in ground water,
surface water, and finished water of selected community water systems in
the United States, 2002-2005: U.S. Geological Survey Data Series 268 ,30
pp.

Chiribau, C.B., Sandu, C., Fraaije, M., Schiltz, E., and Brandsch. 2004.
A novel γ-N-methylaminobutyrate demethylating oxidase involved in
catabolism of the tobacco alkaloid nitotine by *Anthrobacter
nicotinovorans* pAO1. Eur. J. Biochem,Vol 271, pp 4677-4684.

Chiribau, C.B., Mihasan, M. Ganas., P., Igloi, G.L., Artenie, V., and
Brandasch, R. 2006. Final Steps in the catabolism of nicotine-
Deamination versus demethylation of γ-N-methylaminobutyrate. FEBS
Journal, Vol. 273, pp. 1528-1536.

Hochstein, L.I. and Rittenberg, S.C 1959. The Bacterial Oxidation of
Nicotine- I. Nicotine oxidation by cell-free preparations. The Journal
of Biological Chemistry, Vol 234(1), pp 151-155.

Hochstein, L.I. and Rittenberg, E.C. 1959. The Bacterial Oxidation of
Nicotine- II. The isolation of the first oxidative product and its
identification as (1)-6-hydroxynicotine. The Journal of Biological
Chemistry, Vol 234(1), pp 156-160.

Kolpin, D.W., Furlong, E.T., Meyer, M.T. Thurman, E.M., Zaugg, S.D.,
Barber, L.B., Buxton, H.T. 2002. Pharmaceuticals, Hormones, and Other
Organic Wastewater Contaminants in U.S. Streams, 1999-2000: a National
Reconnaissance . Environ. Sci. Technol. Vol. 36, pp 1202-1211.

Lam, M.W., Young, C.J., Brain, R.A., Johnson, D.J, Hanson, M.A., Wilson,
C.J., Richards, S.M.,, Solomon, K.R., and Mabury, S.A. 2004. Env. Tox.
Chem. Vol 23(6), pp .

Lee, K.E., Barber, L.B., Furlong, E.T., Cahill, J.D., Kolpin, D.W.,
Meyer, M.T., and Zaugg, S.D. 2004. Presence and distribution of organic
wastewater compounds in wastewater, surface, ground, and drinking
waters, Minnesota, 2000-2002: U.S.Geological Survey Scientific
Investigation Report 2004-5138, 47pp.

Mihasan M., Chiribau, C-B., Friedrich, T., Artenie, V., and Brandsch, R.
2007.. An NAD(P)H-nicotine blue oxidoreductase is part of the nicotine
regulon and may protect *Arthrobacter nicotinovorans* from oxidative
stress during nicotine catabolism Applied and Environmental
Microbiology. Vol. 73 (April), pp.2479-2485.

Peterson, E.J., Choi, A., Dahan, D.S., Lester, H.A., and Dougherty, D.A.
2002. J. Am. Chem. Soc. A Perturbated pKa at the Binding Site of the
Nicotinic Acetylcholine Receptor: Implications for Nicotine Binding,
Vol. 124, pp.12662-12663.

Seeman, J.I. 2005. Using "Basic Principles" to Understand Complex
Science: Nicotine Smoke Chemistry and Literature Analogies. Journal of
Chemical Education, Vol. 82 (10), pp 1577-1583

Sangster, A.W. and Stuart, K.L. 1965. Ultraviolet Spectra of Alkaloids.
Chen. Rev., Vol 29, pp.69-130.

[]{#__RefHeading___Toc173552932 .anchor}**APPENDIX B. Aquatic Exposure
Model (GENEEC2) Output**

**Nicotine 2-inch Bandwidth**

RUN No. 12 FOR Nicotine ON Ornamental \* INPUT VALUES \*

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

RATE (#/AC) No.APPS & SOIL SOLUBIL APPL TYPE NO-SPRAY INCORP

ONE(MULT) INTERVAL Kd (PPM ) (%DRIFT) ZONE(FT) (IN)

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

.044( .066) 6 60 2376.0\*\*\*\*\*\*\* GRANUL( .0) .0 .0

FIELD AND STANDARD POND HALFLIFE VALUES (DAYS)

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

METABOLIC DAYS UNTIL HYDROLYSIS PHOTOLYSIS METABOLIC COMBINED

(FIELD) RAIN/RUNOFF (POND) (POND-EFF) (POND) (POND)

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

37.50 2 N/A .00- .00 75.00 75.00

GENERIC EECs (IN NANOGRAMS/LITER (PPTr)) Version 2.0 Aug 1, 2001

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

PEAK MAX 4 DAY MAX 21 DAY MAX 60 DAY MAX 90 DAY

GEEC AVG GEEC AVG GEEC AVG GEEC AVG GEEC

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

162.98 108.38 29.03 10.28 6.90

**Nicotine 6-inch Bandwidth**

RUN No. 16 FOR Nicotine 6in ON Ornamental \* INPUT VALUES \*

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

RATE (#/AC) No.APPS & SOIL SOLUBIL APPL TYPE NO-SPRAY INCORP

ONE(MULT) INTERVAL Kd (PPM ) (%DRIFT) ZONE(FT) (IN)

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

.132( .724) 6 2 2376.0\*\*\*\*\*\*\* GRANUL( .0) .0 .0

FIELD AND STANDARD POND HALFLIFE VALUES (DAYS)

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

METABOLIC DAYS UNTIL HYDROLYSIS PHOTOLYSIS METABOLIC COMBINED

(FIELD) RAIN/RUNOFF (POND) (POND-EFF) (POND) (POND)

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

37.50 2 N/A .00- .00 75.00 75.00

GENERIC EECs (IN MICROGRAMS/LITER (PPB)) Version 2.0 Aug 1, 2001

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

PEAK MAX 4 DAY MAX 21 DAY MAX 60 DAY MAX 90 DAY

GEEC AVG GEEC AVG GEEC AVG GEEC AVG GEEC

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

1.80 1.20 .32 .11 .08

**Nicotine 12-inch Bandwidth**

RUN No. 112 FOR Nicotine ON Ornamental \* INPUT VALUES \*

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

RATE (#/AC) No.APPS & SOIL SOLUBIL APPL TYPE NO-SPRAY INCORP

ONE(MULT) INTERVAL Kd (PPM ) (%DRIFT) ZONE(FT) (IN)

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

.264( .393) 6 60 2376.0\*\*\*\*\*\*\* GRANUL( .0) .0 .0

FIELD AND STANDARD POND HALFLIFE VALUES (DAYS)

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

METABOLIC DAYS UNTIL HYDROLYSIS PHOTOLYSIS METABOLIC COMBINED

(FIELD) RAIN/RUNOFF (POND) (POND-EFF) (POND) (POND)

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

37.50 2 N/A .00- .00 75.00 75.00

GENERIC EECs (IN NANOGRAMS/LITER (PPTr)) Version 2.0 Aug 1, 2001

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

PEAK MAX 4 DAY MAX 21 DAY MAX 60 DAY MAX 90 DAY

GEEC AVG GEEC AVG GEEC AVG GEEC AVG GEEC

\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--

977.87 650.29 174.17 61.69 41.40

[]{#__RefHeading___Toc173552933 .anchor}**APPENDIX C. ECOTOX Open
Literature Study Summaries and Bibliography**

***[Freshwater Animals]{.underline}***

**Chemical Name:** Nicotine

**CAS No:** 54-11-5

**ECOTOX Record Number and Citation:** Edsall, C. C. (1991). Acute
Toxicities to Larval Rainbow Trout of Representative Compounds Detected
in Great Lakes Fish. *Bull.Environ.Contam.Toxicol.* 46: 173-178.
EcoReference No.: 138

**Purpose of Review (DP Barcode or Litigation):** D341224

**Date of Review:** June 2007

**Summary of Study Findings:**

The acute 96-hour toxicity of nicotine to larval rainbow trout was
assessed in a static study. Fry 13-21 days old (post-hatch), in the
yolk-sac stage just before swim-up, were used in the toxicity tests.
Test concentrations ranged from 1.0 to 10.0 mg/L. The purity of the test
compound was at least 95%. Nicotine was dissolved in deionized water.
Temperature, dissolved oxygen, and mortality were monitored and recorded
every 24 hours for 96 hours. Complete mortality was observed in the 5 to
10 mg/L nicotine treatments; there were no mortalities in the 1.0 and
2.7 mg/L treatment groups. That is, there were no partial kills over the
range of concentrations tested. The approximate LC~50~ is 4.0 mg/L. The
live fry showed no signs of stress.

This study was performed by the U.S. Fish and Wildlife Service.

**Description of Use in Document (QUAL, QUAN, INV):** Qualitative

**Rationale for Use:** The LC50 of 4.0 mg a.i./L cannot be used to
calculate risk quotients because there is uncertainty in this endpoint
since the exposures were not analytically verified. However, this study
will used to qualitatively describe the potential acute risks of
nicotine to freshwater fish.

**Limitations of Study:**

1\. There is uncertainty in the calculated acute 96-hour LC50 since
there were no partial kills.

2\. Concentrations were not analytically verified; thus, actual exposure
could have been considerably lower than the nominal treatments suggest.

**Primary Reviewer:** Colleen Flaherty, Biologist (ERB3)

**\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--**

**Chemical Name:** Nicotine

**CAS No:** 54-11-5

**ECOTOX Record Number and Citation:** Perry, C. M. and Smith, S. B.
(1988). Toxicity of Six Heterocyclic Nitrogen Compounds to *Daphnia
pulex*. *Bull.Environ.Contam.Toxicol.41(4):604-608 / In: Prog.Abstr.29th
Conf.Int.Assoc.Great Lakes Res., May 26-29, 1986, Searborough, Ont.,
Canada* 604-608. EcoReference No.: 13161**\
**

**Purpose of Review (DP Barcode or Litigation):** D341224

**Date of Review:** June 2007

**Summary of Study Findings:**

The acute 48-hour toxicity of nicotine to *Daphnia pulex* was assessed
in a static study. Culturing and bioassays methods used were ASTM
(1980), U.S. EPA (1975), and the contaminant toxicology project at the
National Fisheries Research Center -- Great Lakes. The purity of the
test compound was at least 97%. Nicotine was dissolved in water.

Toxicity tests were conducted for 48 hours with 10 neonates (\<24 hours
old) in five treatment concentrations plus a control. Each treatment was
at least 50% of the next highest treatment. Test animals were not fed
during the experiment. After 48 hours, immobilization was assessed. The
reported EC50 was 0.242 (standard error = 0.02) mg/L.

**Description of Use in Document (QUAL, QUAN, INV):** Qualitative

**Rationale for Use:** The EC50 of 0.242 mg a.i./L cannot be used to
calculate risk quotients because there is uncertainty in this endpoint
since the exposures were not analytically verified. However, this study
will used to qualitatively describe the potential acute risks of
nicotine to freshwater invertebrates.

**Limitations of Study:**

1\. Raw data for mortality (immobilization) were not provided.

2\. Concentrations were not analytically verified; thus, actual exposure
could have been considerably lower than the nominal treatments suggest.

3\. The range of treatment concentrations was not described.

**Primary Reviewer:** Colleen Flaherty, Biologist (ERB3)

**\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--**

**Chemical Name:** Nicotine

**CAS No:** 54-11-5

**ECOTOX Record Number and Citation:** Savino, J. F. and Tanabe, L. L.
(1989). Sublethal Effects of Phenanthrene, Nicotine, and Pinane on
*Daphnia pulex*. *Bull.Environ.Contam.Toxicol.* 42: 778-784.
EcoReference No.: 390**\
**

**Purpose of Review (DP Barcode or Litigation):** D341224

**Date of Review:** June 2007

**Summary of Study Findings:**

A static-renewal, 16-day chronic toxicity test was conducted to assess
the effects of nicotine on *Daphnia pulex* growth and reproduction.
Culturing and bioassays methods used were ASTM (1980) and U.S. EPA
(1982).

Test nicotine concentrations were 0 (control), 0.02, 0.07, 0.12, 0.18,
and 0.24 mg/L. The purity of the test compound was at least 97%.
Nicotine was dissolved in water. Treatments were renewed three times a
week during the 16-day study. Test concentrations were analytically
verified at 1 hour, 48 hours, and 72 hours after preparation to simulate
exposure in test media at the beginning and end of the renewal cycle.

The recovery rates for nicotine from gas chromatography analysis are
presented in the table below. At 48 and 72 hours after preparation, the
concentrations had dropped substantially (\< 25% nominal), possibly due
to adsorption to glassware, volatility, and uptake by the daphnids.

  -------------- ---------- --------- --------- ---------- --------- ---------
  **Chemical**   **%                            **%                  
                 Recovery                       Recovery             
                 in Water                       in Test              
                 Only**                         Media**              

                 **1 hr**   **48      **72      **1 hr**   **48      **72
                            hrs**     hrs**                hrs**     hrs**

  Nicotine       57         24        9         89         3         0
  -------------- ---------- --------- --------- ---------- --------- ---------

Mortality effects are summarized in the table below. The study author
claims that nicotine significantly reduced growth and fecundity of
daphnids at nominal concentrations from 0.02 -- 0.24 mg/L. The LOAEC for
length was 0.07 mg/L, and the LOAEC for fecundity was 0.18 mg/L. It is
unclear if the NOAEC was 0.02 mg/L or \<0.02 mg/L.

  --------------------------- --------------------- ---------------------
  **Nominal Concentration     **Mean Mortality      **Range Mortality
  (mg/L)**                    (%)**                 (%)**

  0                           10                    0-20

  0.02                        6                     0-13

  0.07                        4                     0-7

  0.12                        10                    0-20

  0.18                        20                    13-37

  0.24                        66                    33-100
  --------------------------- --------------------- ---------------------

This study was performed by the U.S. Fish and Wildlife Service.

**Description of Use in Document (QUAL, QUAN, INV):** Qualitative

**Rationale for Use:** Given the poor recovery of the test substance,
the exposures are very uncertain. Since these are nominal concentrations
and nicotine was undetectable 72 hours after dosing, the actual NOAEC is
likely to be considerably less than 0.02 mg/L. Risk quotients cannot be
calculated using these data; however, this study will be used
qualitatively to characterize the potential chronic risk of nicotine to
freshwater invertebrates.

**Limitations of Study:**

1\. The recovery of the test compound was very poor, and consistent
exposures were not maintained.

2\. The study author claims that nicotine significantly reduced growth
and fecundity of daphnids at nominal concentrations from 0.02 -- 0.24
mg/L. The LOAEC for length was 0.07 mg/L, and the LOAEC for fecundity
was 0.18 mg/L. It is unclear if the NOAEC was 0.02 mg/L or if it was not
determined (*i.e.,* \< 0.02 mg/L).

3\. The study duration was only 16 days instead of 21 days as required
in the daphnid lifecycle study (Guideline 72-4(b)).

**Primary Reviewer:** Colleen Flaherty, Biologist (ERB3)

**\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--**

**Chemical Name:** Nicotine

**CAS No:** 54-11-5

**ECOTOX Record Number and Citation:** Passino-Reader, D. R., Berlin, W.
H., and Hickey, J. P. (1995). Chronic Bioassays of Rainbow Trout Fry
with Compounds Representative of Contaminants in Great Lakes Fish.
*J.Gt.Lakes Res.* 21: 373-383. EcoReference No.: 16362**\
**

**Purpose of Review (DP Barcode or Litigation):** D341224

**Date of Review:** June 2007

**Summary of Study Findings:**

Sixty-day bioassays on the effects of nicotine on the survival, growth,
and behavior of rainbow trout fry were conducted in a constant-flow,
temperature-controlled water system. The standard procedures of ASTM
(1988) and USEPA (1986) were followed. Two bioassays with nicotine were
conducted with rainbow trout fry from time of hatch until past swim-up,
a period of 60 days, in a gravity/constant flow-through,
temperature-controlled water system. The purity of the nicotine test
substance was \>98%; stock solutions were made with deionized, reverse
osmosis water. Treatment concentrations for nicotine were not
analytically verified during this study; the study author described the
difficulty of maintaining nominal concentrations of nicotine in large
test systems and referred to the chemical analyses of Savino and Tanabe
(1989; Ecotox ref 390).

Nicotine concentrations in a geometric progression from 0.06 to 1.0 mg/L
resulted in no significant effects on survivorship, weight, or length.
At nicotine concentrations of 1.4 to 6.0 mg/L, complete mortality
occurred at 6 mg/L, and the estimated 60-day LC50 was 5 mg/L. The median
lethal time (LT50) for fry exposed to 6.0 mg/L nicotine was 22.0 and
19.8 days in replicate tanks. Length and weight decreased linearly with
increasing concentration in the range of 1.4 to 4.2 mg/L. The NOAEC and
LOAEC for length and weight were 2.9 and 4.2 mg/L, respectively.
However, these endpoints are very uncertain, given the uncertainty in
the exposure concentrations.

**Description of Use in Document (QUAL, QUAN, INV):** Qualitative

**Rationale for Use:** The actual exposures in this bioassay are very
uncertain. In a similar study (Eco ref. 390), nicotine was undetectable
72 hours after dosing. Given the uncertainty in the effects endpoints
determined in this study, risk quotients cannot be calculated using
these data. However, this study will be used qualitatively to
characterize the potential chronic risk of nicotine to freshwater fish.

**Limitations of Study:**

1\. Exposures are uncertain; see summary for ecotox ref 390.

2\. Reproductive endpoints were not assessed.

**Primary Reviewer:** Colleen Flaherty, Biologist (ERB3)

**\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--**

***[Estuarine/Marine Animals]{.underline}***

**Chemical Name:** Nicotine

**CAS No:** 54-11-5

**ECOTOX Record Number and Citation:** Borlongan, I. G., Coloso, R. M.,
Mosura, E. F., Sagisi, F. D., and Mosura, A. T. (1998 ). Molluscicidal
Activity of Tobacco Dust Against Brackishwater Pond Snails (*Cerithidea
cingulata* Gmelin). *Crop Prot.* 17: 401-404. EcoReference No.: 83544

**Purpose of Review (DP Barcode or Litigation):** D341224

**Date of Review:** June 2007

**Summary of Study Findings:**

The acute toxicity of nicotine (tobacco dust) to brackishwater pond
snails (*Cerithidea cingulata* Gmelin) was assessed under controlled
laboratory conditions. The test substance contained 2.8% nicotine. The
experiments were conducted in circular basins lined with soil and filled
with brackishwater. There were nine treatments and four replicates for
each stage or size range. Mortality was determined after 24, 48, and 72
h after exposure.

The 72-hour LC~50~ for the juveniles, sub-adult, and adult snails were
30, 87, and 166 kg/ha (0.75, 2.17, and 4.15 lbs a.i./A), respectively.

**Description of Use in Document (QUAL, QUAN, INV):** Qualitative

**Rationale for Use:** Treatments were described in terms of application
rate and were not analytically verified; thus, the actual exposure
concentrations are unknown. Risk quotients cannot be calculated with the
data provided in this study.

**Limitations of Study:**

1\. Exposures were not analytically determined.

2\. Test organisms were collected from the field; thus, previous
exposure is unknown.

**Primary Reviewer:** Colleen Flaherty, Biologist (ERB3)

**\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--**

***[Amphibians]{.underline}***

**Chemical Name:** Nicotine

**CAS No:** 54-11-5

**ECOTOX Record Number and Citation:** Dawson, D. A., Fort, D. J.,
Smith, G. J., Newell, D. L., and Bantle, J. A. (1988). Evaluation of the
Developmental Toxicity of Nicotine and Cotinine with Frog Embryo
Teratogenesis Assay: Xenopus. *Teratog.Carcinog.Mutagen.* 8: 329-338.
EcoReference No.: 83889

**Purpose of Review (DP Barcode or Litigation):** D341224

**Date of Review:** June 2007

**Summary of Study Findings:**

Early embryos of *Xenopus laevis* were exposed for 96 hours to nicotine
or a primary metabolite, cotinine, in two separate static renewal tests
with and without addition of the metabolic activation system (MAS). This
summary will only discuss the tests without the MAS.

Groups of 20 embryos were placed in plastic Petri dishes containing
graduated concentrations of nicotine or cotinine dissolved in FETAX
solution. For each compound, 11-24 concentrations were tested with two
dishes per concentration. Control embryos were placed in FETAX solution.
Treatment and control dishes contained a total of 8 ml of solution,
including 100 U/ml of penicillin and 0.1 mg/ml streptomycin to control
bacterial growth. Test solutions were refreshed at 24, 48, and 72 hours.

The average LC~50~ (of two tests) for nicotine and cotinine were 136 and
4340 mg/L, respectively. Several malformations were reported for
nicotine-exposed frogs. Nicotine exposure beginning at 0.25 mg/L induced
contorted posture (lateral body flexure) and incomplete development of
the underside of the mouth. Gill hyperplasia was also noted. All embryos
were malformed above 0.8 mg/L nicotine. At 1.0 mg/L, skeletal kinking
was observed along with incomplete mouth development. Above 90 mg/L the
head and brain were reduced in size and the mouth was poorly developed,
if present at all. The gut was poorly coiled and the heart swollen, and
generalized pericardial and fin edema were noted. The eyes were reduced
in size and incompletely developed at levels higher than 110 mg/L
nicotine. The average EC~50~ for these malformations was 0.45 mg/L
nicotine.

Malformations were also noted in the cotinine-exposed frogs. Frogs
exposed to 600 mg/L or more of cotinine displayed skeletal kinking and
improper gut coiling. All embryos were malformed at \>1000 mg/L. At
levels above 1500 mg/L cotinine, the brain was reduced in size, the
mouth was incompletely formed, the eyes were abnormally shaped, and
pericardial and fin edema were observed. The heart was a straight tube
at levels \>3000 mg/kg. The average EC~50~ for these effects was 720
mg/L cotinine.

**Description of Use in Document (QUAL, QUAN, INV):** Qualitative

**Rationale for Use:** This study is useful to qualitatively
characterize the potential effects of nicotine to frogs; however, test
results from this study cannot be used to calculate risk quotients.

**Limitations of Study:**

1\. Exposure concentrations were not analytically verified; thus, there
is uncertainty regarding the actual exposures of the test organisms.

2\. Nicotine was dissolved in FETAX solution, which contained other
chemicals (*i.e.,* antibiotics).

3\. Test substance purity was not reported.

4\. There were no negative controls (without antibiotics added to
solution).

**Primary Reviewer:** Colleen Flaherty, Biologist (ERB3)

**\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--**

***[Mammals]{.underline}***

**Chemical Name:** Nicotine

**CAS No:** 54-11-5

**ECOTOX Record Number and Citation:** Ajarem, J. S. and Ahmad, M.
(1998). Prenatal Nicotine Exposure Modifies Behavior of Mice Through
Early Development. *Pharmacol.Biochem.Behav.* 59: 313-318. EcoReference
No.: 84721

**Purpose of Review (DP Barcode or Litigation):** D341224

**Date of Review:** June 2007

**Summary of Study Findings:**

This study investigated the effects of prenatal nicotine exposure on
development and behavior of mice. Pregnant dams were given daily
subcutaneous injections of normal saline (control) or 0.5 mg/kg bw
nicotine dissolved in normal saline for 9-10 days. Nicotine treatment
significantly reduced postnatal body weight gain, and delayed eye
opening, the appearance of body hairs, and sensory motor reflexes.
However, motor activity was stimulated in early adulthood of pups
prenatally exposed to nicotine.

**Description of Use in Document (QUAL, QUAN, INV):** Qualitative

**Rationale for Use:** Since this is not an oral or dietary exposure
test (i.e., exposure is via injection), these data cannot be used
quantitatively to estimate risk in this ecological risk assessment.
However, this study does suggest that nicotine can affect the normal
life processes of mammals and can be used to qualitatively characterize
the potential risk.

**Limitations of Study:**

1\. Exposures were administered via injections.

**Primary Reviewer:** Colleen Flaherty, Biologist (ERB3)

**\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--**

**Chemical Name:** Nicotine

**CAS No:** 54-11-5

**ECOTOX Record Number and Citation:** Johns, J. M., Louis, T. M.,
Becker, R. F., and Means, L. W. (1982). Behavioral Effects of Prenatal
Exposure to Nicotine in Guinea Pigs. *Neurobehav.Toxcol.Teratol.* 4:
365-369. EcoReference No.: 84597

**Purpose of Review (DP Barcode or Litigation):** D341224

**Date of Review:** June 2007

**Summary of Study Findings:**

The offspring of nicotine-treated and control guinea pig dams were
compared on spontaneous alternation and response to a novel alley as
neonates, and spontaneous alternation and a black-white discrimination
problem and reversal as adults. These tasks were selected because they
have been previously shown to be sensitive indicators of brain damage.

Thirty guinea pigs served as subjects: 15 were the offspring of 6 dams
that were injected twice daily throughout pregnancy with 3 mg/kg (SC)
nicotine suspended in saline, and 15 were the offspring of 5 dams
injected twice daily with saline. All offspring were tested for
spontaneous alternation both as neonates beginning at 10 days of age,
and as adults, beginning at 60 days of age. All animals were tested for
response to a novel alley at age 32 days, and 20 offspring (10 treated
and 10 control) were tested on black-white discrimination and reversal
beginning at age 85 days.

The guinea pigs exposed to nicotine prenatally demonstrated severe
behavioral impairments. Treatment offspring alternated at chance or
below chance levels while the controls alternated at normal levels. Most
of the nicotine-treated guinea pigs failed to enter the novel alley
while 80% of the control animals entered. Treated animals were severely
impaired compared to the control subjects on the reversal and
discrimination problems.

**Description of Use in Document (QUAL, QUAN, INV):** Qualitative

**Rationale for Use:** Since this is not an oral or dietary exposure
test (i.e., exposure is via injection), these data cannot be used
quantitatively to estimate risk in this ecological risk assessment.
However, this study does suggest that nicotine can affect the normal
life processes of mammals and can be used to qualitatively characterize
the potential risk.

**Limitations of Study:**

1\. Exposures were administered via injections.

**Primary Reviewer:** Colleen Flaherty, Biologist (ERB3)

**\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--**

**Chemical Name:** Nicotine

**CAS No:** 54-11-5

**ECOTOX Record Number and Citation:** Kita, T., Nakashima, T., Shirase,
M., Asahina, M., and Kurogochi, Y. (1988). Effects of Nicotine on
Ambulatory Activity in Mice. *Jpn.J.Pharmacol.* 46: 141-146.
EcoReference No.: 84604

**Purpose of Review (DP Barcode or Litigation):** D341224

**Date of Review:** June 2007

**Summary of Study Findings:**

Seven-week-old mice were injected with nicotine (0.1, 0.5, and 1.0
mg/kg) or a control solution (saline) and then placed into an
ambulo-cage. Activity counts were recorded during a 180-minute period.
In the saline control group, activity was the highest in the first
period (20 minutes) and gradually decreased thereafter. Mice treated
with the highest nicotine dose (1.0 mg/kg) demonstrated significantly
decreased ambulatory activity in a dose-dependent manner from 5 to 60
minutes after the administration. Mice dosed with 0.5 mg/kg nicotine
showed depressed activity for 40 minutes after administration. On the
contrary, the lowest nicotine dose, 0.1 mg/kg, stimulated activity in
the first 20 minutes. Further analysis of the data revealed that all
nicotine treatments resulted in an initial increase in activity, and
then the ataxic phase developed.

**Description of Use in Document (QUAL, QUAN, INV):** Qualitative

**Rationale for Use:** Since this is not an oral or dietary exposure
test (i.e., exposure is via injection), these data cannot be used
quantitatively to estimate risk in this ecological risk assessment.
However, this study does suggest that nicotine can affect the normal
life processes of mammals and can be used to qualitatively characterize
the potential risk.

**Limitations of Study:**

1\. Exposures were administered via injections.

**Primary Reviewer:** Colleen Flaherty, Biologist (ERB3)

**\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--**

**Chemical Name:** Nicotine

**CAS No:** 54-11-5

**ECOTOX Record Number and Citation:** Romero, R. D. and Chen, W. J. A.
(2004). Gender-Related Response in Open-Field Activity Following
Developmental Nicotine Exposure in Rats. *Pharmacol.Biochem.Behav.* 78:
675-681. EcoReference No.: 84725

**Purpose of Review (DP Barcode or Litigation):** D341224

**Date of Review:** June 2007

**Summary of Study Findings:**

This study examined the effects of developmental nicotine exposure on
rat offspring somatic growth and behavioral performance in an open-field
test. Female rats were implanted with nicotine (35 mg for 21-day
release) or placebo pellets on gestational day 8. A normal control group
with no pellet implant was included in the study design. There was no
significant difference in offspring body weight across the treatments.
The amount of activity, measured by the total number of crossings in the
open-field test, revealed less activity in male offspring and an
increase in female offspring activity as a function of testing day. The
increase in female ambulatory activity was observed in the placebo and
normal control, but not in the nicotine treatment group. This suggests
that the control subjects adapted to the testing apparatus and were less
anxious or afraid whereas the nicotine-exposed subjects failed to adapt
to the test system, learn the contextual cues, or retrieve the learned
information.

**Description of Use in Document (QUAL, QUAN, INV):** Qualitative

**Rationale for Use:** Since this is not an oral or dietary exposure
test (i.e., exposure is via an implanted nicotine pellet), these data
cannot be used quantitatively to estimate risk in this ecological risk
assessment. However, this study does suggest that nicotine can affect
the normal life processes of mammals and can be used to qualitatively
characterize the potential risk.

**Limitations of Study:**

1\. Exposures were administered via an implanted nicotine pellet.

**Primary Reviewer:** Colleen Flaherty, Biologist (ERB3)

**\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\-\--**

**[Mammalian Toxicity References (from the Health Effects
Division)]{.underline}**

Fung YK. 1988. Postnatal behavioural effects of maternal nicotine
exposure in rats. J Pharm Pharmacol. 40(12):870-2.

Johns JM; Walters PA; Zimmerman LI . 1993. The effects of chronic
prenatal exposure to nicotine on the behavior of guinea pigs (Cavia
porcellus). J Gen Psychol; 120(1):49-63.

Peters MA; Ngan LL. 1982. The effects of totigestational exposure to
nicotine on pre- and postnatal development in the rat. Arch Int
Pharmacodyn Ther; 257(1):155-67.

Peters DA; Taub H; Tang S. 1979. Postnatal effects of maternal nicotine
exposure.

Neurobehav Texaco; 1(3):221-5.

Saad AY; Gartner LP; Hiatt JL. 1990. Teratogenic effects of nicotine on
palate formation in mice. Biol Struct Morphog; 3(1):31-5.

Saad AY; Gartner LP; Hiatt JL. 1991. Teratogenic effects of nicotine on
first molar odontogenesis in the mouse. Acta Morphol Hung, 39(2):87-96.

Temocin S; Erenmemisoglu A; Suer C; Beydagi H. 1993. Effect of nicotine
on swimming exercise in rats. Jpn J Physiol; 43(4):567-70.

Williams CM; Kanagasabai T. 1984. Maternal adipose tissue response to
nicotine administration in the pregnant rat: effects on fetal body fat
and cellularity. Br J Nutr; 51(1):7-13.

Witschi H; Lundgaard SM; Rajini P; Hendrickx AG; Last JA. 1994. Effects
of exposure to nicotine and to sidestream smoke on pregnancy outcome in
rats. Toxicol Lett 71(3):279-86.

NICOTINE

Papers that Were Accepted for ECOTOX

**[NOTE]{.underline}**: The following studies were not reviewed for this
assessment because the effects described in the study cannot be
quantitatively linked to an assessment endpoint.

**Acceptable for ECOTOX and OPP**

Adler, I. D. and Attia, S. M. (2003). Nicotine is not Clastogenic at
Doses of 1 or 2 mg/kg Body Weight Given Orally to Male Mice .
*Mutat.Res.* 542: 139-142.

EcoReference No.: 84389\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Amano, H., Goshima, Y., Akema, N., Ueda, H., Kubo, T., and Misu, Y.
(1989). Effects of Acute Nicotine on Catecholamine Turnover in Various
Rat Brain Regions. *J.Pharmacobio.-Dyn.* 12: 18-23.

EcoReference No.: 84601\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Ashakumary, L. and Vijayammal, P. L. (1996). Effect of Nicotine on
Antioxidant Defence Mechanisms in Rats Fed a High-Fat Diet .
*Pharmacology* 52: 153-158.

EcoReference No.: 84594\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Attaway, C. M., Compton, D. M., and Turner, M. D. (1999). The Effects of
Nicotine on Learning and Memory: A Neuropsychological Assessment in
Young and Senescent Fischer 344 Rats. *Physiol.Behav.* 67: 421-431.

EcoReference No.: 84418\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Aubert, J. F., Burnier, M., Waeber, B., Nussberger, J., and Brunner, H.
R. (1987). Nicotine-Induced Release of Vasopressin in the Conscious Rat:
Role of Opioid Peptides and Hemodynamic Effects. *J.Pharmacol.Exp.Ther.*
243: 681-685.

EcoReference No.: 84391\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Battistel, M., Plebani, P., Di, Jocic, M., LIPPE, and Holzer, P. (1993).
Chronic Nicotine Intake Causes Vascular Dysregulation in the Rat Gastric
Mucosa. *Gut* 34: 1688-1692.

EcoReference No.: 84692\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Best, J. B. and Morita, M. (1991). Toxicology of Planarians.
*Hydrobiologia* 227: 375-383.

EcoReference No.: 9653\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: MOR,REP; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Birnbaum, S. C., Kien, N., Martucci, R. W., Gelzleichter, T. R.,
Witschi, H., Hendrickx, A. G., and Last, J. A. (1994). Nicotine- or
Epinephrine-Induced Uteroplacental Vasoconstriction and Fetal Growth in
the Rat. *Toxicology* 94: 69-80.

EcoReference No.: 84611\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: REP,PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Blackburn, C. W., Peterson, C. A., Hales, H. A., Carrell, D. T., Jones,
K. P., Urry, R. L., and Peterson, C. M. (1994). Nicotine, but not
Cotinine, has a Direct Toxic Effect on Ovarian Function in the Immature
Gonadotropin-Stimulated Rat. *Reprod.Toxicol.* 8: 325-331.

EcoReference No.: 84373\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Bowen, D. J., Eury, S. E., and Grunberg, N. E. (1986). Nicotine\'s
Effect on Female Rats\' Body Weight Caloric Intake and Physical
Activity. *Pharmacol.Biochem.Behav.* 25: 1131-1136.

EcoReference No.: 84404\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Broulik, P. D. and Jarab, J. (1993). The Effect of Chronic Nicotine
Administration on Bone Mineral Content in Mice. *Horm.Metab.Res.* 25:
219-221.

EcoReference No.: 84648\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Bryson, R., Biner, P. M., McNair, E., Bergondy, M., and Abrams, O. R.
(1981). Effects of Nicotine on Two Types of Motor Activity in Rats.
*Psychopharmacology* 73: 168-170.

EcoReference No.: 84524\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Bui, L. M., Keen, C. L., and Dubick, M. A. (1995). Comparative Effects
of 6-Week Nicotine Treatment on Blood Pressure and Components of the
Antioxidant System in Male Spontaneously Hypertensive (SHR) and
Normotensive Wistar Kyoto (WKY) Rats. *Toxicology* 98: 57-65.

EcoReference No.: 78779\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Bui, L. M., Keen, C. L., and Dubick, M. A. (1994). Influence of 12-Week
Nicotine Treatment and Dietary Copper on Blood Pressure and Indices of
the Antioxidant System in Male Spontaneous Hypertensive Rats.
*Biol.Trace Elem.Res.* 46: 67-78.

EcoReference No.: 79451\
Chemical of Concern: Cu,NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM,ACC; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN),OK(Cu).

Calore, E. E., Da Rosa, A. R., Perez, N. M., and Vilela de Almeida, L.
(2003). Effects of Nicotine Administration in Developing Muscle Fibers
of Rats Offspring. *Ecotoxicol.Environ.Saf.* 55: 152-156.

EcoReference No.: 84410\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Carr, L. A., Rowell, P. P., and Pierce, W. M. Jr. (1989). Effects of
Subchronic Nicotine Administation on Central Dopaminergic Mechanisms in
the Rat. *Neurochem.Res.* 14: 511-515.

EcoReference No.: 84639\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Chen, W. A. and Edwards, R. B. (2003). Prenatal Nicotine Exposure does
not Cause Purkinje Cell Loss in the Developing Rat Cerebellar vermis.
*Neurotoxicol.Teratol.* 25: 633-637.

EcoReference No.: 84718\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL,REP; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Chen, W. J. A., Edwards, R. B., Romero, R. D., Parnell, S. E., and Monk,
R. J. (2003). Long-Term Nicotine Exposure Reduces Purkinje Cell Number
in the Adult Rat Cerebellar Vermis. *Neurotoxicol.Teratol.* 25: 329-334.

EcoReference No.: 84719\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,BEH,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Chen, Y. P. and Squier, C. A. (1990). Effect of Nicotine on
7,12-Dimethylbenz(a)anthracene Carcinogenesis in Hamster Cheek Pouch .
*J.Natl.Cancer Inst.* 82: 861-864.

EcoReference No.: 84528\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Chowdhury, P. (1990). Endocrine and Metabolic Regulation of Body Mass by
Nicotine: Role of Growth Hormone. *Ann.Clin.Lab.Sci.* 20 : 415-419.

EcoReference No.: 84739\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,GRO,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Chowdhury, P., Ami, M., Hosotani, R., and RAYFORD (1991).
Meal-Stimulated Exocrine Pancreatic Secretion and Release of GI Peptides
in Normal and Nicotine-Treated Rats. *Regul.Pept.* 33: 11-20.

EcoReference No.: 84582\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Chowdhury, P., Hosotani, R., Chang, L., and Rayford, P. L. (1990).
Metabolic and Pathologic Effects of Nicotine on Gastrointestinal Tract
and Pancreas of Rats. *Pancreas* 5: 222-229.

EcoReference No.: 84653\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BEH,BCM,CEL; [Rejection Code]{.underline}: LITE
EVAL CODED(NCTN).

Colzani, R., Fang, S. L., Alex, S., and Braverman, L. E. (1998). The
Effect of Nicotine on Thyroid Function in Rats. *Metabolism* 47:
154-157.

EcoReference No.: 84615\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,GRO,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Corbet, S. A., Tiley, C., Moorhouse, T., Giam, C., Pursglove, S., Raby,
J., and Rich, M. (2000). Surface Films as Mosquito Larvicides:
Partitioning the Mode of Action. *Entomol.Exp.Appl.* 94: 295-307.

EcoReference No.: 61057\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Cruz, F. C., Delucia, R., and Planeta, C. S. (2005). Differential
Behavioral and Neuroendocrine Effects of Repeated Nicotine in Adolescent
and Adult Rats. *Pharmacol.Biochem.Behav.* 80: 411-417.

EcoReference No.: 84411\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Crystal, G. J., Bedran de Castro, M. T. B., and Downey, H. F. (1989).
Regional Hemodynamic Responses to Nicotine in Conscious and Anesthetized
Dogs: Comparative Effects of Pentobarbital and Chloralose.
*Proc.Soc.Exp.Biol.Med.* 191: 396-402.

EcoReference No.: 85505\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Daeninck, P. J., Messiha, N., and Persaud, T. (1991). Intrauterine
Development in the Rat Following Continuous Exposure to Nicotine from
Gestational Day 6 Through 12. *Anat.Anz.* 172: 257-261.

EcoReference No.: 84695\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: REP; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Domino, E. F. (2001). Nicotine induced behavioral locomotor
sensitization. *Progress in Neuro-Psychopharmacol.Biol.Psychiatr.* 25:
59-71.

EcoReference No.: 84371\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Dubick, M. A., Palmer, R., Lau, P. P., Morrill, P. R., and Geokas, M. C.
(1988). Altered Exocrine Pancreatic Function in Rats Treated with
Nicotine. *Toxicol.Appl.Pharmacol.* 96: 132-139.

EcoReference No.: 84384\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BCM,CEL, [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Flemmer, D. D. and Dilsaver, S. C. (1990). Constant Exposure to Darkness
Produces Supersensitivity to Nicotine. *Pharmacol.Biochem.Behav.* 35:
523-526.

EcoReference No.: 84383\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Forrest, C. R., Pang, C. Y., and Lindsay, W. K. (1987). Dose and Time
Effects of Nicotine Treatment on the Capillary Blood Flow and Viability
of Random Pattern Skin Flaps in the Rat. *Br.J.Plast.Surg.* 40: 295-299.

EcoReference No.: 84707\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Fraser, R., Clark, S. A., Day, W. A., and Murray, F. E. M. (1988).
Nicotine Decreases the Porosity of the Rat Liver Sieve: A Possible
Mechanisms for Hypercholesterolaemia. *Br.J.Exp.Pathol.* 69: 345-350.

EcoReference No.: 84165\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN),OK(ALL CHEMS).

French, R. C. (1988). Vacuole Induction by Nicotine in Teliospores of
Uromyces vignae Puccinia punctiformis and Puccinia helianthi.
*Mycologia* 80: 455-459.

EcoReference No.: 84465\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: POP; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Gaddnas, H., Pietila, K., Piepponen, T. P., and Ahtee, L. (2001).
Enhanced Motor Activity and Brain Dopamine Turnover in Mice During
Long-Term Nicotine Administration in the Drinking Water.
*Pharmacol.Biochem.Behav.* 70: 497-503.

EcoReference No.: 84723\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Glavin, G. B. and Lagrotteria, L. (1982). Nicotine and Ascorbic Acid
Effects on Cold-Restraint Ulcers in Rats. *Experientia* 38: 603-604.

EcoReference No.: 84744\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Godin, C. S. and Crooks, P. A. (1986). In Vivo Depletion of
S-Adenosyl-L-Homocysteine and S-Adenosyl-L-Methionine in Guinea Pig Lung
After Chronic S-(-)-Nicotine Administration. *Toxicol.Lett.* 31: 23-29.

EcoReference No.: 84713\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Gomita, Y., Suemaru, K., Furuno, K., and Araki, Y. (1989).
Nicotine-Induced Tail-Tremor and Drug Effects.
*Pharmacol.Biochem.Behav.* 34: 817-821.

EcoReference No.: 84620\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Gomita, Y., Suemaru, K., Furuno, K., and Araki, Y. (1991). Tail-Tremor
Induced by Exposure to Cigarette Smoke in Rats.
*Pharmacol.Biochem.Behav.* 40: 453-455.

EcoReference No.: 84397\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Grunwald, F., Schrock, H., and Kuschinsky, W. (1991). The Influence of
Nicotine on Local Cerebral Blood Flow in Rats. *Neurosci.Lett.* 124:
108-110.

EcoReference No.: 84651\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Haikala, H. (1989). Dual Effects of Nicotine on Neuroleptic-Induced
Changes of Striatal Dopamine Metabolism in Mice. *Pharmacol.Toxicol.*
64: 334-339.

EcoReference No.: 84395\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN),NO COC(MTL).

Haikala, H. and Ahtee, L. (1988). Antagonism of the Nicotine-Induced
Changes of the Striatal Dopamine Metabolism in Mice by Mecamylamine and
Pempidine. *Naunyn-Schmiedeberg\'s Arch.Pharmacol.* 338: 169-173.

EcoReference No.: 84655\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Harrison, P. K., Falugi, C., Angelini, C., and Whitaker, M. J. (2002).
Muscarinic Signalling Affects Intracellular Calcium Concentration During
the First Cell Cycle of Sea Urchin Embryos. *Cell Calcium* 31 : 289-297.

EcoReference No.: 83930\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN),OK(ALL CHEMS).

Hashimoto, T., Yoneda, M., Shimada, T., Kurosawa, M., and Terano, A.
(2004). Intraportal Nicotine Infusion in Rats Decreases Hepatic Blood
Flow Through Endothelin-1 and both Endothelin A and Endothelin B
Receptors. *Toxicol.Appl.Pharmacol.* 196: 1-10.

EcoReference No.: 84715\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Hildebrandt, I. J., Termeer, J. L., Toledo, D., and Cohen, R. W. (2003).
Characterization of Chronic Nicotine Exposure on the Survival of the
Spastic Han-Wistar Rat. *Nicotine Tobacco Res.* 5: 827-836.

EcoReference No.: 84593\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Hock, C. E. and Passmore, J. C. (1985). Mechanisms Mediating Canine
Renal Vasoconstriction Induced y Nicotine Infusion. *Life Sci.* 37:
1997-2004.

EcoReference No.: 84375\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Holgert, H., Hoekfelt, T., Hertzberg, T., and Lagercrantz, H. (1995).
Functional and Developmental Studies of the Peripheral Arterial
Chemoreceptors in Rat: Effects of Nicotine and Possible Relation to
Sudden Infant Death Syndrome. *Proc.Natl.Acad.Sci.U.S.A.* 92: 7575-7579.

EcoReference No.: 84576\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,BCM,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Homayounfar, H., Jamali-Raeufy, N., Sahebgharani, M., and Zarrindast, M.
R. (2005). Adenosine Receptor Mediates Nicotine-Induced Antinociception
in Formalin Test. *Pharmacol.Res.* 51: 197-203.

EcoReference No.: 84530\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN),OK(ALL CHEMS).

Huang, L. Z., Hsiao, S. H., Trzeciakowski, J., Frye, G. D., and
Winzer-Serhan, U. H. (2006). Chronic Nicotine Induces Growth Retardation
in Neonatal Rat Pups. *Life Sci.* 78: 1483-1493.

EcoReference No.: 84799\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Huang, Y. H., Brown, A. R., Costy-Bennett, S., Luo, Z., and Fregosi, R.
F. (2004). Influence of Prenatal Nicotine Exposure on Postnatal
Development of Breathing Pattern. *Respir.Physiol.Neurobiol.* 143: 1-8.

EcoReference No.: 84714\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: REP,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Iba, M. M., Scholl, H., Fung, J., Thomas, P. E., and Alam, J. (1998).
Induction of Pulmonary CYP1A1 by Nicotine. *Xenobiotica* 28: 827-843.

EcoReference No.: 84456\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Idia, M., Iida, H., Dohi, S., Takanaka, M., and Fujiwara, H. (1998).
Mechanisms Underlying Cerebrovascular Effects of Cigarette Smoking in
Rats In Vivo. *Stroke* 29: 1656-1665.

EcoReference No.: 85628\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Jacobsen, T. (1995). Acute Toxicity of 16 Water-Soluble Chemicals to the
Fungus Geotrichum candidum Measured by Reduction in Glucose Uptake.
*Toxicol.In Vitro* 9: 169-173.

EcoReference No.: 18069\
Chemical of Concern: CuS,HgCl2,PL,NCTN,PAQT,SFL,As;
[Habitat]{.underline}: A; [Effect Codes]{.underline}: PHY; [Rejection
Code]{.underline}: LITE EVAL CODED(NCTN,CuS).

Janhunen, S. and Ahtee, L. (2004). Comparison of the Effects of Nicotine
and Epibatidine on the Striatal Extracellular Dopamine.
*Eur.J.Pharmacol.* 494: 167-177.

EcoReference No.: 84806\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Janhunen, S., Linnervuo, A., Svensk, M., and Ahtee, Liisa (2005).
Effects of Nicotine and Epibatidine on Locomotor Activity and
Conditioned Place Preference in Rats. *Pharmacol.Biochem.Behav.* 82:
758-765.

EcoReference No.: 84419\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Johnson, S. K., Carlson, K. M., Lee, Jane, Burr, L. E., and Wagner, G.
C. (2003). Effects of Nicotine on Target Biting and Resident-Intruder
Attack. *Life Sci.* 73: 311-317 .

EcoReference No.: 84786\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Kashkin, V. A. and De Witte, P. (2005). Nicotine Increases
Microdialysate Brain Amino Acid Concentrations and Induces Conditioned
Place Preference. *Eur.Neuropsychopharmacol.* 15: 625-632.

EcoReference No.: 84741\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Katsura, M., Ohkuma, S., Xu, J., Hibino, Y., Tsujimura, A., and
Kuriyama, K. (1998). Continuous Treatment with Nicotine Increases
Diazepam Binding Inhibitor (DBI) and Its mRNA in the Mouse Brain.
*Mol.Brain Res.* 55: 345-349.

EcoReference No.: 84785\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Katyare, S. S. and Shallom, J. M. (1988). Altered Cerebral Protein
Turnover in Rats Following Prolonged In Vivo Treatment with Nicotine.
*J.Neurochem.* 50: 1356-1363.

EcoReference No.: 84511\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Kavitharaj, N. K. and Vijayammal, P. L. (1999). Nicotine Administration
Induced Changes in the Gonadal Functions in Male Rats. *Pharmacology
(Basel)* 58: 2-7.

EcoReference No.: 84399\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Kennedy, L. L., Aguwa, C. C., Rives, J. E., and Bernard, D. G. (2001).
Involvement of Cholinergic Mechanisms in the Central Control of
Respiration in the Cane Toad, Bufo marinus. *Comp.Biochem.Physiol.A*
128: 837-849.

EcoReference No.: 83778\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Kimmel, E. C. and Diamond, L. (1984). The Role of Nicotine in the
Pathogenesis of Pulmonary Emphysema. *Am.J.Respir.Crit.Care Med.* 129:
112-117.

EcoReference No.: 84669\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: MOR,PHY,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Kiritsy-Roy, J. A., Mousa, S. A., Appel, N. M., and Van Loon, G. R.
(1990). Tolerance to Nicotine-Induced Sympathoadrenal Stimulation and
Cross-Tolerance to Stress: Differential Central and Peripheral
Mechanisms in Rats. *Neuropharmacology* 29: 579-590.

EcoReference No.: 84462\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Kita, T., Nakashima, T., and Kurogochi, Y. (1985). Effects of Oral
Administration of Nicotine on Circadian Rhythms of Ambulatory Activity
and Drinking in Rats. *Jpn.J.Pharmacol.* 39: 554-557.

EcoReference No.: 84602\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Kita, T., Okamoto, M., Kubo, K., Tanaka, T., and Nakashima, T. (1999).
Enhancement of Sensitization to Nicotine-Induced Ambulatory Stimulation
by Psychological Stress in Rats. *Progress in
Neuro-Psychopharmacol.Biol.Psychiatr.* 23: 893-903.

EcoReference No.: 84581\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Kita, T., Okamoto, M., and Nakashima, T. (1992). Nicotine-Induced
Sensitization to Ambulatory Stimulant Effect Produced by Daily
Administration into the Ventral Tegmental Area and the Nucleus Accumbens
in Rats. *Life Sci.* 50: 583-590.

EcoReference No.: 84466\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Knofler, R., Takada, Y., Ihara, H., Urano, T., and Takada, A. (1995).
Effects of Nicotine and Electric Footshock on Peripheral Serotonergic
Measures and on Platelet Aggregation in Whole Blood of Rats. *Life Sci.*
57: 363-369.

EcoReference No.: 84782\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Ko, J. K. S. and Cho, C. H. (2005). The Diverse Actions of Nicotine and
Different Extracted Fractions from Tobacco Smoke Against Hapten-Induced
Colitis in Rats. *Toxicol.Sci.* 87: 285-295.

EcoReference No.: 80601\
Chemical of Concern: NCTN,CF; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN),OK(ALL CHEMS).

Kubo, K., Kita, T., Narushima, I., Tanaka, T., Nakatani, T., and
Nakashima, T. (2003). Nicotine-Induced Inflammatory Decreasing Effect on
Passive Skin Arthus Reaction in Paraventricular Nucleus-Lesioned Wistar
Rats. *Pharmacol.Toxicol.* 92: 125-130.

EcoReference No.: 84467\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,PHY,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Kuo, D. Y., Lin, T. B., Huang, C. C., Duh, S. L., Liao, J. M., and
Cheng, J. T. (1999). Nicotine-Induced Hyperlocomotion is not Modified by
the Estrous Cycle, Ovariectomy and Estradiol Replacement at
Physiological Level. *Chin.J.Physiol.* 42: 83-88.

EcoReference No.: 84159\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Lapchak, P. A., Araujo, D. M., Quirion, R., and Collier, B. (1989).
Effect of Chronic Nicotine Treatment on Nicotinic Autoreceptor Function
and Tritiated N Methylcarbamylcholine Binding Sites in the Rat Brain.
*J.Neurochem.* 52: 483-491.

EcoReference No.: 84586\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Lapin, E. P., Maker, H. S., Sershen, H., and Lajtha, A. (1989). Action
of Nicotine on Accumbens Dopamine and Attenuation with Repeated
Administration. *Eur.J.Pharmacol.* 160: 53-59.

EcoReference No.: 84536\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,BCM,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Larose, P., Ong, H., Januszewicz, P., Cantin, M., and Du Souich, P.
(1988). Characterization of the Effect of Nicotine on Vasopressin and
Atrial Natriuretic Factor in the Rabbit. *J.Pharmacol.Exp.Ther.* 244:
1093-1097.

EcoReference No.: 84706\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Lau, P. P., Dubick, M. A., Yu, G., Morrill, P. R., and Geokas, M. C.
(1990). Dynamic Changes of Pancreatic Structure and Function in Rats
Treated Chronically with Nicotine. *Toxicol.Appl.Pharmacol.* 104:
457-465.

EcoReference No.: 84385\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Lee, T. H., Jang, M. i-Hyeon, Shin, M. C., Lim, B. V., Choi, H. H., Kim,
H., Kim, E. e-Hwa, and Kim, Chang-J. u. (2002). Nicotine Administration
Increases Serotonin Synthesis and Tryptophan Hydroxylase Expression in
Dorsal Raphe of Food-Deprived Rats. *Nutr.Res.* 22: 1445-1452.

EcoReference No.: 84588\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Leikola-Pelho, T., Heinamaki, J., Laakso, I., and Ahtee, L. (1990).
Chronic Nicotine Treatment Changes Differentially the Effects of Acute
Nicotine on the Three Main Dopamine Metabolites in Mouse Striatum.
*Naunyn-Schmiedeberg\'s Arch.Pharmacol.* 342: 400-406.

EcoReference No.: 84590\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Lesiuk, N. M. and Drewes, C. D. (1999). Autotomy Reflex in a Freshwater
Oligochaete, Lumbriculus variegatus (Clitellata: Lumbriculidae).
*Hydrobiologia* 406: 253-261.

EcoReference No.: 51471\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: BEH,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Levin, E. D., Ellison, G. D., Salem, C., Jarvik, M., and Gritz, E.
(1988). Behavioral Effects of Acute Hexamethonium in Rats Chronically
Intoxicated with Nicotine. *Physiol.Behav.* 44: 355-359.

EcoReference No.: 84803\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Londonkar, R. L., Sonar, A., Patil, S., and Patil, S. B. (2000).
Nicotine Delays Puberty in Male Rat. *Pharm.Biol.* 38: 291-297.

EcoReference No.: 84641\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BCM,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Londonkar, R. L., Srinivasreddy, P., Somanathreddy, P., and Patil, S. B.
(1998). Nicotine Induced Inhibition of the Activities of Accessory
Reproductive Ducts in Male Rats. *J.Ethnopharmacol.* 60: 215-221.

EcoReference No.: 84532\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,CEL,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Lupien, J. R. and Bray, G. A. (1988). Nicotine Increases Thermogenesis
in Brown Adipose Tissue in Rats. *Pharmacol.Biochem.Behav.* 29: 33-37.

EcoReference No.: 84403\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,GRO,BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Maritz, G. S. (1988). Effect of Maternal Nicotine Exposure on Growth In
Vivo of Lung Tissue of Neonatal Rats. *Biol.Neonate* 53 : 163-170.

EcoReference No.: 84585\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Maritz, G. S. and Burger, B. (1992). The Influence of Maternal Nicotine
Exposure on Neonatal Lung Carbohydrate Metabolism. *Cell Biol.Int.* 16:
1229-1236.

EcoReference No.: 84737\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Maritz, G. S. and Windvogel, S. (2003). Is Maternal Copper
Supplementation During Alveolarization Protecting the Developing Rat
Lung Against the Adverse Effects of Maternal Nicotine Exposure? A
Morphometric Study. *Exp.Lung Res.* 29: 243-260 .

EcoReference No.: 79008\
Chemical of Concern: Cu,NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN),NO MIXTURE(Cu).

Maritz, G. S., Woolward, K. M., and Du Toit, G. (1993). Maternal
Nicotine Exposure During Pregnancy and Development of Emphysema-Like
Damage in the Offspring. *S.Afr.Med.J.(S.Afr.Med.Tydskr.)* 83: 195-199.

EcoReference No.: 84372\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Marks, M. J., Romm, E., Bealer, S. M., and Collins, A. C. (1985). A Test
Battery for Measuring Nicotine Effects in Mice.
*Pharmacol.Biochem.Behav.* 23: 325-330.

EcoReference No.: 84607\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Martin, B. J. and Wehner, J. M. (1988). Influence of Genotype on
Nicotine-Induced Increaes of Plasma Corticosterone in Mice as a Result
of Acute Nicotine Pretreatment. *Pharmacol.Biochem.Behav.* 30:
1065-1070.

EcoReference No.: 84402\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Martin, J. C. and Martin, D. C. (1981). Voluntary Activity in the Aging
Rat as a Function of Maternal Drug Exposure.
*Neurobehav.Toxicol.Teratol.* 3: 261-264.

EcoReference No.: 84645\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Mayhan, W. G. and Patel, K. P. (1997). Effect of Nicotine on
Endothelium-Dependent Arteriolar Dilatation In Vivo. *Am.J.Physiol.*
272: H2337-H2342.

EcoReference No.: 84600\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

McNair, E. and Bryson, R. (1983). Effects of Nicotine on Weight Change
and Food Consumption in Rats. *Pharmacol.Biochem.Behav.* 18: 341-344.

EcoReference No.: 84622\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Meyer, D. C. and Carr, L. A. (1988). The Effects of Perinatal Exposure
to Nicotine on Plasma LH Levels in Prepubertal Rats.
*Neurotoxicol.Teratol.* 9: 95-98.

EcoReference No.: 84464\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,REP,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Miller, R. R. Jr., Heckel, C. D., Koss, W. J., Montague, S. L., and
Greenman, A. L. (2001). Ethanol- and Nicotine-Induced Membrane Changes
in Embryonic and Neonatal Chick Brains. *Comp.Biochem.Physiol.C* 130:
163-178.

EcoReference No.: 62901\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Mitchell, J. A. and Hammer, R. E. (1985). Effects of Nicotine on
Oviducal Blood Flow and Embryo Development in the Rat.
*J.Reprod.Fertil.* 74: 71-76.

EcoReference No.: 84534\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Mitchell, J. A., Hammer, R. E., and Goldman, H. (1981). Effects of
Nicotine on Uterine Blood Flow and Intrauterine Oxygen Tension in the
Rat. *J.Reprod.Fertil.* 63: 163-168.

EcoReference No.: 84406\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Monheit, A. G., Van Vunakis, H., Key, T. C., and Resnik, R. (1983).
Maternal and Fetal Cardiovascular Effects of Nicotine Infusion in
Pregnant Sheep. *Am.J.Obstet.Gynecol.* 145: 290-296.

EcoReference No.: 84671\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Morgan, M. M. and Ellison, G. (1987). Different Effects of Chronic
Nicotine Treatment Regimens on Body Weight and Tolerance in the Rat.
*Psychopharmacology* 91: 236-238.

EcoReference No.: 84516\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Mundy, W. R. and Iwamoto, E. T. (1988). Nicotine Impairs Acquisition of
Radial Maze Performance in Rats. *Pharmacol.Biochem.Behav.* 30: 119-122.

EcoReference No.: 84400\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Muneoka, K., Nakatsu, T., FUJI, Ogawa, T., and Takigawa, M. (1999).
Prenatal Administration of Nicotine Results in Dopaminergic Alterations
in the Neocortex. *Neurotoxicol.Teratol.* 21: 603-609.

EcoReference No.: 84724\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,REP,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Nagata, M. and Osumi, Y. ( 1990). Central Inhibition of Gastric Motility
by Intravenously Administered Nicotine in Rats. *Jpn.J.Pharmacol.* 52:
397-403.

EcoReference No.: 84603\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Newman, M. B., Shytle, R. D., and Sanberg, P. R. (1999). Locomotor
Behavioral Effects of Prenatal and Postnatal Nicotine Exposure in Rat
Offspring. *Behav.Pharmacol.* 10: 699-706.

EcoReference No.: 84163\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: REP,GRO,MOR,BEH; [Rejection Code]{.underline}: LITE
EVAL CODED(NCTN).

Ngah, W. Z. W., Jarien, Z., Rajikin, M. H., and Shamaan, N. A. (1991).
Effect of Nicotine on Glutathione Metabolizing Enzymes and Some
Conjugation Enzymes in the Rat. *Asia Pac.J.Pharmacol.* 6: 55-62.

EcoReference No.: 74292\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN),NO COC(MLT).

Okada, S., Shimizu, T., and Yokotani, K. (2003). Extrahypothalamic
Corticotropin-Releasing Hormone Mediates (-)-Nicotine-Induced Elevation
of Plasma Corticosterone in Rats. *Eur.J.Pharmacol.* 473: 217-223.

EcoReference No.: 84749\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Okamoto, M., Kita, T., Okuda, H., Tanaka, T., and Nakashima, T. (1992).
Effects of Acute Administration of Nicotine on Convulsive Movements and
Blood Levels of Corticosterone in Old Rats. *Jpn.J.Pharmacol.* 60:
381-384.

EcoReference No.: 84605\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,BEH,PHY,ACC; [Rejection Code]{.underline}: LITE
EVAL CODED(NCTN).

Olson, J. W. and Crooks, P. A. (1985). Effect of Nicotine and
N\'Nitrosonornicotine on Rat Lung and Trachea Ornithine Decarboxylase
Activity. *Carcinogenesis (Lond.)* 6: 1517-1520.

EcoReference No.: 84469\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,MOR; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Passino-Reader, D. R. (1993). Rainbow Trout Larvae Compared with Daphnia
pulex Response in Contaminant Bioassays. Copy of a Research Information
Bulletin (RIB). Draft (Personal Communication). *October 19 Letter to
R.Spehar, U.S.EPA, Duluth, MN* 5 p.

EcoReference No.: 67566\
Chemical of Concern: PAH,NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: MOR,REP,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN),OK(ALL CHEMS).

Passmore, J. C., Jimenez, A. E., and Pierce, W. M. (1991). Cardiac
Output and the Blood Pressure Increase in Deoxycorticosterone
Acetate-Salt Hypertension After Nicotine Infusion.
*Clin.Exp.Hypertens.A* 13: 83-102.

EcoReference No.: 84729\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Patel, K. P., Zhang, P. L., and Mayhan, W. G. (1995). Blunting of Renal
Excretory Responses to Acute Volume Expansion by Nicotine: Role of Renal
Nerves. *J.Pharmacol.Exp.Ther.* 274: 1174-1181.

EcoReference No.: 84704\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Patil, S., Patil, S., Bhaktaraj, B., and PATIL (1999). Effect of Graded
Doses of Nicotine on Ovarian and Uterine Activities in Albino Rats.
*Indian J.Exp.Biol.* 37: 184-186.

EcoReference No.: 84634\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Pawlik, W. W., Jacobson, E. D., and Banks, R. O. (1985). Actions of
Nicotine on Renal Function in Dogs. *Proc.Soc.Exp.Biol.Med.* 178:
585-590.

EcoReference No.: 84805\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Pelissier, A. L., Gantenbein, M., and Bruguerolle, B. (1998).
Nicotine-Induced Perturbations on Heart Rate, Body Temperature and
Locomotor Activity Daily Rhythms in Rats. *J.Pharm.Pharmacol.* 50:
929-934.

EcoReference No.: 84388\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BEH,MOR; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Peng, B., Tomashefsky, P., and Nagler, H. M. (1990). The Cofactor
Effect: Varicocele and Infertility. *Fertil.Steril.* 54: 143-148.

EcoReference No.: 84522\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,MOR,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Pennypacker, K. R., Hong, Douglass, J., and McMillian, M. K. (1992).
Constitutive Expression of AP-1 Transcription Factors in the Rat
Adrenal: Effects of Nicotine. *J.Biol.Chem.* 267: 20148-20152.

EcoReference No.: 85501\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Pessoa, R. F., Castro, N. G., and Noel, F. (2005). Binding of
\[3H\]MK-801 in Subcellular Fractions of Schistosoma mansoni: Evidence
for Interaction with Nicotinic Receptors. *Biochem.Pharmacol.* 69:
1509-1516.

EcoReference No.: 83780\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Peters, D. A. V. (1984). Prenatal Nicotine Exposure Increases Adrenergic
Receptor Binding in the Rat Cerebral Cortex.
*Res.Commun.Chem.Pathol.Pharmacol.* 46: 307-318.

EcoReference No.: 84470\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,BEH,GRO,REP; [Rejection Code]{.underline}: LITE
EVAL CODED(NCTN).

Peters, D. A. V. and Tang, S. (1982). Sex-Dependent Biological Changes
Following Prenatal Nicotine Exposure in the Rat.
*Pharmacol.Biochem.Behav.* 17: 1077-1082.

EcoReference No.: 84392\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: REP,BCM,BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Peters, M. A. and Ngan, L. L. E. (1982). The Effects of Totigestational
Exposure to Nicotine on Pre- and Postnatal Development in the Rat.
*Arch.Int.Pharmacodyn.* 257: 155-167.

EcoReference No.: 84738\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BEH,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Pietila, K., Laakso, I., and Ahtee, L. (1995). Chronic Oral Nicotine
Administration Affects the Circadian Rhythm of Dopamine and
5-Hydroxytryptamine Metabolism in the Striata of Mice.
*Naunyn-Schmiedeberg\'s Arch.Pharmacol.* 353: 110-115.

EcoReference No.: 84398\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Pietilae, K., Salminen, O., Leikola-Pelho, T., and Ahtee, L. (1996).
Tolerance to Nicotine\'s Effects on Striatal Dopamine Metabolism in
Nicotine-Withdrawn Mice. *Eur.J.Pharmacol.* 318: 17-22.

EcoReference No.: 84750\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BEH,BCM,CEL; [Rejection Code]{.underline}: LITE
EVAL CODED(NCTN).

Radek, R. J. (1993). Effects of Nicotine on Cortical High Voltage
Spindles in Rats. *Brain Res.* 625: 23-28.

EcoReference No.: 84458\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Rao, A. P. and Patil, S. B. (1992). Effect of Nicotine on the
Spermatogenic Activities of Testis in Albino Mice. *Indian
J.Comp.Anim.Physiol.* 10: 1-6.

EcoReference No.: 84457\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Ravindra, Patil, S. R., Patil, S. R., and Patil, S. B. (1998). Effect of
Increasing Dose of Nicotine on Estrous Cycle in Albino Rats. *Geobios
(Jodhpur)* 25: 105-108.

EcoReference No.: 84584\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Reddy, S., Londonkar, R., RAVINDRA, Reddy, S., and Patil, S. B. (1998).
Testicular Changes due to Graded Doses of Nicotine in Albino Mice.
*Indian J.Physiol.Pharmacol.* 42: 276-280.

EcoReference No.: 84731\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BCM,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Riesenfeld, A. and Oliva, H. (1988). Effects of Nicotine on the
Fertility, Cytology and Life Span of Male Rats. *Acta Anat.* 131:
171-176.

EcoReference No.: 84672\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,REP; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Robert, M. E., Leung, F. W., and Guth, P. H. (1986). Nicotine and
Smoking do not Decrease Basal Gastric Mucosal Blood Flow in Anesthetized
Rats. *Dig.Dis.Sci.* 31: 530-534 .

EcoReference No.: 84158\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Roberts, B. L. and Dorough, H. W. (1984). Relative Toxicities of
Chemicals to the Earthworm Eisenia foetida. *Environ.Toxicol.Chem.* 3:
67-78.

EcoReference No.: 40531\
Chemical of Concern:
ACP,ADC,BMY,BNZ,Captan,CBD,CBF,CBL,Cd,CH3I,CPY,CTC,CuS,CYP,DCTP,DDT,DMM,DU,ES,FML,FNF,FNV,IDM,MBZ,MLN,MOM,NCTN,NHN,PAH,PAQT,Pb,PMR,PMSM,PPB,PPX,PRN,TBO,TFN,TPM;
[Habitat]{.underline}: T; [Effect Codes]{.underline}: MOR; [Rejection
Code]{.underline}: LITE EVAL CODED(NCTN,CBF,ADC,MOM,PPB,CuS,CYP).

Robinson, S. F., Marks, M. J., and Collins, A. C. (1996). Inbred Mouse
Strains Vary in Oral Self-Selection of Nicotine. *Psychopharmacology*
124: 332-339.

EcoReference No.: 83910\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN),OK(ALL CHEMS).

Robinson, S. F., Pauly, J. R., Marks, M. J., and Collins, A. C. (1994).
An Analysis of Response to Nicotine Infusion Using an Automated
Radiotelemetry System. *Psycopharmacology* 115: 115-120.

EcoReference No.: 84512\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Rogers, D. T. and Iwamoto, E. T. (1993). Multiple Spinal Mediators in
Parenteral Nicotine-Induced Antinociception. *J.Pharmacol.Exp.Ther.*
267: 341-349.

EcoReference No.: 84390\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Rowell, P. P. and Clark, M. J. (1982). The Effect of Chronic Oral
Nicotine Administration on Fetal Weight and Placental Amino Acid
Accumulation in Mice. *Toxicol.Appl.Pharmacol.* 66: 30-38.

EcoReference No.: 84405\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: REP,BCM,ACC; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Roy, T. S., Andrews, J. E., Seidler, F. J., and Slotkin, T. A. (1998).
Nicotine Evokes Cell Death in Embryonic Rat Brain During Neurulation.
*J.Pharmacol.Exp.Ther.* 287: 1136-1144.

EcoReference No.: 84710\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,REP; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Roy, T. S. and Sabherwal, U. (1998). Effects of Gestational Nicotine
Exposure on Hippocampal Morphology. *Neurotoxicol.Teratol.* 20: 465-473.

EcoReference No.: 84412\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Saxon, D. J., Diamond, L., and Gillespie, M. M. (1984). Chronic
Administration of Water Soluble Tobacco Smoke Extract or Nicotine Fails
to Influence Porcine Coronary Aftery Reactivity. *Toxicology* 32: 85-91.

EcoReference No.: 84632\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Schwid, S. R., Hirvonen, M. D., and Keesey, R. E. (1992). Nicotine
Effects on Body Weight: A Regulatory Perspective. *Am.J.Clin.Nutr.* 55:
878-884.

EcoReference No.: 84670\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Scott, G. C., Pickett, J. A., Smith, M. C., Woodcock, C. M., Harris, P.
G. W., Hammon, R. P., and Koetecha, H. D. (1984). Seed Treatments for
Controlling Slugs in Winter Wheat. *In: 1984 Br.Crop Prot.Conf.- Pests
and Dis.* 1: 133-138.

EcoReference No.: 79951\
Chemical of Concern:
DZM,RTN,MNK,ADC,MCB,BMN,NP,AMZ,CBF,TMP,BMNO,THO,DM,THM,DNB,NCTN;
[Habitat]{.underline}: T; [Effect Codes]{.underline}: MOR,PHY,POP;
[Rejection Code]{.underline}: LITE EVAL CODED(DZM),OK(ALL CHEMS).

Seidenberg, J. M., Anderson, D. G., and Becker, R. A. (1986). Validation
of an In Vivo Developmental Toxicity Screen in the Mouse.
*Teratog.Carcinog.Mutagen.* 6: 361-374.

EcoReference No.: 56184\
Chemical of Concern: Co,Ni,Se,NaAs,Cd,Zn,DM,AMSV,NCTN;
[Habitat]{.underline}: T; [Effect Codes]{.underline}: GRO,MOR,REP;
[Rejection Code]{.underline}: LITE EVAL CODED(NCTN,AMSV,TRV-NaAs).

Sershen, H., Reith, M. E. A., Banay-Schwartz, M., and Lajtha, A. (1982).
Effects of Prenatal Administration of Nicotine on Amino Acid Pools,
Protein Metabolism, and Nicotine Binding in the Brain. *Neurochem.Res.*
7: 1515-1522.

EcoReference No.: 84689\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,REP,PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Shallom, J. M. and Katyare, S. S. (1985). Altered Synaptosomal ATPase
Activity in Rat Brain Following Prolonged In Vivo Treatment with
Nicotine. *Biochem.Pharmacol.* 34: 3445-3449.

EcoReference No.: 84646\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: ACC,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Shoaib, M., Schindler, C. W., Goldberg, S. R., and Pauly, J. R. (1997).
Behavioural and Biochemical Adaptations to Nicotine in Rats: Influence
of MK801, an NMDA Receptor Antagonist. *Psychopharmacology* 134:
121-130.

EcoReference No.: 84526\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,BCM,BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Siren, A. L. and Feuerstein, G. (1990). Cardiovascular Effects of
Anatoxin-A in the Conscious Rat. *Toxicol.Appl.Pharmacol.* 102: 91-100.

EcoReference No.: 84386\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Slawecki, C. J. and Ehlers, C. L. (2002). Lasting Effects of Adolescent
Nicotine Exposure on the Electroencephalogram, Event Related Potentials,
and Locomotor Activity in the Rat. *Dev.Brain Res.* 138: 15-25.

EcoReference No.: 84726\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,BEH,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Slawecki, C. J., Gilder, A., Roth, J., and Ehlers, C. L. (2003).
Increased Anxiety-Like Behavior in Adult Rats Exposed to Nicotine as
Adolescents. *Pharmacol.Biochem.Behav.* 75: 355-361.

EcoReference No.: 84727\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Slotkin, T. A., Cho, H., and Whitmore, W. L. (1987). Effects of Prenatal
Nicotine Exposure on Neuronal Development: Selective Actions on Central
and Peripheral Catecholaminergic Pathways. *Brain Res.Bull.* 18:
601-611.

EcoReference No.: 84779\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Slotkin, T. A., Greer, N., Faust, J., Cho, H., and Seidler, F. J.
(1986). Effects of Maternal Nicotine Injections on Brain Development in
the Rat: Ornithine Decarboxylase Activity, Nucleic Acids and Proteins in
Discrete Brain Regions. *Brain Res.Bull.* 17: 41-50.

EcoReference No.: 84521\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: MOR,REP,GRO,BCM,CEL; [Rejection Code]{.underline}:
LITE EVAL CODED(NCTN).

Sorensen, C. A., Raskin, L. A., and Suh, Y. (1991). The Effects of
Prenatal Nicotine on Radial-Arm Maze Performance in Rats.
*Pharmacol.Biochem.Behav.* 40: 991-993.

EcoReference No.: 84394\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,REP; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Sparks, J. A. and Pauly, J. R. (1999). Effects of Continuous Oral
Nicotine Administration on Brain Nicotinic Receptors and Responsiveness
to Nicotine in C57Bl/6 Mice. *Psychopharmacology* 141: 145-153.

EcoReference No.: 84712\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Swislocki, A. L. M. (2003). Smokeless Nicotine Administration does not
Result in Hypertension or a Deterioration in Glucose Tolerance or
Insulin Sensitivity in Juvenile Rats. *Metabolism* 52: 67-72.

EcoReference No.: 84463\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Tabassian, A. R., Nnylen, E. S., Lukacs, L., Cassidy, M. M., and Becker,
K. L. (1990). Cholinergic Regulation of Hamster Pulmonary Neuroendocrine
Cell Calcitonin. *Exp.Lung Res.* 16: 267-278.

EcoReference No.: 84682\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Takada, A., Urano, T., Yoshida, M., and Takada, Y. (1996). Comparison of
Changes in Serotonergic Measures in Whole Blood or Plasma and Brain in
Rats Given Nicotine and/or Stresses. *Pol.J.Pharmacol.* 48: 173-177.

EcoReference No.: 84654\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Takada, Y., Urano, T., Ihara, H., and Takada, A. (1995). Changes in the
Central and Peripheral Serotonergic System in Rats Exposed to
Water-Immersion Restrained Stress and Nicotine Administration.
*Neurosci.Res.* 23: 305-311.

EcoReference No.: 84802\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,ACC; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Takita, M., Taniguchi, T., Zhu, J., Piao, H. L., Tsai, T. Y., and
Muramatsu, I. (1999). Effects of Chronic Treatment with (+)-Nicotine on
the Stress-Induced Hypertension and Downregulation of Central Nicotinic
Receptors in Rats: Comparative Study with (-)-Nicotine. *Gen.Pharmacol.*
33: 29-33.

EcoReference No.: 84613\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Tanus-Santos, J. E., Sampaio, R. C., Hyslop, S., Franchini, K. G., and
Moreno, H. Jr. (2000). Endothelin ETA Receptor Antagonism Attenuates the
Pressor Effects of Nicotine in Rats. *Eur.J.Pharmacol.* 396: 33-37.

EcoReference No.: 84748\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Tseng, C. J., Appalsamy, M., Robertson, D., and Mosqueda-Garcia, R.
(1993). Effects of Nicotine on Brain Stem Mechanisms of Cardiovascular
Control. *J.Pharmacol.Exp.Ther.* 265: 1511-1518.

EcoReference No.: 84698\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Valenca, S. S., De Souza da Fonseca, A., Da Hora, K., Santos, R., and
Porto, L. C. (2004). Lung Morphometry and MMP-12 Expression in Rats
Treated with Intraperitoneal Nicotine. *Exp.Toxicol.Pathol.* 55:
393-400.

EcoReference No.: 84376\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Visanji, N. P., Mitchell, S. N., O\'Neill, M. J., and Duty, S. (2006).
Chronic Pre-treatment with Nicotine Enhances Nicotine-Evoked Striatal
Dopamine Release and \[alpha\]6 and \[beta\]3 Nicotinic Acetylcholine
Receptor Subunit mRNA in the Substantia Nigra pars Compacta of the Rat.
*Neuropharmacology* 50: 36-46.

EcoReference No.: 84413\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Wang, J. and Lung, M. A. ( 1988). The Nasal Vascular and Airway
Responses to Intracarotid Injection of Nicotine in the Dog.
*Med.Sci.Res.* 16: 237-238.

EcoReference No.: 84673\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Wang, N. S., Chen, M. F., Schraufnagel, D. E., and Yao, Y. T. (1984).
The Cumulative Scanning Electron Microscopic Changes in Baby Mouse Lungs
Following Prenatal and Postnatal Exposures to Nicotine. *J.Pathol.* 144:
89-100.

EcoReference No.: 84587\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Wang, N. S., Schraufnagel, D. E., and Chen, M. F. (1984). The Effect of
Maternal Oral Intake of Nicotine on the Growth and Maturation of Fetal
and Baby Mouse Lungs. *Lung* 161: 27-38.

EcoReference No.: 84374\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Wang, S. L., Feng, J., Correa, A., Brigham, M., and Wu-Wang, C. Y.
(1996). Effects of In Vivo Treatments of Nicotine and Benzo\[a\]pyrene
on the Epidermal Growth Factor Receptor in Hamster Buccal Pouch.
*Toxicology* 107: 31-38.

EcoReference No.: 84630\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,GRO,ACC; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Wielgus, J. J., Corbin Downey, L., Ewald, K. W., Hatley, M. E., Wilson,
K. C., and Yeilding, R. H. (2004). Exposure to Low Concentrations of
Nicotine During Cranial Nerve Development Inhibits Apoptosis and Causes
Cellular Hypertrophy in the Ventral Oculomotor Nuclei of the Chick
Embryo. *Brain Res.* 1000: 123-133.

EcoReference No.: 84733\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Winders, S. E., Wilkins II, D. R., Rushing, P. A., and Dean, J. E.
(1993). Effects of Nicotine Cycling on Weight Loss and Regain in Male
Rats. *In: 1st Int.Behav.Neurosci.Conf., May 21-24, 1992, San Antonio,
TX, Pharmacol.Biochem.Behav.* 46: 209-213 .

EcoReference No.: 84468\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Witschi, H., Lundgaard, S. M., Rajini, P., Hendrickx, A. G., and Last,
J. A. (1994). Effects of Exposure to Nicotine and to Sidestream Smoke on
Pregnancy Outcome in Rats. *Toxicol.Lett.* 71: 279-286.

EcoReference No.: 84387\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: REP,GRO,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Woodman, O. L. (1991). Coronary Vascular Responses to Nicotine in the
Anesthetized Dog. *Naunyn-Schmiedeberg\'s Arch.Pharmacol.* 343: 65-69.

EcoReference No.: 84408\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,CEL; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Yamada, H., Nakamura, T., and Oguri, K. (1998). Induction of Rat Hepatic
Cytochromes P450 by Toxic Ingredients in Plants: Lack of Correlation
Between Toxicity and Inductive Activity. *J.Toxicol.Sci.* 23: 395-402.

EcoReference No.: 84461\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,GRO,MOR; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Yoshida, T., Yoshioka, K., Hiraoka, N., and Kondo, M. (1990). Effect of
Nicotine on Norepinephrine Turnover and Thermogenesis in Brown Adipose
Tissue and Metabolic Rate in MSG Obese Mice. *J.Nutr.Sci.Vitaminol* 36:
123-130.

EcoReference No.: 84642\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BEH,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Yoshimura, R., Xu, L., Sun, B., and Tank, A. W. (2004). Nicotinic and
Muscarinic Acetylcholine Receptors are Essential for the Long-Term
Response of Tyrosine Hydroxylase Gene Expression to Chronic Nicotine
Treatment in Rat Adrenal Medulla. *Mol.Brain Res.* 126: 188-197.

EcoReference No.: 84728\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Zarrindast, M. R., Sadegh, M., and Shafaghi, B. (1996). Effects of
Nicotine on Memory Retrieval in Mice. *Eur.J.Pharmacol.* 295: 1-6.

EcoReference No.: 84538\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Zbuzek, V. K. and Zbuzek, V. (1991). Effect of Chronic Nicotine
Treatment and Its Withdrawal on the Vasopressinergic System in Rats.
*J.Neuroendocrinol.* 3: 107-112.

EcoReference No.: 84623\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BCM; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Zbuzek, V. K. and Zbuzek, V. (1999). Effect of Pre- and Postnatal
Nicotine Exposure on Vasopressinergic System in Rats. *Dev.Brain Res.*
112: 229-235.

EcoReference No.: 84414\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,GRO,REP; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

Zhu, J., Takita, M., Konishi, Y., Sudo, M., and Muramatsu, I. (1996).
Chronic Nicotine Treatment Delays the Developmental Increase in Brain
Muscarinic Receptors in Rat Neonate. *Brain Res.* 732: 257-260.

EcoReference No.: 84415\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,GRO; [Rejection Code]{.underline}: LITE EVAL
CODED(NCTN).

**Acceptable for ECOTOX but not OPP**

Ando, K., Miyata, H., Hironaka, N., Tsuda, T., and Yanagita, T. (1993).
The Discriminative Effects of Nicotine and Their Central Sites in Rats.
*Jpn.J.Psychopharmacol.* 13: 129-136.

EcoReference No.: 84592\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: NO
CONTROL,ENDPOINT(NCTN).

Balabanova, S. (1993). Effect of Nicotine on the Immunoreactive
Calcitonin in Cerebrospinal Fluid of Sheep. *Neuroendocrinol.Lett.* 15:
431-436.

EcoReference No.: 85058\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Buccafusco, J. J. and Yang, X. (1993). Mechanism of the Hypertensive
Response to Central Injection of Nicotine in Conscious Rats. *Brain
Res.Bull.* 32: 35-41.

EcoReference No.: 84740\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Cadoni, C. and Di Chiara, G. (2000). Differential Changes in Accumbens
Shell and Core Dopamine in Behavioral Sensitization to Nicotine.
*Eur.J.Pharmacol.* 387: R23-R25.

EcoReference No.: 84527\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,BEH; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Calleja, M. C., Persoone, G., and Geladi, P. (1994). Comparative Acute
Toxicity of the First 50 Multicentre Evaluation of In Vitro Cytotoxicity
Chemicals to Aquatic Non-Vertebrates. *Arch.Environ.Contam.Toxicol.* 26:
69-78.

EcoReference No.: 13669\
Chemical of Concern:
24DXY,HCCH,MLN,WFN,PCP,Ba,CTC,PL,SFL,NCTN,LPS,PAQT,As,Cu,CuS,Hg,TI,CF,PCP,AMSV;
[Habitat]{.underline}: A; [Effect Codes]{.underline}: MOR,PHY;
[Rejection Code]{.underline}: NO CONTROL(ALL CHEMS).

Chao, S. L., Dennehy, T. J., and Casida, J. E. (1997). Whitefly
(Hemiptera: Aleyrodidae) Binding Site for Imidacloprid and Related
Insecticides: A Putative Nicotinic Acetylcholine Receptor.
*J.Econ.Entomol.* 90: 879-882.

EcoReference No.: 83896\
Chemical of Concern: NCTN,IMC,ACT; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: MOR,BEH; [Rejection Code]{.underline}: OK(ALL
CHEMS),OK TARGET(NCTN).

Chen, Y. P., Johnson, G. K., and Squier, C. A. (1994). Effects of
Nicotine and Tobacco-Specific Nitrosamines on Hamster Cheek Pouch and
Gastric Mucosa. *J.Oral Pathol.Med.* 23: 251-255.

EcoReference No.: 85124\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,GRO; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Chowdhury, P., Doi, R., Chang, L. W., and Rayford, P. L. (1993). Tissue
Distribution of Tritiated Nicotine in Rats. *Biomed.Environ.Sci.* 6:
59-64.

EcoReference No.: 84331\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: ACC; [Rejection Code]{.underline}: NO CONTROL(NCTN).

Cruz, S. L., Fernandez-Guasti, A., and Villarreal, J. E. (1994).
Cardiovascular Effects of Different Schedules of Nicotine Administration
on Spinal Rats: Influence of Pentobarbital. *Eur.J.Pharmacol.* 258:
39-45.

EcoReference No.: 84537\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Dennis, E. B. and Edwards, C. A. (1963). Phytotoxicity of Insecticides
and Acaricides. II. Flowers and Ornamentals. *Plant Pathol.* 12: 27-36.

EcoReference No.: 40669\
Chemical of Concern: MLN,DMT,PRN,DZ,DLD,AND,DDT,FLAC,NCTN,PPHD,ETN;
[Habitat]{.underline}: T; [Effect Codes]{.underline}: PHY; [Rejection
Code]{.underline}: NO ENDPOINT,CONTROL(ALL CHEMS).

DeNoble, V. J., Dragan, Y. P., and Carron, L. (1982). Behavioral Effects
of Intraventricularly Administered (\--)-Nicotine on Fixed Ratio
Schedules of Food Presentation in Rats. *Psychopharmacology* 77:
317-321.

EcoReference No.: 84514\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: NO
ENDPOINT,CONTROL(NCTN).

Dominiak, P., Fuchs, G., Von Toth, S., and Grobecker, H. (1985). Effects
of Nicotine and Its Major Metabolites on Blood Pressure in Anaesthetized
Rats. *Klin.Wochenschr.* 63: 90-92.

EcoReference No.: 84918\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO
CONTROL,ENDPOINT(NCTN).

Elbert, A. and Nauen, R. ( 2000). Resistance of Bemisia tabaci
(Homoptera: Aleyrodidae) to Insecticides in Southern Spain with Special
Reference to Neonicotinoids. *Pest Manag.Sci.* 56: 60-64.

EcoReference No.: 63840\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Rejection
Code]{.underline}: TARGET(NCTN).

Faraday, M. M., Elliott, B. M., Phillips, J. M., and Grunberg, N. E.
(2003). Adolescent and Adult Male Rats Differ in Sensitivity to
Nicotine\'s Activity Effects. *Pharmacol.Biochem.Behav.* 74: 917-931.

EcoReference No.: 85084\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

File, S. E., Cheeta, S., and Kenny, P. J. (2000). Neurobiological
Mechanisms by Which Nicotine Mediates Different Types of Anxiety.
*Eur.J.Pharmacol.* 393: 231-236.

EcoReference No.: 84747\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Foster, S. P., Denholm, I., and Thompson, R. (2003). Variation in
Response to Neonicotinoid Insecticides in Peach-Potato Aphids, Myzus
persicae (Hemiptera: Aphididae). *Pest Manag.Sci.* 59: 166-173.

EcoReference No.: 82050\
Chemical of Concern: IMC,ACT,NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: MORT,BEH,POP; [Rejection Code]{.underline}:
TARGET(NCTN).

Furvya, T., Kojima, H., and Syono, K. (1971). Regulation of Nicotine
Biosynthesis by Auxins in Tobacco Callus Tissues. *Phytochem* 10:
1529-1532.

EcoReference No.: 30201\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Giri, B. S. and Chowdary, Y. (1992). Inhibition of Nuclear Migration by
Nicotine and Its Reversion by Magnesium. *Indian J.Exp.Biol.* 30:
644-645.

EcoReference No.: 83890\
Chemical of Concern: NCTN,Mg; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: POP; [Rejection Code]{.underline}: NO ENDPOINT(ALL
CHEMS).

Gurkalo, V. K. and Volfson, N. I. (1982). Nicotine Influence upon the
Development of Experimental Stomach Tumors. *Arch.Geschwulstforsch.* 52:
259-265.

EcoReference No.: 84022\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: MOR,PHY; [Rejection Code]{.underline}: NO
MIXTURE(NCTN).

Halm, M. P., Chichery, M. P., and Chichery, R. (2002). The Role of
Cholinergic Networks of the Anterior Basal and Inferior Frontal Lobes in
the Predatory Behaviour of Sepia officinalis. *Comp.Biochem.Physiol.A*
132: 267-274.

EcoReference No.: 83786\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: NO ENDPOINT(ALL
CHEMS).

Ingole, I. V., Ghosh, S. K., and Anbalagan, J. (1994). The Effect of
Nicotine on the Cerebrum of Developing Chick Embryo: A Light Microscopic
Study. *J.Anat.Soc.India* 43: 143-148.

EcoReference No.: 84912\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,CEL; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Izumi, H. and Karita, K. ( 1993). Reflex Vasodilatation in the Cat Lip
Elicited by Stimulation of Nasal Mucosa by Chemical Irritants.
*Am.J.Physiol.* 265: R733-R738.

EcoReference No.: 84735\
Chemical of Concern: NCTN,CPS; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO
CONTROL,ENDPOINT(NCTN).

Koley, J., Majumder, C., Saha, J. K., and Koley, B. N. (1987). Gastric
Relaxation of Cardiac Origin Produced by Nicotine. *Med.Sci.Res.* 15:
1517-1518.

EcoReference No.: 85059\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Lang, M., Huemmer, B., and Hahn, H. L. (1988). Effect of Multiple
Applications of Nicotine on Mucus Secretion and on Circulatory and
Ventilatory Variables. *In: M.J.Rand and K.Thurau (Eds.), ISCU
(Int.Counc.of Sci.Unions) Press Symp.Ser., Vol.9, The Pharmacology of
Nicotine, Staellite Symp.of the 10th Int.Congr.of Pharmacol., Sept.4-6,
1987, Gold Coast, Queensland, Australia, Press Ltd., Oxford, England*
178-179.

EcoReference No.: 85259\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Lilius, H., Hastbacka, T., and Isomaa, B. (1995). A Comparison of the
Toxicity of 30 Reference Chemicals to Daphnia magna and Daphnia pulex.
*Environ.Toxicol.Chem.* 14: 2085-2088.

EcoReference No.: 16385\
Chemical of Concern: 24DXY,MLN,CuS,NCTN,PNB,As; [Habitat]{.underline}:
A; [Effect Codes]{.underline}: PHY,MOR; [Rejection Code]{.underline}: NO
CONTROL(ALL CHEMS),LITE EVAL CODED(OW-TRV-Cu).

Lilius, H., Isomaa, B., and Holmstrom, T. (1994). A Comparison of the
Toxicity of 50 Reference Chemicals to Freshly Isolated Rainbow Trout
Hepatocytes and Daphnia magna. *Aquat.Toxicol.* 30: 47-60.

EcoReference No.: 16756\
Chemical of Concern:
24DXY,HCCH,MLN,WFN,CF,Cu,CuS,PAQT,PL,SFL,LPS,PCP,CTC,BA,Hg,Ti,NCTN;
[Habitat]{.underline}: A; [Effect Codes]{.underline}: PHY,MOR,CEL;
[Rejection Code]{.underline}: NO CONTROL(ALL CHEMS).

Lind, R. J., Hardick, D. J., Blagbrough, I. S., Potter, B. V. L.,
Wolstenholme, A. J., Davies, A. R. L., Clough, M. S., Earley, F. G. P.,
Reynolds, S. E., and Wonnacott, S. (2001). \[3H\]-Methyllycaconitine: A
High Affinity Radioligand that Labels Invertebrate Nicotinic
Acetylcholine Receptors. *Insect Biochem.Mol.Biol.* 31: 533-542.

Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Rejection
Code]{.underline}: TARGET(NCTN).

Marchi, B., Trielli, F., Falugi, C., Corre, M. C., and Fenaux, L.
(1996). Cholinomimetic Drugs may Affect Growth and Metamorphosis of the
Sea Urchin Larva. *Oceanol.Acta* 19: 287-291.

EcoReference No.: 19060\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: GRO,MOR; [Rejection Code]{.underline}: NO
ENDPOINT(ALL CHEMS).

Maritz, G. S. and Woolward, K. (1992). Effect of Maternal Nicotine
Exposure on Neonatal Lung Elastic Tissue and Possible Consequences.
*S.A.M.J.(S.Afr.Med.J.)* 81: 517-519.

EcoReference No.: 85250\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL,GRO; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Marks, M. J., Miner, L., Burch, J. B., Fulker, D. W., and Collins, A. C.
(1984). A Diallel Analysis of Nicotine-Iduced Hypothermia.
*Pharmacol.Biochem.Behav.* 21: 953-959 .

EcoReference No.: 84617\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Marks, M. J., Romm, E., and Collins, A. C. (1987). Mouse Brain ATPase
Activities After Chronic Nicotine Infusion. *Biochem.Pharmacol.* 36:
3318-3320.

EcoReference No.: 84332\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Mikolajczyk, T., Chyb, J., Sokolowska-Mikolajczyk, M., Breton, B.,
Monden, K., and Epler, P. (1998). Nicotine Stimulated GtH2 Secretion In
Vivo in Male Common Carp (Cyprinus carpio); Potentiation of GnRH Action
and Possible Interaction with Dopaminergic System. *Aquat.Living
Resour.* 11: 155-161.

EcoReference No.: 83893\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: NO ENDPOINT(ALL
CHEMS).

Mikolajczyk, T., Epler, P., Sokolowska-Mikolajczyk, M., Breton, B.,
Chyb, J., and Cwioro, E. (1996). The Effects of Nicotine in Combination
with Antidopaminergic Drugs on the Maturational Gonadotropin (GTH2)
Secretion from Perifused Common Carp (Cyprinus carpio L.) Pituitary
Glands. *Pol.Arch.Hydrobiol.* 43: 373-378.

EcoReference No.: 84012\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Mori, K., Okumoto, T., Kawahara, N., and Ozoe, Y. (2001). Interaction of
Dinotefuran and Its Analogues with Nicotinic Acetylcholine Receptors of
Cockroach Nerve Cords. *Pest Manag.Sci.* 58: 190-196.

EcoReference No.: 69381\
Chemical of Concern: DNF,NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: MOR,CEL; [Rejection Code]{.underline}: NO
CONTROL(DNF),TARGET(NCTN).

Msolla, P., Mmbuji, W. E. O., and Kasuku, A. A. (1987). Field Control of
Bovine Parasitic Otitis. *Trop.Anim.Health Prod.* 19: 179-183.

EcoReference No.: 81199\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: POP; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN),COC(DBAC).

Nagata, M. and Osumi, Y. ( 1989). Dual Effects of
Intracerebroventricularly Applied Nicotine on Gastric Motility in Rats.
*In: 16th Annu.Meet.of the Experimental Ulcer Assoc.of Jpn., Dec.2,
1988, Kyoto, Japan, Scan.J.Gastroenterol.Suppl.* 24: 124-126 .

EcoReference No.: 83894\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO ENDPOINT(ALL
CHEMS).

Nakashima, T., Kubo, K., and Kurogochi, Y. (1990). Influence of Nicotine
on Allergic Cutaneous Reactions in Rats. *Res.Commun.Subst.Abuse* 11:
199-202.

EcoReference No.: 85641\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Nauen, R., Ebbinghaus-Kintscher, U., and Schmuck, R. (2001). Toxicity
and Nicotinic Acetylcholine Receptor Interaction of Imidacloprid and Its
Metabolites in Apis mellifera (Hymenoptera: Apidae). *Pest Manag.Sci.*
57: 577-586.

EcoReference No.: 62997\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Rejection
Code]{.underline}: TARGET(NCTN).

Nauen, R., Ebbinghaus, U., and Tietjen, K. (1999). Ligands of the
Nicotinic Acetylcholine Receptor as Insecticides. *Pestic.Sci.* 55:
608-610.

EcoReference No.: 66578\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Rejection
Code]{.underline}: TARGET(NCTN).

Nauen, R., Stumpf, N., and Elbert, A. (2002). Toxicological and
Mechanistic Studies on Neonicotinoid Cross Resistance in Q-Type Bemisia
tabaci (Hemiptera: Aleyrodidae). *Pest Manag.Sci.* 58: 868-875.

EcoReference No.: 69746\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Rejection
Code]{.underline}: TARGET(NCTN).

Ngah, W. Z. W., Jarien, Z., Rajikin, M. H., and Shamaan, N. A. (1991).
Effect of Nicotine on Glutathione Metabolizing Enzymes and Some
Conjugation Enzymes in the Rat. *Asia Pac.J.Pharmacol.* 6: 55-62.

EcoReference No.: 74292\
LITE Eval Status: NO COC(MLT)\
ECOTOX Status: UR\
Citation Source: TOXNET 4/04 (MLT) NCTN; [Effect Codes]{.underline}: T

Nishiuchi, Y. (1980). Toxicity of Formulated Pesticides to Fresh Water
Organisms LXXII. *The Aquiculture (Suisan Zoshoku)* 27: 238-244 (JPN).

EcoReference No.: 6701\
Chemical of Concern:
ACP,MOM,Naled,PPG,CPYM,AMZ,PPG,TVP,PIM,ES,FLAC,PHSL,NCTN,HPT,RTN,DDT,CHD,DLD;
[Habitat]{.underline} : A; [Effect Codes]{.underline}: MOR; [Rejection
Code]{.underline}: NO FOREIGN.

Nishiwaki, H., Nakagawa, Y., Kuwamura, M., Sato, K., Akamatsu, M.,
Matsuda, K., Komai, K., and Miyagawa, H. (2003). Correlations of the
Electrophysiological Activity of Neonicotinoids with Their Binding and
Insecticidal Activities. *Pest Manag.Sci.* 59: 1023-1030.

EcoReference No.: 83927\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY,MOR; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Perez de la Mora, M., Mendez-Franco, J., Salceda, R., Aguirre, J. A.,
and Fuxe, K. (1991). Neurochemical Effects of Nicotine on Glutamate and
GABA Mechanisms in the Rat Brain. *Acta Physiol.Scand.* 141: 241-250.

EcoReference No.: 85244\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM; [Rejection Code]{.underline}: NO COC(NCTN).

Picciotto, M. R., Zoli, M., Rimondin, R., Lena, C., Marubio, L. M.,
Pich, E. M., Fuxe, K., and Changeux, J. P. (1998). Acetylcholine
Receptors Containing the beta2 Subunit are Involved in the Reinforcing
Properties of Nicotine. *Nature (Lond.)* 391: 173-177.

EcoReference No.: 85103\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH,CEL; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Rauch, N. and Nauen, R. (2003). Identification of Biochemical Markers
Linked to Neonicotinoid Cross Resistance in Bemisia tabaci (Hemiptera:
Aleyrodidae). *Arch.Insect Biochem.Physiol.* 54: 165-176.

EcoReference No.: 82266\
Chemical of Concern: IMC,TMX,ACT,NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,MOR; [Rejection Code]{.underline}: TARGET(NCTN).

Rise, M., Prengler, M., and Arad, S. M. (1998). Characterization of a
Nicotine-Resistant Mutant of Chlorella emersonii. *J.Plant Physiol.*
152: 583-585.

EcoReference No.: 84460\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: CEL,BCM; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Romani, G., Pallini, S., and Beffagna, N. (1998). Down-Regulation of the
Plasmalemma H+ Pump Activity by Nicotineinduced Intracellular
Alkalinization: A Balance Between Base Accumulation, Biochemical pH-Stat
Response and Intracellular pH Increase. *Plant Cell Physiol.* 39:
169-176.

EcoReference No.: 85243\
Chemical of Concern: NCTN; [Habitat]{.underline}: A; [Effect
Codes]{.underline}: ACC; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Rosenbruch, M., Kniepen, J., Weishaupt, C., D.J.Benford, Blaauboer, B.
J., and Reinhardt, C. A. (1993). The Early Chick Embryo as a Model to
Evaluate Cardiovascular Effects of Adrenaline and Nicotine.
*Int.Workshop on In Vitro Toxicology, Oct.5-9, 1992, Domaine de Seillac,
France*.

EcoReference No.: 84919\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,PHY; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Schafer, E. W. (1972). The Acute Oral Toxicity of 369 Pesticidal,
Pharmaceutical and Other Chemicals to Wild Birds.
*Toxicol.Appl.Pharmacol.* 21: 315-330.

EcoReference No.: 38655\
Chemical of Concern:
Ziram,AN,BZO,BZC,Captan,THM,ZINEB,CYT,SFL,MAL,MRX,ACL,MLN,ABT,CBZ,MCB,CBL,CMPH,HCCH,EN,AND,ES,NP,TCF,CPY,DDVP,PPHD,DCTP,DS,PRT,DMT,AZ,PSM,ETN,DEM,DZ,FNTH,MP,NCTN;
[Habitat]{.underline}: T; [Effect Codes]{.underline}: MOR; [Rejection
Code]{.underline}: NO CONTROL(ALL CHEMS),NO COC(4AP).

Scott, G. C., Pickett, J. A., Smith, M. C., Woodcock, C. M., Harris, P.
G. W., Hammon, R. P., and Koetecha, H. D. (1984). Seed Treatments for
Controlling Slugs in Winter Wheat. *In: 1984 Br.Crop Prot.Conf.- Pests
and Dis.* 1: 133-138.

EcoReference No.: 79951\
Chemical of Concern:
DZM,RTN,MNK,ADC,MCB,BMN,NP,AMZ,CBF,TMP,BMNO,THO,DM,THM,DNB,NCTN;
[Habitat]{.underline}: T; [Effect Codes]{.underline}: MOR,PHY,POP;
[Rejection Code]{.underline}: LITE EVAL CODED(DZM),NO ENDPOINT(NCTN).

Sefcovic, P. and Hricova, D. (1972). Effect of Nicotine on Tissue
Cultures of Nicotiana Tabacum L. *Biologia* 27: 771-774.

EcoReference No.: 28445\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: POP,GRO; [Rejection Code]{.underline}: NO
ENDPOINT(ALL CHEMS).

Shah, P. V., Fisher, H. L., Sumler, M. R., Monroe, R. J., Chernoff, N.,
and Hall, L. L. (1987). Comparison of the Penetration of 14 Pesticides
Through the Skin of Young and Adult Rats. *J.Toxicol.Environ.Health* 21:
353-366.

EcoReference No.: 84377\
Chemical of Concern:
NCTN,ATZ,CAPTAN,CBL,CBF,CPY,DSMA,FOLPET,MSMA,PRN,PCB,PMR;
[Habitat]{.underline}: T; [Effect Codes]{.underline}: ACC; [Rejection
Code]{.underline}: NO CONTROL(ALL CHEMS).

Slawecki, C. J. and Ehlers, C. L. (2003). The Effects of
Corticotropin-Releasing Factor on the Cortical EEG are Reduced Following
Adolescent Nicotine Exposure. *Neuropeptides* 37: 66-73.

EcoReference No.: 84579\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: GRO,BCM; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Sobey, C. G., Dusting, G. J., and Woodman, O. L. (1989). Reflex
Epicardial Coronary Vasoconstriction Elicited by Nicotine in
Anesthetized Dogs. *Naunyn-Schmiedeberg\'s Arch.Pharmacol.* 339:
464-468.

EcoReference No.: 85174\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: PHY; [Rejection Code]{.underline}: NO CONTROL(NCTN).

Vadlamani, N. L., Pontani, R. B., and Misra, A. L. (1983). Effect of
Chronic Nicotine Pre-treatment on Phencyclidine (PCP) Disposition in the
Rat. *Arch.Int.Pharmacodyn.Ther.* 265: 4-12.

EcoReference No.: 84396\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BCM,BEH; [Rejection Code]{.underline}: NO
MIXTURE(NCTN).

Valand, V. M., Patel, J. R., and Patel, N. C. (1992). Bioefficacy of
Insecticides Against Citrus Leaf Miner Phyllocnistis citrella Stainton
on Kagzi Lime. *Indian J.Plant Prot.* 20: 212-214.

EcoReference No.: 83224\
Chemical of Concern: NCTN,DEM,ES,FVL,FPP,AZD,DMT; [Habitat]{.underline}:
T; [Effect Codes]{.underline}: POP; [Rejection Code]{.underline}:
OK(NCTN,DEM,ES,FVL,FPP,AZD,DMT,OK TARGET-NCTN),NO COC(MCPP1).

Verma, A. N., Sandhu, G. S., and Saramma, P. U. (1967). Relative
Efficacy of Different Insecticides as Contact Poisons to the Adults of
Singhara-d Beetle Galerucella-birmanica Coleoptera Chrysomellidae
trapa-bispinosa-d Mevinphos Carbaryl Bidrin Nicotine Sulfate Parathion
Diazinon Phosphamidon DDT Malathion te. *J.Res.Punjab Agric.Univ.* 4:
415-419.

EcoReference No.: 55198\
Chemical of Concern: CBL,DZ,MLN,NCTN; [Habitat]{.underline}: T;
[Rejection Code]{.underline}: TARGET(MLN,DZ,NCTN).

Wang, H., Cui, W. Y., and Liu, C. G. (1996). Characteristics of
Behavioral and Electroencephalogram. Convulsions Induced by Nicotine and
Its di(+)Tartrate. *Zhongguo Yaolixue Yu Dulixue Zazhi* 10: 169-172.

EcoReference No.: 85276\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: MOR,PHY; [Rejection Code]{.underline}: NO
CONTROL(NCTN).

Weidner, M., Martins, R., Mueller, A., Simon, J., and Schmitz, H.
(2005). Uptake, Transport and Accumulation of Nicotine by the Golden
Potho (Epipremnum aureum): The Central Role of Root Pressure. *J.Plant
Physiol.* 162: 139-150.

EcoReference No.: 85245\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: ACC; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

White, J. M. (1988). Behavioral Interactions Between Nicotine and
Caffeine. *Pharmacol.Biochem.Behav.* 29: 63-66.

EcoReference No.: 85040\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: BEH; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Whiting, A. G. and Murray, M. A. (1946). Histological Responses of Bean
Plants to Nicotine and to Wounding. *Bot.Gaz.* 108: 192-216.

EcoReference No.: 43155\
Chemical of Concern: NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: CEL; [Rejection Code]{.underline}: NO
ENDPOINT(NCTN).

Wong, J. Y. F., Ross, S. A., McColl, C., Massalas, J. S., Powney, E.,
Finkelstein, D. I., Clark, M., Horne, M. K., Berkovic, S. F., and Drago,
J. (2002). Proconvulsant-Induced Seizures in alpha4 Nicotinic
Acetylcholine Receptor Subunit Knockout Mice. *Neuropharmacology* 43:
55-64.

EcoReference No.: 83040\
Chemical of Concern: 4AP,NCTN; [Habitat]{.underline}: T; [Effect
Codes]{.underline}: MOR,PHY; [Rejection Code]{.underline}: NO
ENDPOINT,CONTROL(4AP).

**MRIDs**

**[Nicotine]{.underline}**

72-1       Acute Toxicity to Freshwater Fish

  ------------ ----------------------------------------------------------
  **MRID**     **Citation Reference**

               

  107188       McCann, J. (1971) ?Nicotine Alkaloid: Bluegill\|: Test No.
               382. (U.S. Agricultural Research Service, Pesticides
               Regulation Div., Animal Biology Laboratory; unpublished
               study; CDL:130313-A)
  ------------ ----------------------------------------------------------

81-1       Acute oral toxicity in rats

  ------------ ----------------------------------------------------------
  **MRID**     **Citation Reference**

               

  92216        Jasper, R.L. (1964) (Pharmacology Laboratory Reports on
               Insecti- cides for Use on Animals and Plants).
               (Compilation of reports by U.S. Pharmacology Laboratory;
               unpublished study; CDL: 102296-A)

  92128001     Kelley, J. (1990) Wilbur-Ellis Company Phase 3 Summary of
               MRID 00071037. Acute Oral Toxicity Study with Black Leaf
               40 in Male and Female Albino Rats: IBT Project No. A9166.:
               7 p.

  92128002     Kelley, J. (1990) Wilbur-Ellis Company Phase 3 Summary of
               MRID 00071038. Acute Oral Toxicity Study on Black Leaf 40
               in Albino Rats: IBT Project No. A8604.: 7 p.
  ------------ ----------------------------------------------------------

121-1       Phytotoxicity

  ------------ ----------------------------------------------------------
  **MRID**     **Citation Reference**

               

  75990        Carmel Chemical Corporation (1963) Greenhouse Insecticide
               Devel- opment: Project No. 100. (Unpublished study
               received Sep 25, 1963 under 5011-56; CDL:231159-A)
  ------------ ----------------------------------------------------------

[Tobacco Dust]{.underline}

71-1       Avian Single Dose Oral Toxicity

  ------------ ----------------------------------------------------------
  **MRID**     **Citation Reference**

               

  42625501     Pedersen, C.; Helsten, B. (1992) Tobacco Dust: 14-day
               Acute Oral LD50 Study in Bobwhite Quail: Lab Project
               Number: 131-001-03. Unpublished study prepared by Bio-Life
               Associates, Ltd. 41 p.
  ------------ ----------------------------------------------------------

72-1       Acute Toxicity to Freshwater Fish

  ------------ ----------------------------------------------------------
  **MRID**     **Citation Reference**

               

  42625503     Ward, T.; Boeri, R. (1992) Acute Toxicity of Tobacco Dust
               to the Rainbow Trout, Oncorhynchus mykiss. Unpublished
               study prepared by Resource Analysts, Inc. 15 p.
  ------------ ----------------------------------------------------------

72-2       Acute Toxicity to Freshwater Invertebrates

  ------------ ----------------------------------------------------------
  **MRID**     **Citation Reference**

               

  42625502     Ward, T.; Boeri, R. (1992) Acute Toxicity of Tobacco Dust
               to the Daphnid, Daphnia magna. Unpublished study prepared
               by Resource Analysts, Inc. 15 p.
  ------------ ----------------------------------------------------------

[^1]: These data have not been reviewed by the Agency.

[^2]: Memo from Anthony F. Maciorowski (EFED) to Kathryn Davis (SRRD).
    Subject: Nicotine Data Waiver Requests. D206472. Dated September 8,
    1994.

[^3]: Volunteer cancellation has been petitioned in May 2007 for a third
    nicotine product, Bonide Tobacco Dust (0.5% nicotine; product number
    4-340), which is not included in this assessment.

[^4]: For more information, see
    <http://iaspub.epa.gov/srs/srs_proc_qry.navigate?P_SUB_ID=159855>

[^5]: Since 1995, nicotine and nicotine salts have been listed in the
    Toxic Release Inventory (TRI) and is associated with disposal or
    release from the tobacco industry. In 2005, the major releases were
    associated with fugitive air emissions (total 20,038 lbs), point
    source air emissions (total 317,115 lbs), land treatment (93,204
    lbs), and surface water discharges (755 lbs). North Carolina and
    Virginia were the states with the highest amount disposed or
    released). Chemicals listed in the TRI must report to the Agency
    their release to the Agency on an annual basis.

[^6]: The EPI (Estimation Programs Interface) EPI Suite™ is a Windows^®^
    based suite of physical/chemical property and environmental fate
    estimation models developed by the EPA's Office of Pollution
    Prevention Toxics and Syracuse Research Corporation (SRC). EPI
    Suite^™^ uses a single input to run the following estimation models:
    KOWWIN™, AOPWIN™, HENRYWIN™, MPBPWIN™, BIOWIN™, BioHCWIN, PCKOCWIN™,
    WSKOWWIN™, WATERNT™, BCFWIN™, HYDROWIN™, KOAWIN and AEROWIN™, and
    the fate models STPWIN™, WVOLWIN™, and LEV3EPI™. EPI Suite is a
    screening level tool used to assess the environmental fate and
    exposure of a chemical in the environment. For more information, see
    **Appendix A**.

[^7]: Peterson, E.J., Choi, A., Dahan, D.S., Lester, H.A., and
    Dougherty, D.A. 2002. J. Am. Chem. Soc. A Perturbated pKa at the
    Binding Site of the Nicotinic Acetylcholine Receptor: Implications
    for Nicotine Binding, Vol. 124, pp.12662-12663.

[^8]: Seeman, J.I. 2005. Using "Basic Principles" to Understand Complex
    Science: Nicotine Smoke Chemistry and Literature Analogies. Journal
    of Chemical Education, Vol. 82 (10), pp 1577-1583.

[^9]: GENEEC is a Tier 1 simulation model that estimates the peak
    concentration which occurs on the day of the single large rainstorm
    event as well as multiple day averages over periods of 4, 21, 60 and
    90 days in a receiving water body (farm pond) of 1 ha, 2-m deep
    adjacent to a 10 ha treated site.. The magnitude of the peak
    concentration is the result of how fast the chemical dissipates
    (degradation; partitioning) in the field. The multiple day averages
    over periods of four to 90 days reflect the dissipation (such as
    degradation and partitioning) of the chemical that takes place in
    the water body. These peak and average concentrations are then
    compared with the appropriate toxicity tests for aquatic plants and
    animals.
    (<http://www.epa.gov/oppefed1/models/water/#geneec2>[)]{.underline}

[^10]: The EPI (Estimation Programs Interface) EPI Suite™ is a
    Windows^®^ based suite of physical/chemical property and
    environmental fate estimation models developed by the EPA's Office
    of Pollution Prevention Toxics and Syracuse Research Corporation
    (SRC). EPI Suite^™^ uses a single input to run the following
    estimation models: KOWWIN™, AOPWIN™, HENRYWIN™, MPBPWIN™, BIOWIN™,
    BioHCWIN, PCKOCWIN™, WSKOWWIN™, WATERNT™, BCFWIN™, HYDROWIN™, KOAWIN
    and AEROWIN™, and the fate models STPWIN™, WVOLWIN™, and LEV3EPI™.
    EPI Suite is a screening level tool used to assess the environmental
    fate and exposure of a chemical in the environment. For more
    information, see **Appendix A**.

[^11]: Memo from Anthony F. Maciorowski (EFED) to Kathryn Davis (SRRD).
    Subject: Nicotine Data Waiver Requests. D206472. Dated September 8,
    1994.

[^12]: Sangster, A.W. and Stuart, K.L. 1965. Ultraviolet Spectra of
    Alkaloids. Chen. Rev., Vol 29, pp.69-130.

[^13]: Lam, M.W., Young, C.J., Brain, R.A., Johnson, D.J, Hanson, M.A.,
    Wilson, C.J., Richards, S.M.,, Solomon, K.R., and Mabury, S.A. 2004.
    Env. Tox. Chem. Vol 23(6), pp .

[^14]: The Henderson-Hasselback equation is used to calculate the
    percent distribution of protonated and non-protonated in a dilute
    aqueous solution, where the input parameters are the pKa and the pH
    of the aqueous solution.

    Tobacco is not a dilute solution, and therefore, "Tobacco pH" is not
    used quantitatively to calculate the percent distribution of the
    protonated and non-protonated form of nicotine in a solid or
    heterogeneous matrix. Given that the source of nicotine is "tobacco
    dust" (as defined by the Agency) and the variability in nicotine
    content, the actual exposure of nicotine is uncertain. For this
    reason, the exposure assessment assumes that all of the nicotine in
    the tobacco dust becomes bioavailable and that the available
    nicotine is pure ("neat') nicotine.

[^15]: ^?^ In tobacco smoking, nicotine is transferred to air
    efficiently as a result of thermal conversion to the non-protonated
    form and at temperatures below those required for extensive thermal
    decomposition of the nicotine ring system. This transport is not
    fully explained by the pH of an aqueous extract of tobacco.

    Seeman, J.I. 2005. Using "Basic Principles" to Understand Complex
    Science: Nicotine Smoke Chemistry and Literature Analogies. Journal
    of Chemical Education, Vol. 82 (10), pp 1577-1583

[^16]: Hochstein, L.I. and Rittenberg, S.C 1959. The Bacterial Oxidation
    of Nicotine- I. Nicotine oxidation by cell-free preparations. The
    Journal of Biological Chemistry, Vol 234(1), pp 151-155.

    Hochstein, L.I. and Rittenberg, E.C. 1959. The Bacterial Oxidation
    of Nicotine- II. The isolation of the first oxidative product and
    its identification as (1)-6-hydroxynicotine. The Journal of
    Biological Chemistry, Vol 234(1), pp 156-160.

[^17]: ^?^ "Nicotine blue" is formed spontaneously from
    2,4,6-trihydroxypyridine (an identified metabolite in the catabolic
    pathway of nicotine) in the presence of oxygen.

    Baitsch, D., Sandu, Brandsch, R., and Igloi, G.L. 2001. Gene Cluster
    on pAO1 Of Arthobacter nicotinovorans involved in Degradation of the
    Plant Alkaloid Nicotine: Cloning, Purification, and Characterization
    of 2,6-Dihydroxypyridine 3-Hydroxylase. Journal of Bacteriology.
    Sept. 2001, pp.5262-5267.

    Mihasan M., Chiribau, C-B., Friedrich, T., Artenie, V., and
    Brandsch, R. 2007.. An NAD(P)H-nicotine blue oxidoreductase is part
    of the nicotine regulon and may protect Arthrobacter nicotinovorans
    from oxidative stress during nicotine catabolism Applied and
    Environmental Microbiology. Vol. 73 (April), pp.2479-2485.

[^18]: ^?^ Chiribau, C.B., Sandu, C., Fraaije, M., Schiltz, E., and
    Brandsch. 2004. A novel γ-N-methylaminobutyrate demethylating
    oxidase involved in catabolism of the tobacco alkaloid nitotine by
    Anthrobacter nicotinovorans pAO1. Eur. J. Biochem,Vol 271, pp
    4677-4684.

    Chiribau, C.B., Mihasan, M. Ganas., P., Igloi, G.L., Artenie, V.,
    and Brandasch, R. 2006. Final Steps in the catabolism of nicotine-
    Deamination versus demethylation of γ-N-methylaminobutyrate. FEBS
    Journal, Vol. 273, pp. 1528-1536.

[^19]: Bradley, P.M., Barber, L.B., Kolpin, D.W., McMahon, P.B., and
    Chapell, F.H. 2007. Biotransformation of Caffeine, Cotinine, and
    nicotine in stream sediments: Implications for use as wastewater
    indicators. Environmental Toxicology and Chemistry, Vol. 26 (6), pp.
    1116-1121.

[^20]: http://www.census.gov/main/www/cen2000.html

[^21]: Dated December 28, 2006

[^22]: ^?^ Kolpin, D.W., Furlong, E.T., Meyer, M.T. Thurman, E.M.,
    Zaugg, S.D., Barber, L.B., Buxton, H.T. 2002. Pharmaceuticals,
    Hormones, and Other Organic Wastewater Contaminants in U.S. Streams,
    1999-2000: a National Reconnaissance. Environ. Sci. Technol. Vol.
    36, pp 1202-1211.

    Lee, K.E., Barber, L.B., Furlong, E.T., Cahill, J.D., Kolpin, D.W.,
    Meyer, M.T., and Zaugg, S.D. 2004. Presence and distribution of
    organic wastewater compounds in wastewater, surface, ground, and
    drinking waters, Minnesota, 2000-2002: U.S.Geological Survey
    Scientific Investigation Report 2004-5138, 47pp.

    Carter, J.M., Delzer, G.C., Kingsbury, J.A., and Hopple, J.A. 2007.
    Concentration data for anthropogenic organic compounds in ground
    water, surface water, and finished water of selected community water
    systems in the United States, 2002-2005: U.S. Geological Survey Data
    Series 268 ,30 pp.

[^23]: The nicotine chemicals listed in the TRI are contained in the
    document entitled "Toxic Release Inventory- List of Toxic Chemicals
    within the Nicotine and Salts Category"; EPA 745-R-99-010.

[^24]: These data have not been formally reviewed by the Agency. Source:
    National Institute for Occupational Safety and Health.
    <http://www.cdc.gov/niosh/rtecs/qs501bd0.html>. Accessed July 9,
    2007.

[^25]: http://www.cdc.gov/niosh/rtecs/qs501bd0.html#T

[^26]: EFED Response to Request for New Uses for Imidacloprid, Updated
    Environmental Risk Characterization. September 18, 2001. DP
    Barcodes: D261135; D261168; D265457; D262132; D263672; D263728;
    D263343;D258313; D273584; D273582

[^27]: These data have not been reviewed by the Agency.

[^28]: Memo from Anthony F. Maciorowski (EFED) to Kathryn Davis (SRRD).
    Subject: Nicotine Data Waiver Requests. D206472. Dated September 8,
    1994.

[^29]:
    > Lyman, W.J., Reehl, W.F., and Rosenblatt, D.H. Handbook of
    > Chemical Property Estimation Methods, McGraw-Hill Books, New York,
    > 1982