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Significant New Alternatives Policy Program 

Solvent Cleaning Sector

Risk Screen on Substitute for CFC-113, Methyl Chloroform, and HCFC-225cc/cb

for Electronics, Precision, and Metals Cleaning

Substitute: Trans-1-chloro-3,3,3-trifluoroprop-1-ene (SolsticeTM 1233zd(E))

This risk screen does not contain Clean Air Act (CAA) Confidential Business Information (CBI) 

and, therefore, may be disclosed to the public.

1. INTRODUCTION

Ozone-depleting substances (ODS) are being phased out of production in response to a series of 

diplomatic and legislative efforts that have taken place over the past two decades, including the Montreal 

Protocol and the Clean Air Act Amendments of 1990 (CAAA). The U.S. Environmental Protection 

Agency (EPA), as authorized by Section 612 of the CAAA, administers the Significant New Alternatives 

Policy (SNAP) Program, which identifies acceptable and unacceptable substitutes for ODS in specific 

end-uses based on assessment of their health and environmental impacts. 

EPA’s decision on the acceptability of a substitute is based on the findings of a screening assessment of 

potential human health and environmental risks posed by the substitute in specific applications. EPA has 

already screened a large number of substitutes in many end-uses within all of the major ODS-using 

sectors, including refrigeration and air conditioning, solvent cleaning, foam blowing, aerosols, fire 

suppression, adhesives, coatings and inks, and sterilization. The results of these risk screens are presented 

in a series of Background Documents that are available in EPA's docket.

The purpose of this risk screen is to supplement EPA’s Background Document on the solvent sector (EPA 

1994) (hereinafter referred to as the Background Document). This risk screen evaluates the potential use 

of trans-1-chloro-3,3,3-trifluoroprop-1-ene (hereafter referred to as SolsticeTM 1233zd(E)) as a substitute 

for CFC-113, methyl chloroform (commonly also referred to as 1,1,1-trichloroethane, TCA, or MCF), and 

HCFC-225ca/cb, as a solvent for electronics, precision cleaning and metals cleaning (e.g., vapor 

degreasing and defluxing and flushing) end-uses . Table 1 presents information on the proposed 

substitute. 

Table 1. Composition of SolsticeTM 1233zd(E)

Constituent Chemical Formula CAS Number Percent of Total

(by weight)a

trans-1-Chloro-3,3,3-trifluoroprop-1-ene C3H2ClF3 102687-65-0 >99%

Potential Impurities (maximum concentration)
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The potential risks associated with use of substitutes as solvents have been examined at length in the 

Background Document. The reader is referred to this reference for a detailed discussion of the 

methodologies used to conduct this risk screen. This risk screen addresses flammability risk, and an 

occupational exposure assessment was performed to ensure that manufacture and disposal of the proposed 

substitute in the solvent end-use does not pose unacceptable risk to workers. Modeling was performed at 

the end-use to examine potential exposures for those using SolsticeTM 1233zd(E) for the intended 

application. Lastly, a general population exposure assessment was performed to ensure that the proposed 

substitute would not pose an unacceptable risk to the population at large. 

Section 2 of this report summarizes the results of the risk screen for the proposed substitute. The 

remainder of the report is organized into the following sections:

• Section 3: Atmospheric Assessment

• Section 4: Flammability Assessment

• Section 5: Potential Health Effects

• Section 6: Occupational Exposure Assessment

• Section 7: End-Use Exposure Assessment

• Section 8: General Population Exposure Assessment

• Section 9: Volatile Organic Compound Assessment 

• Section 10: References

2. SUMMARY OF RESULTS

SolsticeTM 1233zd(E) is recommended for SNAP approval as a solvent in the electronics, precision, and 

metals cleaning end-uses. EPA's risk screen indicates that the use of the proposed substitute will be less 

harmful to the atmosphere than the continued use of CFC-113, MCF, and HCFC-225ca/cb. Given that 

proper training will be given to personnel; appropriate personal protective equipment (PPE) (e.g., 

respiratory protection, goggles, impervious gloves, chemical-resistant clothing and boots) according to 

Section VIII of the Occupational Safety & Health Administration (OSHA) Technical Manual (OSHA 

1994) are available; and proper procedures according to the MSDS (including provisions of adequate 

ventilation) will be used during manufacturing, end-use, and disposal, no significant toxicity risk to 

workers, end-users, or the general population is anticipated. In addition, based on characteristics of the 

proposed substitute and precautions listed in the MSDS for SolsticeTM 1233zd(E), the proposed substitute 

is not expected to present a significant flammability concern during manufacture or end-use.

3. ATMOSPHERIC ASSESSMENT

This section presents an assessment of the potential risks to atmospheric integrity posed by the use of 

SolsticeTM 1233zd(E) in the solvent sector. The ozone depletion potential (ODP), global warming 

potential (GWP), and atmospheric lifetime (ALT) of the proposed substitute are presented in Table 2. As 

compared to CFC-113, MCF, and HCFC-225ca/cb, SolsticeTM 1233zd(E) is substantially less harmful to 

the ozone layer, has less climate impact, and a shorter atmospheric lifetime. SolsticeTM 1233zd(E) does 

however, have a non-zero ODP, but at 0.00024 to 0.00034, this value is very small when considering 

continued used of HCFC-225ca/cb. A recent report by Wang et al. (2011) modeled the potential of 

SolsticeTM 1233zd(E) to affect the amount of ozone in the global atmosphere. This report found that the 

release of all SolsticeTM 1233zd(E), assuming it were substituted for all compounds it might replace and 

that the entire amount used were released, would have a statistically zero impact on global atmospheric 

ozone (less than 0.01% change in total column ozone). Thus, EPA believes that use of SolsticeTM 

1233zd(E) would result in substantially less harm to the climate and ozone layer than the continued use of 

ozone-depleting substances. 
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Table 2. Atmospheric Impacts of SolsticeTM 1233zd(E) Compared to Other Solvents

Refrigerant ODP GWP ALT

SolsticeTM 1233zd(E) 0.00024-0.00034a,c 4.7-7a,b 26 daysc

CFC-113 0.85d 6,130e 85 yearsd

HCFC-225ca 0.02d 122e 1.9 yearsd

HCFC-225cb 0.03d 595e 5.9 yearsd

Methyl Chloroform 0.16d 146e 5 yearsd

a Assuming emissions occur in the major regions of the world where blowing agents that SolsticeTM 1233zd(E) might replace are 

significantly used. Wang et al. (undated).

b Wang et. al. (2011)

c SolsticeTM 1233zd(E) SNAP Submission (Anonymous Submitter 2011b) d WMO (2011)

e IPCC 4th Assessment Report (Forster et al. 2007)

4. FLAMMABILITY ASSESSMENT

SolsticeTM 1233zd(E) is not flammable at ambient temperatures and atmospheric pressure, but can ignite 

when mixed with air under pressure and exposed to strong ignition sources (Anonymous Submitter 

2011b). In accordance with the MSDS, SolsticeTM 1233zd(E) is to be kept away from all sources of 

ignition and high heat. Because proper precautions specified by the proposed substitute’s MSDS are 

followed, SolsticeTM 1233zd(E) does not pose a significant flammability risk.

5. POTENTIAL HEALTH EFFECTS

To assess potential health risks from exposure to this proposed substitute, the submitter developed an 

acceptable exposure limit (AEL) of 300 ppm1 for SolsticeTM 1233zd(E) (Anonymous Submitter 2012). 

The AEL represents the maximum 8-hour time weighted average (TWA) at which a worker can be 

exposed regularly without adverse effects. EPA also developed a short-term exposure limit (STEL) of 

900 ppm over a 15-minute period, based on the submitter’s 300 ppm AEL value.2

According to the MSDS, exposure to SolsticeTM 1233zd(E) may be harmful by ocular or dermal 

absorption, inhalation, or ingestion. Exposures of SolsticeTM 1233zd(E) to the eyes will cause serious eye 

irritation and may cause frostbite. In case of ocular exposure, the MSDS for SolsticeTM 1233zd(E)

recommends that person(s) should immediately flush the eyes, including under the eyelids, with water for 

15 minutes; should frostbite occur, the water should be lukewarm, not hot. Medical attention should be 

sought if irritation develops or persists. Exposures of SolsticeTM 1233zd(E) to the skin may cause 

irritation or frostbite. In the case of dermal exposure, the MSDS for SolsticeTM 1233zd(E) recommends 

that person(s) should immediately wash the affected area with water and remove all contaminated 

clothing; should frostbite occur, person(s) should bathe (not rub) the affected area with lukewarm, not hot, 

water. If water is not available, cover the affected area with a clean, soft cloth. Medical attention should 

be sought if irritation develops or persists. SolsticeTM 1233zd(E) vapors are heavier than air and can cause 

suffocation by reducing oxygen available for breathing, causing asphyxiation at high concentrations. If 

person(s) are exposed to high concentrations, the person(s) will likely not realize that he/she is 

suffocating, but may experience central nervous system effects, such as drowsiness and dizziness. If 

SolsticeTM 1233zd(E) is inhaled, person(s) should be immediately removed and exposed to fresh air. The 

MSDS recommends that if breathing is difficult, person(s) should be given oxygen, provided a qualified 

 1

An AEL of 1,000 ppm was independently developed for SolsticeTM 1233zd(E) based on review of available toxicity studies (see 

Appendix A). However, as the AEL value of 300 ppm determined by the submitter is more conservative, for purposes of this risk 

screen, 300 ppm was used to evaluate the potential exposures.

2 STEL was estimated as three to five times the AEL, which is an established method of estimating a short-term exposure by the 

American Conference of Governmental Industrial Hygienists. [STEL] = [AEL] x 3 = 300 ppm x 3 = 900 ppm.
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operator is present, and medical attention be sought. SolsticeTM 1233zd(E) is not likely to be hazardous 

by ingestion; however, in case of ingestion, the MSDS recommends providing the person(s) with a cup of 

water, if fully conscious, consulting a physician immediately, and to not induce vomiting without medical 

advice. EPA’s review of the human health impacts of this proposed substitute is contained in the public 

docket for this decision. 

When good industrial hygiene practices and the PPE and engineering control (e.g., ventilation) 

recommendations outlined in the MSDS for SolsticeTM 1233zd(E) , as summarized in Sections 6 and 7 of 

this risk screen, are followed, these potential health effects are unlikely to occur. 

6. OCCUPATIONAL EXPOSURE ASSESSMENT AT MANUFACTURE

Occupational exposure to SolsticeTM 1233zd(E) may occur during manufacture of the proposed substitute 

as a solvent. For electronics and precision cleaning, SolsticeTM 1233zd(E) will be used primarily for 

degreasing carried out in an enclosed vapor degreaser. For metals cleaning, SolsticeTM 1233zd(E) is 

expected to be used as a flushing agent, and may also be used for degreasing. Flushing solvent can be 

dispensed from a self-contained unit (e.g., an aerosol can), pressurized cylinder, or tank. As indicated by 

the submitter, occupational exposure will vary depending on the type of equipment being used, nature of 

the operation, and other factors such as the degree of ventilation. 

For use of SolsticeTM1233zd(E) as a degreasing solvent in enclosed vapor degreasing units, occupational 

exposure during manufacture is not expected, as filling occurs at the end-use, as summarized in Section 7. 

The submitter indicates that aerosol can filling operations, including those for use of SolsticeTM 

1233zd(E) as a flushing agent, are highly automated and involve little or no direct contact with the 

components charged into the can. Actual monitoring data for manufacture of SolsticeTM 1233zd(E)

aerosol cans are not available; however, the submitter measured the loss rate of SolsticeTM 1233zd(E)

from an open (i.e., uncapped) aerosol can at room temperature to be 0.5 grams per minute and that a 

medium-speed aerosol line fills between 150 to 250 cans per minute.

Concentrations are expected to be well below the AEL (300 ppm) for SolsticeTM 1233zd(E), and the 

manufacturing process is highly automated with minimal emissions anticipated. Thus, exposure during 

the manufacture of aerosol cans for flushing is not expected to present a significant toxicity concern to 

workers. See the risk screen for SolsticeTM 1233zd(E) in aerosol solvents for more detailed information 

(document EPA-HQ-OAR-2003-0118-0316).

For engineering control and PPE recommendations, the proposed substitute’s MSDS and OSHA 

guidelines (OSHA 1994) should be referenced. Adequate ventilation should always be established during 

any handling, or storage of SolsticeTM 1233zd(E). Engineering controls should include vapor-in air 

detection systems and local exhaust ventilation during use of SolsticeTM 1233zd(E), and filling operations 

for vapor degreasing units are to occur only at stations with exhaust ventilation facilities. In addition, an 

eye wash and safety shower should be near the manufacturing facility and ready for use. In general, use of 

PPE is recommended, such as respiratory protection (a NIOSH-approved positive-pressure supplied-air 

respirator is recommended where there is insufficient ventilation; self-contained breathing apparatuses 

should be used for rescue and maintenance work in storage tanks), chemical splash goggles or face shield,

impervious gloves, and chemical-resistant boots and clothing consistent with OSHA guidelines

(Anonymous Submitter 2011b, OSHA 1994). Contaminated clothing is not to leave the workplace and 

must be cleaned before reuse (Anonymous Submitter 2011b). Because these PPE and ventilation 

requirements are followed, the risk of exposure to SolsticeTM 1233zd(E) is unlikely to pose a risk to 

workers during manufacture.
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In addition, there is a risk of generation of toxic decomposition products such as hydrogen fluoride, 

hydrogen chloride, carbonyl halides, carbon monoxide, and carbon dioxide when SolsticeTM 1233zd(E) is 

exposed to fire. Containers of SolsticeTM 1233zd(E) should not be allowed to contact open flames, 

glowing metal surfaces, or electrical heating elements. Because proper handling and disposal guidelines 

are followed in accordance with good industrial hygiene and manufacturing practices and the MSDS for 

SolsticeTM 1233zd(E), there is no significant risk to workers during the manufacture of SolsticeTM 

1233zd(E).

7. END-USE EXPOSURE ASSESSMENT

SolsticeTM 1233zd(E) is proposed for use as a solvent for electronics, precision, and metals cleaning enduses; SolsticeTM 1233zd(E) is not intended to be used for residential or direct consumer use. For 

electronics and precision cleaning, SolsticeTM 1233zd(E) will be used for degreasing, which will typically 

be performed in an enclosed vapor degreaser, rather than an open-top vapor degreaser. A degreaser is a 

rectangular unit containing one or more sumps which are filled with solvent. One of the sumps is heated 

to boil the solvent and generate a layer of solvent vapor above the liquid. Degreasers are fitted with 

cooling coils around the upper perimeter of the sidewalls to condense solvent vapors, maintain the vapor 

zone, and minimize emissions. Dirty parts are immersed in the liquid sumps and then raised into the vapor 

zone where solvent vapors give a final rinse before the parts are removed from the unit. In flushing 

applications for metals and precision cleaning, solvent is forced through a part or device to remove 

contaminants. SolsticeTM 1233zd(E) will be most likely be used to flush refrigeration systems and gauges, 

lines, and regulators of oxygen-handling equipment. 

For use of SolsticeTM 1233zd(E) as a degreasing solvent in enclosed vapor degreasing units, occupational 

exposure during manufacture will be limited to the filling of containers of solvent and filling of 

degreasing equipment by the end user. The submitter anticipates that there will be very low emissions 

from these operations. In addition, the submitter and a major degreaser manufacturer have developed a 

system for transferring solvent directly into the solvent cleaning equipment that keeps the solvent 

enclosed by using hoses and a valve to control transfer of the solvent from the storage container. This will 

minimize emissions, compared to opening the solvent container.

Note that this substitute boils at 19 °C and at room temperature. Therefore, EPA recommends using this 

substitute in equipment designed to minimize solvent losses, emissions and worker exposure. Examples 

of such equipment include containers with connected hoses and valves that allow for direct transfer of the 

solvent to cleaning equipment without opening of the storage container, and enclosed or low-emission 

cleaning equipment.

A surrogate compound, HFC-245fa, was used to assess the potential end-use exposures to SolsticeTM 

1233zd(E). Air concentrations found during the air monitoring studies with HFC-245fa (described below 

in sections 7.1 and 7.2) are a reasonable estimate of potential air concentrations of SolsticeTM 1233zd(E), 

due to similar boiling points of the two compounds and the lower vapor pressure of SolsticeTM 

1233zd(E).Therefore, EPA believes that this is a reasonable surrogate compound to use for this 

assessment that will result in protective estimates of exposure. 

Table 3. Boiling Point and Vapor Pressure for SolsticeTM 1233zd(E) and HFC-245fa Analog

Parameter SolsticeTM 1233zd(E) Proposed 

Substitute HFC-245fa Analog

Boiling Point 19 °C (66 °F) 15.3 °C (59.5 °F)

Vapor Pressure 205 kPa (at 54 °C) 245 kPa (at 54 °C)

Source: Anonymous Submitter 2011a, Anonymous Submitter 2011c
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7.1 Degreasing

For this degreasing exposure analysis, EPA reviewed the submitter’s exposure assessment, which 

monitored the loss of solvent from a low-emission open-top vapor degreaser containing HFC-245fa run 

continuously for ten days. The loss rate was estimated to be 342 mL of HFC-245fa per 24-hour day, 

which is equivalent to approximately 147 grams of HFC-245fa, released every 8 hours (equivalent to one 

worker’s shift). Vapor degreasers are often run on a continual basis to minimize emissions in facilities 

that frequently use degreasers. The submitter assumed the same loss rate (147 grams of SolsticeTM 

1233zd(E) every 8-hours), which is equivalent to an exposure of less than 8 ppm on an 8-hr TWA, 

assuming the degreasing takes place in a 2000 ft3 workshop (20x10x10 feet) with a ventilation rate of 6 

ACH.3 As this exposure concentration is substantially lower than the submitter’s AEL value of 300 ppm, 

use of SolsticeTM 1233zd(E) in electronics and precision cleaning does not pose a significant risk to endusers.

7.2 Flushing

For flushing operations, the submitter administered two separate monitoring tests using HFC-245fa as a 

flushing agent. In the first assessment, a service technician flushed an automobile refrigerant condenser 

with 2 pounds of HFC-245fa. The process was repeated 4 times over a period of 80 minutes. Ventilation 

in the 20x30x8 foot laboratory was limited to the normal building air supply (i.e., no additional 

ventilation was provided4

). The submitter assumed that the losses of HFC-245fa are equivalent to the 

anticipated emissions for SolsticeTM 1233zd(E), so under this first exposure assessment, the timeweighted average exposure level for SolsticeTM 1233zd(E) was estimated to be approximately 84 ppm

over 80 minutes, or 14 ppm on an 8-hr TWA.

In the second monitoring test, two operators flushed 10 automobile air conditioning units over the course 

of 5 hours with 2 pounds each of HFC-245fa. Ventilation in the 50x100x20 feet garage was limited to the 

normal automobile garage air supply (i.e., no additional ventilation was provided5

).Under this second 

assessment, the time-weighted average exposure levels for SolsticeTM 1233zd(E) were estimated to range 

from 80 to 97 ppm over 5 hours, or 50 to 61 ppm on an 8-hr TWA, assuming the same losses as with 

HFC-245fa. 

Based on the two HFC-245fa monitoring test results, the anticipated occupational exposure levels for 

SolsticeTM 1233zd(E) would be significantly lower than its 300 ppm AEL. Because these exposures are 

under the AEL, and end-users of SolsticeTM 1233zd(E) are subject to following certain guidelinesincluding training to properly use and handle the proposed substitute, that employee work habits are 

conducted in accordance with good industrial hygiene practices, and that the solvent is only used in areas 

with adequate ventilation or local exhaust ventilation, EPA believes that SolsticeTM 1233zd(E) does not 

pose a significant concern to workers at end-use.

8. GENERAL POPULATION EXPOSURE ASSESSMENT

This section screens potential risks to the general population from exposure to releases of SolsticeTM 

1233zd(E) to ambient air, surface water, and solid waste. SolsticeTM 1233zd(E) is not expected to cause 

significant risks to the environment and human health in the general population when manufactured or 

used for solvent cleaning end-uses.

 3 Air change rates for factories vary, but recommended levels range from 6-15 ACH (Anonymous Submitter 2011a). 4 According to ASHRAE 62: Ventilation for Acceptable Indoor Air Quality, laboratories must have a minimum ventilation rate 

of 0.18 cubic feet per minute (cfm) per square feet of space. For a 20x30x8 foot lab, the ventilation rate should be at least 1.4

ACH

5 According to ASHRAE 62, automobile service garages must have a minimum ventilation rate of 1.5 cfm/ft2

. For a 50x100x20 

foot garage, the ventilation rate should be approximately 90 ACH.
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8.1 Ambient Air

Electronics, precision, and metals cleaning are emissive end-uses. As such, it is important to minimize 

emissions to ambient air when possible. However, significant releases to air are not expected, because 

accidental release measures, handling and storage, engineering controls, and waste disposal are followed 

per the proposed substitute’s MSDS. Other releases to the air during electronics, precision, and metals 

cleaning end-uses are expected to occur indoors with good manufacturing practices, such as employment 

of capture and destruction technologies, and therefore not expected to cause exposure to the general 

population.

8.2 Surface Water

As indicated by the submitter, SolsticeTM 1233zd(E) has only slight solubility in water (1.9 g/l) and use of 

SolsticeTM 1233zd(E) as a solvent is not anticipated to result in releases to surface water. At manufacture, 

the solvent will be handled indoors with waste or accidental spills collected with a non-combustible 

absorbent material, such as sand or vermiculite. Vapor degreasers are fitted with water separators so that 

water and SolsticeTM 1233zd(E) can be separated after the solvent is condensed. The volume of SolsticeTM 

1233zd(E) remaining in the water after separation is low. In addition, disposal of SolsticeTM 1233zd(E) is 

subject to local, state, and federal regulations, which ensure that releases of SolsticeTM 1233zd(E) and 

water contaminated with SolsticeTM 1233zd(E) are not be dumped into sewers, on the ground, or into any 

body of water, but rather taken to a wastewater treatment facility or disposed of properly. 

8.3 Solid Waste

Industrial sites using SolsticeTM 1233zd(E) as a solvent in electronics, precision, and metals cleaning can 

collect spent solvent for disposal or reclaim the spent solvent through distillation. To minimize waste, 

EPA recommends that reclamation of the proposed substitute through distillation be performed to the 

extent feasible. Residuals from the distillation process and spent solvent collected in containers (and 

therefore not collected as wastewater and sent to a wastewater treatment facility) are generally considered 

to be a RCRA hazardous waste (40 CFR Part 261.31). Although the constituents of SolsticeTM 1233zd(E)

are not listed as RCRA hazardous wastes, applications of SolsticeTM 1233zd(E) may generate hazardous 

wastes. Solvents used for electronics, precision, and metals cleaning end-uses generally pick up metals 

and other chemical compounds during the process, which causes the spent solvent to display 

characteristics of hazardous waste (ignitability, corrosivity, reactivity, or toxicity); any solid waste 

displaying these characteristics or having a specific waste code (e.g., F001) is considered to be a 

hazardous waste (40 CFR Part 261). Waste regulated as RCRA hazardous wastes is subject to the 

requirements of the Subtitle C program (including storage, treatment, and disposal requirements), which 

EPA believes are sufficient to control human health and environmental risks. In addition, the SolsticeTM 

1233zd(E) MSDS recommends that solid materials used during end-use or clean-up procedures that may 

be absorbed with SolsticeTM 1233zd(E) (e.g., rags, paper towels, non-combustible absorbent material) are 

disposed of in accordance with federal, state, and local environmental regulations.

9. VOLATILE ORGANIC COMPOUND (VOC) ASSESSMENT

EPA has proposed to exempt SolsticeTM 1233zd(E) as a volatile organic compound (VOC) under the 

CAA (40 CFR 51.100(s)), but this determination is not yet final. Thus, emissions of SolsticeTM 

1233zd(E) should be controlled. SolsticeTM 1233zd(E) continues to be regulated as a VOC for purposes 

of regulations in state implementation plans for local air quality unless and until EPA issues a final rule 

exempting it. 

An assessment was performed to compare the annual VOC emissions from use of SolsticeTM 1233zd(E)

in the solvent cleaning end-uses in one year to other anthropogenic sources of VOC emissions. Because a 

breakdown of production by end-use was not provided by the submitter, this assessment assumes that 163

MT of SolsticeTM 1233zd(E), the maximum production for all solvent end-uses (i.e., aerosol solvent; 
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metals, electronics and precision cleaning; and adhesives, coatings, and inks) is released. This quantity is

approximately equal to 7.0 x 10-3 percent of the VOC emissions from the solvent sector in the United 

States,6 or approximately 1.3 x 10-3 percent of all anthropogenic VOC emissions in the U.S.7

For solvent cleaning applications that take place in less-controlled environments, such as a fume hood or 

other ventilated areas which would vent the solvent to the atmosphere, an emissions rate of 74%8 was 

applied to the entire intended U.S. annual production of SolsticeTM 1233zd(E) for solvent use in order to 

estimate the maximum impact on VOC emissions across this end-use. Assuming 74% of the submitter’s 

intended annual production is entirely released, approximately 121 MT of VOCs would be emitted, which 

is approximately equal to 5.2 x 10-3 percent of the VOC emissions from the solvent sector in the United 

States, or only approximately 9.9 x 10-4 percent of all anthropogenic VOC emissions in the U.S.

Although these VOC emissions for the metals, electronics and precision cleaning end-uses are already 

several orders of magnitude lower than other anthropogenic emissions, it is likely that emissions would be

even lower than estimated here, as this assessment uses very conservative assumptions. First, the amount 

of SolsticeTM 1233zd(E) produced encompasses the production for other end-uses, included adhesives, 

coatings and aerosol solvents. Secondly, it assumes that all applications of SolsticeTM 1233zd(E) in the

metals, electronics and precision cleaning end-uses occur in a less-controlled environment, where 74% of 

production would be emitted to the atmosphere. However, much of the use of SolsticeTM 1233zd(E) is 

anticipated to occur indoors, where emissions would be less than 10%. Further, these emissions could be

further reduced, and in some facilities may already be so reduced. For example, capture and destruction 

technologies for fugitive emissions are employed in larger facilities, and hence emissions during use of 

SolsticeTM 1233zd(E) may be even less than 10%.

In addition, the majority of solvent use of ODS substitutes (approximately 69%), including aerosol 

solvents, adhesives and coatings, and solvent cleaning, is made up of solvents that are considered to be 

VOCs, such as trichloroethylene and a number of other non-halogenated organic solvents (ICF 2012b). 

Because SolsticeTM 1233zd(E) has a small intended production for solvent uses compared to the size of all 

solvent uses, emissions from SolsticeTM 1233zd(E) for its intended end-uses would not have a significant 

impact on VOC emissions compared to other alternatives and compared to VOC emissions. Further, 

emissions from SolsticeTM 1233zd(E) for its intended end-uses would not have a significant impact on 

total VOC emissions from all solvents, including solvent cleaning.

 6 Based on the 2010 annual VOC emissions for solvent utilization for the U.S. (EPA 2011). The solvent utilization sector includes 

consumer and commercial solvent use, degreasing, dry cleaning, graphic arts, industrial surface coating and solvent use, and nonindustrial surface coating.

7 Based on the 2010 annual total VOC emissions for the U.S. (EPA 2011). 8 Leak rate for non-medical aerosols from EPA’s Vintaging Model (VM IO file_v4.4_11.17.11).
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10. REFERENCES

American Society of Heating, Refrigeration, and Air-Conditioning Engineers, Inc. (ASHRAE). 2007. 

Standard 62.1-2007: Ventilation for Acceptable Indoor Air Quality. 

Anonymous Submitter. 2012. SolsticeTM 1233zd(E) Material Safety Data Sheet. February 28, 2012.

Anonymous Submitter. 2011a. Response to Incomplete SNAP Submission for SolsticeTM 1233zd(E). 

October 20, 2011.

Anonymous Submitter. 2011b. SNAP Submission to EPA for SolsticeTM 1233zd(E). May 13, 2011. 

Anonymous Submitter. 2011c. Next Generation (LGWP) of Blowing Agents for Spray Foam 

Applications. 

EPA 2011. National Emissions Inventory Air Pollutant Emissions Trends Data. Accessed 23 December 

2011. Updated October 2011. Available at: 

<http://www.epa.gov/ttn/chief/trends/index.html#tables>

EPA 1994. Significant New Alternatives Policy Technical Background Document: Risk Screen on the 

Use of Substitutes for Class I Ozone-depleting Substances: Refrigeration and Air Conditioning. 

Stratospheric Protection Division. March, 1994.

Forster, P., V. Ramaswamy, P. Artaxo, T. Berntsen, R. Betts, D.W. Fahey, J. Haywood, J. Lean, D.C. 

Lowe, G. Myhre, J. Nganga, R. Prinn, G. Raga, M. Schulz and R. Van Dorland. 2007. Changes 

in Atmospheric Constituents and in Radiative Forcing. In: Climate Change 2007:The Physical 

Science Basis. Contribution of Working Group I to the Fourth Assessment Report of the 

Intergovernmental Panel on Climate Change [Solomon, S., D. Qin, M. Manning, Z. Chen, M. 

Marquis, K.B. Averyt, M.Tignor and H.L. Miller (eds.)]. Cambridge University Press, 

Cambridge, United Kingdom and New York, NY, USA.

ICF International. 2012a. Determination of an Acceptable Exposure Limit (AEL) for SolsticeTM 

1233zd(E). Prepared for U.S. Environmental Protection Agency. January 23, 2012.

ICF International. 2012b. Solvent Sector VOC Analysis for ODS Substitutes. Prepared for EPA’s 

Stratospheric Ozone Protection Division. March 9, 2012

OSHA. 1994. General description and discussion of the levels of protection and protective gear. 1910.120 

App B. Last updated August 22, 1994. Available at: 

http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_table=STANDARDS&p_id=9767

Wang D., Olsen S., Wuebbles D. Undated. “Three-Dimensional Model Evaluation of the Global 

Warming Potentials for tCFP.” Department of Atmospheric Sciences. University of Illinois, 

Urbana, IL. Draft Report. 

Wang D., Olsen S., Wuebbles D. 2011. “Preliminary Report: Analyses of tCFP’s Potential Impact on 

Atmospheric Ozone.” Department of Atmospheric Sciences. University of Illinois, Urbana, IL. 

September 26, 2011. 

WMO (World Meteorological Organization), 2011. Scientific Assessment of Ozone Depletion: 2010, 

Global Ozone Research and Monitoring Project—Report No. 52, 516 pp., Geneva, Switzerland, 

2011.
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10

WMO (World Meteorological Organization), 2007. Scientific Assessment of Ozone Depletion: 2006, 

Global Ozone Research and Monitoring Project—Report No. 50, 572 pp., Geneva, Switzerland, 

2007.

Wuebbles, D.J., and K.O. Patten, Three-dimensional modeling of HCFC-123 in the atmosphere: 

Assessing its potential environmental impacts and rationale for continued use, Environ. Sci. 

Technol., 43 (9), 3208-3213, doi: 10.1021/es802308m, 2009.
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11

Appendix A:

Determination of an Acceptable Exposure Limit (AEL) 

for Trans-1-chloro-3,3,3-trifluoropro-1-ene

Recommended AEL: 1,000 ppm (8-hour time-weighted average)

Basis and Endpoints: NOAEC: 4,000 ppm (no significant cardiac histopathology observed 

at this concentration)

Critical Study: 90-day inhalation toxicity study in the rat (TNO, 2011b)

Protocol: Whole-body inhalation, 6 hours/day, 5 days/week for 13 weeks 

Concentrations: 0; 4,000; 10,000; 15,000 ppm

NOAEC9: 4,000 ppm (identified by independent pathologist’s report 

[Experimental Pathology Laboratories, 2011])

NOAEC [HEC10]: 4,000 ppm x 6 hr/8 hr = 3,000 ppm

Uncertainty Factors:

 

3 – animal to human extrapolation

I. Introduction

Trans-1-chloro-3,3,3-trifluoropro-1-ene has been proposed as a replacement for PFCs, HCFCs, HFCs and 

other halogenated compounds. It is intended for several end-uses, including as a foam blowing agent, a 

refrigerant in chillers, a solvent for cleaning in equipment and aerosol cans, and a carrier solvent in 

adhesives, coatings, and inks. Several studies assessing the potential toxicity of trans-1-chloro-3,3,3,-

trifluoroprop-1-ene have been conducted and include the following: 14-day, 28-day, and 90-day 

inhalation study in rats, developmental inhalation studies in rats and rabbits, a cardiac sensitization assay 

in beagle dogs, and genotoxicity studies in bacteria and mammalian cells in culture. The results of these 

studies are discussed in the following sections to inform an assessment of the potential health risks from 

human exposure to the study compound in the intended end use.

II. Summary of Toxicity Studies

The toxicological studies reviewed for the determination of acceptable exposure limit for trans-1-chloro3,3,3-trifluoropro-1-ene are summarized below in Table 1. Subclinical effects in the heart (monocyte 

infiltration) associated with inhalation exposure to trans-1-chloro-3,3,3-trifluoropro-1-ene were 

considered to be the most sensitive human health effects for derivation of an AEL; therefore inhalation 

studies are described and discussed in detail. The remaining studies are considered supporting studies.

 9 No Observable Adverse Effect Concentration (NOAEC) 10 Human Equivalent Concentration (HEC)
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Table 1. Trans-1-chloro-3,3,3-trifluoropro-1-ene Toxicological Studies 

Inhalation Studies Doses NOAEL/ NOAECa Effects

4-hour acute (nose-only, 

rat)

96,000, 156,000

and 120,000 ppm

LC50, 120,000 ppm 

(combined sexes)

Mortality at ≥96,000 ppm for 

some animals.

Tremors at all exposures.

14-day (whole body, rat) 0, 2,000, 7,500, and

20,000 ppm

2,000 ppm (study rpt.)

7,500 (peer review)b

Min. focal mononuclear cell 

infiltrate (heart)

28-day with 14-day 

recovery (nose-only, rat)

0, 2,000, 4,500, 7,500,

and 10,000 ppm

4,500 ppmc (study report)

10,000 ppm (peer review)

Increased serum K+ in 7,500-

ppm males; reversible 

changes in hematology/clinical 

chemistry parameters

90-day study (noseonly, rat)0, 4,000, 10,000 and 

15,000 ppm

<4,000 (study rpt.) 

4,000 ppm (peer review)

Min. focal mononuclear cell 

infiltrate (heart)

Rat fetotoxicity 0, 4,000, 10,000, and 

15,000 ppm 10,000 ppm Dilated urinary bladders in the fetuses

Rabbit fetotoxicity 0, 2,500, 10,000, and 

15,000 ppm 15,000 ppm No adverse effects noted

Cardiac sensitization, 

dog

25,000, 35,000, and 

50,000 ppm Negative at 25,000 ppm Tremors present during 

exposure ≥35,000 ppm

Supporting Studies Doses NOAEL/ NOAECa Effects

In vitro bacterial reverse 

mutation assay

Max concentration of 

905,000 ppm NA Negative

In vivo unscheduled 

DNA synthesis 0, 7,500, and 10,000 ppm NA Negative

In vitro chromosomal 

aberration assay 469, 783, or 1,305 μg/mL NA Negative

In vivo micronucleus 

test (rats)

0, 2,000, 4,500, 7,500, 

and 10,000 ppm NA Negative

In vivo micronucleus 

test (mice) 0, 50,000 ppm NA Negative

aAll values NOAEL/NOAEC unless otherwise reported; NA- not applicable bNOAEC values were revised in a pathology peer review report (Experimental Pathology Laboratories, 2011) c

NOAEC identified in study report based on clinical chemistry, not histopathology results

NOAEL = No observed adverse effect level (commonly used for oral or dermal exposures)

NOAEC = No observed adverse effect concentration (commonly used for inhalation exposures)

III. Development of the AEL for Trans-1-chloro-3,3,3-trifluoropro-1-ene

Several repeated dose inhalation studies assessing the toxic potential of trans-1-chloro-3,3,3-trifluoropro1-ene were available for development of an AEL. The NOAEC of 4,000 ppm from the 90-day inhalation 

study was chosen as the point of departure for derivation of an AEL for trans-1-chloro-3,3,3-trifluoropro1-ene as this was the study with the longest exposure duration with an identifiable NOAEC.

The AEL was calculated in the following manner:

4000 ppm x (6/8) ÷ 3 = 1000 ppm
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A human equivalent concentration (HEC) was determined by adjusting the NOAEC of 4,000 ppm by the 

ratio of rat exposure duration per day to that of an occupational worker (6 hours/8 hours). The guidelines 

for developing Reference Concentration (RfC) values (U.S. EPA, 1994) were followed, which obviated 

the need to apply a full uncertainty factor (UF) of 3 for pharmacokinetic differences between rats as the 

animal model of study and humans (a UF of 3 for pharmacodynamic [PD] differences between the two 

species has been applied). Because subchronic studies are typically used to develop AEL values, an 

additional UF was not added to account for study duration. Similarly, the database is considered 

comprehensive, and no additional UF was added to account for database limitations. 

IV. Inhalation Studies for Trans-1-chloro-3,3,3-trifluoropro-1-ene

Four-Hour Acute Inhalation Study

In an acute 4-hour nose-only inhalation study, male and female Crl:SD (outbred) rats (5/sex) were 

exposed to three different concentrations of trans-1-chloro-3,3,3-trifluoropro-1-ene (purity 99.99%) and 

observed for 14 days post-exposure (when possible) (TNO, 2009b). In the first group (Group A), animals 

were exposed to 100,000 ppm trans-1-chloro-3,3,3-trifluoropro-1-ene. Because of the low toxicity 

observed, a second group (Group B) of 5/sex were exposed to a target concentration of 125,000 ppm, that 

actually reached 156,000 ppm due to technical difficulties; all animals died during exposure. A third 

group (Group C) of 5 animals/sex were then exposed to a target concentration of 120,000 ppm in an effort 

to provide more data to identify the LC50. 

One female from Group A died on the first day of the recovery period. Other animals, male and female, 

showed clinical signs during recovery, but did not die prematurely. Necropsy did reveal gray discolored 

lungs in these animals, which was coincident with petechiae in some cases. Three males and two females 

of Group C died during exposure. Necropsy of these animals revealed reddish lung, with some also being 

enlarged. In two female animals of group B, red spots were also found on the thymus. In animals 

sacrificed on schedule following recovery, no other abnormalities were found. The combined-sex LC50

was reported as 120,000 ppm.

Fourteen-Day Inhalation Study

In a dose-range-finding inhalation study, Crl:CD(SD) rats (5/sex/group) were exposed via whole body 

inhalation to 0, 2,000, 7,500, or 20,000 ppm trans-1-chloro-3,3,3-trifluoropro-1-ene (99.9% pure) (TNO, 

2008). Exposures were for 6 hr/day, 5 days/week, in a 2-week period (10 total exposure days). Clinical 

signs, body weights and food consumption data were collected. Blood samples were drawn at the end of 

each week for hematology and clinical chemistry analyses. Gross necropsies were performed on all 

animals following the last exposure, and organs were collected and weighed; select organs, including the 

respiratory tract, were processed and analyzed histologically. 

The only clinical sign reported during exposure was restlessness, particularly in the highest concentration; 

the effect resolved after 15 minutes of exposure. Test-compound exposure did not induce changes in 

body weights or food consumption. Regarding changes in hematology, the study author noted an increase 

in prothrombin time in females at 20,000 ppm, and an increase in absolute and relative numbers of 

monocytes and absolute number of neutrophils in males at 20,000 ppm. Increases in serum liver enzymes 

(ALAT11 and ASAT in males at 20,000 ppm) were not confirmed by any changes in liver histopathology, 

and were thus considered not toxicologically relevant. Histology revealed multifocal mononuclear cell 

infiltrates, identified by the study laboratory in animals at 7,500 and 20,000 ppm. However, following an 

independent pathological review of the histology slides of the heart tissues (Experimental Pathology 

Laboratories, 2011), many of the lesions originally identified by the study laboratory as ‘very slight’ or 

‘slight focal myocardial mononuclear cell infiltrate’ were downgraded to being ‘not remarkable’ or were 

 11 ALAT, alanine transaminase; ASAT, aspartate transaminase
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noted to be located only in the apex of the heart. According to the reviewing pathologist, lesions in the 

apex and/or adjacent portions of the right and left ventricles and interventricular septum of the heart in 

rats with such lesions are “typical of rodent spontaneous cardiomyopathy”. Given the revised heart data, 

the reviewing pathologist increased the NOAEC to 7,500 ppm from the original NOAEC of 2,000 ppm 

identified by the study laboratory. ICF accepts the revised value as reasonable, given the expertise of the 

reviewing pathologist.

Four-Week Inhalation Study

In a longer-term, nose-only, multi-exposure study, five groups of Sprague-Dawley rats were exposed 6 

hrs/day, 5 days/week in a 28-day period (e.g., 20-21 exposure days) to the following nominal 

concentrations of trans-1-chloro-3,3,3-trifluoropro-1-ene (99.9% pure): 0, 2,000, 4,500, 7,500, and 

10,000 ppm (TNO, 2009c). Additional groups of 5/sex were exposed to either 0 or 10,000 ppm in an 

identical fashion, followed by a 14-day recovery period. The following parameters were measured in 

accordance with harmonized guidelines for this repeat-exposure study: clinical signs and mortality, food 

consumption and body weights, hematology and clinical chemistry, gross necropsy, organ weights, and 

histopathology of select organs.

The study compound did not induce any changes in clinical signs or mortality, food consumption, body 

weights, urinalysis parameters, gross necropsy, or organ weights. In contrast to the histopathological 

findings from the 14-day study, no mononuclear cell infiltration was noted in cardiac tissue or any other 

selected organ. An increase in the relative number of basophils was observed in males at 10,000 ppm, 

which resolved in the 14-day recovery period. Changes in clinical chemistry parameters were limited to 

decreased creatinine in males at 10,000 ppm, which also was resolved at the end of the recovery period; 

and an increase in serum potassium in males at both 7,500 and 10,000 ppm, which were still slightly 

elevated after recovery, although not statistically significant. The study authors identified a NOAEC of 

4,500 ppm, arguing that the increased potassium levels at exposure concentrations of 7,500 and 10,000 

ppm were related to treatment. The pathologist who performed the peer review of the cardiac 

histopathology slides for this study identified a NOAEC of 10,000 ppm, based on the confirmed absence 

of compound-induced subcellular damage to cardiac tissue. ICF concurs with the reviewing pathologist, 

and would add that although the increased potassium level observed at ≥7,500 ppm may be indicative of 

subclinical cardiotoxicity, it cannot be attributed to the test compound alone, and may also be attributed to 

spontaneous cardiomyopathy. The NOAEC for this study is set, therefore, at 10,000 ppm.

Ninety-Day Inhalation Study

In a 90-day inhalation study, male and female Sprague-Dawley (Crl:CD[SD]) rats (10/sex/concentration) 

were exposed to trans-1-chloro-3,3,3-trifluoropro-1-ene (99.9% purity) via nose-only exposure at 

nominal vapor concentrations of 0, 4,000, 10,000, or 15,000 ppm for 6 hrs/day for 5 days/week for 13 

weeks, for a total of 65 exposure days (TNO, 2011b). Rats were observed twice daily for clinical signs of 

toxicity, morbidity, and mortality. Body weights and mean food consumption were recorded weekly. 

Ophthalmoscopic examinations were conducted at study initiation and during the last week of exposure. 

Blood samples and urine samples were taken just prior to or the last day prior to sacrifice at the end of the 

exposure period and were analyzed for hematology, clinical chemistry and urinalysis parameters 

consistent with harmonized guidelines. Gross necropsies were performed on all surviving animals after 

the scheduled exposure period, and select organs were removed, weighed, and prepared for 

histopathology. 

Mortality, Clinical Effects, Eye Effects, Food Consumption, Body Weights

Exposure to the study compound did not induce early mortality or any observed clinical abnormalities. In 

addition, exposure did not affect body weight gain, food consumption, or changes in eye condition.
***** FINAL May 1, 2013 *****

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Hematology/Clinical Chemistry

Effects on hematological parameters were limited. For example, exposure to the study compound had no 

effect on red blood cell parameters and affected white blood cells only by decreasing the relative number 

of lymphocytes in males at 15,000 ppm. Clinical chemistry effects were inconsistent between the sexes. 

Plasma of ASAT and ALAT were increased at statistically significant levels in males at 15,000 ppm, but 

not in females, while glucose and urea were increased in females at 10,000 and 15,000 ppm, but not in 

males. High-concentration females also exhibited significantly increased potassium and decreased 

triglyceride levels. No other changes in measured analytes were noted.

Organ Weights

A treatment-related decrease in absolute and relative heart weights was noted in male rats, which reached 

statistical significance at the highest concentration (15,000 ppm). In addition, relative liver weights in the 

males were significantly increased at the highest concentration, whereas by contrast, the relative liver 

weights of high-concentration females were significantly decreased. No other changes in organ weights 

were noted.

Macroscopic and Microscopic Findings

Exposure to trans-1-chloro-3,3,3-trifluoropro-1-ene did not induce any changes in the appearance of 

organs or tissues in the bodies of rats evaluated following gross necropsy. In general, no compoundinduced effects were noted upon analyses of histopathology findings, with the exception of minimal 

multifocal mononuclear cell infiltrate in the heart. The study authors also identified inflammation 

occurring coincident with the infiltrates in some animals. The report of the peer review pathologist 

indicated that the vast majority of these findings were downgraded to categories of ‘not remarkable’ or 

were localized to the apex, the lower left ventricle, or the atrium. Again, because of the downgrading of 

incidences of infiltration reported in the original study, and the identification of some mononuclear 

infiltration as ‘spontaneous cardiomyopathy,’ the NOAEC was identified as the lowest exposure 

concentration of 4,000 ppm (the study authors reported the NOAEC as less than 4,000 ppm). ICF 

concurs, and as such, the NOAEC used in the determination of an AEL for trans-1-chloro-3,3,3-

trifluoropro-1-ene is 4,000 ppm.

Developmental Toxicity

Trans-1-chloro-3,3,3-trifluoropro-1-ene was evaluated in two developmental toxicity studies, one in rats 

and one in rabbits. 

Rat Prenatal Developmental Study

In the prenatal developmental study in rats (TNO, 2011a), groups of 24 mated female Wistar 

(Crl:WI[WU]) rats were exposed 6 hrs/day to concentrations of 0, 4,000, 10,000 or 15,000 ppm trans-1-

chloro-3,3,3-trifluoropro-1-ene (99.9% pure) from gd 6 to gd 19. Health parameters measured during the 

in-life phase of the study included mortality, clinical signs, maternal body weight, and food consumption. 

A Caesarean section was performed following the gestation period, and females and fetuses were 

evaluated via gross necropsy, and for standard reproductive toxicity endpoints (e.g., gestation index, 

fecundity index, fetus sex ratio, number of corpora lutea, implantation sites, pre- and post-implantation 

loss, live and dead fetuses, and resorptions). Fetuses, placentas and reproductive organs were weighed, 

and fetuses were processed for soft tissue and skeletal abnormalities and malformations.

Exposure to the study compound did not affect any of the measured parameters with the exception of the 

following: a statistically significant increase in the incidence of dilated urinary bladders was observed in 

the fetuses of the highest concentration group (15,000 ppm). Because the incidence of this anomaly 

increased with higher concentrations, it is considered treatment-related. No signs of maternal toxicity 

were noted at any concentration. The NOAEC for developmental effects is established at 10,000 ppm; 

the NOAEC for maternal toxicity is established at 15,000 ppm.
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Rabbit Prenatal Developmental Study

Trans-1-chloro-3,3,3-trifluoropro-1-ene was further evaluated in a developmental study in the New 

Zealand white rabbit. Huntingdon (2010) exposed 22 pregnant dams/group during organogenesis (gd 6 to 

gd 28) to trans-1-chloro-3,3,3-trifluoropro-1-ene (~100% purity) via whole-body inhalation for 6 hrs/day 

at concentrations of 0, 2,500, 10,000 or 15,000 ppm. During the in-life phase of the study, the rabbits 

were observed for mortality, clinical signs, body weights, and food consumption. Following euthanasia 

of the dams at gd 29, the following parameters were measured: gross necropsy, number of corpora lutea 

and implantation sites, uterine and placental weights, fetus number (live), fetal weights (live only), sex

ratio, external defects, soft tissue abnormalities, and skeletal abnormalities and state of ossification. 

Exposure to the test compound did not affect any changes in any of the measured parameters of the 

exposed dams or the fetuses. Therefore, the NOAEC for both maternal and fetal toxicity is established at 

15,000 ppm in this rabbit developmental assay.

Cardiac Sensitization

In a muzzle-only, acute exposure study in young beagle dogs (WIL Laboratories, 2011), 6 male dogs 

were exposed successively to 25,000 ppm (2.5%), 35,000 ppm (3.5%) and 50,000 ppm (5%) trans-1-

chloro-3,3,3-trifluoropro-1-ene (99.978% pure) up to a maximum period of 33 minutes, with 48 hours of 

rest occurring in between exposure periods. Control (air) exposures occurred in the same dogs. For the 

cardiac sensitization, epinephrine challenge (bolus) injections, ranging from 2 to 8 μg/kg, were started 

approximately 5 minutes following the initiation of exposure and continued until termination of exposure. 

Only 2 of the 6 dogs were exposed to the highest trans-1-chloro-3,3,3-trifluoropro-1-ene concentration, 

because behavioral responses (vocalization, injected sclera, excessive salivation, tremors, convulsions 

and/or reddened gums) prompted the termination of exposure; only one of the dogs at this concentration 

was given an epinephrine challenge.

No test-related deaths occurred in the study. Clinical findings similar to those at 50,000 ppm were noted 

after exposure to 35,000 ppm (tremors, injected sclera, reddened ears and excessive salivation). No signs 

of cardiac sensitization were noted at 25,000 or 35,000 ppm; however, the presence of ECG noise in two 

animals exposed to 35,000 ppm precluded the complete characterization of cardiac sensitization at this 

concentration. As noted previously, exposure was terminated at the highest concentration and assessment 

of cardiac sensitization was not possible at this level. Therefore, the results indicate that the study 

compound does not cause cardiac sensitization at 25,000 ppm.
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V. Supporting Genotoxicity Studies for Trans-1-chloro-3,3,3-trifluoropro-1-ene

Reverse Mutation Assay

Trans-1-chloro-3,3,3-trifluoropro-1-ene was tested in an Ames bacterial reversion assay in Salmonella 

typhimurium (S. typhimurium) strains TA98, TA100, TA1535, TA1537, and E. coli strain WP2 uvrA

(BioReliance, 2011). This assay measures the ability of the compound to induce either base pair or frame 

shift mutations in DNA in the presence or absence of S9 fraction from induced rat livers (metabolizing 

enzymes including mixed function oxidases). The study compound was analyzed for mutagenicity twice 

at concentrations ranging from 73,400 to 905,000 ppm at 25°C for either 24 or 48 hours. Appropriate 

negative and positive controls were analyzed in the same assays. Mutagenicity was not induced in any 

strain, but toxicity was seen in all strains at the top two concentrations of 513,000 and 905,000 ppm. A 

confirmatory assay was performed using slightly reduced exposure concentrations of 17,200; 24,500; 

73,400; 147,000; 196,000; 269,000; 513,000 and 905,000 ppm at 25°C. Cytotoxicity was observed at the 

top two concentrations, as in the previous assays. No positive responses were observed with strains TA98 

and TA1537 in the absence of S9 activation and with strains TA98 and WP2uvrA in the presence of S9 

activation. The compound is categorized as negative in this genotoxicity assay.

Unscheduled DNA Synthesis

Trans-1-chloro-3,3,3-trifluoropro-1-ene was tested for the ability to induce unscheduled DNA synthesis 

in liver hepatocytes isolated from male rats exposed to 7,500 and 10,000 ppm of the study compound in a 

28-day study (TNO, 2009c). The control group was the same one used in the 28-day study and positive 

control animals were dosed with standard toxicants per harmonized guidelines. Trans-1-chloro-3,3,3-

trifluoropro-1-ene did not induce unscheduled DNA synthesis under the conditions used in the 28-day 

study, and is categorized as negative for this genotoxicity assay.

Micronuclei

Two studies were performed to assess the ability of trans-1-chloro-3,3,3-trifluoropro-1-ene to include 

micronuclei in bone marrow, as described below. 

Micronuclei Rat Study

Trans-1-chloro-3,3,3-trifluoropro-1-enewas tested for the ability to induce micronuclei in bone marrow 

cells isolated from male rats exposed to 0, 2,000, 4,500, 7,500 and 10,000 ppm of the study compound in 

a 28-day study (TNO, 2009c). Appropriate positive control animals were dosed with standard toxicants 

per harmonized guidelines. The test compound did not induce chromosomal damage or induce 

micronuclei in the bone marrow under the conditions used in this study (the maximum concentration 

administered, 10,000 ppm is equivalent to >12,000 mg/kg, much higher than the 1,000 mg/kg limit dose 

recommended by EPA) and is judged negative for this genotoxicity assay.

Micronuclei Mice Study

Trans-1-chloro-3,3,3-trifluoropro-1-enewas tested in an independent assay for its ability to induce 

micronuclei in the bone marrow of male mice (TNO, 2009a). Two groups of 10 male, albino mice (CD-1 

strain, Charles River) were exposed for a single, nose-only, 4-hour exposure to trans-1-chloro-3,3,3-

trifluoropro-1-ene (99.99% pure) at concentrations of 0, or 50,000 ppm. Five male mice were injected 

intraperitoneally with 0.75 mg/kg-bw of mitomycin C as a positive control for comparison. Bone marrow 

cells were isolated from half the exposure group at 24 hours and the remainder at 48 hours post-exposure; 

bone marrow smears were prepared and analyzed for the presence of polychromatic and normochromatic 

erythrocytes and micronuclei in a manner consistent with established guidelines. The test compound did 

not induce micronuclei or any other form of chromosomal damage and was thus judged negative in this 

genotoxicity assay. 
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18

Chromosomal Aberration

Trans-1-chloro-3,3,3-trifluoropro-1-ene (99.98% pure) was tested in a chromosomal aberration assay with 

human lymphocytes cultured in vitro (Huntingdon, 2011). Human lymphocytes, isolated from whole 

blood samples provided by volunteers, were treated with trans-1-chloro-3,3,3-trifluoropro-1-ene for either 

3 or 21 hours at doses of 469, 783, or 1,305 μg/mL in the presence or absence of metabolic activation (S9 

from the livers of chemically-induced rats). Trans-1-chloro-3,3,3-trifluoropro-1-ene was determined to be 

negative in this genotoxicity assay, as the cells treated with trans-1-chloro-3,3,3-trifluoropro-1-ene in all 

experiments did not cause an increase in the frequency of cells exhibiting structural chromosomal 

aberrations compared with negative controls at any concentration tested.

References

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Report. BioReliance, Rockville, MD, performed for Anonymous Submitter. 27 April 2011.

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Anonymous Submitter, USA, 27 May 2009.

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