Mckee 2015

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ISSN: 1040-8444 (print), 1547-6898 (electronic)
Crit Rev Toxicol, 2015; 45(4): 273–365

© 2015 Informa Healthcare USA, Inc. DOI: 10.3109/10408444.2015.1016216
REVIEW ARTICLE
Characterization of the toxicological hazards of hydrocarbon solvents

Richard H. Mckee1, M. David Adenuga1, and Juan-Carlos Carrillo2
1ExxonMobil Biomedical Sciences, Inc., Annandale, NJ, USA and 2Shell International BV, The Hague, The Netherlands
Critical Reviews in Toxicology Downloaded from informahealthcare.com by Universitaet Zuerich on 05/06/15
Abstract
Hydrocarbon solvents are liquid hydrocarbon fractions derived from petroleum processing Keywords
streams, containing only carbon and hydrogen atoms, with carbon numbers ranging from ap-
acute CNS effects, benzene, GHS
proximately C5–C20 and boiling between approximately 35–370°C. Many of the hydrocarbon
classification, hydrocarbon solvent, HPV,
solvents have complex and variable compositions with constituents of 4 types, alkanes (nor-
n-hexane, naphthalene, occupational
mal paraffins, isoparaffins, and cycloparaffins) and aromatics (primarily alkylated one- and two-
exposure limits, REACH, UVCB
ring species). Because of the compositional complexity, hydrocarbon solvents are now identi-
fied by a nomenclature (“the naming convention”) that describes them in terms of physical/
chemical properties and compositional elements. Despite the compositional complexity, most History
hydrocarbon solvent constituents have similar toxicological properties, and the overall toxi-
cological hazards can be characterized in generic terms. To facilitate hazard characterization, Received 15 October 2014
the solvents were divided into 9 groups (categories) of substances with similar physical and Revised 26 January 2015
chemical properties. Hydrocarbon solvents can cause chemical pneumonitis if aspirated into Accepted 3 February 2015
the lung, and those that are volatile can cause acute CNS effects and/or ocular and respiratory Published online 9 April 2015
For personal use only.
irritation at exposure levels exceeding occupational recommendations. Otherwise, there are

few toxicologically important effects. The exceptions, n-hexane and naphthalene, have unique
toxicological properties, and those solvents containing constituents for which classification
is required under the Globally Harmonized System (GHS) are differentiated by the substance
names. Toxicological information from studies of representative substances was used to fulfill
REACH registration requirements and to satisfy the needs of the OECD High Production Vol-
ume (HPV) initiative. As shown in the examples provided, the hazard characterization data can
be used for hazard classification and for occupational exposure limit recommendations.
Abbreviations: ACGIH, American College of Governmental Industrial Hygienists; AIHA, American

Industrial Hygiene Association; ALT, Alanine amino transferase; AML, Acute myelogenous leu-
kemia; API, American Petroleum Institute; AST, Aspartate amino transferase; ASTM, American
Society of Testing and Materials; BW, Body Weight; C, Carbon; oC, Degrees Centigrade; CARB,
California Air Resources Board; CAS, Chemical Abstract Services; CLP, Classification and Labelling
Procedures; Cm, Centimeter; CNS, Central Nervous System; CPSC, Consumer Product Safety Com-
mission; CTE, Chronic toxic encephalopathy; CYP, Cytochrome P; DNEL, Derived no-effect level;
ECETOC, European Centre for Ecotoxicity and Toxicology of Chemicals; ECHA, European Chemicals
Agency; EEC, European Economic Community; EINECS, European Inventory of Existing Chemical
Substances; EN, Ethylnaphthalene; EPA, Environmental Protection Agency (US); ET, Ethylbenzene;
EU, European Union; F, Mole fraction; GGV, Group guidance value; GHS, Globally Harmonized
System (of Chemical Classification); HPV, High production volume; Hr, Hour; HSDB, Hazardous
Substances Data Bank; IP, Intraperitoneal; IUPAC, International Union of Pure and Applied Chem-
istry; JP-8, Jet Fuel 8; Kg, Kilogram; LC, Lethal Concentration; LOAEL, Lowest observed adverse
effect level; Mg, Milligram; Min, Minute; MN, Methylnaphthalene; n – Normal; nmol, Nanomole;
NOAEL, No observed adverse effect level; OECD, Organization for Economic Cooperation and
Development; OEL, Occupational Exposure Limit; PBPK, Physiologically-Based Pharmacokinetic
(Model); RBC, Red blood cell; RCP, Reciprocal Calculation Procedure; RD, Respiratory depression;
REACH, Registration, Evaluation, Authorisation and Restriction of Chemicals; RTECS, Registry of
Toxic Effects of Chemical Substances; TEB, Triethylbenzene; TLVⓇ, Threshold Limit ValueⓇ; TMB,
Trimethylbenzene; TSCA, Toxic Substances Control Act; μg, Microgram; μmol, Micromole; UVCB,
Unknown or variable composition, complex reaction products or biological materials; VOC, Vola-
tile organic carbon; VM&P, Varnish Makers’ and Painters’ (naphtha); WBC, White blood cell; WEEL,
Workplace Environmental Exposure Limit
Address for correspondence: Richard H. McKee, ExxonMobil Biomedical

Sciences, Inc., 1545 U.S. Highway 22 East, Annandale, NJ -3059, USA.
Tel: (1) 908-730-1037. E-mail: richard.h.mckee@exxonmobil.com
274 R. H. Mckee et al. Crit Rev Toxicol, 2015; 45(4): 273–365
Table of Contents Exposure to hydrocarbon solvents and occupational

Abstract ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 273 exposure limits�� 305
Introduction �� 274 Exposure to hydrocarbon solvents �� 305
Scope and purpose of the document�� 274 Occupational exposure limits �� 306
Historical approaches to characterize the hazards of Conclusions�� 307
hydrocarbon solvents�� 276 Appendix . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 308
Toxicokinetic properties of saturated constituents of Acknowledgments �� 355
hydrocarbon solvents�� 280 Declaration of Interest�� 355
Absorption �� 281 References ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 355
Absorption of inhaled hydrocarbon solvent constituents�� 281
Absorption of ingested aliphatic hydrocarbon Introduction
solvent constituents�� 281
Percutaneous absorption of aliphatic hydrocarbon Scope and purpose of the document
solvent constituents�� 281
Systemic distribution of aliphatic hydrocarbon
The present document summarizes information on the physical/
solvent constituents�� 282 chemical properties and toxicological hazards of hydrocarbon
Metabolism and excretion of aliphatic hydrocarbon solvents and provides examples of the ways in which the infor-
solvent constituents�� 283 mation on hazard characterization can be used for hazard clas-
C5–C9 hydrocarbon solvent constituents �� 283 sification and to set occupational exposure limits. Many of the
C9–C14 hydrocarbon solvent constituents�� 283
C14–C20 hydrocarbon solvent constituents �� 284
toxicological studies were published separately, but the results
Summary of toxicokinetic properties of aliphatic hydrocarbon are summarized herein and referenced in the appendices.
solvent constituents�� 284 Hydrocarbon solvents are liquid hydrocarbon fractions that
Metabolism of aromatic constituents of hydrocarbon solvents�� 286 are primarily produced by the distillation of petroleum feed
Metabolism of one-ring aromatics �� 286 stocks or their synthetic analogs (e.g., Fischer-Tropsch derived
Metabolism of aromatic/naphthenic molecules�� 286
Metabolism of naphthalene (a two-ring aromatic)�� 287
materials), sometimes followed by additional processing steps
Metabolism of alkylated two-ring aromatics (alkylated such as solvent extraction, hydrodesulfurization, or hydroge-
naphthalenes) �� 287 nation.1 Most hydrocarbon solvents are complex substances
Summary of information on metabolism of aromatic with variable compositions and are best described as UVCB2
constituents of hydrocarbon solvents �� 287 (unknown and variable composition) substances, but some are
Potential interactions among hydrocarbon solvent constituents
single constituent (mono-constituent) substances. The complex
and other solvents �� 287

Common toxicological properties of hydrocarbon solvents and variable nature of these solvents is the consequence of their
and their constituents �� 289 manufacturing processes. In short, most hydrocarbon solvents
Common acute effects of hydrocarbon solvents�� 289 are complex substances containing a large number of individual
Acute CNS effects�� 289 hydrocarbon molecules, that is, molecules containing only car-
Ocular and upper respiratory tract irritation �� 290
Dermal irritation �� 290
bon and hydrogen atoms, with carbon numbers ranging from
Chemical pneumonitis�� 290 approximately C5–C20 and having boiling points in the range
Common effects of repeated exposures to hydrocarbon of approximately 35–370°C. Although the principle constitu-
solvents ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 291 ents are alkanes and aromatics, the hydrocarbon solvents are
Liver enlargement�� 291 commonly described as “aliphatic” (straight chain, i.e., normal
Kidney effects�� 291
Hematological changes �� 291
paraffinic solvents, or branched, i.e., iso-paraffinic) solvents or
Persistent CNS effects�� 292 “alicyclic” (or “naphthenic”), that is, solvents containing pri-
Hydrocarbon solvent constituents with unusual toxicological marily cycloparaffinic constituents, when the constituents are
properties �� 295
Constituents of concern that may be present at levels  1%�� 295
n-Hexane�� 295
1The following definitions from R.J. Lewis (Ed.), Hawley’s Condensed
Naphthalene �� 296
Constituents of concern that may be present at levels  1%�� 296 Chemical Dictionary (12th Ed. Van Nostrand Reinhold Co. 1993) are
Diethyl and triethylbenzenes�� 296 relevant to the terms used in this report:
Biphenyl�� 297 Solvent – A substance capable of dissolving another substance to form a
Constituents of concern that may be present at levels  1%�� 297 uniform dispersed mixture at the molecular or ionic-size level.
Benzene�� 297 (a) Organic Solvent – An organic (carbon-based) substance capable of
Polycyclic aromatic hydrocarbons �� 298 dissolving another substance.
Hydrocarbon solvent categorization and nomenclature . .. . .. . .. . .. . 298 (b) Hydrocarbon – An organic compound consisting exclusively of the
Hydrocarbon solvent categories �� 298 elements carbon and hydrogen.
Nomenclature for hydrocarbon solvents – the naming (c) Hydrocarbon Solvent – Chemical compounds composed of carbon
convention �� 299 and hydrogen and capable of dissolving another substance.
Summary of the Toxicological Information �� 300 2As explained by OECD, a number of complex substances that contain
Aromatic solvents (C9–C12)�� 300 a diverse range of materials are defined as ‘Substances of unknown or
Aliphatic/Aromatic solvents (C9–C20)�� 301 variable composition, complex reaction products or biological materials
Light Aliphatic solvents (C5–C9)�� 301 (UVCB). The common characteristics of UVCB substances are that they
Heavy Aliphatic solvents (C9–C20) �� 302 contain numerous (but typically closely related) components with defined
Use of Toxicological Hazard Characterization Information for carbon numbers or distillation ranges and cannot be represented by sim-
Hydrocarbon Solvents and Their Constituents for Hazard ple chemical structures; they are not intentional mixtures of chemicals;
Classification and for Occupational Exposure they are of natural origin and cannot be separated into their constituent
Recommendations �� 303 chemical species; the concept of “impurities” does not apply; and they
Evaluation of hydrocarbon solvents for classification following the are produced according to a performance specification related to their
recommendations of the Globally Harmonized System �� 303 physical and chemical properties (OECD 2007, p. 67).
DOI 10.3109/10408444.2015.1016216 Hydrocarbon solvent toxicology 275
primarily saturated hydrocarbons. The solvents are referred to range of 153–194°C, an aromatic content of approximately 15%,
as “aromatic” when composed primarily of aromatic constitu- and has predominant carbon numbers in the range of C9–C11.
ents. However, hydrocarbon solvents may contain any or all of The overall composition of solvents of this type has remained
these types of constituents in varying amounts. relatively constant for the past 40 years (Carrillo et al. 2014).
Hydrocarbon solvents are manufactured to meet technical In more recent years, solvents have become more highly
specifications which usually pertain to physical and/or chemical refined and more narrowly defined in order to keep pace with
properties, so solvents of particular types and end uses may evolving end use applications and other factors including tech-
have compositional differences. Hydrocarbon solvents are pri- nological developments, regulatory changes, and increased
marily derived from petroleum refining streams, for example, knowledge of the hazards of solvents and their constituents.
“naphtha” or “kerosene”, but, because of their more special- Among the early but influential regulations with implications
ized applications, may be more highly refined with narrower for solvent composition were those from the early 1960s includ-
boiling ranges and may have restrictions on some specific con- ing “Rule 66” in the Los Angeles area and Regulation 3 in San
stituents, for example, benzene. The more narrow carbon range Francisco that limited the aromatic content of solvents to pre-
and constituent composition is reflected in the solvent names serve air quality in California. These regulations started a trend
(see Hydrocarbon solvent categorization and nomenclature). toward the development of solvents with lower levels of aro-
One reason for the need for detail on substance definition is matic constituents. A more recent trend has been the develop-
that terms such as “hydrocarbon”, “hydrocarbon solvent”, or ment of solvents with higher molecular weight and lower vola-
“petroleum solvent” are often used in a more generic sense, tility to reduce evaporative emissions. For example, the U.S.
making it difficult to link reported outcomes to specific agents. Environmental Protection Agency (1998) established limits on
In one historical example, a series of papers was published that the volatile organic carbon (VOC) content of certain consumer
discussed the possible link between exposure to hydrocarbon products such as aerosol coatings and household cleaners. These
solvents and the development or exacerbation of Goodpasture rules exempted low vapor pressure substances (vapor pressures
syndrome. In the first of these papers (Bierne and Brennan  0.1 mm Hg at 20°C) because these higher-boiling, lower-
1972), the authors used the term “hydrocarbon solvent” as a volatility solvents were not expected to contribute significantly
generic reference to organic solvents of all types, and to fuels to evaporative emissions. Reports of unusual toxicological
as well. Subsequent authors have continued to follow this findings of specific constituents, including the neurological
terminology, such that in a review article published in 1992 effects of n-hexane, the association between benzene exposure
(Bombassei and Kaplan 1992), the title refers to “hydrocarbon and the development of acute myelogenous leukemia (AML),
exposure”, but the identified agents included solvents (some and, more recently, the demonstration that exposure to naphtha-
of which are specifically identified as chlorinated solvents), lene causes nasal tumors in rats have led to efforts to produce
fuels, and complex mixtures including smoke and exhaust. In solvents with lower levels of constituents with unusual toxico-
a more recent example, Sabbath et al. (2014) reported cogni- logical properties. There have also been restrictions on applica-
tive deficits in workers that had been exposed to a variety of tions and developments in hazard communications, including
agents including “petroleum solvents”, but the example of a classification and labeling statements, to avoid inappropriate
“petroleum hydrocarbon” was hydrazine, which is not a hydro- solvent uses and to limit the potential for exposure to hazard-
carbon, not a solvent, and not derived from petroleum.3 ous constituents. Published information indicates that occupa-
The scope of this document is limited to hydrocarbon sub- tional exposures to hydrocarbon solvents have been declining
stances manufactured for solvent uses. It does not cover organic (Caldwell et al. 2000, Lundberg et al. 1995), at least in part
solvents that are not hydrocarbons (e.g., alcohols, ethers, ketones, because of changes in attitudes to health and safety, along with
acetates, chlorinated solvents) or hydrocarbon substances such the influence of regulators, employers, and the general public.
as fuels that are not intended for solvent uses. Also not covered These developments have also led to changes in the way in
are some hydrocarbons such as toluene and xylene that may be which hydrocarbon solvents are identified and described. In the
present at low levels in some complex hydrocarbon solvents. 1970s, with the advent of the U.S. Toxic Substances Control Act
Toluene and xylene can be used as solvents but are primarily (TSCA), refinery streams were given Chemical Abstract Services
used as chemical intermediates. As an example of a complex (CAS) numbers to allow them to be included in inventories of
hydrocarbon solvent, consider “Stoddard solvent”, which was existing substances. There was a similar process in Europe, under
developed for use in the dry cleaning industry in the 1920s, which these streams were given EINECS (European Inventory
had a distillation range of approximately 150–200°C, and had a of Existing Chemical Substances) numbers (European Economic
maximum aromatic content of 25% (ASTM D-484-71, 1971). Community 1990) and included in existing substance inventories
Because Stoddard solvent and similar hydrocarbon solvents are to comply with the European Directive 67/548/EC. Under these
derived from petroleum feed stocks by distillation, the aromatic systems, hydrocarbon solvents were identified by the same CAS
content of the solvents is related to the characteristics of the numbers as the refinery streams from which they were derived.
crude oils from which they are derived, but are normally in the As shown in Table 1, and using Stoddard solvent4 and similar
range of 15%–20%. Carpenter et al. (1975c) described one sam-
ple of Stoddard solvent as typical of commercial manufacture 4Stoddard solvent is the name that was applied to solvents boiling in the range of
and supplied by a major American producer. It has a distillation approximately 150–200oC and containing up to 25% aromatics. The more com-
monly used names for solvents of this type are type 1 mineral spirits in the United
States and white spirit in Europe. The use of these more common terms in the
3“Petroleum solvent” is a term that describes the liquid hydrocarbon frac- literature has created confusion as the common terms tend to be used as broad
tions obtained from petroleum, for use as solvents in industrial processes descriptors and in some cases refer either to wider groups of solvents or conditions
and commercial formulations (ASTM 2010). of use (i.e., house painting).
Table 1. Chemical Abstract System (CAS) and International Union of tains 5 solvents of this type with differing distillation ranges, as
Pure and Applied Chemistry (IUPAC) descriptions that have been used to reflected by their carbon number distributions.
describe Stoddard solvent and similar hydrocarbon solvents.
The REACH and HPV processes required the identification
CAS Number CAS Name IUPAC Name and review of all available hazard information, including previ-
8052-41-3 Stoddard solvent A colorless refined petroleum ously unpublished toxicology studies on these substances. For
distillate that is free from rancid or HPV purposes, the information was compiled and summarized.
objectionable odors and that boils
in a range of approximately 148.8
However, the REACH process had a broader scope as it also
to 204°C. entailed additional data evaluation steps including the develop-
64742-82-1 Naphtha A complex combination of ment of “Derived No-Effect Levels” (DNELs), which are the
(petroleum), hydrocarbons obtained from a highest exposure levels to which an individual could be exposed
hydrodesulfurized catalytic hydrodesulfurization
heavy process. It consists of without undue risk of harm.5 Registrants were required to com-
hydrocarbons having carbon pare the DNEL for each substance to the predicted exposures
numbers predominantly in the associated with each registered use, to ensure that risks from
7–12 range and boiling in the range
exposure were controlled by the recommended risk management
of approximately 90 to 230°C.
647541-92-0 Naphtha A complex combination of procedures. In addition, registrants were required to review the
(petroleum) hydrocarbons obtained as the substance classifications and to update them in order to comply
solvent-refined raffinate from a solvent extraction with the European adaptation of the Globally Harmonized System
heavy process. It consists predominantly
of aliphatic hydrocarbons having (GHS), that is, the Classification, Labelling and Packaging (CLP)
carbon numbers predominantly requirements. In short, the REACH registration process required
in the 7–12 range and boiling in not only a critical review of all of the available information on
the range of approximately 90 to hazards but also required risk assessments and reviews of the risk
230°C.
64742-48-9 Naphtha A complex combination of communication and risk management procedures, to ensure that
(petroleum), hydrocarbons obtained by treating all registered substances are safe for their intended uses.
hydrotreated heavy a petroleum fraction with hydrogen Having gone through these data compilation and assessment
in the presence of a catalyst. It processes, it seemed appropriate to make the overall conclu-
consists of hydrocarbons having
carbon numbers predominantly sions publicly available and to also publish the key information
in the 6–13 range and boiling in that formed the basis for these conclusions. The search strategy
the range of approximately 65 to for publicly available information was based on the category
230°C.
64742-88-7 Solvent naphtha A complex combination of
and substance definitions listed in Table 2 and used these
(petroleum) hydrocarbons obtained from the generic descriptors along with the readily identifiable constitu-
medium aliphatic distillation of crude oil or natural ents of the solvents. Also used as search terms were common
gasoline. It consists predominantly trade names for the solvents and constituents. The search was
of saturated hydrocarbons having
carbon numbers predominantly conducted using the chemical names, common names, and
in the 9–12 range and boiling in CAS registry numbers of the various substances. Bibliographic
the range of approximately 140 to databases that were searched using both CAS registry numbers
220°C. and common names included Toxcenter (previously Toxline),
Chemical Abstracts, and Medline. Additionally, searches were
substances as an example, there are several CAS numbers that conducted by CAS registration numbers only of the Hazard-
could be applied to solvents of this type, but all are relatively non- ous Substance Data Bank (HSDB) and the Registry of Toxic
specific. Aside from CAS number 8052-41-3, the CAS definitions Effects of Chemical Substances (RTECS). Because the data
identify the source (petroleum), the boiling range, and the car- were compiled for use in HPV and REACH registration sub-
bon number range, but provide little compositional information. missions, previously unpublished information from company
The wider boiling ranges and carbon numbers of the petroleum sources as well as documents from trade organizations were
substances (i.e., the feed stocks for hydrocarbon solvent manufac- also available. For unpublished information from company
ture) may be appropriate for fuels for which compositions are less sources, references are provided to accessible summaries when
constrained, but are less useful for hydrocarbon solvents which these are available. When information has not been formally
have more narrowly defined product specifications. Thus, to more published but is in the public domain, references are provided.
precisely describe its products, the hydrocarbon solvent industry
developed a naming convention with more specific information Historical approaches to characterize the hazards
on the principal constituents of the solvents including, particu- of hydrocarbon solvents
larly hazardous constituents (Table 2), which was used to register Among the lower molecular weight solvents, the potential
substances in Europe in compliance with the new chemical con- number of constituents is limited, but as molecular weight
trol legislation REACH (Registration, Evaluation, Authorisation, increases, the number of isomers increases exponentially
and Restriction of Chemicals) (European Commission 2006).
The naming convention was also used identify hydrocarbon sol-
5As described in the REACH Guidance (ECHA 2012), a DNEL is the
vents for which hazard information was supplied to the OECD
in response to the High Production Volume (HPV) initiative. level of exposure above which humans should not be exposed to. DNELs
are normally calculated by identifying a NOAEL from an animal study
As shown in Table 2, “Stoddard solvent-like” solvents belong in and applying assessment factors (ECETOC 2010). DNELs are normally
“Category 3” and are generically described as C9–C14 aliphatic calculated for workers (occupational) and the population at large, and
solvents with 2–25% aromatic constituents, but the category con- consider exposure by oral, dermal, and inhalation routes (ECHA 2012).
Table 2. Hydrocarbon solvent categories and substance identification by the naming convention with cross-referencing to identification by Chemical
Abstract Services (CAS) Registry number.
Primary distinguishing
characteristic of
Primary distinguishing substances within the
Category/Substance name CAS number(s) Carbon number rangea characteristic of category category
Category 1
C9 Aromatics
Hydrocarbons, 64742-82-1 C8-–C10  99% aromatics None
C9 aromatics 64742-95-6
Category 2b
C10-–C12 Aromatics
Hydrocarbons, 64742-94-5 C9-–C11  99% aromatics  1% naphthalene
C10 aromatics,
 1% naphthalene
Hydrocarbons, 64742-94-5 C9-–C11  99% aromatics  1% naphthalene
C10 aromatics,
 1% naphthalene
Hydrocarbons, 64742-94-5 C10-–C13  99% aromatics  1% naphthalene

C10-–C13 aromatics,
 1% naphthalene
Hydrocarbons, 64742-94-5 C10-–C13  99% aromatics  1% naphthalene
C10-–C13 aromatics,
 1% naphthalene
Category 3
C9-–C14 Aliphatics
(2-–25% Aromatics)
Hydrocarbons, 64742-81-0 C9-–C11 10-–25% aromatics  2% aromatics
C9-–C10, NICc, aromatics (2-–25%) 64742-82-1
C8-–C12, NIC, aromatics (2-–25%) 64742-82-1
C9-–C12, NIC, aromatics (2-–25%) 64742-82-1

C10-–C13, NIC, aromatics (2-–25%) 64742-82-1
C11-–C14, NIC, aromatics (2-–25%)
Category 4
C14–C20 Aliphatics
(2–30% Aromatics)
Hydrocarbons, 64741-44-2 C11–C17 10–29% aromatics  2% aromatics
C14–C18, NIC, aromatics (2–30%) 64742-81-0
64742-13-8
Hydrocarbons, 64742-80-9 C16–C20 10–28% aromatic  2% aromatics
C16–C20, NIC, aromatics (2–30%) 68915-96-8
Hydrocarbons, C11–C20, NIC, 64741-44-2 C11–C20 3–25% aromatics  2% aromatics
aromatics (2–30%) 64742-13-8
64742-80-9
Category 5
C5 Aliphatic
Normal pentane 109-66-0 C5 Pentane C5 aliphatic
Isopentane 78-78-4 C5 Isopentane isomers C5 aliphatic
Cyclopentane 287-92-3 C5 Cyclopentane C5 aliphatic
Hydrocarbons, C5, 78-78-4 C5 Pentane isomers C5 aliphatic
n-alkanes, isoalkanes 109-66-4
64742-49-0
Category 6
C6 Aliphatics
Normal hexane 110-54-3 C6 Normal hexane Normal hexane
Isohexane 107-83-5 C5–C6 Hexane isomers  5% n-hexane
64742-49-0
Hydrocarbons, C6, NIC, 5–80% 64742-49-0 C6–C7 Mixed hexane/heptane
n-hexane isomers
92112-69-1  5% n-hexane
Hydrocarbons, C5–C7, n-alkanes, 64742-49-0 C5–C7 Mixed C5–C7 isomers  5% n-hexane
isoalkanes,  5% n-hexane
Hydrocarbons, C5–C7, isoalkanes, 64742-49-0 C6–C7 Mixed C5–C7 isomers  5% n-hexane
cyclics,  5% n-hexane
92062-12-2
Category 7
C7–C9 Aliphatics
Hydrocarbons, C7, NIC 64742-49-0 C7 Heptane isomers  5% n-hexane
92045-53-9
(continued)
Table 2.(continued)
characteristic of
Hydrocarbons, C6–C7, NIC,  5% 142-82-5 C6–C7 Mixed hexane/heptane  5% n-hexane
n-hexane isomers
64741-84-0
64742-49-0
64742-89-8
Hydrocarbons, C6–C7, NIC,  5% 64742-49-0 C6–C7 Mixed hexane/heptane  5% n-hexane
n-hexane isomers
Hydrocarbons, C7–C9, NIC 64741-84-0 C7–C9 Mixed C7–C9 isomers  5% n-hexane
64742-48-9
64742-49-0
Hydrocarbons C6–C10, n-alkanes, 90622-50-7 C6–C10 Mixed C6–C10  5% n-hexane
isoalkanes, isomers
 5% n-hexane
Hydrocarbons C7–C9 isoalkanes 90622-56-3 C7–C9 Mixed C7–C9 isomers  5% n-hexane
Hydrocarbons, C7–C8 cyclics 64742-48-9 C7–C9 Mixed C7–C9  5% n-hexane
cycloparaffins
Normal heptane 142-82-5 C7 n-heptane  5% n-hexane
Normal octane 111-65-9 C8 n-octane  5% n-hexane
Iso-octane 540-84-1 C8 Mixed octane isomers  5% n-hexane
90622-56-3
Nonane 111-84-2 C9 n-nonane  5% n-hexane
Category 8
C9–C14 Aliphatics ( 2% Aromatics)
Hydrocarbons C9–C10, NIC, 2% 64742-48-9 C9–C10 Mixed isomers  2% aromatics
aromatics 48-9
Hydrocarbons C9–C11, NIC,  2% 64742-48-9 C9–C11 Mixed Isomers  2% aromatics
aromatics
Hydrocarbons C9–C11, isoalkanes, 64742-48-9 C9–C11 Mixed Isomers  2% aromatics
cyclics,  2% aromatics

Hydrocarbons C10–C13, NIC,  2% 64742-47-8 C10–C14 Mixed isomers  2% aromatics
aromatics
64742-48-9
93924-07-3
90622-58-5
aromatics alkanes
cyclics,  2% aromatics isoalkanes,
cycloparaffins
aromatics alkanes
cyclics,  2% aromatics isoparaffins,
cycloparaffins
Hydrocarbons, C10–C12 isoalkanes, 64741-65-7 C10–C12 Mixed Isomers  2% aromatics
 2% aromatics isoparaffins
90622-57-4
93924-07-3
64771-72-8
93924-07-3
Hydrocarbons, C9–C11 cyclics,  2% 64742-48-9 C9–C11 Mixed Isomers  2% aromatics
aromatics cycloparaffins
Nonane 111-84-2 n-C9 Monoconstituent  2% aromatics
Decane 124-18-5 n-C10 Monoconstituent  2% aromatics
Undecane 1120-21-4 n-C11 Monoconstituent  2% aromatics
Dodecane 112-40-3 n-C12 Monoconstituent  2% aromatics
(continued)
characteristic of
Tridecane 629-50-5 n-C13 Monoconstituent  2% aromatics
Isododecane 64741-65-7 Iso-C12 Mixed isomers  2% aromatics
Category 9
C14–C20 Aliphatics ( 2%
Aromatics)
Hydrocarbons C14–C18, NIC,  2% 64742-47-8 C14–C18 Mixed isomers  2% aromatics
aromatics
aromatics
aromatics
Hydrocarbons, 90622-46-1 C14–C18 Mixed Isomers  2% aromatics
C14–C18, n-alkanes, isoalkanes,  2% 90622-53-0
aromatics
aromatics
C16–C20, n-alkanes, isoalkanes,  2%
aromatics
C13–C16, isoalkanes, cyclics,  2% 64742-47-8
aromatics
Hydrocarbons, C14–C19, isoalkanes, 64742-46-7 C14–C19 Mixed Isomers  2% aromatics
C14–C17, n-alkanes,  2% aromatics 64771-72-8 C14–C17 Mixed Isomers  2% aromatics
90622-47-2
C14–C20, n-alkanes,  2% aromatics 64771-72-8 C14–C20 Mixed Isomers  2% aromatics
Tetradecane 629-59-4 C14 monoconstituent  2% aromatics
Pentadecane 629-62-9 C15 monoconstituent  2% aromatics
Hexadecane 544-76-3 C16 monoconstituent  2% aromatics

n-Heptadecane 629-78-7 C17 monoconstituent  2% aromatics
Octadecane 593-45-3 C18 monoconstituent  2% aromatics
Nonadecane 629-92-5 C19 monoconstituent  2% aromatics
Icosane 112-95-8 C20 monoconstituent  2% aromatics
aAccording to the rules of the naming convention, the carbon number range must include 80% of the constituents, but constituents present at lower levels
are not necessarily reflected in the name of the substance name.
bCategory 2 was defined as C10-–C12 aromatics for REACH purposes, based on the rules of the naming convention rules. However, it was re-designated
as C10-–C13 aromatics in the OECD HPV program.

cNIC  Normal paraffins, isoparaffins, cycloparaffins (cyclics).
and it becomes impractical, if not impossible, to identify all the boundary conditions that define the limits of similarity and
of the constituents of the more complex solvents. Further, a the concentrations of uniquely hazardous constituents. This is
detailed analytical characterization of any particular solvent is the basis for substance categorization, described in more detail
at best only representative, since the majority of the solvents below. When toxicologically important effects were identified
are complex, with variable compositions that depend on the in the studies of representative materials, the findings usually
manufacturing processes and the characteristics of the petro- led to further studies to identify the causative agents and to
leum-derived feed stocks from which they are manufactured. investigate the underlying mechanisms. One of the advantages
This inherent compositional variability is not problematic of the naming convention as a means of describing hydrocar-
from a technical perspective, as the end uses are dictated by bon solvents and the category approach to hazard character-
the physical/chemical properties of the solvents. On the other ization is that the substance descriptions are more constrained,
hand, from a toxicological perspective, it is recognized that the boundary conditions are better defined, the constituents are
compositional variability may present difficulties characteriz- more clearly identified, and the concentrations of particularly
ing the hazards of these complex substances. Accordingly, the hazardous constituents are more clearly specified, than is the
petroleum and hydrocarbon solvent industries have relied on case with the CAS descriptors.
tests of representative substances to characterize the hazards The first systematic attempt to characterize the hazards
of “similar” substances, with an underlying principle being of hydrocarbon solvents, primarily as a basis for occupa-
that constituents of similar structures have similar toxicologi- tional exposure advice, was a project initiated in the 1960s
cal properties. There are a few exceptions such as n-hexane by the American Petroleum Institute (API) and conducted by
and naphthalene, but these exceptions are uncommon, and Carpenter and associates at the Chemical Hygiene Fellowship
their toxicological properties are often associated with of the Carnegie-Mellon Institute of Research (now Carnegie-
structures that lead to the production of metabolites that are Mellon University). As described in the first paper in the series
constituent-specific. However, when relying on representative (Carpenter et al. 1975a), the test samples were supplied by
substance information, it is always necessary to be mindful of manufacturers and met the technical specifications that were
then in effect for the specific substances. Each substance was observed adverse effect levels (LOAELs)7 were not always
then tested for acute and repeated dose (13-week) inhalation experimentally defined. An overall conclusion of the study
toxicity in Harlan-Wistar rats and beagle dogs. The studies pre- with Stoddard solvent (Carpenter et al. 1975c) was that the
dated the development of international guidelines and Good human and animal data supported an occupational exposure
Laboratory Practice regulations, but were conducted to the limit of 200 ppm (1050 mg/m3), which was the recom-
best scientific standards of the times. In the repeated dose stud- mendation of the American Conference of Governmental
ies, the animals were exposed to vapors of test material at one Industrial Hygienists (ACGIH®) at the time the studies
of three graded levels or to an air control with the exposure were conducted.8
concentrations measured at least once/day. At the end of the Following Carpenter, there were investigations of the effects
13-week exposure period, blood samples were taken for hema- of these solvents at higher levels to better characterize margins
tological and clinical chemistry measurements, and the ani- of exposure, and also to better understand the underlying pro-
mals were then sacrificed for gross evaluation and microscopic cesses leading to the effects reported, particularly those in the
examination of the major organ systems. Other tests that were kidneys of male rats. Further studies of the effects of hydro-
conducted included the assessment of upper respiratory tract carbon solvents led to the observation that the kidney effects
irritation in mice using the method described by Alarie (1966), described by Carpenter were produced by some hydrocarbon
and in many cases, there were also studies in which volunteers solvent constituents including branched paraffins (Phillips and
were exposed for short periods of time to graded doses of test Egan 1984a, Phillips and Egan 1984b, Phillips and Cockerell
material to help identify levels that would be considered tolera- 1984), who referred to these changes as “light hydrocarbon
ble for occupational exposure. The results of these studies were nephropathy”. In later studies, these kidney changes were
published in a series of 17 papers that appeared between 1975 shown to be consistent with an a2u-globulin-mediated pro-
and 1978 (Carpenter et al. 1975a, 1975b, 1975c, 1975d, 1975e, cess, that is male rat-specific and not relevant to humans (US
1975f, 1975g, 1975h, 1976a, 1976b, 1976c, 1976d, 1976e, EPA 1991, Swenberg and McKeeman 1998).
1977a, 1977b, 1977c, 1978), and for many years comprised The objectives of the present report are to summarize the
much of the toxicological information on these substances and physical/chemical properties of these solvent groups and
provided the principal data underpinning occupational expo- the toxicological properties of solvents and constituents; to
sure recommendations. provide a detailed discussion of the more generic properties
Among other things, these studies showed that high of the solvents and the unique properties of some specific
acute exposures to volatile hydrocarbons could cause ocu- constituents; to explain the process used to group similar
lar and respiratory tract irritation and acute central nervous substances (categorization), and the new nomenclature for
system (CNS) depression, but there was little evidence hydrocarbon solvents; and finally, to show how these data can
of toxicologically relevant systemic effects. For example, be used to characterize toxicological hazards for purposes of
in the study of Stoddard solvent (Carpenter et al. 1975c), hazard classification and development of occupational expo-
exposure of rats to 1400 ppm (8200 mg/m3)6 for 8 h was sure limits.
associated with eye irritation, bloody exudate around the
nostrils, and slight loss of coordination. In repeated expo- Toxicokinetic properties of saturated constituents
sure studies at vapor concentrations of 84, 190, or 330 ppm of hydrocarbon solvents
(480, 1100, and 1900 mg/m3), the only notable pathological A fundamental requirement for the use of representative
observation was a change in the kidneys of male rats. In substances to characterize the hazards of a larger group of
volunteer studies, 150 ppm (850 mg/m3) was considered substances is that the substances in the category should have
to produce only slight eye irritation whereas at 470 ppm properties that are similar or at least vary in a predictable way.
(2663 mg/m3), the degree of eye irritation was considered To begin to address the issue of whether hydrocarbon solvents
to be unacceptable. One limitation of these studies was that have properties that are sufficiently similar to justify a repre-
they were intended for “safety” assessments and examined sentative substance approach, it is reasonable to start with the
the potential for effects at levels approximating the levels toxicokinetic properties of the various constituents. As described
at which humans were being exposed. The volunteers were below, there are two general types of hydrocarbon solvent con-
exposed for a short duration, and therefore the potential stituents: saturated components (alkanes, i.e., normal paraffins,
effects of repeated exposure could not be assessed, whereas isoparaffins, and cycloparaffins) and aromatics. Although there
the toxicological studies in animals assessed the poten- are metabolic differences between the saturated constituents and
tial for effects of repeated exposures but were not always the aromatics, the major difference is that the aromatics without
conducted at the highest possible exposure levels. In most alkyl side chains (benzene, naphthalene) are metabolized by ring
cases, the toxicological studies in animals did not identify oxidation, leading to uniquely toxic metabolites as described in
human-relevant effects at the highest levels tested, so the more detail below.
no observed adverse effect levels (NOAELs) and lowest
7For purposes of this paper, a NOAEL is the highest dose or concentra-
6In some studies, the exposure levels were given as mg/m3 and in oth- tion of a test material that can be given to an animal without causing
ers, as parts per million (ppm). To avoid confusion, in this report, the adverse effects. A LOAEL is the lowest concentration of a test material
exposure levels are reported in the units given by the original authors, but that causes an adverse effect.
conversion factors are provided in the appendices, and, to the extent pos- 8There was an ACGIH TLV® of 500 ppm (2625 mg/m3) from 1948 to
sible, the data were converted, usually from ppm to mg/m3, to facilitate 1969. This value was reduced to 200 ppm (1050 mg/m3) in 1970 and to
comparisons among studies. 100 ppm (525 mg/m3), the present value, in 1976.
Absorption (C10 cycloparaffin), 53–67% of the administered material was

eliminated in the urine, with the total amount of eliminated mate-
Absorption of inhaled hydrocarbon solvent constituents
rial depending on the isomeric configuration (Elliott et al. 1966).
Due to of their low molecular weight and high volatility, Importantly, absorption rates of complex aliphatic hydrocarbon
inhaled pentanes are not well absorbed (Dahl et al. 1988) solvents were similar to those of the individual constituents. In a
and are eliminated primarily by exhalation (Filser et al. 1983, study with pigs, Tulliez (1986) measured the absorption of a min-
unpublished data summarized by Galvin and Marashi 1999c). eral oil (60% n-paraffins, 7% isoparaffins and 33% cycloparaffins)
Higher molecular weight, less volatile hydrocarbons are relatively with an average carbon number of C16 (i.e., a carbon number
well absorbed, metabolized to water-soluble forms and excreted range that overlaps that of higher molecular weight hydrocarbon
as urinary metabolites. Sprague-Dawley rats were exposed via solvents). In this study, test material was given daily by dietary
short term inhalation (8 hours/day for 14 days) to saturated air administration for 10 consecutive days. The absorption rate of
concentrations of C9–C13 normal alkanes to evaluate short 88% of ingested dose was higher than the 53% estimated from
term toxicity (Nilsen et al. 1988). Blood/air and brain/air parti- the empirical relationship of Albro and Fishbein. The higher
tion coefficients provided evidence that C9–C11 normal alkanes absorbed fraction of the complex C16 hydrocarbon substance, in
were more readily absorbed and distributed to the CNS than the comparison to the predicted amount, is similar to the results from
C12–C13 substances. Mean brain hydrocarbon concentrations
studies by Tulliez and Bories (1978), in which it was shown that

of 239.7, 45.4 and  0.5 mg/kg were noted for C10, C11, and absorption rate is inversely proportional to exposure dose over
C13 normal alkanes respectively. These values correlated with a 200-fold range. Hence, the absorbed fraction of the complex
the decreasing trend in mean steady state air concentrations with C16 hydrocarbon substance was probably higher than expected
increasing carbon number, a reflection of the differences in vapor because it had been administered at low dietary levels.
pressure. Data from Zahlsen et al. (1992, 1993) indicate that In summary, the percentages of n-, iso- and cycloalkanes
normal- and isoparaffinic hydrocarbons with carbon numbers in with the same total number of carbon atoms that are absorbed
the range of C8–C10 reach similar steady state concentrations in the gastrointestinal tract are similar, and thus, for con-
in the brain, but the levels of cycloparaffinic hydrocarbons reach stituents of these types, absorption seems to be influenced
maximum concentrations at C8 and then decline with increasing by carbon numbers and doses but not by the structures of the
molecular weight. Accordingly, for hydrocarbons with carbon constituent hydrocarbons.
numbers  C12, exposure by the inhalation route is expected
to be limited because of their low vapor pressures, and may be

Percutaneous absorption of aliphatic hydrocarbon
further limited by blood/brain barrier effects (Hau et al. 2001).
solvent constituents
Absorption of ingested aliphatic hydrocarbon solvent constituents Under most circumstances, volatile hydrocarbon solvent constit-
Hydrocarbon solvent constituents are reasonably well absorbed uents are poorly absorbed through the skin, in part because they
if ingested, but the extent of absorption decreases with increasing are more likely to evaporate than to be absorbed. Absorption
molecular weight. In a study to characterize oral absorption of rates were measured for n-pentane, 2-methylpentane, n-hexane,
various saturated and unsaturated aliphatic hydrocarbons, male n-heptane, and n-octane using excised abdominal skin flaps from
rats were fed alkanes over a wide range of molecular weights, rats (Tsuruta 1982). In these studies, one ml of test substance was
and relative absorption rates were evaluated (Albro and Fishbein added to the upper chamber of a diffusion cell, and the amount
1970). It was shown that the principal site of absorption for of test substance that had diffused into the lower chamber con-
ingested hydrocarbons was the small intestine and that there were taining 0.9% NaCl was determined by gas chromatography after
no differences in the absorption rates for n-, iso- and cycloparaf- several hours of shaking. The absorption rate was inversely
fins with the same carbon numbers (P  0.9; two-tailed t-test). It related to the number of carbon atoms, with pentane having the
was concluded that the carbon number was the most important highest percutaneous absorption rate (0.519 nmoles/min/cm2
determinant of absorption with an inversely proportional rela- of skin), and n-octane the lowest (0.0817  10 3 nmoles/min/
tionship between the carbon number and the fraction absorbed, cm2). Similar penetration rates were noted for n-hexane and its
represented by the empirical formula: 2-methylpentane isomer, at 0.0118 and 0.0209 nmoles/min/cm2
respectively. Based on the hexane studies, the overall rate of der-
Percentage absorption  115.9–3.94 × (number of carbon atoms) mal absorption for n-alkanes is  0.1 mg/hr/cm2. In a separate
study, it was shown that  5% of topically applied n-hexadecane
Using this formula, the approximate percentages of absorbed penetrated fresh intact porcine skin, regardless of the vehicle
hydrocarbons for solvents in differing carbon ranges are: through which it was applied (Brown et al. 1994).
∼ 92% absorption for a C6 aliphatic hydrocarbon solvent, The results of rat skin studies were extended in studies of
∼ 70% absorption for a C9–C14 aliphatic hydrocarbon solvent, human skin to evaluate the permeability coefficients of larger
∼ 50% absorption for a C14–C20 aliphatic hydrocarbon solvent carbon chain constituents (decane, C10; undecane, C11, and
dodecane, C12) to quantify percutaneous absorption of a related
Across the entire range, the percentage of hydrocarbon substance, jet fuel (JP-8), in humans following dermal contact.9
absorbed decreases from  90% (for C6 constituents) to approxi-
mately 37% (for C20 constituents). Consistent with the predic- 9The technical specifications for JP-8 jet fuel include a distillation range
tions, following oral administration to rats (0.5 mg/kg bw), the of approximately 205°C–300°C and a maximum aromatic content of
fraction of absorbed pristane (C19) was about 45% (Le Bon 25%. Thus, jet fuel is similar to Stoddard solvent but with somewhat
1988). In another oral study in rabbits with radiolabeled decalin higher molecular weight constituents.
Ten healthy adult volunteers (five males and five females) with cycloparaffins or aromatics across a C6–C10 carbon number
no prior occupational exposure to jet fuel were recruited for range. For example, after 3 days of exposure by inhalation to
the study. One ml of undiluted JP-8 fuel was applied to 10 cm2 100 ppm (581 mg/m3), the mean distribution of normal decane
of the forearm of each volunteer for 0.5 h, using sealed expo- (nC10) was 6.8/45.9/60.2/1230 mmol/kg in blood/liver/brain/
sure chambers to limit evaporation. The permeability coeffi- fat. The corresponding mean distribution of t-butylcyclo-
cients (cm/h) of decane, undecane, and dodecane, which were hexane (C10 cycloparaffin) was 12.9/21.9/60.2/1363 mmol/kg.
calculated based on levels of JP-8 components in blood gener- In a similar study by the same group (Zahlsen et al. 1993),
ally decreased with increasing carbon number. The exception tissue distributions of hydrocarbons with dissimilar structures
was undecane, which had a lower permeability coefficient than but similar carbon number ranges (C8–C10) were measured
dodecane [decane  6.5  10 6, undecane  4.5  10 7, and in male Sprague-Dawley rats following exposure to 100 ppm,
dodecane  1.6  10 6] (Kim. et al. 2006). 12 hours/day for 3 consecutive days. Overall, as expected,
In summary, data from studies with humans, animals, and tissue concentrations of C8–C10 isoalkanes increased with
excised skin have all shown that C5–C12 alkanes are poorly increasing carbon number, because of the increase in octanol/
absorbed following dermal administration. As the percuta- water partition coefficients with increasing molecular weight.
neous absorption rates varied inversely with the number of Whole-body distribution of 14C-heptadecane and 14C-
carbon atoms in the molecules, it can be reasonably assumed octadecane was investigated in rats (Tulliez and Bories 1978).
that the percutaneous absorption rates for aliphatic constituents Both hydrocarbons were detected in the liver within 6 h of
with  12 carbons would be even lower than those measured for single oral doses. However, most of the absorbed hydrocarbons
the C10–C12 constituents (although the C12  constituents have partitioned into the adipose tissue after 12 h, with a marked
lower vapor pressures, so the contact periods could be longer). decline in liver hydrocarbon content by 7 days (Tulliez and
Bories 1978). In a similar study, male and female rats were
Systemic distribution of aliphatic hydrocarbon fed a diet containing 280 mg of n-heptadecane per kg of feed
solvent constituents for 12 months. Rapid accumulation of heptadecane increased
Concentrations in blood of n- and cycloalkanes with equiva- for the first 4 months before reaching steady state. Adipose
lent carbon numbers were similar to each other but lower than tissue concentrations of heptadecane were 80 and 272 mg/kg in
the values for the corresponding aromatic molecules (which males and females respectively, while levels in the liver were
are more water soluble) (Eide and Zahlsen 1996). In studies 2.5 and 1.9 mg/kg respectively (Tulliez et al. 1975c). A similar
involving exposure by inhalation for 12 h, tissue distribu- distribution pattern was observed in rats fed a diet containing
tions varied slightly by dose; there was a linear increase in 0.1% eicosane for 7 days (Tulliez and Bories 1975a, 1975b).
tissue concentrations of a C9 cycloalkane (1,2,4-trimethyl- Analysis of hydrocarbon tissue content by day 10 showed that
cyclohexane) whereas quadratic relationships were found for 7.2% of the total hydrocarbon ingested had been retained in the
a n-nonane and 1,2,4-trimethylbenzene (Eide and Zahlsen carcass. As shown for the normal C17 and C18 hydrocarbons,
1996). Levels of tissue uptake for cycloparaffins appeared eicosane partitioned preferentially to the adipose tissue (317
to be marginally higher than those for n-alkanes when the mg/g in adipose tissue compared to 3 mg/g in liver). There were
experiments were conducted at low exposure levels, but the no differences in absorption and distribution patterns when the
reverse was true at vapor concentrations exceeding 250 ppm experiment was extended over a 6-month period; 6.6% of total
(Eide and Zahlsen 1996). Levels of aromatics in blood and hydrocarbon ingested was detected in the carcass. Liver and
fat were higher than those of the corresponding aliphatics at adipose tissue concentrations were 17.2 and 1172 mg/g respec-
both low and high exposure levels, but levels of aromatics in tively (Tulliez and Bories 1975a, 1975b).
the CNS were similar to those of n-paraffins and lower than Le Bon (1988) evaluated the distribution kinetics of pris-
those of cycloparaffins (Eide and Zahlsen 1996). In other tane (branched C19 paraffin). Male Wistar rats were given a
studies, however, systemic absorption of n-alkanes appeared single oral dose (0.5 mg/kg bw) of 3H-pristane, and the distri-
to be lower than levels of cycloparaffins over the C6-C9 bution of radioactivity in tissues was measured after 7 days.
carbon number range, but higher for C10 alkanes than for the The highest concentrations of radioactivity were found in the
corresponding cycloparaffins (Zahlsen et al. 1992, 1993). liver and adipose tissue, followed by the spleen, kidney, heart,
Compared to aromatics, similar brain/blood, fat/blood and and lungs. Only 0.5% of the administered dose was detected in
fat/brain ratios for n-nonane and 1,2,4-trimethylcyclohexane the liver (Le Bon 1988).
indicated strong similarities in biological affinities to proteins Tulliez and Bories (1979) fed rats a diet of dodecylcyclo-
and/or lipids, solubility, and tissue distribution at high expo- hexane (C18) for 3 months, followed by control diet for 5 months.
sure concentrations (500–1000 ppm) (Zahlsen et al. 1990). In Hydrocarbon levels in the liver increased over time before reach-
a study to evaluate tissue distribution of C6–C10 n-paraffins, ing steady state after 2 months. Similar to n-heptadecane (C17)
aromatics and cycloparaffins, rats were exposed 12 hours/day and n-octadecane (C18), 7% of ingested dodecylcyclohexane was
for 3 consecutive days to each of these constituents at 100 ppm. retained in the carcass after 8 months. Liver and adipose tissue lev-
Aromatic hydrocarbons were more soluble with higher con- els were 30 and 1134 mg/g respectively after the 3-month exposure
centrations in the blood and moderate tissue accumulation in period (Tulliez and Bories 1979). Tissue distribution of dodecyl-
the brain, liver, and kidneys (Zahlsen et al. 1992). In contrast, cyclohexane in rats 24 h after the administration of a single oral
in repeated inhalation studies, cycloparaffins and n-paraffins dose (100 mg/kg bw) was similar to that observed in the heptade-
showed lower blood concentrations but higher concentra- cane and octadecane studies. The highest levels of radioactivity
tions in the brain than aromatics, with n-paraffins showing a were found in the adipose tissue, liver, kidneys, lungs, and blood.
lower potential for accumulation in adipose tissues than either Adipose tissue concentrations were 4–5 times higher than those
in the liver and 100 times greater than those observed in blood a shoe factory who were exposed to these substances under
(Tulliez and Bories 1979). occupational conditions (Perbellini et al. 1981). The profile of
Although the data are limited, there is no evidence to sug- urinary metabolites was identical to that found in the rat study
gest that there would be large differences in the tissue distri- from Frontali et al. (1981).
butions of n-paraffins, isoparaffins or cycloparaffins following Olson et al. (1986) evaluated the metabolism of n-octane
exposure. Overall, regardless of structure, aliphatic hydrocar- (C8 n-alkane) in F-344 rats. Male and female rats (12 and 14/
bons partition to tissues with a higher adipose content. The group respectively) were given 1.4 g/kg bw of n-octane by oral
preferential disposition of aliphatic hydrocarbons in adipose gavage on alternate days over a 2-week period. Urine samples
tissue compared to blood was observed both with differing were collected within the first 48 h of dosing and analyzed for
routes of exposure (inhalation and oral) and over a range of the presence of urinary metabolites. GC analysis showed that
carbon numbers (C6–C20), which encompasses nearly all of n-octane was primarily metabolized to the 2- and 3- mono-
the aliphatic constituents of hydrocarbon solvents. alcohols, with 2-octanol as the major urinary metabolite in
females, compared to 5-oxohexanoic acid in males. The four
Metabolism and excretion of aliphatic hydrocarbon key metabolites identified in the urine were similar in males and
solvent constituents females, with differences in the relative amounts; in male rats,
C5–C9 hydrocarbon solvent constituents 2-octanol, 3-octanol, 5-oxohexanoic acid, and 6-oxohexanoic
acid were identified in relative amounts of 7:12:15.5:1. The
To study the metabolism of n-pentane (C5 n-paraffin) and relative amounts in the female rat urine were 2.0:1.7:0.7:1.
isopentane (C5 isoparaffin), male mice were exposed to 5% The presence of the 5- and 6-oxohexanoic acid metabolites
concentrations of n-pentane and isopentane in ambient air for is evidence of oxidative metabolism at the end carbons. In
1 h, followed by analyses of blood and liver tissues for the a similar gavage study of 2-methylheptane (C8 iso-alkane),
presence of metabolites (Chiba and Oshida 1991). Both hydro- metabolism strongly favored the formation of alcohols and
carbons were metabolized in the liver to alcohols and ketones, their carboxylic acid oxidation products, similar to the results
with 2-pentanol, 3-pentanol, and 2-pentanone detected as the obtained with n-octane (Serve et al. 1991). Major urinary
principal metabolites of n-pentane. The initial metabolites of metabolites in order of relative abundance were 2-methyl-2-
isopentane isomers were the corresponding alcohols, 3-methyl- ,5-heptanediol  2-methyl-5-heptanolactone  2-methylhep-
2-butanol and 2-methyl-2-butanol. The same metabolites were tanoic acid  2-methyl-1,2-heptanediol, all of which were
detected under in vitro conditions when normal- and isopen-
found in urine as sulfated and glucuronidated conjugates.

tane were incubated with mouse liver microsomes, and in vivo In an in vivo study of the metabolism of cyclohexane, adult
when male ICR mice were pre-fed with phenobarbital at 80 rabbits were given single gavage doses (0.3–400 mg/kg bw).
mg/kg bw to stimulate CYP450 enzyme induction in the liver. Nearly all (95%) radioactivity was recovered in expired air as
To determine the metabolic pathway for C5–C7 aliphatic 14C carbon dioxide and unchanged cyclohexane, and in the urine
hydrocarbons, Frontali et al. (1981) exposed rats to n-hexane (50–90% of the orally administered dose) primarily as cyclo-
– 500 or 2500 ppm [1786 or 8929 mg/m3] (30 weeks), 1500 hexanol with lower amounts of the trans-1,2-cyclohexane-diol
or 5000 ppm [5358 or 17858 mg/m3] (14 weeks), 1500 or (Elliott et al. 1959). As described earlier for n- and iso-octane,
2500 ppm [5172 or 8621 mg/m3] (30 weeks); cyclohexane – the alcohol and diol metabolites were excreted in the urine as
3000 ppm [9091 mg/m3] (30 weeks); n-pentane – 1500 ppm glucuronide conjugates. To determine what role methylation of
[6250 mg/m3] (30 weeks); n-heptane – 1500 ppm [5357 mg/m3] the cyclohexane ring could play in in vivo metabolism, Elliott
(14 weeks); 2-methylpentane – 1500 ppm [5357 mg/m3] (14 et al. (1965) administered 14C-methylcyclohexane (2–2.5
weeks); and 3-methylpentane – 1500 ppm [5357 mg/m3] (14 mmols/kg bw) to rabbits by dietary administration. As shown
weeks). The animals were exposed 9–10 hours/day for 5–6 for cyclohexane, 80% of 14C-methylcyclohexane-derived
days/week. Animals exposed to 2500 [8929 mg/m3] (30 weeks) radioactivity was excreted in expired air and urine (65% in
or 5000 ppm [17858 mg/m3] (14 weeks) of n-hexane devel- urine and 15% in expired air), with 0.5% in feces and 4–5%
oped peripheral axonal degeneration, but no neurological present in the carcass. A third of the radioactivity in expired
effects were observed in the studies of cyclohexane, n-heptane, air was 14C-carbon dioxide and the remainder was unchanged
n-pentane, or branched hexane isomers. These data provided methylcyclohexane. Consistent with the metabolic profile for
evidence that the n-hexane metabolic pathway involves the cyclohexane, 7 methylcyclohexanols were detected in the urine
production of a toxic metabolite, 2,5-hexanedione. as glucuronide conjugates with trans-4-methylcyclohexane as
Rat urinary metabolites were mainly glucuronidated forms of the major urinary metabolite (14.7%). Perbellini et al. (1981)
alcohols and included 2-methyl-2-pentanol following 2-methyl- evaluated the lung uptake and metabolism of cyclohexane in 22
pentane exposure, and 3-methyl-2-pentanol and 3-methyl-3-pen- workers at a shoe factory who were exposed occupationally to
tanol following 3-methylpentane exposure. Although 2-hexanol cyclohexane (17–2484 mg/m3; 5–720 ppm). Urinary concen-
was found to be the major urinary metabolite of n-hexane in trations of cyclohexanol, the principal cyclohexane metabolite,
rats, the other metabolites identified included 3-hexanol, gam- correlated with blood levels of cyclohexane and accounted for
ma-valerolactone, 2,5-dimethylfuran, and 2,5-hexanedione, a 0.1–0.2% of inhaled cyclohexane (Perbellini et al. 1981).
gamma di-ketone metabolite (Frontali et al. 1981).
A similar study was conducted to investigate the urinary
C9–C14 hydrocarbon solvent constituents
metabolites associated with n-hexane (32–500 mg/m3;
9–140 ppm), 2-methylpentane (11–250 mg/m3; 3–70 ppm), and In a study to evaluate the metabolism of tert-butylcyclohexane
3-methylpentane (10–204 mg/m3; 3–57 ppm) in 41 workers at (a C10 cycloparaffin with a branched alkyl side group attached
to a cyclohexane ring), F-344 rats were given 0.8 g/kg hydro- pristane, 8.3% of the radiolabel remained in the carcass after
carbons by oral gavage every 2 days for 14 days. Urine samples 7 days ( 75% as pristane metabolites), primarily in adipose
were collected during the first 48 h of exposure to determine uri- tissue and liver. Characterization of tissue metabolites by thin
nary metabolites. As shown for the straight chain and branched layer chromatography, gas chromatography, and mass spec-
chain alkanes, t-butylcyclohexane underwent oxidative metabo- trometric analysis led to the identification of four metabolites:
lism primarily on the cyclohexane ring. Urinary metabolites pristan-1-ol, pristane-2-ol, pristanic acid, and 4, 8, 12-trimeth-
resulting from metabolic oxidation of the cyclohexane ring were yltridecanoic acid, demonstrating that metabolism of branched
mostly C4-derived cyclohexanols, similar to the metabolic pat- hydrocarbons also involves terminal (w) oxidation followed by
tern described previously for the cyclohexyl ring of cyclohexane the classic fatty acid b-oxidation process (Le Bon et al. 1988).
and methylcyclohexane. The alkyl side chains were mostly To investigate the metabolism of cycloparaffins, Tulliez and
metabolized to carboxylic acid compounds (2-methyl-2-cyclo- Bories (1979) followed the metabolism of 3H-labeled dodecyl-
hexylpropanoic acid) and alcohols (2-methyl-2-cyclohexyl-1,3- cyclohexane (cyclic C18 paraffin). The parent molecule was
propanediol) (Henningsen et al. 1987). Similar to the metabolic not detected in the urine despite extensive urinary excretion of
profile for tert-butylcyclohexane, the metabolism of cis- and 3H, suggesting almost complete metabolism of absorbed hydro-
trans- isomers of decalin in rabbits primarily involves the in carbon. Seven percent of absorbed dodecylcyclohexane remained
vivo hydroxylation of the C-2 position on the cyclohexyl ring to in the carcass unchanged (primarily in the lipid compartments),
yield secondary alcohols (mainly 2-decalol) (Elliott et al. 1966). while the rest was metabolized via an w-oxidation process to
Methylation of the C-2 position on the cyclohexyl ring did not cyclohexyldodecanoic acid, which was incorporated into neutral
change the metabolic profile as trans- and cis-2-methyldecalin lipids and phospholipids. The authors reported that the aliphatic
were metabolized to isomers of secondary alcohols (trans-cis-6- chain of the cycloparaffin underwent the normal fatty acid
or 7-methyl-2-decalol, cis-6 methyl-2-decalol and cis-7-methyl- degradation pathway while the cyclohexyl rings were mostly
2-decalol) (Robertson and Champion 1970). eliminated in urine as cyclohexylacetic acid (Tulliez 1986). Ring
hydroxylation was also shown to occur, leading to the generation
C14–C20 hydrocarbon solvent constituents of free hydroxylated cyclohexyl acids that were then eliminated
in the urine as conjugated metabolites (Tulliez 1986). However,
Tulliez and Bories (1978) evaluated the metabolism of low
a small proportion of the cyclohexyl ring structures were shown
doses of a radiolabeled straight chain n-C17 alkane (heptade-
to have undergone w–oxidation to w-cyclohexyl fatty acids
cane) in male Wistar rats. Animals were given a small amount
(Tulliez and Bories 1979). The authors concluded as follows:

of 14C-heptadecane (10 and 30 mCi; approximately 1 and 200
“our results clearly show that absorption, storage, and oxidation
mg n-heptadecane) by dietary administration. This was ingested
of the aliphatic chain of dodecylcyclohexane occurs in the same
in approximately 15 min, and the rats were then switched to
way as for the n-paraffin”. The authors also concluded that total
control diets. The kinetics of absorption and metabolism were
hydrocarbon absorption was related to the administered dose but
followed in rats sacrificed at 6, 12, 24, 48 h, and 7 days after
not to chemical structure, and that there were no differences in
ingestion of the marker compound (Tulliez and Bories 1978).
absorption or metabolism between cycloparaffins and n-paraffins
Much of the n-heptadecane was absorbed, although low levels
with similar carbon numbers (Tulliez and Bories 1979).
of radioactivity found as n-heptadecane in the feces provided
In studies of the metabolism of hydrocarbons with even
evidence that some of the ingested hydrocarbon was excreted
longer chains, Halladay et al. (2002) gave 1.8 g/kg bw of
unchanged. Despite the presence of radioactive 14C in the urine,
eicosanylcyclohexane (cyclic C26 aliphatic hydrocarbon) to
no heptadecane was detected, suggesting that the absorbed por-
Sprague-Dawley rats as single oral doses. The major metabo-
tion of the hydrocarbon was almost completely metabolized and
lites identified (12-cyclohexyldodecanoic acid and 10-cyclo-
excreted in the urine as water-soluble metabolites. Radioactivity
hexyldecanoic acid) appeared to be products of w-oxidation of
in the form of n-heptadecane accumulated in the liver during the
alkyl side chains. Thus, there is a common metabolic pattern
first 6 h, and then decreased rapidly, whereas labeled lipids in the
for aliphatic hydrocarbons across the range of carbon numbers
carcass accumulated progressively. About 7% of the 14C label
(C5–C20) found in hydrocarbon solvents.
was found as n-heptadecane in the lipid fraction, an indication
that a small portion of the hydrocarbon was retained unchanged.
Summary of toxicokinetic properties of aliphatic
The rest of the ingested hydrocarbon appeared to undergo
hydrocarbon solvent constituents
w-oxidation to heptadecanoic acid, which was then incorporated
into neutral lipids and phospholipids of similar chain length fol- There is good evidence that C5–C20 alkanes have similar
lowing the normal fatty acid degradation pathway (Tulliez and absorption, distribution, metabolism, and excretion proper-
Bories 1978). Similarly, a study with 14C labeled octadecane ties. It is also apparent that the pharmacokinetic properties of
(nC18) and hexadecane (nC16) showed that both hydrocarbons these molecules are more dependent on carbon chain length
were converted directly to fatty acids of the same chain length than differences in chemical structure. Alkanes in the C5–C20
via an w-oxidation pathway in rats (McCarthy 1964). range are absorbed in the small intestine from which they are
In a similar study, the metabolic fate of pristane (a C19 satu- distributed to tissues including liver, kidneys, adipose tissue,
rated isoalkane) in rats was evaluated following a single oral and brain. Absorption is inversely proportional to carbon chain
dose (Le Bon et al. 1988). Approximately 66% of the material length, with C6 hydrocarbons being absorbed about 3 times as
was excreted unchanged in the feces. About 14% of ingested efficiently as C20 hydrocarbons. As carbon numbers increase,
pristane was excreted in the urine, largely as water-soluble there is a corresponding increase in the fraction of hydrocar-
metabolites. In a study of orally administered, radiolabeled bons that are excreted unchanged in the feces.
Figure 1. Metabolic pathway of n-, iso-, and cycloparaffins in mammals (EFSA 2012).
The tissue distribution pattern for paraffins is similar regard- appear to be metabolized via an identical but abridged pro-
less of chemical structure. Following oral exposure, n-, iso- cess. Biotransformation by hepatic cytochrome P450 enzymes
and cycloparaffinic hydrocarbons preferentially partition to yields primarily hydroxylated metabolites that can be further
adipose tissue in comparison to blood and liver. Overall, there oxidized to the diols of corresponding chain lengths, carboxy-
is a common metabolic pathway for all classes of alkanes lic acids, ketones, and/or hydroxyketones at much lower levels.
(Figure 1). For absorbed long-chain paraffins, the first meta- These metabolites are eliminated as CO2 in expired air and as
bolic step is the w-oxidation of the alkyl chain leading to the sulfated and glucuronidated conjugates in urine.
formation of hydroxylated metabolites (mainly fatty alcohols) Overall, the similarities in the toxicokinetic/metabolic
and fatty acids of similar chain length. These fatty acids then profiles of the C5–C20 aliphatic hydrocarbons (i.e., systemic
undergo a b-oxidation process, leading to the formation of absorption, preferential distribution to adipose tissue, and
products that are either incorporated in the biosynthesis of common metabolic and excretion pathways) indicate that
fatty acids and lipids (Kolattukudy and Hankin 1966, Albro these substances are likely to have similar toxicological prop-
and Fishbein, 1970), exhaled as CO2, or excreted in the urine erties.10 Studies with complex aliphatic solvents consistently
as hydroxylated cycloacids (naphthenes only), either as free or
10One exception to the general rule that metabolism of saturated paraffins
conjugated metabolites. In some cases, the initial w-oxidation
leads to intermediates with similar toxicological properties is n-hexane.
step occurs on the branched chains of iso-paraffins, leading to Although the initial step (CYP 450-mediated hydroxylation to the mono-
the formation of water-soluble tertiary alcohols that are primar- alcohols, 1-, 2- and 3-hexanol) is the same as it is for all other normal
ily excreted in the urine without undergoing b-oxidation. For paraffins, there is a specific n-hexane metabolite, 2,5-hexane dione, that
cycloparaffins, direct oxidation of the cyclohexyl ring occurs to is unusually toxic but is not formed at toxicologically important levels
form cyclic alcohols (cyclanols) before the w-oxidation step. by hexane isomers, cyclohexane, n-pentane or n-heptane (Egan et al.
1980; Frontali et al. 1981; Ono et al. 1981; Takeuchi et al. 1980). The
When oxidation of the alkyl chain predominates, metabo- 2,5-hexane dione metabolite is responsible for the reproductive and neu-
lism of the cycloparaffins is identical to that for n- and isopar- rotoxic effects of n-hexane (IPCS 1991). (For more detailed discussion
affins. Hydrocarbons with shorter chain lengths (C5–C9) see Appendix C.II.10.)
show absorption, distribution, metabolism, and excretion dimethylbenzoic acid (Laham and Potvin 1989). Other TMB
profiles similar to those of the mono-aliphatic constituents, metabolites detected in the urine included mercapturic acids
suggesting that not only do aliphatic constituents of similar (via GSH conjugation of dimethylbenzyl alcohols), trimeth-
carbon chain length have similar toxicokinetic profiles, but ylphenols, and glucuronide and sulfate conjugates (Huo et al.
that interactions of C5–20 aliphatic hydrocarbons that could 1989, Mikulski and Wiglusz 1975, Wiglusz et al. 1975).
affect their metabolism, disposition within the body, or rates Walde and Scheline (1983) studied the metabolism of 100
of excretion are unlikely. Though most of the toxicokinetic mg of radiolabeled p-butyltoluene in male Wistar rats and
studies were conducted using the oral route of administration, guinea pigs following administration via the oral and inha-
it is anticipated that following absorption, metabolism, and lation routes (trace concentrations for 24 h). There were no
excretion of mono-constituent aliphatic hydrocarbons would differences in metabolic profiles that were related to the route
be similar. In other words, the route of exposure is important of exposure, as over 83% of radioactivity was excreted in the
when considering systemic dose, but has little impact on the urine or feces (3.5/1 ratio in the first 3 days) within 10 days,
fate of absorbed hydrocarbons. regardless of the route of administration. The principal urinary
metabolites in rats were (p-butylbenzoic acid and 2-p-carboxy-
Metabolism of aromatic constituents phenyl-2-methyl-propan-1-ol), and the urinary metabolites in
of hydrocarbon solvents guinea pigs (hippuric conjugate of p-butylbenzoic acid) were

Metabolism of one-ring aromatics similar to those from the metabolism of trimethylbenzenes,
indicating a preference for side chain oxidation as the first
Alkylated one-ring aromatics are metabolized by conjugation metabolic step.
of the alkyl side chains. As one example, trimethylbenzene
(TMB) has a similar metabolic profile in rats and humans.
Metabolism of aromatic/naphthenic molecules
The principal urinary metabolites include mercapturic acid,
sulfate, and glucuronide conjugates of 2,4-, 2,5- and 3,4-di- Serve et al. (1989) conducted a series of studies to identify the
methylbenzyl alcohols, dimethylbenzoic acids, and dimethyl- potential metabolic pathway of one-ring aromatics combined
hippuric acids, all products of side chain oxidation and trim- with a saturated ring structure (such as cyclopentane-indane
ethylphenols (ring-oxidized products) which together account or cyclohexane-tetralin). Male and female F344 rats (6/sex)
for 81% of the administered dose (Huo et al. 1989). Of these, were given 485 mg/kg bw/day of tetralin by oral gavage on
3,4-dimethylhippuric acid was the major metabolite (30% of alternate days for 14 days. Urine samples were collected 24
the dose) followed by 2,4-dimethylbenzyl alcohol (13%) and and 48 h after administration of the first dose on day 1, and
2,5-dimethylbenzyl alcohol (12%) in the form of sulfate and again 24 h after the last dose on day 14. With the exception
glucuronide conjugates (Huo et al. 1989), indicating that side of minor differences, the metabolic profiles for both male
chain oxidation is the major route of 1,2,4-TMB metabolism. and female rats were similar. In both male and female rats,
More than 99% of the administered dose was eliminated in the the principal urinary metabolites after 24 h, 48 h, and 15 days
urine within 24 h (Huo et al. 1989). For 1,3,5-TMB, 78% of the were 2-hydroxy-1-tetralone, 4-hydroxy-1-tetralone, 1-tetralol,
administered oral dose was excreted as 3,5-dimethylhippuric and 2-tetralol, similar to metabolites found following decalin
acid, with an additional 16% excreted in the form of glucuronide administration (see C9–C14 hydrocarbon solvent constituents).
and sulfate conjugates (Mikulski and Wiglusz 1975). All metabolites were derived from oxidation of the saturated
In humans, 22% of inhaled 1,2,4-TMB was excreted in the cyclohexane ring, with the tetralones being di-substituted
urine as hippuric acid metabolites within 24 h of a 2-hour expo- moieties with dehydrogenation of one hydroxyl group. Elliott
sure to 25 ppm (Jarnberg et al. 1997a, 1997b). As in the rats, and Hanam (1968) gave 5.1 mmoles/kg bw of 14C-tetralin to
3,4-dimethylhippuric acid was the major hippuric acid metab- adult albino rabbits (3/group) by oral gavage and evaluated the
olite (82% of urinary hippuric acids) indicating a role for steric metabolic profile in urine. Most (87–90%) of the radioactivity
hindrance in the formation of specific metabolites (Jarnberg was excreted in the urine within 48 h of exposure, with a fur-
et al. 1997a, 1997b). Hippuric acid metabolites made up 11% ther 0.5–3.7% excreted in urine 24 h later. Excretion of tetra-
of the total urinary metabolites of inhaled 1,2,3-TMB, with 2,3- lin in feces was very low, accounting for between 0.6% and
dimethylhippuric acid (82%) as the predominant hippuric acid 1.8% of the administered radioactivity. No radioactivity was
metabolite. In humans, the metabolic profile for 1,3,5-TMB detected in the breath, indicating very low accumulation in the
was similar to the metabolic profiles of 1,2,3-TMB and 1,2,4- lungs. Tissue retention was negligible, suggesting extensive
TMB. Following exposure to 30 ppm (approximately 150 mg/ absorption and metabolism. The major urinary metabolites
m3) for 8 h, urinary hippuric acid metabolites accounted for (in aggregate 56% of all metabolites) were glucuronide con-
only 3% of an inhaled dose; 73% of absorbed 1,3,5-TMB was jugates, primarily 1-tetralol (52.4%), 2-tetralol (25.3%), and
metabolized and excreted in the urine as 3,5-dimethylbenzoic 4-hydroxy-1-tetralone (6.1%). Cis and trans-1,2-diols were
and 3,5-dimethylhippuric acid metabolites (Kostrzewski et al. also detected in trace amounts (Elliott and Hanam 1968). In
1997, Kostrewski and Wiaderna-Brycht 1995). humans, 1-tetralol was the most common urinary metabolite
Metabolic profiles for the trimethylbenzenes in rabbits following ingestion of Cuprex (an ectoparasiticide-containing
were similar to those in rats and humans. The major urinary tetralin) (Drayer and Reiderberg 1973).
metabolites following oral 1,2,4-TMB administration were To evaluate the metabolic pathway of indane, male and
2,4-dimethylbenzoic acid and 3,4-dimethylhippuric acid female F-344 rats (9/sex) were given 0.25 ml/kg bw by oral
(Cerf et al. 1980), while 68% of an oral dose of 1,3,5-TMB gavage every other day for 14 days, and urine samples were
was excreted in urine as dimethylhippuric acid and 9% as collected and analyzed (Serve et al. 1989). The principal
urinary metabolites of indane were 2- and 3-hydroxy-1-in- a possible indication of reduced absorption with increasing
danone, cis- and trans-1,2-indandiol, 1- and 2-indanone, and length of alkyl side chains. The metabolites of MN that were
1- and 2-indanol, which were all products of oxidation of the detected in the urine included 1-naphthylcarboxylic acid and
cyclopentyl ring, and there were no apparent gender-related 1-hydroxy-2-methylnaphthalene. The major urinary metabo-
differences. Trace amounts of 5-indanol, a product of oxida- lites for EN were 1-(1-naphthalenyl)-ethanone, 1-(2-hydroxy-
tion of the aromatic ring, were identified in urine samples of 1-naphthalenyl)-ethanone, and 1-(1-hydroethyl)-naphthalene.
both males and females but at concentrations 27- to 54-fold Side chain oxidation accounts for approximately 50–85% of
less than 3-hydroxy-1-indanone (Serve et al. 1989). 2-methylnaphthalene metabolism. Evidence for this pathway is
also provided by studies showing that 2-naphthuric acid is the
Metabolism of naphthalene (a two-ring aromatic) most abundant metabolite of 2-methylnaphthalene (accounting
for 55–76% of total metabolites excreted in the urine within
The metabolism of naphthalene differs from that of other the first 24 h of exposure, in rats and guinea pigs) (Grimes and
two-ring aromatics in that it involves ring oxidation (due to Young 1956, Melancon et al. 1982, Teshima et al. 1983). As
the lack of alkyl side chains) to the epoxide intermediate (1,2- shown in Figure 3, 2-naphthuric acid is the conjugation prod-
naphthalene oxide) followed by rearrangement to naphthols uct of 2-naphthoic acid and glycine. In contrast to naphthalene,
(predominantly 1-naphthol) or hydration to the dihydrodiol by
only 15–20% of 2-methylnaphthalene metabolism occurs via

epoxide hydrolase. The epoxide metabolite is a target for GSH ring epoxidation at the 3,4-, 5,6-, and 7,8- positions (Breger
S-transferases to form mercapturic acids that are eliminated et al. 1983, Melancon et al. 1985).
in the urine (Figure 2). Although the metabolic pathways for Hoke and Zellerhoff (1998) reported that the metabolism
naphthalene are similar in humans, rats, and mice, mice are of longer-chain alkylated naphthalenes such as diisopropy-
more susceptible than rats to acute naphthalene-induced club lnaphthalene (C18 aromatic) proceeded primarily through
cell injury. This difference in species sensitivity is most likely CYP450-mediated oxidation of the isopropyl side chain, lead-
explained by the differences in the rates and stereoselectivity ing to glucuronide conjugation of the hydroxylated metabo-
of the epoxide intermediates formed as the first step in naph- lites and ultimately urinary excretion (more than 40% of the
thalene metabolism. In mice, a specific epoxide enantiomer administered dose). About 1.5% of the administered dose was
(1R, 2S-naphthalene oxide) appears to be formed, with higher eliminated in the urine as dihydrodiol metabolites, providing
rates in lung microsomes (Buckpitt et al. 1992). In contrast to some evidence that the potential for metabolism of alkylated
mice, hamsters, rats, and rhesus monkeys preferentially form
naphthalenes through ring oxidation decreases with increasing

the 1S, 2R-naphthalene oxide enantiomer, with much lower length of alkyl side chains.
rates of epoxide formation in lung microsomes (Buckpitt et al.
1992, 1995). Human lung microsomes expressing CYP2F1 also Summary of information on metabolism of aromatic
showed a preference for the 1S, 2R enantiomer, suggesting that constituents of hydrocarbon solvents
the metabolism of naphthalene in humans is more similar to that Aromatics such as benzene and naphthalene that have no alkyl
of rats than mice (Lanza et al. 1999). In separate experiments, it side chains are metabolized via a process in which the principal
was shown that the rate of naphthalene metabolism (as a mea- metabolic pathway involves oxidation of the ring forming the
sure of glutathione conjugation) by human CYP2F enzyme was corresponding epoxide. In contrast, the alkylated aromatics,
about 3000 times slower than that of the mouse CYP2F homolog particularly those most commonly found in hydrocarbon sol-
(Lanza et al. 1999, Shultz et al. 1999). In carcinogenicity stud- vents (alkylated one-ring, two-ring or one-ring aromatics, with
ies, mice showed chronic lung inflammation in response to or without attached alicyclic rings), are metabolized primarily
naphthalene exposures at levels as low as 10 ppm (53 mg/m3), by oxidation of the alkyl side chain(s), leading to the production
whereas rats exposed to naphthalene at 60 ppm (316 mg/m3) of the corresponding alcohols which are excreted in the urine as
had no lung parenchymal damage (NTP 1992, 2000). glucuronide and/or sulfate conjugates. The two-ring aromatics
follow the same pattern, although a small amount (20% or less)
Metabolism of alkylated two-ring aromatics (alkylated
of the metabolism may involve ring oxidation leading to epox-
naphthalenes)
ide formation, as is observed with 1- and 2-methylnaphthalene.
Naphthalene and alkylated naphthalenes have different meta- However, the longer the alkyl side chain, the less likely it is that
bolic profiles, and this may have toxicological consequences aromatic ring epoxidation will occur. With moieties consist-
(ATSDR 2005, Buckpitt et al. 1986, Buckpitt and Franklin ing of a benzene ring fused to an alicyclic ring (tetralins and
1989, NTP 2000). Unlike naphthalene, for which ring epoxi- indanes), metabolism primarily occurs on the saturated ring,
dation to the arene oxide is exclusively the first metabolic step, leading to production of alcohols, diols, and ketones, which are
side chain oxidation is the most common initial metabolic the products of dehydration of hydroxyl groups.
step for alkylated naphthalenes, with ring epoxidation as a
process of minor metabolic fate. To determine the metabolic Potential interactions among hydrocarbon solvent
fate of 1-methylnaphthalene (MN) and 1-ethylnaphthalene constituents and other solvents
(EN), male Wistar rats were given single intraperitoneal (IP) The evidence to date suggests that toxicologically important
doses (10 mg/kg bw) of tritiated MN and EP (Sapota et al. interactions involving hydrocarbons are usually the outcome
1996). More than 65% of the administered dose of MN was of metabolic interference. As one example, the effects of
eliminated in the urine within 72 h, with only 5% eliminated n-hexane can be enhanced by co-exposure to substances that
in the feces. In contrast, 24% of the EN was found in the inhibit the breakdown and elimination of the toxic 2,5-hexane-
feces with 72% of the administered dose eliminated in urine, dione intermediate by competing for metabolizing enzymes.
Figure 2. Metabolism of naphthalene. (Buckpitt et al. 2002, Waidyanatha et al. 2002, ATSDR 2005).
There is evidence for an interaction between n-hexane and ing toxicological characteristics other than those of n-hexane
methyl ethyl ketone (Altenkirch et al. 1977, 1978) in which and naphthalene as previously discussed, it seems unlikely
the effects of n-hexane are exacerbated. There are also studies that interactive effects would have toxicologically important
in which the effects of benzene have been modified by co- consequences. That this is true is demonstrated empirically by
exposure to other aromatics to either enhance or inhibit the evidence that the results of studies of complex solvents, sum-
formation of reactive metabolites (Bird et al. 2010, Medinsky marized in the appendices, are similar to those of mono-con-
et al. 1994, Tardif et al. 1992, Travis et al. 1992). However, the stituents or solvents with narrowly restricted carbon number
effects of co-exposures on metabolism are unlikely except at ranges or are comprised of specific hydrocarbon types. The
relatively high exposure levels, associated with experimental characteristics that have the greatest influence on the differing
conditions at which metabolizing enzymes are saturated (Kwai properties of hydrocarbon solvents are those related to physi-
et al. 2000, Ogata et al. 1993). Further, as indicated above, cal/chemical properties.
aside from some well-characterized exceptions, the constitu- It should be noted that the aromatic hydrocarbons can induce
ents of hydrocarbon solvents have similar and relatively subtle their own metabolism (Zahlsen et al. 1990, 1992), therefore in
toxicological effects. Accordingly, as there is little evidence experimental studies, the rates of elimination of aromatic con-
that the metabolites of hydrocarbon solvents have distinguish- stituents increase with increasing dose and time of treatment
Figure 3. Metabolism of 2-methylnaphthalene (major pathway is indicated by thick arrows) (ATSDR 2005).
(McKee et al. 2010). The aliphatic constituents do not appear some detail in this section, but are not discussed in detail in the
to induce their own metabolism, but if metabolism is induced summaries of toxicology studies included as appendices.
by co-exposure to aromatics, the rates of elimination of the
aliphatic constituents are also increased (Jarnberg et al. 1997, Common acute effects of hydrocarbon solvents
Lammers et al. 2007). Acute CNS effects
Among the earliest observations of workers exposed to vola-
Common toxicological properties of hydrocarbon tile hydrocarbons was evidence of CNS effects (Battig et al.
solvents and their constituents
1956, Riala et al. 1984). In early studies to characterize the
There are a number of effects that are common to many of the potential for overexposure, Patty and Yant (1929) documented
hydrocarbon solvent constituents, including acute effects that the effects of short-term, high-level exposures to volatile
are linked to the physical and chemical properties of these con- hydrocarbons including n-pentane, n-hexane, and n-heptane.
stituents. These include acute CNS effects, upper respiratory Exposure to n-heptane at 0.5% (approximately half of the
tract irritation, and chemical pneumonitis. There are also some lower flammability limit) for 15 min caused marked vertigo
changes that are commonly observed in repeated exposure stud- and loss of coordination, with feelings of hilarity lasting for
ies, some of which are adaptive and others are species-specific. up to 30 min after exposure. Accidental exposure for longer
These include liver weight increases, kidney changes in male periods to even higher levels of volatile hydrocarbons, as for
rats, and in some studies, small but statistically significant example in situations involving confined space entry, has led
reductions in hematological parameters. As these effects are to profound CNS depression and even death. It was apparent
common to many hydrocarbon solvents, they are described in from these observations that even relatively short exposures at
high levels could produce effects that might impede the ability hydrocarbons and other substances to produce ocular and
of workers to escape potentially hazardous situations. Thus, respiratory irritation at levels approximating the recommended
the potential for hydrocarbon solvents and their constituents to occupational exposure limits and reported that aromatics
cause acute CNS effects is one factor that should be considered (toluene, xylene) were more irritating than aliphatic solvents
in setting occupational exposure limits. Accordingly, short- (specifically hexane and Stoddard solvent). Nau et al. (1966)
term exposure studies with volunteers (Patty and Yant 1929, used volunteer studies to assess the potential for C9–C12
Nau et al. 1966, Nelson et al. 1943) have often included both aromatic solvents to be identified by odor and/or to produce
questionnaires and performance testing to elicit information on ocular and respiratory irritation. The results were then used
acute CNS effects. There have also been studies to character- in the development of occupational exposure limits. Carpen-
ize the acute CNS effects of hydrocarbon solvents and their ter et al. (1975a–h, 1976a–e, 1977a–c, 1978) systematically
constituents in laboratory animals (Bowen and Balster 1998, investigated the irritant potential for vapors of hydrocarbon
Lammers et al. 2007, 2009, 2011, McKee et al. 2010, 2011, solvents and volatile petroleum products in volunteers and
Nilsen et al. 1988). These studies indicate that the potential for used this information in making recommendations for occupa-
aliphatic and aromatic constituents to cause acute CNS effects tional exposure limits. Volunteer studies continue to be used
is associated with concentrations of hydrocarbons in the CNS. to assure that occupational exposure limits are not associated
Although there is evidence of distribution to the CNS during with discomfort (Ernstgard et al. 2009a, 2009b, Jones et al.
exposure, the hydrocarbon constituents quickly achieve steady- 2006, Juran et al. 2014, Lammers et al. 2007, 2009).
state and are then rapidly eliminated from the brain after expo-
sure with half-times in the range of 2 h (Hissink et al. 2007, Dermal irritation
2009). Because the aliphatic constituents become increasingly
In most cases, hydrocarbon solvents evaporate from the skin
lipophilic with increasing molecular weight, concentrations in
relatively rapidly, without consequence, although repeated
the brain increase with increasing carbon number to approxi-
exposures may cause dryness from defatting of the skin. How-
mately C10. However, aliphatic constituents with carbon
ever, if evaporation is impeded, as for example by the use of
numbers greater than C10 have such low vapor pressures that
occlusive patches, then skin contact can lead to severe dermal
it is difficult to sustain exposure levels sufficient to cause acute
irritation and skin damage. For this reason, skin protection is
CNS effects under experimental conditions (Nilsen et al. 1988,
normally advised when these solvents are used. When evaluat-
Bowen and Balster 1998). Further, changes in partition coef-
ing the results of studies involving dermal application in both
ficients for alkanes with carbon numbers  C10 suggest blood/

human volunteers and in experimental animals, it should be
brain barrier effects that inhibit the uptake of larger molecules
noted that the outcomes of dermal irritation tests (as well as
(Hau et al. 2001). The behavior of aromatic constituents is less
studies of percutaneous absorption) may be influenced by
well characterized, but, as with the aliphatic constituents, the
the procedures followed, and the potential influence of study
potential for exposure by inhalation to hydrocarbons with more
design should always be taken into consideration in evaluat-
than 10 carbons is limited by considerations of vapor pressure.
ing the results. Although the majority of the dermal irritation
In summary, acute CNS effects are associated with hydro-
studies are unpublished, summarized information can be found
carbon solvents and their constituents with carbon numbers in
in a number of references (Amoruso et al. 2008, Jacobs et al.
the range of approximately C6–C10. The potential for acute
1987, Johnson et al. 2012, Mullin et al. 1990) as well as in
CNS effects increases with increasing carbon number over that
summary information compiled as part of the HPV initiative
range, most likely due to increasing octanol/water partition
(OECD 2008, 2010, 2012a, 2012b, 2012c, 2012d). In general,
coefficients favoring distribution to the CNS. For hydrocarbons
low molecular weight hydrocarbon solvents are more irritating
with carbon numbers  C10, the limited likelihood for exposure
to the skin than their higher molecular weight counterparts.
by inhalation, associated with the reductions in vapor pressure
and apparently by blood/brain barrier effects make acute CNS
Chemical pneumonitis
effects unlikely, as shown empirically by Bowen and Balster
(1998) and Nilsen et al. (1988). Volunteer studies confirm that If hydrocarbon solvents are taken in to the lung in the liquid
exposures at currently recommended levels11 are not associated state (aspirated), they may cause chemical pneumonitis, a
with acute CNS effects (Ernstgard et al. 2009a, 2009b, Jones potentially serious condition that requires medical attention.
et al. 2006, Juran et al. 2014, Lammers et al. 2007, 2009). As described by Gerarde (1959), this condition appears to
be a local reaction in the lung rather than a consequence of
systemic absorption. The importance of this distinction is that
Ocular and upper respiratory tract irritation
if hydrocarbon solvents are accidentally ingested (or inhaled
The early literature also contains reports of upper respiratory as aerosols), it is better to allow them to be eliminated than
irritation associated with the use of hydrocarbon solvents to induce vomiting to void the stomach. Chemical pneumoni-
and constituents including C9 aromatic solvents (Battig et al. tis appears to be primarily a physical/chemical effect related
1956) and naphthalene (Robbins 1951). In volunteer studies, to the viscosity and solvency properties of the solvents. The
Nelson et al. (1943) investigated the potential for selected potential for hydrocarbon solvents to be aspirated is normally
associated with accidents or intentional misuse, as hydrocarbon
11Recommended
solvents are not intended for uses leading to intentional inges-
occupational exposure levels for hydrocarbon solvent
constituents and recommended guidance values used to calculate occu- tion. It should be noted that inhalation of solvents in aerosol
pational exposure limits for complex hydrocarbon solvents are shown in form, even at relatively high levels, did not cause chemical
Table 7. pneumonitis in experimental studies in rats (Gerarde 1963).
Common effects of repeated exposures to matic solvents (Carrillo et al. 2013, Phillips and Egan 1984a,
hydrocarbon solvents 1984b) and were referred to as “light hydrocarbon nephropa-
thy” in some of the early publications. It was noted that the
Liver enlargement
observations were similar to those described by Alden et al.
Liver weight increases in rats are a common finding in repeated (1984) in a report of a study of decalin (a C10 cycloparaffin) in
dose studies of hydrocarbon solvents and other organic sub- which the renal effects were associated with the induction of
stances that undergo metabolism to facilitate excretion. When a specific urinary protein (a2u-globulin). Evidence that long-
the liver weight increases are not associated with pathological term exposure to motor gasoline could cause kidney tumors
changes or elevations of liver enzyme markers (alanine amino in male rats (MacFarland et al. 1984), along with similar
transferase, ALT, or aspartate amino transferase, AST), they observations in studies of jet fuel (summarized in Mattie and
are interpreted as adaptive changes to increased metabolic Sterner 2011) led to further studies to characterize the struc-
demands (Ennulat et al. 2010, Goldberg 1966, Maronpot et al. tural requirements for substances associated with these effects.
2010, Schulte-Hermann 1974). This interpretation is sup- Iso-paraffinic hydrocarbons including 2,2,4-trimethylpentane
ported by evidence from studies that include recovery groups were the hydrocarbon solvent constituents most commonly
in which the liver weights return to control levels if the rats are associated with the induction of a2u-globulin (Aranyi et al.
held without treatment for several weeks. In one of the earliest
1986, Halder et al. 1985, Short et al. 1986), but these effects
papers on this subject, Goldberg (1966) noted that in rats fed have also been observed in studies of cycloparaffinic constitu-
high protein diets, there was evidence of liver enlargement with- ents (Alden et al. 1984, Johannsen and Levinskas 1987). Nor-
out corresponding changes in levels of liver-specific enzymes mal paraffins and aromatics are not commonly associated with
(other than enzymes involved in the urea cycle, Argyris 1971). a2u-globulin induction, although there is some evidence of
Other physiological events reported to induce liver enlargement weak induction in studies of n-butylbenzene (Izumi et al. 2005)
in rats include hormones induced during pregnancy (Gershbein and isopropylbenzene (Cushman et al. 1995, NTP 2009).
1958, Ochs et al. 1986, White and Gershbein 1987) and inflam- The renal effects which are unique to male rats are the con-
mation (Kunz et al. 1966). In these circumstances, liver enlarge- sequence of a process by which substances of diverse structures
ment was not associated with pathological changes, and the liver bind to a2u-globulin. The complexes are reabsorbed in the
weights returned to control levels when the conditions causing the proximal tubules where they accumulate in the lysosomes of
liver enlargement were terminated. Based on the results of studies renal epithelial cells. This leads to a cytotoxic and regenerative
of 11 drug candidate substances, Amacher et al. (1998) concluded process which, if sustained, can ultimately lead to the devel-
that liver weight increases of up to 50% with no more than slight opment of renal carcinoma (Swenberg et al. 1989). However,
elevations in levels of ALT or mild pathological changes should as this process has an absolute requirement for a2u-globulin
be interpreted as adaptive responses. Ennulat et al. (2010) [citing which is most commonly found in male rats but not in other spe-
Greaves (2007) and Schulte-Hermann et al. (1974)] concluded cies including humans, these effects and their consequences are
that the most common histological change associated with liver regarded as male-rat-specific and not relevant to human health
enzyme induction was hepatocellular hypertrophy. In a recent (US EPA 1991, Swenberg and Lehman-McKeeman 1998).
review, Maronpot et al. (2010) identified the characteristics of It should be noted that the documents that contain the con-
adaptive liver enlargement as dose-dependency, a self-limited clusions about the lack of relevance to humans also include
response and reversibility. In many of the repeated dose studies criteria by which to assess the plausibility that the underlying
of hydrocarbon solvents, there is evidence of dose-dependent mechanism for renal effects, and especially renal cancer, is
increases in liver weight accompanied by hepatocellular hyper- an a2u-globulin-mediated process (Swenberg and Lehman-
trophy, but increased levels of enzyme markers of liver damage or McKeeman 1998, U.S. EPA 1991). Many of the studies on
other histological changes are seldom observed. When recovery hydrocarbon solvents pre-dated the development of these
studies are conducted, the liver weights return to control values. criteria, and complete evidence in support of the mechanism
In short, increases in liver weights without pathological changes is not available in all cases. However, there have been two
and/or elevated levels of liver enzyme markers are most likely recent studies by the U.S. National Toxicology Program,
adaptive responses to increased metabolic demands, and should one on Stoddard solvent (NTP 2004) and the other on deca-
not be considered as adverse findings. lin (NTP 2005), in which chronic exposure to the test sub-
stances resulted in increased levels of kidney tumors in male
rats. Each of these studies included very detailed mechanistic
Kidney effects
investigations that provided evidence that the tumors were, in
A common observation in repeated dose studies of hydro- fact, the consequence of a2u-globulin-mediated processes. In
carbon solvents is a characteristic effect in male rat kidneys. the summarized results of studies on hydrocarbon solvents,
Among the earliest observations were those by Carpenter et al. provided in the appendices, kidney effects in male rats were
(1975f, 1975g, 1977b) who reported increased frequencies of noted, attributed to a2u-globulin induction, and were not fur-
moderate tubular degeneration in the kidneys of male rats and ther discussed unless there was some reason to believe that
attributed these changes to an exacerbation of spontaneous there is an alternative explanation for the data.
age-related lesions (chronic progressive nephropathy). The
changes were not observed in kidneys of female rats or dogs.
Hematological changes
In studies to further characterize these effects, it was noted that
these changes tended to be more commonly observed in studies In some studies, small but statistically significant reductions
of isoparaffinic hydrocarbons than in normal paraffinic or aro- in hematological parameters have been observed (Carrillo
et al. 2013). These changes, which are more commonly found 2003).13 The non-acute CNS changes that have been attributed
in male than female rats, are characterized by reductions in to long-term solvent exposure are non-specific and similar to
hemoglobin content, packed cell volumes, and/or red blood those of aging and alcoholism. A comprehensive review of this
cell (RBC) counts. The underlying causes are unknown, but topic is beyond the scope of this current paper, but of direct
there are several possibilities. The effects may be simply bio- relevance is a re-evaluation of the evidence linking exposure
logical variations, as the differences are normally within nor- to hydrocarbon solvents with chronic neurological effects.
mal physiological ranges and not associated with histological Of particular importance is white spirit, as that substance spe-
changes in the bone marrow or spleen. This explanation seems cifically has been implicated in the development of chronic
unlikely as the changes are often dose-responsive and reversed toxic encephalopathy (IPCS 1996).
in recovery studies. An alternative hypothesis is that the The concern was based on studies of painters exposed to
changes are secondary to other treatment-related effects such solvents through the use of paints in the construction industry in
as renal changes in male rats. Although there is uncertainty Scandinavia, between approximately 1960 and 1980. Elements
about the underlying causes, small reductions in hematological that should be considered in determining the most plausible
parameters are not unique to studies of hydrocarbon solvents; explanation for the outcomes include substance identification,
rather they are common findings in repeated dose studies and the levels and circumstances of exposure, the neurological tests
are considered to lack toxicological significance (Car et al. and their outcomes, and the potential for confounding factors
2006). In the summarized results of studies of hydrocarbon including exposure to other solvents. White spirit, a C9–C11
solvents, hematological changes in male rats are assumed to be aliphatic solvent containing approximately 20% aromatics and
without toxicological significance as long as they are within having a boiling range of approximately 150–200°C (i.e., white
normal physiological limits. It should be noted that the physi- spirit has the same technical requirements as originally defined
ological range for hematological parameters in toxicological for Stoddard solvent, described in detail in Appendix 1B (III)),
studies may vary based on gender (related to size and hormonal was the solvent used in formulating paints used in the largest vol-
differences), age, animal strain, and test methods, among oth- umes in the construction industry in Scandinavia between 1960
ers. Representative physiological ranges for hematological (when it replaced turpentine and linseed oil) and 1980 (when
parameters in untreated Sprague-Dawley rats with a mean age oil-based paints were replaced by water-based paints). However,
of 3.7 months (males) and 3.9 months (females) can be found the house painters also used epoxy paints, which were formu-
in Lillie et al. (1996) and Wolford et al. (1986). Data from lated with solvents including toluene, xylene, aliphatic alcohols,
Sprague-Dawley rats are referenced because they represent the and ketones but apparently not white spirit (Riala et al. 1984).
most commonly used rat strain among those represented in the Although the epoxy paints represented a relatively small frac-
studies discussed in later sections of this paper. tion of total solvent exposure, the use of these paints accounted
for the majority of the incidents of overexposure which were
Persistent CNS effects relatively common during that period of time. As documented
Although acute exposure to volatile hydrocarbon and other by Riala et al. (1984), between 1960 and 1973, feelings of
organic solvents can cause acute CNS effects, and repeated drunkenness (presumably an indication of acute CNS depres-
exposure to some specific solvents including n-hexane and tolu- sion) were estimated to have occurred on average 46 times/year,
ene can cause more profound and persistent effects (Grandjean with the frequency dropping to 27 times/year between 1974
1985), a possible relationship between long-term exposure to and 1978. Between 1960 and 1973, 8% of the painters reported
volatile organic solvents and persistent CNS effects is more experiencing drunkenness 100 or more times/year. The painters
controversial. As summarized by Ridgway et al. (2003), there attributed these episodes to the use of epoxy paints.
have been reports since the 1970s of “chronic toxic encephal- Aside from exposures that occurred during house painting,
opathy” (CTE), particularly among painters.12 Quoting from some of the construction painters were exposed to hydrocar-
Ridgway “This condition has been described as ‘characterized bons and other organic solvents when engaged in other paint-
by a global mental impairment including changes in: (i) cog- ing occupations. In particular, those engaged in spray painting
nitive functions, memory and concentration; (ii) personality, were more highly exposed to solvents than those in the con-
(iii) motivation, vigilance and energy. The clinical picture struction painting industry (Lundberg et al. 1995, Mikkelsen
may be described as a psycho-organic syndrome or a mild et al. 1988). Spray painting is most commonly used to apply
degree of dementia; that is a clinical syndrome of premature paint to metal surfaces in the automotive and appliance indus-
ageing of higher cortical functions” [citing European Com- tries. Solvent systems used for paints in spraying applications
mission 1997]. Contributing to the controversy is a lack of are complicated and include aromatic hydrocarbons, ketones,
consensus on the basic elements of this condition: there is no acetates, and chlorinated solvents; white spirit is not well
common terminology (Baker and Seppalainen 1986) nor are suited to this technical application. As documented by
there consistent diagnostic criteria (Triebig and Hallermann Elofsson et al. (1980), there was apparently some exposure
2001, van der Hoek et al. 2001); no animal model (Bruckner to white spirit in the car painting industry but not in indus-
and Warren 2001, ECETOC 1996); and no consistent patho- trial painting. In addition, spray painters were exposed at
logical changes in humans following long-term exposure to higher levels than construction painters. Fidler et al. (1987)
solvents, other than some specific examples (Ridgway et al. 13Inthe early literature (e.g. Mikkelsen et al. 1988), there were reports of
cerebral atrophy based on imaging techniques. These findings were not
substantiated in a similar study (Lundberg et al. 1995), and a principal
12Other names include organo-psycho syndrome (OPS), painters’ syn- conclusion of Ridgway et al. (2003) was that the imaging techniques that
drome, and solvent dementia. had been used in these early studies were insensitive and unreliable.
estimated that the breathing zone concentrations experienced and worse in 7, with the largest differences associated with the
by spray painters were 4 to 10 times those experienced by test for figure classification (which the authors considered to be
brush or roller painters, and that even though the spray painters a test of visual-logical ability) and the rivet test (a measure of
commonly used respirators, exposures during spray painting psychomotor coordination). Hane et al. (1977) did not find any
exceeded those in roller or brush painting by factors of 2–5. correlation between test performance and measures of expo-
Not surprisingly, industrial spray painters were found to have sure. A similar study was reported by Lindstrom and Wickstrom
both functional and neurobehavioral deficits when compared to (1983) but with a larger number of subjects and a better-defined
unexposed reference groups (Elofsson et al. 1980). The poten- reference group (concrete reinforcement workers). The main
tial for exposures to other solvents and particularly exposures subjective complaint of the painters was memory impairment
of higher intensity should be considered in the assessment of (forgetfulness). The statistical analysis of the psychological
effects in the construction painting industry. In a cohort of 85 tests revealed that the painters performed less well in tests of
painters investigated by Mikkelsen et al. (1988), 82 had been visual-constructive ability, visual memory, and sensorimo-
employed as house painters, but some had also been engaged tor speed and accuracy, but when pre-exposure intellectual
in ship painting, coach and industrial painting, and/or sign level was taken into account, the visual memory test was the
painting and silk screening. The average daily exposures in only statistically significant finding. The authors reported that
these other painting occupations were higher than those expe- psychological test performance was slightly related to various
rienced in house painting. In particular, the estimated daily indices of solvent exposure. A study by Seppalainen and Lind-
average exposures of those that had been engaged in coach strom (1982) was primarily an investigation of effects on the
and industrial painting were almost 6 times higher than those peripheral nervous system and acute CNS effects. The painters
of house painters. Thus, any study of the neurological effects were more likely than the reference group (concrete reinforce-
of white spirit on house painters should consider the potential ment workers) to have reported “feeling sick”, or experiencing
impact of other higher intensity exposures to other solvents “nausea” or a “drunken feeling” (presumably evidence of acute
that were experienced by some of the cohort members. CNS effects), whereas the concrete reinforcement workers
Because of the use of white spirit as a paint solvent in the were more likely to have reported paresthesia of the hands and
construction industry, the studies in which house painters were feet. The authors considered that the average concentration of
evaluated are the most relevant, whereas studies of painters white spirit (40 ppm, 232 mg/m3) was a no-effect level for the
in other occupations and/or those engaged in paint formula- parameters (primarily acute effects) that they investigated.
tion provide little information about white spirit specifically. Studies by Mikkelsen et al. (1988) and Lundberg et al.
However, it should be noted that many of the citations cited as (1995) were conducted after the replacement of white spirit-
evidence of neurological effects from white spirit in the IPCS based paints with water-based paints, and, as these were ret-
(1996) report (as well as subsequent reports which have relied rospective, the potential for recall bias must be considered.
on the IPCS report as a primary information source) were from Mikkelsen et al. (1988) compared a cohort of 85 people (only
occupational investigations in which exposure to white spirit 82 had actually worked as house painters, and a number had
was not documented, or if used, was likely a small contributor worked in other painting occupations as described previously)
to overall exposure. As many of these citations were not specific to a reference group composed of bricklayers. The authors
for white spirit exposure, reliance on this database obscures also developed an exposure metric based on time spent in
the role of white spirit in the development of neurobehavioral specific activities, particularly brush, roller, or spray paint-
effects. Accordingly, the following section is a summary of ing, that is, [(liters/day)] years, and divided subjects into low-,
those specific studies that involved construction painters who medium-, and high-exposure groups.14 The subjects were
worked between 1960 and 1980. One of the first of these stud- given a psychological evaluation and evaluated for neurobe-
ies was an evaluation of 70 individuals suspected of suffering havioral performance, and a smaller group of subjects was
from solvent intoxication (Arlien-Soborg et al. 1979). Of these, further investigated for pathological changes using imaging
20 subjects were excluded on the basis of alternative causality; techniques. Based on the psychological evaluation, the overall
the remaining 50 were given neuropsychological examina- performance of the low-exposure group was better, although
tions. Among these, claims of memory loss, fatigue, inability not statistically different from that of the controls, but higher
to concentrate, and irritability were relatively common. The levels of impairment (the authors called this dementia15) were
results of neuropsychological testing suggested that the major- recorded in the medium and most highly exposed groups. It
ity had slight to moderate neuropsychological impairment.
However, severe dementia was not observed. It should be
noted that these individuals had not been selected randomly; 14According to Mikkelsen et al. (1988), “low”   15(l/d) years;
all of the subjects had been referred for clinical evaluation and “medium”   15 (l/d) years; “high”   30 (l/d) years. SCOEL (2007)
reported that the low-exposure group was equivalent to an average occu-
were assumed to have been experiencing neurological effects.
pational exposure of approximately 40 ppm (230 mg/m3). On that basis,
Further, the results were provided only in summary form, and the average in the most highly exposed group would have been  80 ppm
there was no reference group for the neurobehavioral tests. (460 mg/m3).
Hane et al. (1977) compared the performance of house 15In the original Scandanavian literature, “dementia” was used to describe
painters in a battery of neurobehavioral tests to that of a group some of the relatively mild, intial symptoms of solvent intoxication, but
of unexposed individuals of similar ages. Interestingly, they did in other countries, the term was assumed to refer to a more severe psychi-
atric condition. This led to confusion as some investigators reported that
not find any objective correlates for the subjective complaints they had been unable to replicate the earlier findings because they were
(memory loss, fatigue, etc.). The painters performed better than observing only minor or “subacute” effects in their studies of solvent-
the reference group in 5 of the psychological tests conducted exposed workers.
should be noted that there were no explicit operational cri- One of the most controversial aspects of all of this is the
teria for dementia scoring (Mikkelsen et al. 1988, Appendix clinical significance of the findings. Some authors regard
1, p. 98), and the investigators were aware of the occupations the outcomes as early evidence of irreversible neurological
but not the specific exposure groups of the subjects. The effects whereas others consider them as mild and “subclinical”
results of the psychometric tests were similar to those pre- (Spencer and Schaumburg 2000). Differing views of the rela-
viously reported by Hane et al. (1977) and Seppalainen and tive importance of these findings seem to be related to national
Lindstrom (1982). There were few differences between the compensation laws (see, for example, Spurgeon 2006). How-
painters and bricklayers in general, but the painters in the low- ever, the purpose of this paper is not to discuss the toxicologi-
exposure group tended to perform better than the bricklayers, cal significance of the findings but rather to evaluate whether
and those in the highly exposed group tended to perform less white spirit was the likely causative agent.
well. Painters with high solvent exposure performed more The case for white spirit as the causative agent was most
poorly than painters with low solvent exposure in the majority strongly advanced by Mikkelsen et al. (1988) who attributed
of the 13 tests, but in only one test (symbol digit modalities the effects observed to white spirit exposure, because the sub-
test [p  0.0012]) was the difference statistically significant. jects were mostly house painters and white spirit had been the
The authors also reported that cerebral atrophy was associated solvent most heavily used in construction painting. This view
with solvent exposure as demonstrated by imaging techniques, was then taken forward into the IPCS (1996) report. However,
but in a later review, Ridgway et al. (2003) concluded that the there are two other possible explanations although neither
imaging equipment that was available at the time the investi- appears to have been seriously considered:
gations were conducted was not sufficiently sensitive to allow
1. that the effects were due to repeated episodes of solvent
reliable conclusions to be drawn.
overexposure and more likely due to the use of epoxy paints
Lundberg et al. (1995) compared the neuropsychiatric effects
rather than white spirit, or
in 135 house painters to those in an age-matched group of 71
2. that the effects were due to higher intensity exposures during
carpenters. Like Mikkelsen et al. (1988), the authors divided
spray painting in occupations other than construction paint-
the painters into three groups (low, medium, and high) based on
ing and that the causative agents were solvents other than
the number of months that the painters had been exposed to sol-
white spirit (which is not commonly used in spray painting).
vents at the corresponding occupational exposure limits.16 The
painters in the most highly exposed group tended to complain It is plausible that the chronic effects in painters resulted
about impaired memory, decreased speed, sleeping problems, from repeated episodes of overexposure to solvents in epoxy
and vegetative symptoms, but there were no significant differ- paints, particularly as one of the solvents was toluene which
ences in moods. Among 12 different psychometric tests with can produce profound CNS effects under abuse conditions
29 outcome variables, there was only one test (block design) in (Grandjean 1985). Mikkelsen et al. (1988) did not differentiate
which there was a significant difference between painters and exposures to toluene and xylene from those of white spirit in
carpenters, and in this test it was the carpenters that performed the overall evaluation.
more poorly (p  0.05). In comparisons across the exposure With respect to acute CNS effects specifically, it should
groups, the painters in the low-exposure group performed better be noted that maximum exposures to white spirit during con-
than the carpenters in 23/29 tests and those in the high-exposure struction painting are estimated to have been in the range of
group performed more poorly in 20/29 tests. The frequencies of 250–300 ppm (1375–1650 mg/m3) (Riala et al. 1984). Based
psychiatric disorder were similar in both groups, as were the on volunteer studies (Carpenter et al. 1975c, Nelson et al.
results of the imaging study. The authors cautiously concluded 1943), white spirit might produce ocular and respiratory irri-
that exposure during painting was associated with neurobehav- tation at these levels, but is unlikely to produce acute CNS
ioral deficits, but they did not find test performance to be highly effects that could have been perceived by the painters.
correlated with any of the exposure parameters. As noted above, many of the painters in Mikkelsen’s cohort
From the above, it is apparent that the findings most had been engaged in other types of painting occupations. In par-
commonly reported in house painters were small changes in ticular, 10 of the 85 painters had worked in coach and industrial
psychometric tests which were not strongly associated with painting under conditions in which, as documented by Mik-
long-term solvent exposure. There did not appear to be evi- kelsen, exposures were much higher than those experienced dur-
dence of more profound psychiatric disorders, and reports ing construction painting. As reported by Elofsson et al. (1980),
of pathological changes based on imaging techniques were industrial painters were more profoundly affected than the house
not substantiated. One interpretational problem is that the painters even though exposure to white spirit by these workers
neurobehavioral effects associated with painting were vague, was not documented. It seems likely that the 10 industrial paint-
non-specific, and difficult to differentiate from those related ers workers would have been included among the 33 construction
to other conditions, including aging and alcoholism. Indeed, painters in Mikkelsen’s high-exposure group. It is plausible that
alcohol usage and pre-exposure intellectual level were identi- the small performance deficits in the high-exposure group were
fied as potential confounding factors in some of the studies. due to higher intensity exposures associated with spray paint-
The no-effect concentrations were estimated to have been in ing rather than the long-term, relatively low-level exposures to
the range of 40–90 ppm (230 mg/m3–540 mg/m3). white spirit that were associated with construction painting.
If the effects in house painters were in fact due to either
repeated episodes of acute overexposure or higher intensity
16According to SCOEL (2007), the average exposures in the low-exposure exposures in other painting occupations, that could explain
group were equal to approximately 90 ppm (approximately 540 mg/m3). the difficulties in demonstrating exposure-response relation-
ships, as the contributing events would be more episodic or properties of the majority of the constituents. The distinguish-
concentrated in a small number of individuals. In this respect, ing features are often related to structural elements, leading
it is important to consider the paper by Hooisma et al. (1994) to the production of unusual metabolites. These constituents,
in which it was reported that there was a significant relation- like the other constituents of hydrocarbon solvents, are present
ship between symptoms of solvent exposure with the periods in the feed stocks and are carried through into the final prod-
of heavy exposure but a lack of consistent relationships with ucts at variable levels depending on both constitutive levels
other exposure parameters. and production methods. In some cases, these constituents are
It is also interesting to note that exposures to white spirit only present at low levels in the feed stocks; in other cases,
in the dry cleaning industry seem to have been similar to some constituents are specifically controlled to low levels
those experienced by house painters (Oberg 1969). However, by the manufacturing processes used. There are some cases,
there are no reports of neurological effects in the dry cleaning however, in which n-hexane and naphthalene may be present
industry prior to the replacement of hydrocarbon solvents with at relatively high levels, and these solvents are differentiated
chlorinated solvents. from those solvents that do not contain these constituents at
There is one study which, if taken at face value, supports problematic levels by the naming convention as previously
the white spirit hypothesis: Bazylewicz-Walczk et al. (1990), described. The following section discusses the toxicological
which summarized an investigation of neurological effects hazards of these specific substances.

among workers in the shoe industry in Poland over a period
of approximately 30 years (1960–1990). The exposed workers Constituents of concern that may be present
performed less well than controls in a number of tests, and at levels  1%
those exposed for longer periods of time performed less well n-hexane
than those who had only worked for a few years, so the data
are consistent with a relationship between occupational expo- n-Hexane, which can be a constituent of C6 or C7–C9 aliphatic
sure and neurobehavioral effects. According to the authors, hydrocarbon solvents, has been associated with peripheral
the solvent used was white spirit and the exposures were in neuropathy in humans. As summarized by the World Health
the range of the Polish hygiene standard (500 mg/m3, approxi- Organization (IPCS 1991), the neurological effects of n-hexane
mately 100 ppm) between at least 1977 and 1990. However, were first identified in the late 1960s and early 1970s through
the authors did not provide any chemical description or any occupational studies. These cases were reported to have been
the result of high exposures, primarily during shoe manufac-
analytical information to confirm that the solvent used was

in fact white spirit. This is important because glue solvents, ture. Based on those observations, n-hexane has been largely
particularly those for use in producing rubber footwear, are withdrawn from use in solvents for consumer use, at least in the
typically of lower molecular weight, more volatile aliphatic US and Europe, although it continues to be offered for some
solvents that provide the fast-drying properties that are critical specialized industrial applications. Hydrocarbon solvents with
for that technical application. It seems more likely that “white more than 5% n-hexane are differentiated from other solvents
spirit” was used by the author as a generic descriptor rather by the naming convention, as shown in Table 2.
than as an identifier of the specific solvent used. The toxicological properties of n-hexane differ from those
In summary, it has been reported that exposure to white spirit of other aliphatic hydrocarbons because the effects are medi-
during construction painting results in a chronic neurological ated through a specific metabolite, 2,5-hexanedione, which,
condition. However, based on a critical review of the original because of its unique structure, can interfere with microtubule
literature, it seems more likely that these neurobehavioral formation. The most characteristic toxicological effect of
effects were due to episodes of repeated overexposure during 2,5-hexanedione is a peripheral neuropathy which is charac-
the use of epoxy paints or to higher intensity exposures during terized by a “dying back” process involving the long nerves.
spray painting than to long-term, relatively low-level exposure This neuropathy has been observed in humans and has also
to white spirit. Adding to the plausibility of these alternative been reproduced in animals under experimental conditions
explanations is that repeated exposure to toluene under abuse (the basis for classification of n-hexane as a specific-organ
conditions has been associated with chronic neurological toxicant) (Frontali et al. 1981, Ono et al. 1981, Takeuchi
effects (Grandjean 1985). The alternative explanations are also et al. 1980). There is also evidence that 2,5-hexanedione can
consistent with the observations of Spurgeon et al. (1994) who interfere with spindle fiber formation, inhibiting meiosis, and
reported that there were no effects on cognitive functioning or resulting in testicular atrophy17 (Krasavage et al. 1980). Other
mental health in workers exposed to solvents in paint making hydrocarbon solvent constituents of similar structure or carbon
and concluded that moderate levels of exposure (below current number including n-pentane, other hexane isomers, n-heptane
occupational exposure limits) are not associated with neurolog- and cyclohexane are not metabolized to 2,5-hexanedione
ical effects even if the exposures take place over many years. intermediates and do not cause peripheral neuropathy (Egan
et al. 1980, Frontali et al. 1981, Ono et al. 1981, Takeuchi
et al. 1980).
Hydrocarbon solvent constituents with unusual Inhaled n-hexane is well-absorbed in the lung (Dahl et al.
toxicological properties 1988). Metabolism of n-hexane occurs in the lungs and liver
As noted in previous sections (e.g., C5–C9 hydrocarbon solvent with bioactivation (the primary route of n-hexane metabolism)
constituents, Metabolism of aromatic/naphthenic molecules),
there are some hydrocarbon solvent constituents with toxico- 17Based on the reports of testicular atrophy, n-hexane has been classified
logical properties that distinguish them from the more generic in Europe as toxic to reproduction.
occurring in the liver. The metabolic pathways are detailed in hallucinations. Examination of peripheral blood revealed evi-
Figure 4. In the liver, n-hexane is preferentially hydroxylated dence of erythrocyte fragmentation. Robbins (1951) reported
at C-2 by rat liver microsomes to form 2-hexanol (the rate- eye irritation in workers exposed to naphthalene vapor at levels
limiting step) in a bioactivation pathway (Frommer et al. 1974) exceeding 15 ppm (79 mg/m3). Naphthalene was not carcino-
or 1-hexanol in a detoxification pathway that primarily occurs genic when repeatedly given to animals by oral administration
in the lungs (Crosbie et al. 1997). Conversion of n-hexane to (Schmahl 1955).
2-hexanol appears to be catalyzed by CYP2B1, since use of More recently, the toxicology database on naphthalene was
a CYP2B1 inhibitor reduced the production of 2-hexanol by substantially expanded through the publication of chronic
33% in liver microsomes and 74% in lung microsomes but had toxicity/carcinogenicity studies in mice and rats in which
no effect on the production of 1-hexanol (Crosbie et al. 1997). naphthalene was administered by inhalation (Abdo et al. 1992,
In a toxicity study of n-hexane and its metabolites, Couri 2001, Adkins et al. 1986, NTP 1992, 2000). Of particular
and Milks (1982) showed that the LD50 values for 2-hexanone importance were the statistically significant increases in inci-
and 2,5-hexanedione in rat were virtually identical (2.6 and dence of pulmonary alveolar/bronchiolar adenomas in female
2.7 g/kg respectively), suggesting that 2,5-hexanediol and mice and nasal tumors in rats. The tumors were associated
2-hexanone are rapidly converted to 2,5-hexanedione (5-hy- with inflammation in mouse lung and rat nasal tract, suggest-
droxy-2-hexanone is most likely a short-lived intermediate). By ing that tumor induction may have been related to cytotoxic
comparison, the LD50 for n-hexane was 32 g/kg. As described effects in the affected tissues (see North et al. 2008, for a
in the previous paragraph, n-hexane is metabolized to 1- or detailed summary). Questions have been raised as to whether
2-hexanol, and 2-hexanol is then converted to 2-hexanone the pulmonary effects in rats and mice were species-specific,
(methyl-n-butyl ketone) and 2,5-hexanediol, both of which due in part to differences in pulmonary physiology (Morris
are converted to 5-hydroxy-2-hexanone and 2,5-hexanedione and Buckpitt 2009) and metabolic differences (Buckpitt et al.
as principal metabolites (DiVincenzo et al. 1977, Krasavage 2002), and the underlying mode of action is controversial
et al. 1980). To identify the metabolite responsible for distal (Piccirillo et al. 2012). Rhomberg et al. (2010) concluded that
axonopathy in response to n-hexane exposure, Krasavage and the tumors were most likely the consequence of a cytotoxic
colleagues gave groups of 5 rats similar doses of all of the (or dual cytotoxic/genotoxic) mode of action although a geno-
n-hexane metabolites that had been identified (Figure 4), that toxic mode of action could not be ruled out. As Rhomberg
is, 660 mg/kg/day 2-hexanone, 755 mg/kg/day 2,5-hexane- pointed out, the choice of mode of action is directly related
dione, 675 mg/kg/day 2-hexanol, 765 mg/kg/day 5-hydroxy- to human relevance as humans…“have insufficient metabolic
2-hexanone, and 755 mg/kg/day 2,5-hexanediol, as well as 3 activation to the naphthalene epoxide to deplete GSH or to
doses of commercial n-hexane (570, 1140, and 3980 mg/kg/ create sufficient levels of reactive metabolites so as to produce
day), by oral gavage, for 90 days (Krasavage et al. 1980). All cytotoxicity, genotoxicity, or both, and without such toxicity,
doses (including the lowest dose for n-hexane) were equivalent no carcinogenic risk is induced” (Rhomberg et al. 2010).
to 6.6 mmol/kg. Mid and high doses for n-hexane were 13.2 and
46.2 mmol/kg respectively. Hind limb paralysis observed in Constituents of concern that may be present at
order of decreasing potency was 2,5-hexanedione  5-hydroxy- levels  1%
2-hexanone  2,5-hexanediol  2-hexanone  2-hexanol  Diethyl and triethylbenzenes
n-hexane (Krasavage et al. 1980). Severity of toxicity corre-
lated with peak serum concentrations of 2,5-hexanedione. Only Gagnaire et al. (1990, 1991, 1992) reported that
in rats treated with the high dose of n-hexane, producing serum 1,2-diethylbenzene (1,2-DEB) produced a characteristic neu-
concentrations close to the peak levels of 2,5-hexanedione, was rological effect in mice, whereas the other two diethylbenzene
clear evidence of peripheral nervous system toxicity observed isomers (1,3-DEB, 1,4-DEB) were not neurotoxic. In subse-
(Krasavage et al. 1980). quent studies, 1,2,4-triethylbenzene (1,2,4-TEB) was found to
be neurotoxic, but the 1,3,5 isomer was not (Gagnaire et al.
1993, Tshala-Katumbay et al. 2006). Mechanistic studies by
Naphthalene
these and later investigators (Kim et al. 2001, 2002, Sabri
Naphthalene, an aromatic with 2 fused benzene rings but et al. 2007) showed that 1,2-DEB could be metabolized to
no alkyl side chains, is most commonly found in C10–C12 1,2-diacetylbenzene, which could produce a g-diketone neu-
aromatic solvents at levels up to 10%. When naphthalene is ropathy similar to that produced by n-hexane. Although these
present at levels  1.0%, it is differentiated by the naming con- publications established that the neurotoxic molecules had
vention (Table 2). As summarized by Gerarde (1960), the his- ethyl groups on adjacent carbons and provided a good basis
torical toxicological studies indicated that oral administration for hazard characterization, the majority of the studies utilized
of naphthalene caused cataract formation in rabbits and rats single high-dose administrations, making it difficult to define
and hemolytic anemia in dogs. In humans, exposure to naph- dose-response relationships. However, based on unpublished
thalene vapor was reported to cause eye irritation, headaches, information, the American Industrial Hygiene Association
nausea, and profuse perspiration depending on the exposure (AIHA) produced a Workplace Environmental Exposure Level
levels, and lens opacities were noted in individuals exposed (WEEL) of 28 mg/m3 (5 ppm) (AIHA 2005). According to
to high levels of naphthalene for extended periods of time. information summarized in that document, rats were exposed
In 50 cases involving severe intoxication following repeated by inhalation 6 hours/day and 5 days/week for 3 months at
ingestion of a naphthalene-isopropanol beverage, there were levels of approximately 200, 600, or 1200 mg/m3, to a prod-
symptoms of tremor, restlessness, extreme apprehension, and uct containing diethylbenzene isomers. The overall no-effect
Figure 4. Metabolic pathway for n-hexane in mammals (modified from Couri and Milks 1982).
concentration was 190 mg/m3 which the AIHA converted to non-genotoxic process. In short, there is evidence to suggest
a 28 mg/m3 WEEL through the use of adjustment factors.18 that biphenyl can be irritating to the upper respiratory tract,
It should be noted that this recommendation was based on a but only at levels above those that would be experienced if the
complex isomeric substance, and, therefore, applies to the sum occupational exposure recommendations are respected.
of all of the diethyl (and triethyl) benzene isomers and not to
the neurotoxic isomers specifically. Constituents of concern that may be present at
levels  1%
Biphenyl Benzene
In 1968 the ACGIH® adopted a threshold limit value of As summarized by Gerarde (1960), the historical concern for
1.3 mg/m3 (0.2 ppm) based on reports of adverse effects in benzene was related to the potential for repeated exposure to
humans associated with occupational exposures. The exposure cause severe anemia, and, as a consequence, by 1960, occupa-
conditions were unclear but exposures were reported to have tional exposure levels had been lowered to 25 ppm (81 mg/m3),
ranged from less than 1 mg/m3 to more than 100 mg/m3 (0.15 a level that was not associated with unpleasant odor, upper
→ 77 ppm). Due to difficulties in defining dose-response rela- respiratory irritation, or acute CNS effects. As Gerarde pointed
tionships in humans, the ACGIH® relied on animal data, basing out, the effects of benzene on the bone marrow have a strong
their recommendation on evidence of respiratory irritation in structural dependence as alkylated benzenes, particularly tolu-
mice following exposure to 5 mg/m3 (1 ppm). It should be noted ene and xylene, do not produce similar effects. It is now under-
that in the respiratory irritation study, mice were not exposed stood that the toxicological distinction between benzene and
to biphenyl vapor but rather to biphenyl dust impregnated on alkylated benzene is related to the fact that benzene is metabo-
diatomaceous earth. The relevance of this experimental result lized through a process involving ring epoxidation, whereas the
to the occupational environment is unknown. From the infor- alkylated derivatives are primarily metabolized via glucuroni-
mation summarized the US Environmental Protection Agency dation of the alkyl side chains, as described previously.
(2011), repeated inhalation of biphenyl vapor by mice results A study of cancer in Pliofilm workers (Infante et al. 1977)
in respiratory irritation at levels  25 ppm ( 125 mg/m3) and which associated benzene with AML raised concerns about the
apparently produces similar but less severe effects in rats and adequacy of the occupational exposure recommendations then
rabbits at these and higher exposure levels. In one dietary in effect, and drew attention to commercial products including
study, biphenyl increased the frequency of liver tumors in hydrocarbon solvents and fuels that contained benzene as low-
female BDF1 mice at doses  414 mg/kg/day (Umeda et al. level constituents. The US Consumer Product Safety Commission
2005). However, there was no increase in liver tumors in male (CPSC) commissioned a study by Battelle Columbus to evaluate
BDF1 mice in the study by Umeda et al. (2005), and there was “benzene-critical” solvents to determine whether they contained
no increase in tumors in mice of other strains (Imai et al. 1983, benzene at levels  0.1% (1000 ppm). As documented in the
Innes et al. 1969). Based on a toxicokinetic mode of action study report (Hillman 1978), the solvents most likely to contain
analysis, Klapacz et al. (2014) concluded that the tumors in benzene (hexane, heptane, rubber solvent, lacquer diluent, tolu-
BDF1 female mice were the consequence of a gender-specific, ene) had already been identified by that time, and many produc-
ers had taken steps to reduce benzene levels or remove from the
18According to information provided in the HPVIS data base, the observa- market place products with benzene levels  0.1%. As reported
tion in this study was a blue-gray color in the brain and other tissues. The by McKee et al. (2007), benzene levels in solvents now being
NOAEC was 190 mg/m3 (US Environmental Protection Agency 2009). produced by US manufacturers (i.e., members of the Hydrocar-
bon Solvents Panel) are below approximately 100 ppm (0.01%) Table 3. Benzene levels in hydrocarbon solventsa.
in all hydrocarbon solvents and less than 10 ppm (0.001%) in Type of Solvent/CAS Number Benzene (ppmv)
most (Table 3). It should be noted that there are regulatory Category 1 (C9 Aromatic Solvents)
requirements in both the United States and Europe which require 95-63-6  10
that hydrocarbon solvents (or any other substances) containing 108-67-8  10
 0.1% benzene be classified as carcinogenic. 25550-14-5  1
64742-95-6  1
Category 2 (C10–C13 Aromatic Solvents)
Polycyclic aromatic hydrocarbons 538-93-2  1
1321-94-4  1
Polycyclic aromatic hydrocarbons (PAH), particularly mol- 64742-94-5  1
ecules with 4–7 aromatic rings, have been associated with skin 70693-06-0  1
cancer in humans. This has historically been an occupational 68477-31-6  1
hazard in the petroleum industry, with evidence of elevated Category 3 (C9–C14 aliphatics (2–25%
aromatics) [Stoddard solvent, white spirit,
levels of skin cancer in workers exposed to unrefined lubricant mineral spirits]
base oils and waxes (IARC 1984). Because the upper carbon 8052-41-3  50
number range of hydrocarbon solvents is C20, there is a theo- Category 4 No data available
retical possibility that three- or even four-membered ring aro- [Heavy aliphatic solvents]
Category 5 (C5 Aliphatics)
matics could be present, but compositional analysis indicates [Pentane Solvents]
that aromatics with 3 or more rings are not commonly found. 78-78-4  1
Rather, the aromatic constituents of hydrocarbon solvents are 287-92-3  1
Category 6 (C6 Aliphatics)
primarily alkylated one- and two-ring molecules, and higher
[Hexane Solvents]
molecular weight constituents are often complex naphthenic/ 110-54-3  10
aromatic molecules. This is primarily a consequence of the 64741-84-0  100
manufacturing processes and the physical/chemical properties 64742-89-8  0.5
of the constituents. More specifically, the alkanes have boiling 68410-97-9  1
68476-50-6  1
points that are similar or lower than aromatics with the same Category 7 (C7–C9 Aliphatics)
carbon numbers, and molecules with fewer aromatic rings [Light Aliphatic Solvents – heptanes,
and more side chain carbons have lower boiling points than octanes, nonanes]
111-65-9
molecules with more aromatic rings. For example, eicosane  1
142-82-5  10
(a C20 alkane) has a boiling point of 343°C; tridecylbenzene 540-84-1  1
(C19, one-ring aromatic) has a boiling point of 340–346°C; 8032-32-4 10
but anthracene (C14, three-ring aromatic) has a boiling point 64741-63-5  1
of 343°C. Thus, most of the aromatic molecules with more 64742-89-8  10
70024-92-9  1
than two rings are excluded by the distillation processes that Category 8 (C9–C14 Aliphatics,  2% Aromatics)
are used in the manufacturing processes of these solvents. [De-aromatized aliphatic solvents]
111-84-2  1
112-40-3  1
Hydrocarbon solvent categorization 8052-41-3  1
and nomenclature 64741-65-7  1
64742-47-8  1
Hydrocarbon solvent categories 64742-48-9  1
64742-88-7  1
Hydrocarbon solvents with similar constituents have similar 64771-72-8  1
physical/chemical and toxicological properties. In part, this 68551-16-6  1
is because substances with similar physical/chemical prop- 68551-17-7  1
129813-67-8  1
erties have similar pharmacokinetic properties and follow
Category 9 (C14–C20 Aliphatics,  2%
similar metabolic pathways. The propensity for hydrocarbon Aromatics)
solvent constituents to exhibit unusual toxicological prop- [De-aromatized heavy aliphatic solvents]
erties has been associated with either a distinct metabolic 544-76-3  1
pathway (e.g., naphthalene) or the production of an unusual 629-62-9  1
64741-73-7  1
metabolite (e.g., n-hexane). The theory is supported by the 64771-72-8  1
empirical evidence that the results of toxicological studies of 64742-46-7  1
complex hydrocarbon solvents are similar to those of studies 64742-47-8  1
of individual constituents. Because constituents of complex 68551-19-9  1
68551-20-2  1
hydrocarbon solvents have similar toxicological properties, 90622-46-1  1
it has been possible to use generic approaches for hazard
aThese data were previously published in McKee et al. (2007). As
characterization. Accordingly, the process used by the hydro-
described in that reference, the data were compiled from information
carbon solvent industry to characterize the hazards of the provided by the Hydrocarbon Solvents Panel, from information provided
substances that it produces is to group “similar” solvents into to the U.S. Environmental Protection Agency in response to the EPA Vol-
categories and to use the results of studies of “representa- untary High Production Volume Challenge program (OECD 2008, 2010,
tive” test materials for all of the substances in the respective 2012a, 2012b, 2012c, 2012d).
groups.
As a means of grouping those substances that are “similar”, Another very important consideration is that the constituents
the hydrocarbon solvent industry has utilized the concepts of should have common metabolic profiles with limited opportu-
substance categories and “reasonable worst case” examples. nities for interactions. As described in more detail earlier (see
A category is a group of substances with similar properties, Sections Metabolism and excretion of aliphatic hydrocarbon
such that the results of a study of any category member would solvent constituents and Metabolism of aromatic constituents
be broadly applicable to all other category members (see Clark of hydrocarbon solvents), the aliphatic constituents are rela-
et al. 2013, for an example of a similar approach for petro- tively rapidly metabolized by w-oxidation to similar metabo-
leum products). According to the Organization for Economic lites (primarily alcohols and carboxylic acids with similar
Cooperation and Development (OECD), a chemical category carbon numbers) that are excreted in the urine. The only ali-
is a group of substances with physiochemical and toxico- phatic constituent known to produce an unusual toxic effect is
logical properties that follow a regular pattern as a result of n-hexane, which produces a characteristic neurological effect,
structural similarities, that is, the structural similarities create peripheral neuropathy, as a consequence of its propensity to
a predictable pattern in biological response (OECD 2007). form a uniquely toxic metabolite, 2,5-hexanedione. There are
In fact, hydrocarbon solvents are listed as a specific example no other aliphatic hydrocarbon solvent constituents that form
of complex substances for which a category approach is both 2,5-hexanedione at toxicologically relevant levels, although it
useful and practical (OECD 2007, p. 69). A similar definition is also a metabolite of methyl n-butyl ketone.
is included in the REACH regulation (Annex XI, section 1.5, The aromatic constituents are primarily metabolized by
European Commission 2006). For hazard characterization a process of glucuronidation of the alkyl side chains. The
purposes, hydrocarbon solvents were divided into 9 categories principal exception is naphthalene, which has no alkyl side
(Table 2). This was done originally to facilitate data submis- chains and is metabolized by ring epoxidation, producing
sion to satisfy the needs of the OECD HPV program but the unique metabolites that differentiate naphthalene from the
category structure was also used as a basis for REACH regis- alkylated aromatic constituents.
tration. The underlying categorization strategy was to provide Interactive effects are most commonly the consequence of
a means of grouping hydrocarbon solvents of certain types metabolic interference. The most common example is that the
based on compositional elements including aromatic content, aromatic constituents induce their own metabolism, leading to
carbon number range, and the potential to contain components more rapid excretion of hydrocarbons. However, under condi-
of unusual toxicity. In other words, the category definitions tions at which metabolism is not saturated, which is normally
provided the boundary conditions for hydrocarbon solvents of the case for the exposure situations under which these sub-
particular types and identified certain components that may be stances are used, interactive effects are not expected (Kwai
present at potentially harmful levels in category members. As et al. 2000, Ogata et al. 1993).
a practical matter, it should be noted that there will always be In summary, hydrocarbon solvents are well suited for category
some complex solvents with compositions that span category formation, because, for the most part, the constituents with simi-
boundaries. In such cases, the solvents were assigned to the lar physical and chemical properties also have common meta-
categories that represented the largest fractions of the constit- bolic and toxicological properties. There are a few constituents
uents. For example, n-nonane is a member of the C7–C9 ali- with distinctive properties that can be present at toxicologically
phatic hydrocarbon solvent category, but if the majority of the relevant levels, but these are well characterized and differentiated
constituents of nonane-containing solvents had carbon num- through the naming convention, as described in Section Nomen-
bers  C9, the solvents were included in the C9–C14 aliphatic clature for hydrocarbon solvents – the naming convention.
hydrocarbon solvent categories.
The benefit of a category approach is that results of studies Nomenclature for hydrocarbon solvents – the naming
of a relatively small number of substances can be used to char- convention
acterize the hazards of a larger group of substances. However, Hydrocarbon solvents were originally identified and described
it is important to carefully consider the extent to which the sub- with the same CAS numbers as those used for the refinery
stances on which the conclusions are based are representative streams from which they were derived, and this convention
of the broader group of substances in the category. In a paper continues to be used for regulatory purposes in most parts of
on toxicology testing in the petroleum industry, Scala (1988) the world. Because the CAS numbers were relatively broad and
discussed the concept of a “reasonable worst case” which he generic and hydrocarbon solvents have become more narrowly
defined as a substance that met the technical requirements defined to meet evolving technical requirements, the hydrocar-
but had higher levels of specific constituents of concern than bon solvent industry developed a new nomenclature (the nam-
might normally be found in commercial products. Because ing convention) to more precisely describe its products (Hydro-
hydrocarbon solvents tend to be more narrowly defined, and in carbon Solvent Producers Association 2011). As many of the
some cases, more highly refined than petroleum products, test- hydrocarbon solvents are of complex and variable composition,
ing has commonly been conducted on commercial products a convention was developed in which the complex hydrocarbon
meeting technical requirements, but to the extent possible, substances can be described in the following way:
with relatively high levels of constituents of certain types and
or constituents of concern, particularly n-hexane and naphtha- 1. “Hydrocarbons” is in the first part of the name to make
lene. The “reasonable worst case” approach ensures that the clear the chemical character of these solvents.
potential hazards of commercially available substances are not 2. The carbon number range must include at least 80% of the
underestimated. These examples are discussed in more detail components of the substance as determined by gas chroma-
in the sections on the specific substances in which they occur. tography or an equivalent test method.
3. The types of hydrocarbon structures present or the “PINA” Summary of the Toxicological Information
(paraffins, isoparaffins, naphthenes,19 aromatics) distribu-
The toxicological data on hydrocarbon solvents are discussed
tion is described. The paraffins, isoparaffins, and naphthenes
in more detail in the appendix. However, from the generic
(the aliphatic constituents) are mentioned when present in
information provided in earlier sections and the more complete
the substance at levels between 10% and 80%. The aromatic
toxicological data in the appendix, the following conclusions
contents are given as ranges.
can be drawn:
4. Components that are distinguished by unique toxicologi-
cal properties (and classifications) are specifically men-
Aromatic Solvents (C9–C12)
tioned if present at levels exceeding the classification
limits (using the guidance on concentration limits in the There are two groups of aromatic solvents: Category 1 sol-
EU Dangerous Substance Guidance Directive [EU 1967] vents that contain primarily C9 aromatics (isomers of trim-
as of 2010). ethylbenzene and ethyl toluene), and Category 2 solvents that
contain C10–C12 aromatics (primarily alkylated benzenes,
The naming convention can also be used for hydrocarbon naphthalene, and alkylated naphthalenes). Benzene levels in
solvents that do not meet the UVCB definition (e.g., mono- these solvents are below 1 ppm (Table 3). The constituent of
constituent substances) if they fall within the broad definitions particular concern in these solvents is naphthalene, and the
for the categories. To better understand the increased precision solvents that contain naphthalene at levels  1% are identi-
that was introduced by the naming convention, the following fied by the naming convention. Other constituents of these
examples are provided: solvents that may be of interest include cumene, which can
As shown in Table 4, “regular white spirit” was found to be present in low concentrations in Category 1 solvents, and
contain 81% aliphatic constituents and 19% aromatics, by gas diethylbenzene isomers, indene, and biphenyl that may be
chromatography. Using mass spectrometry, the aliphatic dis- present in low concentrations in Category 2 solvents. Aro-
tribution was found to be 22% normal paraffins, 26% isopar- matic constituents with more than two rings are not present
affins, and 33% naphthenes, giving a PINA distribution of at more than trace levels. The animal data and human experi-
22%/26%/33%/19%.20 ence provide evidence that the principal hazards associated
Accordingly, this substance fits into the category “Hydro- with the use of these solvents are ocular and upper respiratory
carbons, C9–C14, aromatics (2–25%)”, and under the nam- system irritation. These solvents can also cause acute CNS
ing convention becomes Hydrocarbons, C9–C11, n-alkanes,
effects at exposure levels above the thresholds for irritation

iso-alkanes, cyclics, aromatics (2–25%). For comparative (Korsak and Rydzynski 1996, McKee et al. 2010), and chemi-
purposes, the CAS definition for Stoddard solvent is “A col- cal pneumonitis if taken into the lungs as liquids. Occupa-
orless, refined petroleum distillate that is free from rancid or tional exposure recommendations for these solvents are based
objectionable odors and that boils in a range of approximately primarily on the need to minimize discomfort (Battig et al.
148.8 to 204.4°C” although, as discussed previously, hydro- 1956, Carpenter et al. 1977a, Gerarde 1960, Nau et al. 1966).
carbon solvents of this type can be described by other CAS Volunteer studies (Jarnberg et al. 1996, 1997, Jones et al. 2006,
descriptors. Kostrzewski et al. 1997) confirm that at current occupational
As a second example, a complex C6 aliphatic hydrocarbon exposure levels (20–25 ppm, 100–125 mg/m3) there is no evi-
solvent contained 99.6% C6 isomers, 0.2% C5 isomers, and dence for discomfort or acute CNS effects. In repeated dose
0.1% C7 isomers. Using mass spectrometry, the aliphatic dis- studies in animals, there is little if any evidence of systemic
tribution was found to be 47% normal paraffins (i.e., n-hexane), effects or pathological changes, but inflammatory responses
34% isoparaffins, and 18% naphthenes, giving a PINA distri- in the respiratory tract have been observed (Carpenter et al.
bution of 47%/34%/18%/0%. Under the naming convention, 1977a, Clark et al. 1989, Korsak et al. 2000a, 2000b, Nau
this substance is “Hydrocarbons, C6, n-alkanes, cyclics;  5% et al. 1966, Swiercz et al. 2000). In developmental toxicity
n-hexane.” tests, there was no evidence of malformations in studies which
A complete listing of the hydrocarbon solvents which were involved examination of the uterine contents or effects on the
registered in Europe in 2010, in compliance with REACH developing nervous system in studies in which offspring were
requirements, along with the corresponding CAS and EINECs examined at postnatal day 90, but developmental delays were
numbers, is given in Table 2. observed at or above maternally toxic levels (Lehotzky et al.
In summary, the categories provide the boundary condi- 1985a, 1985b, McKee et al. 1990, Saillenfait et al. 2005, Ung-
tions and identify potentially hazardous constituents that may vary et al. 1983, 1985). Reproductive toxicity tests did not
be present in the category members. The naming convention identify any effects on fertility (Lehotzky et al. 1985a, McKee
more precisely describes the specific substances in terms of et al. 1990). In subchronic neurotoxicity tests, there were no
carbon numbers and molecular types, and differentiates those
solvents that contain hazardous constituents at problematic
levels from those that do not. Table 4. Carbon number distribution in “regular white spirit”.
Carbon Number Fractional Composition (%)
C8 1
C9 10
19In the petroleum and hydrocarbon solvent industries, cycloparaffinic C10 40
substances are referred to as “naphthenes”. C11 42
20See Carrillo et al. (2014) for additional historical compositional infor- C12 6
C13 1
mation for white spirit.
persistent changes in motor activity or functional observations liquids. The principal target organ effects of aliphatic/aromatic
and no pathological changes in the nervous system (Douglas solvents and similar substances include liver enlargement
et al. 1993, Gralewicz et al. 1997, Gralewicz and Wiaderna and male rat kidney effects, associated with a2u-globulin
2001, Wiaderna et al. 2002), although there have been reports (Amoruso et al. 2008, Carrillo et al. 2014, Carpenter et al.
of functional changes following the administration of pain 1975c, Jenkins 1971, Rector et al. 1966). The only fetal effects
(Gralewicz et al. 1997, Gralewicz and Wiaderna 2001, Wia- in developmental toxicity studies are developmental delays at
derna et al. 2002). Genetic toxicity studies have mostly been maternally toxic levels (American Petroleum Institute 1977,
negative (Schreiner et al. 1989), although one C9 isomer was 1979a, 1979b, Cooper and Mattie 1996, Jakobsen et al. 1986,
reported as mutagenic when tested under in vitro conditions Schreiner et al. 1997), and, based on studies of related sub-
(Janik-Spiechowicz et al. 1998a, 1998b). In carcinogenicity stances, there is no evidence of impairment of fertility (Mattie
studies, cumene, naphthalene, and methylnaphthalene iso- and Sterner 2011, Schreiner et al. 1997). These solvents are
mers have produced respiratory tract tumors in rodents (Abdo not active in tests for mutagenic activity (Amoruso et al.
et al. 1992, 2001, NTP 1992, 2000, 2009, Murata et al. 1993, 2008, Gochet et al. 1984, McKee et al. 1994), and there are
1997). Cumene, naphthalene, and other structurally similar no reports suggesting that they might be carcinogenic. The
aromatic chemicals (e.g., ethylbenzene, styrene) that cause principal concern for these solvents relates to the potential
similar tumor profiles in rodents are usually inactive in stan- for some of the Category 3 solvents, in particular those with
dard mutagenicity assays. Based on an evaluation of mode of primarily C9–C11 aliphatic constituents, to cause chronic
action, Cruzan et al. (2009) concluded that these tumors arose neurological effects in humans. As discussed in more detail
as a secondary consequence of the generation of cytotoxic in Section Persistent central nervous system effects, exposure
metabolites of CYP2F2 in nasal and lung tissue in mice and to white spirit may have contributed to the neurobehavioral
CYP2F4 in nasal tissues in rats. Rhomberg and colleagues changes observed in house painters (Arlien-Soborg et al. 1979,
(2010) explored the hypothesis-based weight of evidence Hane et al. 1977, Lindstrom and Wickstrom 1983, Lundberg
analysis of the naphthalene data in more detail and essentially et al. 1995, Mikkelsen et al. 1988, Seppalainen and Lindstrom
reached the same conclusions as Cruzan et al. (2009), although 1982), but it seems more plausible that these changes were
they could not definitively rule out the involvement of other associated with periods of overexposure related to the use of
CYPs such as CYP2E1 and/or other biochemical events aromatic-based solvent systems (Riala et al. 1984) or periods
such as GSH depletion (Rhomberg et al. 2010). Overall, the of higher intensity exposure to other types of solvents dur-
underlying cancer mechanism, particularly with regard to the ing industrial spray painting (Elofsson et al. 1980). Notably,
proposed involvement of CYP2F enzymes for cumene, naph- repeated exposure studies in animals have not revealed any
thalene, styrene, and other similar chemicals remains under persistent functional changes or pathological findings (Kulig
investigation, and its relevance to humans is controversial 1990, Nielsen et al. 2006).
(Cruzan et al. 2009, Rhomberg et al. 2010).
Light Aliphatic Solvents (C5-C9)
Aliphatic/Aromatic Solvents (C9–C20)
There are three groups of light aliphatic solvents, Category
There are two groups of aliphatic/aromatic solvents: Category 5 (pentanes), Category 6 (hexanes), and Category 7 (C7–C9
3 (C9–C14 aliphatics, 2–25% aromatics) and Category 4 aliphatics). For most of these solvents, benzene levels are
(C14–C20 aliphatics, 2–25% aromatics). less than 1 ppm but Category 6 and Category 7 solvents that
The aliphatic constituents include normal-, iso-, and contain higher levels of C6 aliphatic constituents may also
cycloalkanes, with carbon numbers ranging from approxi- contain higher levels of benzene, in some cases at levels
mately C8–C20. The aromatic constituents in C9–C14 up to approximately 100 ppm (Table 3). The constituent of
aliphatic (2–25% aromatic) hydrocarbon solvents are pri- particular concern for C5–C9 aliphatic solvents is n-hexane,
marily alkylated benzenes and are similar to those found in which can be found in some hexane and C6–C7 aliphatic sol-
C9 aromatic solvents. Benzene levels in these solvents are vents at toxicologically relevant levels. As described above,
generally below 50 ppm (Table 3). The aromatic constituents prolonged exposure to n-hexane may cause peripheral neu-
in C14–C20 aliphatic (2–30% aromatics) solvents are pri- ropathy (IPCS 1991). However, this specific effect is unique
marily alkylated one-ring and two-ring aromatics along with to n-hexane and not caused by other aliphatic hydrocarbon
complex aromatic/naphthenic structures. Benzene levels are solvents or their constituents (Egan et al. 1980, Frontali et al.
expected to be very low due to the boiling ranges of these 1981, Ono et al. 1981, Takeuchi et al. 1980), and solvents
solvents. Other constituents including naphthalene, indene, containing n-hexane at levels  5% are specifically identified
and diethylbenzenes may also be present in these solvents at by the naming convention. The other aliphatic constituents
levels typically below 1%. These solvents may cause upper have only generic toxicological properties. More specifically,
respiratory irritation, but the potential for acute CNS effects these solvents, particularly with C6–C9 constituents, are
is limited because of their relatively high molecular weights relatively well absorbed following inhalation exposure (Dahl
and low vapor pressures (Carpenter et al. 1975c, 1976c, Cohr et al. 1988, 1989) or oral administration (Albro and Fishbein
et al. 1984, Ernstgard et al. 2009a, 2009b, Gamberale et al. 1970), but are not well absorbed dermally (Tsuruta 1982).
1975, Hastings et al. 1982, Juran et al. 2014, Lammers et al. However, once absorbed, the constituents are relatively rap-
2007, Nelson et al. 1943). Otherwise, these solvents are not idly excreted (Filser et al. 1983, Galvin and Maraschi 1999c,
considered to be acutely toxic (Amoruso et al. 2008), but they Mraz et al. 1998). They are not acutely toxic by oral, der-
can cause chemical pneumonitis if aspirated into the lungs as mal, or inhalation routes of administration (Carpenter et al.
1975d, Galvin and Marashi (1999a, 1999b, 1999c), Hine gible. For C14–C20 aliphatic ( 2% aromatic) hydrocarbon
and Zuidema 1970, Johnson et al. 2012, Kinkead et al. 1985, solvents, exposure by inhalation at more than very low levels
McKee et al. 1998, Swan et al. 1974), but they can cause can only occur under conditions in which aerosol formation
chemical pneumonitis if taken into the lung as liquids. These is possible. Percutaneous absorption of these molecules is
solvents are volatile and may cause ocular and/or respira- also very limited, so the likelihood of substantial systemic
tory tract irritation and acute CNS effects if the exposure dose from dermal contact is also unlikely. Gastrointestinal
levels are sufficiently high (Bowen and Balster 1998, Car- absorption of high molecular weight aliphatic constituents
penter et al. 1975d, Christoph et al. 2000, Lammers et al. can occur, but absorption over this carbon number range
2011, McKee et al. 2011, Patty and Yant 1929, Swann et al. decreases from approximately 60% to 30%. However, even
1974, Virtue 1948). There are specific occupational expo- the highest molecular weight aliphatic hydrocarbon con-
sure limits for n-hexane that are related to its neurological stituents can be absorbed if ingested. Once absorbed, the
properties; for other aliphatic constituents of these solvents, aliphatic constituents are distributed, primarily to adipose
occupational recommendations are set to avoid respiratory tissue, metabolized by w-oxidation, and either excreted or
irritation and/or acute CNS effects. In repeated exposure re-incorporated into other macromolecules. Most of the
studies, liver enlargement is commonly observed; male rat alkanes are excreted relatively rapidly following termination
kidney effects associated with a2u-globulin induction are of exposure, but there may be retention in depot fat.
seen in studies of substances containing isoparaffinic and/or The potential for being absorbed has practical implications
cycloparaffinic constituents, and small but statistically sig- for experimental design. Inhalation toxicity studies are lim-
nificant reductions in hematological parameters have been ited to the maximally attainable vapor concentrations, so few
reported in some studies (Carrillo et al. 2013, Carpenter et al. repeated inhalation toxicity studies have been conducted with
1975d, 1978, Duffy et al. 1991, Galvin and Marashi 1999a, solvents with carbon numbers greater than C10. The primary
1999b, 1999c, Kinkead et al. 1985, Kim et al. 2012, Malley route of human exposure is probably by dermal contact, but
et al. 2000, McKee et al. 1998, Schreiner et al. 1998, Sung the rates of percutaneous absorption are low and the likeli-
et al. 2010). As described above, liver enlargement without hood of producing appreciable systemic doses is limited. Oral
pathological changes or elevated levels of marker enzymes administration is often the recommended route of exposure
(ALT, AST) is considered to be an adaptive response rather for repeated dose studies of these solvents, in order to maxi-
than an adverse effect. The kidney changes in male rats are mize systemic dose. However, there are some technical issues
due to the induction of a2u-globulin and not relevant to associated with repeated oral administration of solvents, and
humans. The underlying processes resulting in reductions in the utility of such data for risk assessment is somewhat lim-
hematological parameters are unknown, but given that the ited as there are few end uses for these solvents that result in
differences are small, within normal physiological limits, ingestion. For any of the toxicity studies of these solvents it is
and reversible, they are not considered to be toxicologically always necessary to consider whether or not the outcomes are
relevant. The only fetal effects in developmental toxicity related to the route of administration and whether or not the
studies were developmental delays at maternally toxic lev- data that might be obtained from any particular study would
els (Daughtrey et al. 1994a, Johnson et al. 2012, Keenan be useful in assessing human risk.
et al. 1991, Kreckmann et al. 2000, McKee et al. 1998), and The above having been said, these solvents are not toxic in
reproductive toxicity tests provided no evidence of impair- acute studies conducted in accordance with current guidelines,
ment of fertility (Bui et al. 1998, Daughtrey et al. 1994a, they are not dermal or ocular irritants, and they do not produce
Kreckmann et al. 2000, Yu et al. 2011). These solvents are allergic contact dermatitis (Amoruso et al. 2008, Carpenter
not active in tests for mutagenic activity (Brooks et al. 1988, et al. 1978, Johnson et al. 2012, Nilsen et al. 1988, OECD
Daughtrey et al. 1994b, Galvin and Marashi 1999a, 1999b, 2012d).
1999c, Kirwin et al. 1980, 1991, Loury et al. 1986, McCar- In repeated dose studies by either oral or inhalation routes,
roll et al. 1981a, 1981b, Mortelmans et al. 1986, Richardson the principal effects are liver enlargement without patho-
et al. 1986). Chronic exposure to commercial hexane resulted logical changes and male rat kidney effects consistent with an
in an increase in liver tumors in female mice, but it did not a2u-globulin-mediated process (Adenuga et al. 2014b, Car-
increase the frequency of tumors in male mice or in rats of rillo et al. 2014, Carpenter et al. 1978, Johannsen and Levins-
either gender (Daughtrey et al. 1999). kas 1987, NTP 2005, OECD 2012d). These solvents are not
mutagenic under in vitro or in vivo conditions (Amoruso et al.
2008, Johnson et al. 2012, NTP 2005, OECD 2012d), and
Heavy Aliphatic Solvents (C9–C20)
they do not produce developmental or reproductive effects
The C9–C14 aliphatic solvents are divided into two cat- (Amoruso et al. 2008, Johnson et al. 2012, Ministry of Health
egories, Category 8 (C9–C14 aliphatic hydrocarbon,  2% and Welfare 1996, OECD 2012d). The C9 and C10 aliphatic
aromatic solvents) and Category 9 (C14–C20 aliphatic constituents have been shown to produce acute CNS effects
hydrocarbon,  2% aromatic solvents), that are differentiated (Carpenter et al. 1978, Lammers et al. 2011, McKee et al.
on the basis of carbon number. There is some potential for 2011, Nilsen et al. 1988), but no effects were reported in stud-
vapor exposure to the lower molecular weight constituents ies with higher molecular weight constituents (Bowen and
of the C9–C14 aliphatic ( 2% aromatic) hydrocarbon sol- Balster 1998, Nilsen et al. 1988). No neurological effects were
vents, particularly the C9 and C10 constituents, but at higher reported in repeated inhalation exposure studies of a C9–C14
molecular weights, the vapor pressures are so low that the aliphatic solvent with approximately 20% aromatic constitu-
likelihood of appreciable vapor exposure becomes negli- ents (Kulig 1990), and there were no neurological effects fol-
lowing repeated dermal administration of hydrodesulfurized described by some of the same CAS numbers that were used
kerosene (Breglia et al. 2014).21 for petroleum-derived substances, but more recently, the nam-
In a carcinogenicity test of Stoddard solvent Type IIC, con- ing convention was introduced to more precisely describe
ducted by the NTP (2004), there was an elevated frequency of solvent composition.
pheochromocytoma in male rats and hepatocellular adenoma Classification of UVCB substances can be particularly dif-
in female mice. The development of the adrenal gland tumors ficult, in part because of the compositional complexity and
may have been secondary to the exacerbation of chronic pro- variation, and, further, because uncertainties in substance
gressive nephropathy in the rats (Nyska et al. 1999). Pheo- identity may confound the hazard characterization process.
chromocytomas are not considered relevant to humans (Greim In fact, one specific objective of the current publication is to
et al. 2009). The NTP considered that the female mouse liver address this uncertainty at least to the extent that it relates
tumors were related to elevated body weights of the female to hydrocarbon solvents by providing more specific compo-
mice in the high-exposure group. sitional information via the naming convention along with
detailed information on toxicological hazards, which can then
be used for classification purposes, as shown by the generic
Use of Toxicological Hazard Characterization
classification example as shown in Table 5.
Information for Hydrocarbon Solvents and Their
Constituents for Hazard Classification and for As summarized in this document and discussed in more
Occupational Exposure Recommendations detail in the appendices, the hydrocarbon solvents have some
generic hazards that require classification, and some con-
In this section, the use of toxicology data as a basis for hazard stituents have other hazards that must be identified if the con-
classification and as a basis for occupational exposure recom- stituents are present at levels at which these hazards might be
mendations is described. expressed. More specifically:
Evaluation of hydrocarbon solvents for classification 1. Aspiration Hazard – Classification for Category 1 aspira-
following the recommendations of the Globally tion hazards [chemicals known to cause human aspiration
Harmonized System toxicity hazards or to be regarded as if they cause human
aspiration toxicity hazard] is required for any substance
One specific aspect of hazard communication is the need to that …”is a hydrocarbon and has a kinematic viscosity
describe the hazards in terms that would be understood by  20.5 mm2/s, measured at 40°C…”. Accordingly, clas-
customers and downstream users. One means of doing this is sification of hydrocarbon solvents as Category 1 aspira-
through substance classification, following the Globally Har- tion hazards is required unless they have kinematic vis-
monized System of Classification and Labeling of Chemicals cosities  20.5 mm2/s at 40°C. Most of the hydrocarbon
(GHS) developed by the United Nations (2011). However, the solvents fall within the classification requirements although
guidance on GHS classification does not define UVCB sub- there may be some high molecular weight aliphatic sol-
stances, nor does it explain how the GHS framework should vents (i.e., C14–C20 solvents) that have viscosities above
be applied to complex substances. Recognizing this limita- the classification limit.
tion, an approach that could be used as a means to classify 2. Acute Toxicity – The need for classification of acute oral,
UVCB substances was proposed, using petroleum substances dermal, or inhalation toxicity is usually based on the out-
as an example (Clark et al. 2013). The guidance provided comes of tests of substances administered in single, rela-
was essentially to divide the substances into categories, to tively high doses, in accordance with specific regulatory
use generic information on representative substances as the protocols. The solvents have acute LD50 values  2000
basis for classification, and to take specific constituents with mg/kg in oral and dermal tests, so data are available dem-
unique properties into account, and can be directly applied to onstrating that classification at the Category 4 level is not
hydrocarbon solvents. In fact, in some respects, this approach required. However, not all substances have been tested
can be more easily applied to hydrocarbon solvents as they at levels  5000 mg/kg that are required to assess the
have fewer compositional elements, cover a smaller range of potential for classification at the Category 5 level. Simi-
carbon numbers, are better defined compositionally, and may larly, hydrocarbon solvents do not meet the requirements
contain lower levels of uniquely hazardous constituents than for classification as acute inhalation toxicants, although it
the corresponding petroleum substances. A particular issue should be noted that the maximally attainable vapor con-
for petroleum substances is that they are described by CAS centrations are limited by the physical properties of the
numbers, which may be relatively broad and generic. As dis- individual solvents. In some cases, the lethal concentra-
cussed in Section Nomenclature for hydrocarbon solvents – tions are simply greater than the maximally attainable
the naming convention, hydrocarbon solvents were originally vapor concentrations. In summary, hydrocarbon solvents
are not classified for acute toxic effects.
21Hydrodesulfurized kerosene is similar to JP-8 jet fuel and can, in some 3. Skin Corrosion/Irritation – Hydrocarbon solvents do not
ways, be considered as the feed stock for C9–C14 aliphatic hydrocarbon produce irreversible skin damage in acute exposure stud-
solvent production. As reported by Schreiner et al. (1997), kerosenes ies and are not classified for corrosion. However, some
are complex aliphatic substances with approximately 20% aromatics. solvents produce irritation at levels exceeding classifica-
The carbon number range is approximately C9–C16 and the distillation
range is approximately 150–290°C. The aromatic constituents are pre- tion limits during the normal 3-day observation period
dominantly alkylated one- and two-ring aromatics with  0.01% aromatic or persisting to the end of the 14-day observation period.
constituents with 3 or more aromatic rings. A decision as to whether any specific solvent should be
Table 5. Hazard classifications for hydrocarbon solvents in accordance with guidance provided by the United Nations (UN 2011).
Category/Substance Aspiration Hazard Acute CNS Effects Skin Irritation Other Considerations
Category 1 –
C9 Aromatics Yes Yes No See Note 1.
Category 2 –
C10–C12 Aromatics
C10 Yes Yes No Classification as a Category 2 carcinogen may be required depending
on the naphthalene content.
C10–C13 Yes No No Classification as a Category 2 carcinogen may be required depending
on the naphthalene content.
Category 3 –
C9–C14 aliphatics,
2–25% aromatics
C8–C12 aliphatics Yes Yes No See Note 2.
C9–C10 aliphatics
C9–C12 aliphatics
C10–C13 aliphatics Yes No No None
C11–C14 aliphatics
Category 4 –
C14–C20 Aliphatics, Yes No No None

2–25% aromatics
Category 5 –
C5 aliphatics Yes Yes No None
Category 6 –
C6 aliphatics Yes Yes Yes Classification on the basis of neurotoxic and reproductive effects
must be considered depending on the n-hexane content. See Note 3.
Category 7 –
C7–C9 aliphatics Yes Yes Yes Classification on the basis of neurotoxic and reproductive effects
must be considered depending on the n-hexane content. See Note 3.
Category 8 –
C9–C14 aliphatics
 C10 Yes Yes No Hydrocarbon solvents with high levels of cyclics are more
irritating to the skin than those containing primarily normal and/or
isoparaffinic constituents.
 C10 Yes No No Hydrocarbon solvents with high levels of cyclics are more
irritating to the skin than those containing primarily normal and/or
isoparaffinic constituents.
Category 9 –
C14–C20 aliphatics Yes (if kinematic No No None
viscosity is
 20.5 mm2/s at
40°C)
1Classification of 1,2,4 trimethyl benzenes as respiratory irritants is required under the European adaptation of GHS (European Commission, 2008).
2Note that classification for neurotoxicity (STOT RE 1 for CNS; H372) is now required under the European adaptation of GHS (European Commission
2008).
3Classification of some C6 and C7 aliphatic hydrocarbons is required under the European adaptation of GHS (European Commission 2008).
classified as Category 2 (irritant), Category 3 (mild irri- inhibit meiosis, but this is unique to n-hexane, and more
tant), or not classified, requires scientific judgment. It properly considered as a reproductive than a genetic effect.
should be noted that solvents that are not acutely irritat- Hydrocarbon solvents have been found to be inactive in the
ing may produce dermatitis under conditions of repeated majority of the more commonly used in vitro and in vivo
skin contact. screening tests for mutagenic properties. Accordingly, hydro-
4. Serious eye damage/eye irritation – Hydrocarbon solvents carbon solvents are not classified for germ cell mutagenicity.
do not produce serious eye damage. They may cause 7. Carcinogenicity – There is no human evidence suggest-
discomfort but do not cause irritant effects at levels suf- ing that hydrocarbon solvents could be carcinogenic in
ficient to require classification. Accordingly, hydrocarbon humans. There have been several carcinogenicity studies of
solvents are not classified for ocular effects. hydrocarbon solvents and their constituents in animals in
5. Respiratory or skin sensitization – There are no reports which increases in tumor frequencies have been reported.
of respiratory sensitization associated with hydrocarbon However, the tumors seem to have been the consequence
solvent use. The potential for allergic contact dermatitis of repeated injury or other non-genotoxic processes, and
has been assessed via toxicological tests in animals and may not be directly relevant to humans. In general, these
in studies with volunteers. There is no indication that observations fall within the category of “Confounding
hydrocarbon solvents can produce skin sensitization. effects of excessive toxicity or localized effects” (paragraph
Accordingly, hydrocarbon solvents are not classified for 3.6.5.3.2.4, United Nations 2011). Benzene is considered
sensitizing properties. to be a Category 1 (human) carcinogen, but classification
6. Germ cell mutagenicity – There is no evidence that hydro- of hydrocarbon solvents on the basis of benzene content is
carbon solvents produce heritable changes in germ cells in not required as levels in solvents are  0.1% (the concen-
humans and animals. There is evidence that n-hexane can tration limit for classification). Naphthalene is considered
to be a Category 2 carcinogen and is identified by the nam- Exposure to hydrocarbon solvents and occupational
ing convention in those solvents in which it is present at exposure limits
concentrations  1%.
Exposure to hydrocarbon solvents
8. Reproductive Toxicity – There is no evidence that hydro-
carbon solvents in general have adverse effects on sexual It seems likely that the principal exposures to hydrocarbon
function and/or fertility. The only constituent that has solvents, per se, are related to the use of the solvents under
been shown to produce such effects is n-hexane (Section occupational conditions or, secondarily, to the use of solvents
n-Hexane) which causes testicular atrophy. Accordingly, in consumer products. For the population at large, exposures to
hydrocarbon solvents in general are not classified for solvents are more difficult to assess as hydrocarbon solvent con-
effects on fertility, but classification should be considered stituents are present in other materials including petroleum prod-
for those solvents containing n-hexane at levels above clas- ucts, and some are produced during combustion processes. Thus,
sification limits (for further information, see Appendix C, there is exposure data for hydrocarbons that may be hydrocarbon
II.8.1).22 In developmental studies, hydrocarbon solvents solvent constituents, but it is often not clear to what extent these
do not produce malformations or severe developmental exposures are due specifically to hydrocarbon solvent production
toxicity. There is evidence in some studies of develop- or use, as opposed to other unrelated processes.
The production of the first commercial hydrocarbon solvent,
mental delays, but these have only been observed at levels

that are also maternally toxic. Accordingly, hydrocarbon Stoddard solvent, began in the 1920s, and solvents of this type
solvents are not classified for developmental toxicity. It dominated the market until the 1960s when more specialized
should be noted that polycyclic aromatic constituents solvents became available. Stoddard solvent was first used in
with more than three rings have been shown to produce the dry cleaning industry with estimated 8-hour exposures in the
developmental effects; however, these constituents are not range of 15–35 ppm (82–192 mg/m3), depending on the volatil-
found at toxicologically important levels in hydrocarbon ity of the solvents (Oberg 1969). Solvents of this type were then
solvents (McKee et al. 2014). used in the manufacture of house paints for indoor application,
9. Specific Target Organ Toxicity, Single Exposure – Hydro- as discussed in Section Persistent central nervous system effects.
carbon solvents and their constituents with carbon num- There is little data on exposure prior to 1980, but it is believed that
bers ranging between C5 and C9 can produce narcotic exposures were close to the occupational exposure limit of 200
effects including lethargy, lack of coordination, righting ppm (approximately 1100 mg/m3) during that period. The occu-
reflex, narcosis, and ataxia, if inhaled at high levels. As pational exposure limits for these solvents were reduced to 100
all of these effects are reversible within 24 h of exposure, ppm (approximately 550 mg/m3) during the 1970s, and by the
and the data are consistent with classification as Category early 1980s, it was estimated that the 8-hour average exposures of
3 for target organ effects. Hydrocarbon solvents and their house painters were approximately 40 ppm (230 mg/m3) (Riala
constituents with carbon numbers  C9 do not produce et al. 1984). Based on personal breathing zone sample results,
CNS effects and do not require classification. the weighted average exposure to Stoddard solvent between
10. Specific Target Organ Toxicity, Repeated Exposure – As 1961and1998 was 467 mg/m3 (Caldwell et al. 2000).
indicated in the Summary section, repeated exposure to Caldwell et al., (2000) surveyed the published information
hydrocarbon solvents may cause liver enlargement, and available as of 1998 and estimated that during the 1970s, the
in some cases, kidney changes in male rats and small but overall exposures were approximately the recommended occu-
statistically significant reductions in hematological param- pational exposure limits (93%) of the respective TLV® values,
eters. None of these effects is regarded as toxicologically with an overall average exposure of approximately 300 mg/m3.
important, and none has been reported to occur at classifi- By the 1980s, the average exposure was estimated to have been
able levels. The only exceptional constituent is n-hexane, approximately 75 mg/m3 or about 16% of the respective TLV®
which causes peripheral neuropathy. Most hydrocarbon values. During the 1990s, the average exposures were again
solvents are not classified for repeated dose toxicity, but approximately 75 mg/m3, but as some of the TLV® values had
classification should be considered for those solvents con- been lowered, the exposures were approximately 37% of the
taining n-hexane, as described in the previous section.23 respective TLVs®. A subsequent unpublished review of the
literature, extending the Caldwell survey to 2005, concluded
that hydrocarbon solvent exposures have been declining at an
average rate of approximately 5%/year.
22There may be national and regional differences in concentration limits. Although hydrocarbon solvents are used in some consumer
Further, it should be noted that in some regions, classification is based applications, the contribution of hydrocarbon solvent use to
strictly on concentration limits, whereas in other parts of the world, clas- non-occupational exposures is difficult to assess because of
sification is not required if experimental evidence is available demon- contributions from other sources. In particular, hydrocarbons
strating that the stipulated effects do not occur at the classification limits.
In fact, there are experimental data showing that a complex hydrocarbon are also emitted during fuel production and use, and also dur-
solvent containing n-hexane at a concentration of approximately 50% ing combustion. As reported by Hodgson and Levin (2003),
did not affect fertility or produce neurological effects in experimental median residential levels of aromatic hydrocarbons are  25
animals. Thus, both national requirements and compositional informa- ug/m3, and levels of aliphatic hydrocarbons are  10 ug/m3.
tion must be taken into account when classifying hydrocarbon solvents Median levels of these constituents in ambient air are  3 ug/
containing n-hexane.
23Note that in Europe, white spirit has a required classification for neu- m3 (US EPA 2004). These levels are far below the no-effect
rotoxic properties based on studies of construction painters, discussed in levels in both the repeated dose studies in animals and the
Section 4.2.4 of the present document. acute exposure studies in volunteers.
Occupational exposure limits In which Fa is the mass fraction of constituent a, Fb is the

mass fraction of constituent b, etc., and OELa is the exposure
As indicated in Section Exposure to hydrocarbon solvents,
limit for constituents a, b, etc.
the majority of exposure to hydrocarbon solvents occurs
In 2009, this procedure was adopted by the ACGIH® as a
under occupational conditions. Because hydrocarbon sol-
means of calculating occupational exposure limits for complex
vents are widely used and often volatile, there is a need
hydrocarbon solvents for which other occupational exposure
for occupational exposure limit recommendations. As
advice was not available. Although the ACGIH TLV® committee
discussed in preceding sections, this has been an ongoing
accepted the guidance value approach for use in calculating occu-
activity for many years, involving occupational experience,
pational exposure limits for complex hydrocarbon solvents, they
toxicology tests in animals, and short-term tests with vol-
required that the TLVs® for specific hydrocarbon solvent con-
unteers. In previous years, recommendations were given for
stituents be respected. For those constituents with TLVs® (Table
generic types of hydrocarbon solvents. However, because of
6), it is necessary to consider concentrations that may be present
the complex nature of these substances, it has at times been
to determine whether or not the calculated occupational exposure
difficult to relate specific substances to specific recommen-
limits ensure that the TLV® for constituents are not exceeded.
dations, and several different national methods have been
Since the completion of the ECETOC report in 1997 which
proposed. As a means of addressing the issues raised by
formed the basis for the later publication by McKee et al. (2005)
the complex and variable nature of hydrocarbon solvents,
and the adoption of the methodology by the ACGIH®, there
methods were developed which could be used to calculate
have been a number of advances including the expansion of the
occupational exposure levels for complex solvents based on
toxicology data base as described elsewhere, changing views
compositional information (ECETOC 1997, McKee et al.
on recommended occupational exposure limits for some hydro-
2005, TRGS 2007, UK HSE 1995). The underlying prin-
carbon solvent constituents, and the identification of some addi-
ciples were similar to those for categorization and hazard
tional hydrocarbon solvent constituents that may require special
classification, that is, constituents of similar physical and
attention. With those points in mind, the following modifica-
chemical properties were grouped, and toxicological infor-
tions are suggested in order to calculate occupational exposure
mation on representative substances (as well as contempo-
limits consistent with the ACGIH® recommendations:
rary occupational exposure recommendations) were used to
define occupational exposure limits. Substances of unusual 1. Withdraw the guidance value from C7–C8 aromatics –
toxicological properties were distinguished, and it was oth- With changes in regulatory recommendations for toluene
erwise assumed that with no evidence to the contrary, con- and ethylbenzene, the previous recommendation cannot be
stituents of similar physical and chemical properties would justified. Further, the levels of these constituents in hydro-
also be toxicologically similar. The occupational exposure carbon solvents are so low that that they can be ignored in
limits for the groups (group guidance values, GGVs) could most cases. It is suggested that the currently recommended
then be used to calculate a unique occupational exposure TLVs® for C7 and C8 aromatic constituents (i.e., toluene,
limit for any complex hydrocarbon solvent as long as some xylenes, ethylbenzene) be used as specific substance values
relatively simple compositional information was available. in any solvent in which the concentrations of any of these
The calculation method itself was based on a reciprocal constituents exceeds 1%.
calculation procedure (RCP) using the compositional infor- 2. Maintain the currently recommended GGV of 1500 mg/m3
mation and GGVs using a reciprocal formula, as shown for C5–C7 aliphatic constituents, but reduce the GGV for
below. C8 aliphatic constituents to 1400 mg/m3, removing the
inconsistency with the ACGIH TLV® for octanes. It is also
Fa/OELa Fb/OELb  Fc/OELc  …..  1/OELcomplex substance recommended that the TLV® for cyclohexane (350 mg/m3)
Table 6. Group Guidance Values (GGVs) and ACGIH TLV® values for specific hydrocarbon solvent constituents (table adapted
from Appendix H of ACGIH TLV® documentation).
Hydrocarbon Solvent Constituent Group Group Guidance Value Constituents with ACGIH TLVs®
C5–C6 Alkanes 1500 mg/m3 Pentane, all isomers – 2950 mg/m3
Hexane isomers – 1760 mg/m3
C7–C8 Alkanes 1500 mg/m3 Heptane, all isomers – 1640 mg/m3
Octane, all isomers – 1401 mg/m3
C5–C6 Cycloalkanes 1500 mg/m3 Cyclopentane – 1720 mg/m3
Cyclohexane – 350 mg/m3
C7–C8 cycloalkanes 1500 mg/m3 Methyl cyclohexane – 1610 mg/m3
C7–C8 Aromatics 200 mg/m3 Toluene – 75 mg/m3
Xylene, all isomers – 434 mg/m3
Ethylbenzene – 87 mg/m3
C9–C15 Alkanes 1200 mg/m3 Nonane – 1050 mg/m3
C9–C15 Cycloalkanes 1200 mg/m3 None
C9–C15 Aromatics 100 mg/m3 Trimethyl benzene isomers – 123 mg/m3
Naphthalene – 52 mg/m3
Cumene – 246 mg/m3
Indene – 24 mg/m3
Biphenyl – 1.3 mg/m3
be used in the formula for any solvents in which the cyclo- Table 7. Revised method for calculating occupational exposure limits
hexane content exceeds 20%24. for complex hydrocarbon solvents on the basis of compositional
information.
3. Maintain the currently recommended GGV of 1200 mg/m3
for C9–C14 constituents. Recent publications (Adenuga Constituents Recommendations Other Considerations
et al. 2014b, Carrillo et al. 2013) provide support for this C5–C7 aliphatics 1500 mg/m3 At concentrations  20%,
recommendation. It should be noted, however, that there (excluding use: 350 mg/m3 for
n-hexane) cyclohexane
is an ACGIH® TLV® of 1000 mg/m3 for nonanes. To C8 aliphatics 1400 mg/m3
maintain consistency with the ACGIH®, an occupational C9–C14 aliphatics 1200 mg/m3 Use 1000 mg/m3 for C9
exposure limit of 1000 mg/m3 should be used for aliphatic aliphatic solvents
solvents in which the concentration of nonanes exceeds C14–C20 1000 mg/m3 5 mg/m3 for aerosols
aliphatics
80%. However, the use of 1200 mg/m3 for complex solvents C7–C8 aromatics At concentrations  1%,
containing C9 aliphatics at concentrations less than 80% use:
would assure that the overall exposure limit of 1000 mg/m3 Toluene  75 mg/m3
for the C9 aliphatic constituents is not exceeded. Ethylbenzene  86 mg/m3
Xylene  434 mg/m3
4. Maintain the currently recommended GGV of 100 mg/m3 C9–C15 aromatics 100 mg/m3 At concentrations  1%
for C9–C15 aromatics. Concentrations of cumene, indene, (excluding use:

and biphenyl are low enough that they will be controlled naphthalene) Cumene  246 mg/m3
Indene  24 mg/m3
to their own occupational exposure limits as long as the
Biphenyl  1.3 mg/m3
overall recommendation of 100 mg/m3 is maintained. Other constituents
Diethyl- and triethylbenzenes are also of concern, but if that should
present, are expected to be at such low levels that overall be separately
monitored to
recommendations of 100 mg/m3 should assure that expo- assure exposure
sures to these constituents are below levels of toxicologi- limits are not
cal concern. exceeded
5. Maintain the previous recommendation that n-hexane and n-hexane 176 mg/m3
Naphthalene 52 mg/m3
naphthalene be treated separately using their own TLVs®. Methylnaphthalene 3 mg/m3
The current values for these constituents are: n-hexane  50 isomers
ppm, 176 mg/m3; naphthalene  10 ppm, 52 mg/m3. The

continued use of these levels as specific substance values in
the calculations would facilitate changes in recommenda-
Conclusions
tions, if there are any changes in the recommended TLVs®
for these constituents. The objectives of this document are to describe hydrocarbon
6. Separately monitor for methylnaphthalene isomers to solvents, summarize the toxicological information, and to
assure that their TLVs® (0.5 ppm, 3 mg/m3) are not then show how this information can be used to characterize
exceeded. and communicate toxicological hazards. As discussed, hydro-
7. Use the GGV of 1200 mg/m3 for constituents with carbon carbon solvents are composed of constituents with carbon
numbers C14–C20, for purposes of calculating the OELs. numbers in the range of C5–C20. Although many of these
Because of the low vapor pressures of these constituents, hydrocarbon solvents have complex and variable composi-
these would not contribute significantly to the overall vapor tions (UVCB), most of the constituents have similar proper-
levels. Under circumstances in which aerosol formation is ties, allowing hazard characterization to be approached on a
possible, the aerosol concentrations should be measured generic basis. To facilitate the hazard characterization process,
separately, and exposures to aerosols should be limited to 5 a more precise nomenclature (the naming convention) has been
mg/m3 as recommended by the ACGIH® for highly refined introduced to more clearly describe the types of constituents
mineral oils. that might be present. Based on the constituent information
and considering the physical/chemical properties, the solvents
Note that for these higher molecular weight solvents, care
were grouped into 9 categories of similar substances. Hazards
should be taken to avoid exceeding the maximally attainable
were characterized on a category basis using the results of
vapor concentration to minimize the likelihood of aerosol for-
studies of representative substances. The hazard characteriza-
mation.
tions were then used for hazard classification and as a basis for
The revised recommendations are shown in Table 7.
occupational exposure recommendations as demonstrated by
examples provided. In short, hydrocarbon solvents have com-
plex and variable compositions, but because of the similari-
24Ifthe cyclohexane levels are below 20%, then the exposure to cyclo- ties in physical/chemical and toxicological properties of most
hexane would not exceed 300 mg/m3, assuming the recommended overall of the constituents, the complexity can be addressed through
exposure limit for the solvent of 1500 mg/m3 is maintained. As exposure generic approaches. There are a few hydrocarbon solvent
to cyclohexane would not exceed its own occupational exposure limit constituents with unusual properties, but these are well char-
(350 mg/m3) if these conditions are observed, and it does not need to be acterized and are accommodated through the differentiation
considered in the calculations. If the concentration of cyclohexane  20%,
then the molar fraction of hexane should be separated from the other ali- provided by the naming convention. More complete informa-
phatic constituents and considered separately, using the TLV® as a group tion on the toxicological hazards of these solvents is provided
guidance value. in the appendices.
Appendices: Summarized toxicological information 9.2.1 Studies to assess the potential for persistent
on hydrocarbon solvents and their constituents, CNS effects in animals . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 326
9.2.2 Studies to assess the potential for persistent
organized on a category basis
CNS effects in humans . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 326
9.2.3 Occupational exposures and recommended
Appendix A. Aromatic hydrocarbon solvents occupational exposure limits ... ... ... ... ... ... ... ... ... ... ... ... 326
II. Toxicological properties of C14–C20 Aliphatic

I. Toxicological properties of C9 aromatic solvents (Category 1
Hydrocarbon Solvents (2–30% Aromatics)
solvents)�� 309
(Category 4 solvents)�� 327
1. Introduction . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 309
1. Introduction . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 327
2. Physical/chemical properties ... ... ... ... ... ... ... ... ... ... ... ... ... 309
2. Physical and chemical properties . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 327
3. Toxicokinetic properties of C9 aromatic solvents . .. . .. . .. . .. . .. . 309
3. Toxicokinetics ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 327
3.1 Absorption . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 309
3.1 Absorption . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 327
3.2 Distribution ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 310
3.2 Metabolism ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 327
3.3 Metabolism and excretion . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 310
3.3 Excretion ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 328
4. Acute toxicological properties . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 310
4. Acute toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 328
5. Repeated dose studies . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 311
5. Repeated dose toxicity ... . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 328
5.1 Repeated inhalation toxicity studies .. . .. . .. . .. . .. . .. . .. . .. . .. 311

6. Genetic toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 329
5.2 Repeated oral toxicity studies ... . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 312
7. Developmental toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 329
6. Genetic toxicity studies ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 313
8. Reproductive toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 329
9. Carcinogenicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 329
10. Neurotoxicity . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 329
10. Neurotoxicity ... . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 315
10.1 Acute neurological effects ... ... ... ... ... ... ... ... ... ... ... ... 315 Appendix C. Volatile aliphatic solvents
10.2 Subchronic neurological studies . .. . .. . .. . .. . .. . .. . .. . .. . .. . 316 I. Toxicological Properties of C5 Aliphatic Solvents (Pentanes)
11. Observations in humans ... ... ... ... ... ... ... ... ... ... ... ... ... ... 316 (Category 5 solvents)�� 329
1. Introduction . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 329
II. Toxicological properties of C10–C12 Aromatic solvents
3. Toxicokinetics ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 329
1. Introduction . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 316 4. Acute toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 330
2. Physical and chemical properties . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 317 5. Repeated dose toxicity ... . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 330
3. Toxicokinetic properties ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 317 6. Genetic toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 331
4. Acute toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 317 7. Developmental toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 331
5. Repeated dose toxicity studies ... . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 317 8. Reproductive toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 331
5.1 Repeated inhalation toxicity studies . .. . .. . .. . .. . .. . .. . .. . .. . 317 9. Carcinogenicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 332
5.2 Repeated oral toxicity studies ... ... ... ... ... ... ... ... ... ... ... 318 10. Neurological studies .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. 332
6. Genetic toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 319 11. Human observations . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 332
8. Reproductive toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 319 II. Toxicological properties of C6 Aliphatic Hydrocarbon Solvents
9. Carcinogenicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 319 (Hexanes) (Category 6 solvents)�� 332
10. Neurotoxicity . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 319
11. Human observations . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 320 1. Introduction . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 332
11.1 Ocular and respiratory irritation .. . .. . .. . .. . .. . .. . .. . .. . .. . .. 320 2. Physical and chemical properties . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 332
11.2 Studies to assess the potential for dermal irritation 3. Toxicokinetics ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 332
and allergic contact dermatitis ... ... ... ... ... ... ... ... ... ... ... 320 3.1 Commercial hexane ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 332
11.3 Occupational exposures and recommended 3.2 Cyclohexane ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 333
occupational exposure limits . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 320 3.2.1 Toxicokinetic studies of cyclohexane in Male F344 rats ... 333
3.2.2 Toxicokinetic studies of cyclohexane in humans . .. . .. . 333
Appendix B. Aliphatic/aromatic hydrocarbon solvents [C9–C20 4. Acute toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 333
Aliphatic hydrocarbon solvents (2–25% aromatics)] 5. Repeated dose toxicity ... . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 334
5.1 Repeated dose toxicity studies of n-hexane ... ... ... ... ... ... 334
I. Toxicological properties of C9–C14 Aliphatic Hydrocarbon 5.1.1 Repeated oral toxicity studies . .. . .. . .. . .. . .. . .. . .. . .. . .. . 334
Solvents (2–25% Aromatics) (Category 3 solvents) �� 320 5.1.2 Repeated inhalation toxicity studies ... ... ... ... ... ... ... 334
1. Background . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 320 5.2 Repeated dose toxicity studies of commercial hexane ... ... 334
2. Physical and chemical properties . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 320 5.2.1 Repeated toxicity studies of commercial
3. Toxicokinetics ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 321 hexane by inhalation ... ... ... ... ... ... ... ... ... ... ... ... ... ... 334
4. Acute toxicity tests ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 321 5.3 Repeated dose toxicity studies of cyclohexane .. . .. . .. . .. . .. 335
5. Repeated dose toxicity ... . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 322 5.4 Repeated dose toxicity studies of methylcyclopentane . .. . 335
5.1 Repeated inhalation toxicity studies of complex C9–C14 6. Genetic toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 335
aliphatic solvents (2–25% Aromatics) Solvents ... ... ... ... ... 322 6.1 Genetic toxicity studies of n-hexane .. . .. . .. . .. . .. . .. . .. . .. . .. 335
6. Genetic toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 323 6.2 Genetic toxicity studies of other C6 aliphatic solvents ... ... 335
7. Developmental toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 323 6.3 Genetic toxicity studies of cyclohexane . .. . .. . .. . .. . .. . .. . .. . 335
8. Reproductive toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 323 7. Developmental toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 335
9. Neurotoxicity studies ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 324 7.1 Developmental toxicity studies of n-hexane .. . .. . .. . .. . .. . .. 335
9.1 Studies of acute CNS effects ... ... ... ... ... ... ... ... ... ... ... ... 324 7.2 Developmental Toxicity Studies of Commercial Hexane ... . 335
9.1.1 Studies of acute CNS effects in animals ... ... ... ... ... ... 324 7.3 Developmental toxicity studies of cyclohexane . .. . .. . .. . .. . 335
9.1.2 Observations of acute CNS effects and 8. Reproductive toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 336
upper respiratory tract irritation in humans . .. . .. . .. . .. . .. . 324 8.1 Reproductive toxicity studies of n-hexane ... . .. . .. . .. . .. . .. . 336
9.2 Studies to assess the potential for persistent CNS 8.2 Reproductive toxicity studies of commercial hexane ... ... 336
effects of C9–C14 aliphatic hydrocarbon 8.3 Reproductive toxicity studies of cyclohexane ... ... ... ... ... 336
solvents (2–25% aromatics) . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 326 9. Carcinogenicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 336
10. Neurological studies .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. 337 10. Observations in humans ... ... ... ... ... ... ... ... ... ... ... ... ... ... 350
10.1 Acute neurological studies ... ... ... ... ... ... ... ... ... ... ... ... 337 10.1 Studies of acute central nervous system effects
10.1.1 Acute neurological effects in animals ... ... ... ... ... ... 337 and upper respiratory irritation ... ... ... ... ... ... ... ... ... ... ... 350
10.1.2 Acute neurological effects in humans ... ... ... ... ... ... 337 10.2 Investigations involving other physiological
10.2 Persistent neurological effects . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 337 and/or clinical parameters ... ... ... ... ... ... ... ... ... ... ... ... ... 351
10.2.1 Persistent neurological effects in animals .. . .. . .. . .. . .. 337 10.3 Occupational exposures and recommended
10.2.1.1 Hypothesized mode of action for n-hexane- occupational exposure limits ... ... ... ... ... ... ... ... ... ... ... ... 351
induced peripheral neuropathy ... ... ... ... ... ... ... ... ... 337
11. Human observations . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 339 IV. Toxicological properties of C14–C20 aliphatic
(< 2% aromatics) hydrocarbon solvents
III. Toxicological properties of C7–C9 aliphatic hydrocarbon
solvents (Category 7 solvents) �� 339 1. Introduction . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 351
1. Introduction . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 339
3. Toxicokinetic properties of C14–C20 aliphatic ( 2% aromatics)
hydrocarbon solvents ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 352
3. Toxicokinetics ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 339
3.1 Absorption . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 352
3.1 Absorption of C7–C9 aliphatic hydrocarbons ... ... ... ... ... 339
3.2 Metabolism, distribution, and excretion ... . .. . .. . .. . .. . .. . .. . 352
3.2 Distribution of C7–C9 aliphatic hydrocarbons ... ... ... ... ... 340
4. Acute toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 353
3.3 Metabolism and elimination of C7–C9 aliphatic

hydrocarbons . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 340
3.4 Pharmacokinetic modeling ... ... ... ... ... ... ... ... ... ... ... ... 340
4. Acute toxicological effects ... ... ... ... ... ... ... ... ... ... ... ... ... ... 340
4.1 Acute inhalation toxicity studies ... ... ... ... ... ... ... ... ... ... 341
4.2 Acute oral and dermal toxicity ... ... ... ... ... ... ... ... ... ... ... 341
10. Neurotoxicity . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 354
4.3 Dermal and ocular irritant effects ... ... ... ... ... ... ... ... ... ... 341
11. Observations in humans ... ... ... ... ... ... ... ... ... ... ... ... ... ... 354
5.1 Repeated inhalation studies of C7–C9 aliphatic
hydrocarbon solvents . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 341
5.2 Repeated dose studies of C7–C9 Aliphatic hydrocarbon
solvent constituents ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 342
7. Developmental toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 343 Appendices: Summarized toxicological information
8. Reproductive toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 343 on hydrocarbon solvents and their constituents,
9. Carcinogenicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 343 organized on a category basis.
10. Neurotoxicity . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 343
10.1 Studies of acute neurological effects ... ... ... ... ... ... ... ... 343 Appendix A. Aromatic hydrocarbon solvents
10.2 Studies of persistent neurological effects ... . .. . .. . .. . .. . .. . 343
11. Occupational exposures and recommended I. Toxicological properties of C9 aromatic solvents (Category
occupational exposure limits ... ... ... ... ... ... ... ... ... ... ... ... ... 344 1 solvents)
Appendix D. Non-Volatile aliphatic hydrocarbon solvents 1. Introduction

“C9 Aromatic solvents” are complex and variable combina-
I. Toxicological properties of C9–C14 Aliphatic Hydrocarbon
Solvents (< 2% aromatics) (Category 8 solvents)�� 344 tions of aromatic hydrocarbons with carbon numbers pre-
1. Introduction . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 344
dominantly in the C9 range (i.e., isomers of trimethylbenzene
2. Physical and chemical properties . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 344 and ethyltoluene) and boiling in the range of approximately
3. Toxicokinetics ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 344 140°C–200°C. These substances are listed on the US EPA
3.1 Absorption . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 344 Toxic Substances Control Act (TSCA) registry under the
3.2 Distribution ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 345 Chemical Abstract Services (CAS) registry number 64742-
3.3 Metabolism ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 345
3.4 Excretion ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 345
95-6, in Europe under the EC number 918-668-5, and for
4. Acute effects ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 346 REACH purposes were registered as “Hydrocarbons, C9 Aro-
4.1 Acute toxicity . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 346 matics” in accordance with the naming convention introduced
4.2 Ocular and dermal irritation, sensitization . .. . .. . .. . .. . .. . .. . 346 by the European Hydrocarbon Solvent Producers Association
5. Repeated dose studies . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 346 (HSPA 2011, Table 2).
5.1 Repeated inhalation toxicity studies .. . .. . .. . .. . .. . .. . .. . .. . .. 346
5.2 Repeated oral toxicity studies ... . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . 346
5.2.1 Results of repeated oral administration studies of 2. Physical/chemical properties
normal paraffins ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 347 The C9 aromatic solvents are colorless liquids at room tem-
5.2.2 Results of repeated oral toxicity studies of
isoparaffinic hydrocarbons .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. . .. 347
perature with pungent odors and relatively low odor thresh-
5.2.3 Results of repeated dose studies of olds. The physical and chemical properties of these solvents
cycloparaffinic hydrocarbons ... ... ... ... ... ... ... ... ... ... ... 348 are summarized in Table 8.
5.2.4 Results of repeated dose studies of complex
C9–C14 aliphatic ( 2% aromatic) hydrocarbon solvents ... 348 3. Toxicokinetic properties of C9 aromatic solvents
5.3 Repeated dermal toxicity studies ... ... ... ... ... ... ... ... ... ... 348
6. Genetic toxicity ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 348 3.1 Absorption
7. Developmental and reproductive toxicity ... ... ... ... ... ... ... ... 348
8. Carcinogenicity studies ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 349 Inhaled C9 alkylbenzenes are readily absorbed in the lung.
9. Neurotoxicity studies ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... ... 350 In an investigation of the uptake and eventual disposition of
Table 8. Physical and chemical properties of C9 aromatic solvents. In a study of the comparative properties of hydrocarbons of
Property Result different molecular types, Zahlsen et al. (1990, 1992) found that
Colorless liquid with pungent
levels of 1,2,3-TMB were higher in blood and perirenal fat than
Physical state at 20°C and 1013 hPa
odor the levels of aliphatic constituents with similar carbon numbers.
Odor Threshold 11 mg/m3 (Nau et al. 1966) In a separate study, 1,2,4-TMB was preferentially distributed
Melting/Freezing Point The pour point is   30°C, to fat as compared to brain, kidney and liver (Eide and Zahlsen
the melting point is  100°C
(ASTM D5950) 1996). As shown by Hissink et al. (2007), inhaled 1,2,4-TMB
Boiling Point 165–180°C (ASTM D86) is taken up quickly into the brain but is also quickly eliminated
Relative Density 0.80–0.95 g/cm3 at 15°C. once exposure is discontinued. This uptake and elimination
Vapor Pressure  1 Kpa at 20°C pattern was observed in other studies, suggesting that the abil-
Flash Point 35°C (DIN 51755)
Self-Ignition Temperature  400°C at 1 atm
ity of aromatic constituents to induce their own metabolism
Surface Tension 28–32 mN/m @ 15°C may be responsible for the more rapid elimination of these
Viscosity 0.7–1.7 mm2/s at 20°C compounds relative to saturated hydrocarbons (Zahlsen et al.
Maximally Attainable Vapor Approximately 10000 mg/m3 1990). Preferential distribution of 1,2,4-TMB to adipose tissue
Concentration (2000 ppm)
Conversion factor (ppm/mg/m3) 1 ppm  ∼ 5 mg/m3
in rats was also observed following oral administration (Huo
Octanol/water partition coefficient 3.4–3.9 et al. 1989). Similar results were observed in a tissue distribu-
Water solubility 40–75 g/l at 25°C tion study of 4-ethyltoluene (4-ET). Male rats were exposed
to 50 ppm (250 mg/m3) or 200 ppm (1000 mg/m3) of 4-ET as
either a single 6-hour exposure or repeatedly (6 hours/day for
5 days) (Swiercz et al. 1996). Blood concentrations of 4-ET
the three TMB isomers, Jarnberg et al. (1996) exposed vol- were 10-fold higher after a single exposure at 200 ppm (1000
unteers to 25 ppm of 1,2,3-; 1,2,4-; or 1,3,5-TMB for 2 h at a mg/m3) than at 50 ppm (250 mg/m3), suggesting saturation of
continuous 50-watt workload to simulate light activity. The metabolism and reduced elimination rates at the higher expo-
25 ppm exposure level was selected to correspond to occu- sure levels. Elimination followed a two-compartment model,
pational exposure recommendations of 20–25 ppm (100–125 and 4-ET was almost entirely cleared from the blood within
mg/m3). Relative respiratory uptake (reported as net uptake 24 h.
plus amount cleared by expiration during exposure) was
approximately 60% for all three isomers, and similar experi-
3.3 Metabolism and excretion

ments with 2 ppm (10 mg/m3) of 1,2,4-TMB indicated that
absorption was similar across the range of concentrations Studies by Huo et al. (1989) showed that side chain oxida-
tested. Volumes of distribution were similar across all three tion is the major pathway by which 1,2,4-TMB is metabolized,
isomers and ranged from 30–39 l/kg. The large volumes of mostly to mercapturic, sulfate, glycine, and glucuronide con-
distribution are consistent with the tendency for trimethyl- jugates of dimethylbenzyl alcohols and dimethylbenzoic acids.
benzenes to be retained in adipose tissue, which is supported More than 99% of the administered dose was eliminated in the
by the long terminal half-lives, as discussed in the next para- urine within 24 h (Huo et al. 1989). For 1,3,5-TMB, 78% of the
graph. administered dose was excreted as 3,5-dimethylhippuric acid
Studies by Kostrzewski et al. (1997) and Jones et al. (2006) with an additional 16% excreted as glucuronide or sulfate con-
provided similar results. Jones reported that elimination fol- jugates (Mikulski and Wiglusz 1975). Based on urinary data,
lowed a two-compartment model with the half-life for the TMB metabolism is similar in humans (Jarnberg et al. 1997),
initial elimination phase of 60 min and a slower second elimi- rats (Huo et al. 1989), and rabbits (Laham and Potvin 1989).
nation phase with a mean half-life of 600 min. Kostrzewski
4. Acute toxicological properties
et al. (1997) reported a three-compartment elimination model,
and Jarnberg et al. (1996) reported four elimination half-lives A summary of the acute toxicity data on complex C9 aromatic
with a terminal phase of 78–120 hours. Breath concentrations solvents, extracted from the REACH registration dossier
of 1,3,5-TMB returned to pre-exposure levels within 24 h of (ECHA 2010a), along with results of individual C9 isomers
exposure, indicating rapid metabolic clearance (Jones et al. (Korsak and Rydzynski 1996), is given in Table 9. As shown
2006). in oral toxicity studies, the complex C9 aromatic solvents
produced lethality at doses exceeding 3000 mg/kg but were
not acutely toxic following dermal administration or by inhala-
3.2 Distribution
tion at maximally attainable vapor concentrations. Similarly,
Based on a model, Meulenberg and Vijverberg (2000) predicted the trimethylbenzene isomers were not toxic by inhalation
that tissue: air partition coefficients for all three TMB isomers at maximally attainable vapor concentrations. However, C9
would be similar in humans and rats, indicating that the TMB solvents can cause acute CNS effects (Korsak and Rydzynski
isomers are likely to have similar tissue distribution patterns 1996, McKee et al. 2010). C9 aromatic solvents did not cause
in both species and that the results of studies in rats would toxicologically relevant levels of skin or eye irritation when
probably be reflective of systemic distribution in humans. tested for these properties in animals, but human observations
Estimated fat/air partition coefficients were at least 10-fold indicate that they can cause ocular and/or upper respiratory
greater than those for other tissues and 100-fold greater than tract irritation if inhaled at high levels (Battig et al. 1956, Nau
the blood/air partition coefficients, indicating a propensity for et al. 1966). There is no evidence that these solvents can cause
distribution to adipose tissue. allergic contact dermatitis in humans or animals.
Table 9. Summarized results of acute toxicity tests of complex C9 aromatic solvents and C9 isomers.
Test Test Substance Methods Resultsa
Acute Oral Complex C9 aromatic solvent Oral gavage (2 males, 2 females) Doses  1, 2, 4, 8 Males – LD50  8 ml/kg (7000 mg/kg)
Toxicity ml/kg Females – LD50  4 ml/kg (3500 mg/kg)
Acute Dermal Complex C9 aromatic solvent Occlusive dermal exposure in male/female New LD50  3160 mg/kg
Toxicity Zealand rabbits (equivalent to OECD 402)
Acute Complex C9 aromatic solvent Rat, male/female, whole body vapor exposure LC50  6193 mg/m3 (analytically
Inhalation (equivalent to OECD 403) determined maximally attainable vapor
Toxicity concentration) [approximately 1240 ppm]
Complex C9 aromatic solvent Rat, male/female, whole body vapor exposure LC50  10,200 mg/m3 (nominal
(equivalent to OECD 403) concentration, approximately 2000 ppm)
1,2,4-trimethylbenzene Rat male/female 12-hour whole body vapor exposure, LC50  10800 mg/m3 (nominal
concentrations  2000 ppm or 10800 mg/m3 concentration, approximately 2000 ppm)
aSome of these data have not been formally published, but the results are summarized in OECD 2012a. The exception is the LC50 value for 1,2,4-trim-
ethylbenzene which was taken from a publication by Cameron et al. (1938).
In subacute inhalation toxicity studies, rats survived expo- rats, dogs, and monkeys. The animals were exposed at levels
sures at levels approximating the maximally attainable vapor ranging from 50 ppm to 1000 ppm (250–5000 mg/m3), for
concentrations. In summary information published by Gage periods ranging from 90 to 150 days, and with daily exposure
(1970), it was reported that in studies of rats exposed 12 times periods ranging from 8 to 23.5 hours/day. Because the proto-
in exposure periods lasting 6 h to 2000 ppm (10000 mg/m3) cols were so different from those now being used and because
(which Gage considered to be a saturated atmosphere), there there are some uncertainties relating to the compositions of
was evidence of ocular and nasal irritation, respiratory diffi- the test materials used in these early studies, generalizations
culty, lethargy, tremors, and reduced body weight gain, but that can be made from these data are limited. However, in
there were no changes in hematological parameters, and no reference to the test most similar to current protocols, the
pathological findings were reported. authors stated that at “…[C]oncentrations of C9-C10, 200
ppm [approximately 1000 mg/m3]. Rats exposed eight hours/
day and five days per week (90 exposures), showed no per-
5. Repeated dose studies
sistent or significant peripheral blood changes, weight gains,

5.1 Repeated inhalation toxicity studies bone marrow or eye lens changes…”
Carpenter et al. (1975g) reported repeated inhalation toxic-
The first reports of toxicological investigations of C9 aromatic
ity studies of “70 solvent” (described as containing primarily
solvents were published in the German literature (Battig et al.
C9 (47%) and C10 (20%) alkylbenzenes) in rats and dogs.
1956, 1958). The investigated solvent, “Fleet-X-DV-99”, was
They observed acute CNS depression in acute studies at
described by the authors as a commercial product containing
810 ppm (approximately 4050 mg/m3) and small but statisti-
80% TMB isomers with the remaining constituents being other
cally significant reductions in body weight gain in repeated
aliphatic and aromatic components. The 1956 paper reported
dose studies at 410 ppm (approximately 2050 mg/m3). The
observations of 27 painters who were exposed to this solvent
overall no-effect concentration was 200 ppm (approximately
under occupational conditions at levels between 10 and 60
1000 mg/m3).
ppm (50–300 mg/m3). Symptoms included headaches, feelings
More recently, two repeated inhalation toxicity studies
of tiredness during and after the shift; bronchitis with expec-
(13 weeks, 12 months) were conducted in male and female
toration and coughing; bleeding of nose and gums with hema-
Wistar rats using study designs similar to OECD 413 (Clark
tomas and reduced coagulation times; and a reduction in the
et al. 1989). The test material contained 42% TMB isomers and
number of erythrocytes. The 1958 paper reported the results of
31% ET isomers.25 In the 13-week study, exposure levels were
toxicological investigations including one study to investigate
1800, 3700, or 7400 mg/m3 (360, 740, or 1480 ppm). Based
hematological profiles. In a 90-day study, rats were injected
on summarized information, there was evidence of increased
with daily doses of 0.5 ml (approximately 2000 mg/kg/day,
liver and kidney weights in the female rats from the mid- and
assuming the rats weighed approximately 250 g); no changes
high-exposure groups and small reductions in hematological
in hematological profiles were observed. In a repeated inhala-
parameters in female rats from all exposure groups. Using
tion toxicity study of approximately 90 days at an exposure
the information from the 13-week study to define exposure
level of 1700 ppm (8500 mg/m3, approximately the maximally
levels for a longer term study, Wistar rats were exposed for
attainable vapor concentration for a solvent of this type), the
12 months at levels of 470, 930, or 1830 mg/m3 (94, 186, or
authors observed a decrease in lymphocytes, an increase in
366 ppm). Liver and kidney weights were elevated in males
neutrophils, effects in liver and kidney, evidence of respiratory
from the high-exposure group, but there were no pathologi-
irritation, and increased levels of phenol in the urine. Based
on the hematological findings in the human subjects, Gerarde
(1960) suggested that Fleet-X-DV-99 may have contained low
25The test material used by Clark et al. (1989) contained 2.3% o-xylene,
levels of benzene as a contaminant.
4.0% propylbenzene, 7.1% 1-methyl-3-ethylbenzene, 16.6% 1-methyl-4-
The first systematic investigation of the toxicological ethylbenzene, 9.4% 1,3,5-trimethylbenzene, 7.2% 1-methyl-2-ethylben-
properties of complex C9 aromatic solvents was by Nau et al. zene, 32.7% 1,2,4-trimethylbenzene, 2.8% 1,2,3-trimethylbenzene, and
(1966) who conducted repeated inhalation toxicity studies in 8.3% C10 alkylbenzenes.
cal changes in these organs. In rats exposed at the high level study included an additional group of animals exposed at the
for 12 months and then held for 4 weeks without treatment highest level for 13 weeks and then held for an additional 4
to assess recovery, the organ weights were similar to control weeks to assess recovery. There were no significant differ-
values, providing evidence that the increased organ weights ences in terminal body weights, and there were no changes
were more likely adaptive responses than adverse findings. in clinical or hematological parameters that were judged to
There was no consistent pattern of hematological effects. be toxicologically important. There were statistically signifi-
There were some statistically significant differences in clini- cant increases in liver, kidney and adrenal weights in the 500
cal values, but these were within normal physiological ranges and 1200 ppm (2500 and 6000 mg/m3) groups, but the only
for the Wistar rat. The authors concluded that the overall no histological findings were changes in the kidneys of male rats
observed adverse effect concentration in the 12-month study consistent with an a2u-globulin-mediated nephropathy. There
was 1830 mg/m3 (366 ppm). were no apparent toxicologically important findings at 1200
Similar results were obtained in repeated dose studies of ppm (6000 mg/m3), the highest concentration tested.
individual TMB and ET isomers. Swiercz et al. (2000) evalu- The NTP (2009) exposed rats and mice to cumene vapor at
ated the repeated dose toxicity of ethyltoluene in a study in concentrations of 0, 62.5, 125, 250, or 1000 ppm (312, 625,
which rats were exposed 5 days/week for 4 weeks, at levels of 1250, or 5000 mg/m3), for 6 hours/day and 5 days/week, for 14
477 or 2337 mg/m3 (95 or 467 ppm). The authors reported that
weeks. All rats survived to scheduled termination, and there

the rats survived the exposure period, that there were no dif- were no significant differences in terminal body weights or in
ferences in terminal body weights, and that the clinical obser- body weight gains in comparison to the control groups. Statis-
vations did not reveal any important toxicological findings. tically significant increases in liver weights were observed in
As the objective of the study was to assess the potential for male rats in all of the exposed groups and in female rats in all
respiratory effects, the pathological investigation was limited but the lowest-exposure (62.5 ppm, 312 mg/m3) group. A sta-
to the lungs. The principal finding was evidence of respira- tistically significant increase in bile acids was found in males
tory irritation in male rats exposed to 2337 mg/m3 (467 ppm); at the end of the exposure period in the 500 and 1000 ppm
there were no reported effects in rats exposed to 477 mg/m3 (2500 and 5000 mg/m3) groups, but there was no evidence of
(95 ppm). liver damage. Other changes in serum chemistry parameters
Korsak et al. (2000a) reported a study in which rats were (reduced levels of sorbitol dehydrogenase, alkaline phos-
exposed to 1,2,3-TMB at levels of 123 mg/m3, 492 mg/ phatase, and alanine aminotransferase) did not appear to be
m3, or 1230 mg/m3 (25, 100, or 250 ppm), 6 hours/day and toxicologically important as the levels of liver enzyme markers
5 days/week for 3 months. There were no unscheduled deaths were reduced, not elevated. Dose-related increases in proximal
and no changes in body weight. There was an approximately tubular hyaline droplet accumulation and a2u-globulin were
10% increase in liver weight in male rats from the high expo- observed in kidneys of male rats (NTP 2009).
sure group and an approximately 10% decrease in absolute In the mouse study, eight females in the 1000 ppm (5000
spleen weight in female rats, but there were no reported patho- mg/m3) group died or were sacrificed prior to scheduled ter-
logical changes in liver or spleen, and levels of liver enzyme mination, but all other animals survived to the end of the expo-
markers were not elevated. The authors reported changes in sure period (NTP 2009). Mortality appeared to be related to
peribronchial lymphatic tissue, but they did not consider these acute toxicity expressed during the first exposure week. Male
effects to have been exposure-related. In summary, there was mice apparently developed a tolerance to cumene by the third
suggestive evidence of respiratory effects in rats from the high day of exposure. Statistically significant reductions in terminal
exposure group, but there were no apparent systemic effects. body weights and body weight gains were observed in male
The no observed effect concentration for systemic effects was mice exposed at levels above 250 ppm (1250 mg/m3), but the
1230 mg/m3 (250 ppm), the highest concentration tested. differences were less than 10% in comparison to control levels.
Korsak et al. (2000b) exposed rats to 1,2,4-TMB at levels of Liver weights in both male and female mice were elevated in
123 mg/m3, 492 mg/m3, or 1230 mg/m3 (25, 100, or 250 ppm), exposure groups of 125 ppm (625 mg/m3) and above. Some
for 6 hours/day, 5 days/week for 3 months. There were no liver necrosis and focal chronic inflammation was noted in
unscheduled deaths and no effects on body weight. There were the livers of male and female mice, but these changes did
no statistically significant changes in organ weight or hemato- not appear to have been dose-related. The investigators con-
logical parameters. The only statistically significant change in sidered the liver findings to have been part of a pathological
clinical chemistry parameters was a small increase in sorbitol continuum.
dehydrogenase. There were no pathological changes. The only In summary, there have been repeated inhalation toxicity
microscopic observations were small but statistically signifi- studies of complex C9 aromatic hydrocarbon solvents as well
cant increases in interstitial infiltrate and alveolar macrophages as studies of many of the individual isomers. As summarized
in the mid and high exposure groups. The no observed effect in Table 10, there was evidence of respiratory irritation in
concentration for systemic effects was 1230 mg/m3 (250 ppm), studies involving exposures at levels equal to or greater than
the highest concentration tested. 500 mg/m3 (100 ppm), but there were no systemic effects in
Cushman et al. (1995) exposed rats to cumene (isopro- any of the studies at levels up to 1830 mg/m3 (366 ppm).
pylbenzene) in an initial study involving exposures for
6 hours/day and 5 days/week at levels of 100, 500, or 1200
5.2 Repeated oral toxicity studies
ppm (500, 2500, or 6000 mg/m3) for 13 weeks, and then in a
second study in which the exposure concentrations were 50, Adenuga et al. (2014a) reported the results of a 90-day
100, 500, or 1200 ppm (250, 500, 2500 or 6000 mg/m3). Each repeated dose toxicity study in rats in which 1,3,5-TMB was
Table 10. Summarized results of repeated inhalation toxicity studies of complex C9 aromatic hydrocarbon solvents and their constituents.
Test Material Length of Study Species No Observed Adverse Effect Concentration References
Complex C9 aromatic solvent 12 months Rat  1800 mg/m3( 360 ppm) Clark et al. (1989)
4-ethyltoluene 4 weeks Rat  2500 mg/m3 ( 500 ppm) Swiercz et al. (2000)
1,2,3-trimethylbenzene 3 months Rat  1230 mg/m3( 246 ppm) Korsak et al. (2000a)
1,2,4-trimethylbenzene 3 months Rat  1230 mg/m3( 246 ppm) Korsak et al. (2000b)
Isopropyl benzene 13 weeks (2 studies) Rat  6000 mg/m3( 1200 ppm) Cushman et al.
(1995)
Isopropyl benzene 14 weeks Rat  4900 mg/m3 ( 980 ppm) NTP (2009)
Isopropyl benzene 14 weeks Mouse  4900 mg/m3 ( 980 ppm) NTP (2009)
administered by gavage at levels of 50, 200, or 600 mg/kg/ 7. Developmental toxicity

day. Another group was treated for 90 days at the high-dose
Ungvary and co-workers (Ungvary et al. 1983, Ungvary and
level and then held for 4 weeks without treatment, to assess
Tatrai 1985) exposed pregnant female CFY rats continuously
recovery. The principal effects noted in the high-dose group
(i.e., 24 hours/day) from days 7–15 of gestation to 120, 200, or

included liver weight increases in both male and female rats
400 ppm (600, 1000, or 2000 mg/m3) of Aromatol,27 a branded
and kidney weight increases in male rats. There were no path-
product meeting the specifications of high flash aromatic naph-
ological findings in these or other organs, and levels of marker
tha. The majority of the animals were sacrificed on gestational
enzymes for liver toxicity were not elevated. There were no
day 21 and the uterine contents were examined, but there
differences in organ weights in animals treated for 90 days at
was an additional group containing dams that were allowed
the high dose and then held for 4 weeks without treatment.
to deliver naturally. The offspring from this additional group
The overall no observed adverse effect level for this study was
were sacrificed and examined on postnatal day 90. Among the
600 mg/kg/day.
animals sacrificed on gestational day 21, the maternal livers
6. Genetic toxicity studies were enlarged but there were no pathological changes in the
livers or other organs of these animals, and there was no evi-
The mutagenic potential of a complex C9 aromatic hydrocar- dence of fetal mortality or major malformations. Developmen-
bon solvent was assessed using a battery of genetic toxicity tal delays including reduced body weights in male fetuses and
tests including Salmonella, a forward mutation test in mam- an increased incidence of skeletal anomalies were observed in
malian cells (Chinese hamster ovary cells, CHO cells), in vitro the 200 and 400 ppm (1000 and 2000 mg/m3) groups. How-
tests for sister chromatid exchange and chromosomal aberra- ever, as there were also significant reductions in maternal
tions in CHO cells, and an in vivo test for chromosome aber- body weight gains in dams from these exposure groups, the
rations in rat bone marrow cells. All tests produced negative developmental delays may have been either direct fetal effects
results (Schreiner et al. 1989). or secondary consequences of the maternal toxicity. There
In tests of individual TMB isomers, 1,2,4-TMB and 1,3,5- were no differences among the offspring that were allowed
TMB were reported to be inactive in Salmonella, but 1,2,3- to deliver normally, providing support for the view that the
TMB was reported as active in the absence but not the pres- developmental delays were secondary effects. The lowest con-
ence of S9 activation. All three TMB isomers were inactive in centration tested, 120 ppm (or approximately 600 mg/m3) was
micronucleus tests in mice (Janik-Spiechowicz et al. 1998a). a no-effect level for both maternal and fetal effects.
The same group also evaluated the genotoxic potential of an In a developmental toxicity study, mice were exposed 6
aromatic solvent Farbasol26 and 4-ethyltoluene in a similar hours/day on days 6–15 of gestation at target exposure levels
battery of tests (Janik-Spiechowicz et al. 1998b). Farbasol and of 100, 500, or 1500 ppm (500, 2500, or 7500 mg/m3) (McKee
4-ethyltoluene were reported as inactive in Salmonella and in et al. 1990). The highest exposure level resulted in the deaths
in vivo micronucleus tests in mice, but small but statistically of 14 of the 32 treated dams. There was an increased incidence
significant increases (1.4-fold increase) in sister chromatid of cleft palate, a stress-related effect in mice, in offspring from
exchange frequency were reported. Cumene was inactive in dams that survived exposure to 1500 ppm (7500 mg/m3). The
Salmonella; under in vivo conditions, it produced small but intermediate exposure level, 500 ppm (2500 mg/m3) which
statistically significant increases in micronucleus tests when was maternally toxic, caused a small but statistically significant
given by intraperitoneal administration, but did not increase reduction in fetal weight but did not produce malformations.
the frequency of micronuclei in erythrocytes in peripheral The lowest exposure level, 100 ppm (approximately 500 mg/m3)
blood following exposure by inhalation (NTP 2009). Given was a no-effect level for both maternal and fetal effects.
the limited evidence of effects in bone marrow assays for Saillenfait et al. (2005) assessed the potential for devel-
micronucleus induction and chromosomal aberrations, the opmental toxicity of 1,2,4- and 1,3,5-TMB in rats exposed
data suggest that neither the complex C9 aromatic hydrocar- at levels of 100, 300, 600, and either 900 or 1200 ppm (500,
bon solvents nor their constituents are likely to be mutagenic 1500, 3000, and either 4500 or 6000 mg/m3). In the study of
under in vivo conditions.
27Aromatol is the brand name for a commercial solvent containing 48%

26Farbasolcontained 46% trimethylbenzene isomers and 40% ethyltolu- trimethylbenzene isomers, 38% ethylbenzene isomers, 9% propylbenzene
ene isomers. isomers and 3% o-xylene.
1,3,5-TMB there were no maternal deaths or treatment-related differences in frequency of productive litters, litter size, off-
clinical findings. Maternal body weight gain was significantly spring weight, or gender ratios between treated groups. There
reduced among dams exposed to 300 ppm (1500 mg/m3) in were no significant differences in time to developmental land-
the late gestational stages and throughout the exposure period marks or in functional observations performed at postnatal day
in groups exposed to 600 or 1200 ppm (3000 or 6000 mg/m3). 21. There were no significant differences in locomotor activity
There was a small but statistically significant reduction in fetal tests conducted on postnatal days 23, 36, or 90. There were
weights in the 600 ppm (3000 mg/m3) group and an increased no significant differences in amphetamine sensitivity tests
(but not significantly different) incidence of incomplete stern- conducted on 37-day-old pups or in studies of learning ability
ebral ossifications in the 1200 ppm (6000 mg/m3) group. There in which 42-day-old pups were assessed. Of note is that the
were no effects on fetal survival and no differences in fre- learning tests involved electric shock. The absence of effects in
quencies of malformations. The overall no-effect levels were these studies is in contrast to the reports of effects in adult rats
100 ppm (approximately 500 mg/m3) for maternal toxicity and in similar tests described below. The overall conclusion from
300 ppm (approximately 1500 mg/m3) for fetal effects. this study was that prenatal exposure of rats to a complex C9
In the 1,2,4-TMB study, maternal weight gain was signifi- aromatic solvent at levels of 600–2000 mg/m3 (120–400 ppm)
cantly reduced in the 600 and 900 ppm (3000 and 4500 mg/ had no effects on postnatal neurological development.
m3) groups, and fetal body weights were significantly lower
As summarized in Table 11, repeated exposure to C9

than control values. However, as in the 1,2,3-TMB study, there aromatic solvents and their constituents had no effects on
was no increase in malformations, and there were no differ- development in rats or mice at approximately 500 mg/m3 (100
ences in fetal viability. The overall all no-effect level was 300 ppm), but at levels between 1000 and 3000 mg/m3 (200 and
ppm (approximately 1500 mg/m3) for both maternal and fetal 600 ppm), there was evidence of maternal toxicity and devel-
toxicity. opmental delays. There was no evidence of selective develop-
Darmer et al. (1997) assessed the potential for developmen- mental effects.
tal toxicity of isopropylbenzene (cumene) in rats and rabbits.
Pregnant female rats were exposed by inhalation on gestational 8. Reproductive toxicity
days 6–15 to isopropylbenzene vapor at concentrations of 100, In a one-generation reproductive toxicity study (Lehotzky
500, or 1200 ppm (500, 2500, or 6000 mg/m3). Pregnant female et al. 1985a, 1985b) it was found that if dams were exposed
rabbits were exposed on gestational days 6–18 at exposure lev- continuously from days 7–15 of gestation to 120, 200 or 400
els of 500, 1200, or 2300 ppm (2500, 6000, or 11,500 mg/m3). ppm (600, 1000, or 2000 mg/m3) and then maintained with-
There were some indications of reduced maternal weight gains out exposure until parturition, there were no significant dif-
of both rats and rabbits, but there were no differences in termi- ferences in birth weight, postnatal gain or postnatal survival.
nal body weights, and there were no fetal effects. In summary, Additionally, there were no apparent effects on nervous system
there was no evidence of developmental toxicity at the highest development.
concentrations tested in either species. A three-generation reproductive toxicity test was conducted
Lehotzky et al. (1985a, 1985b) conducted a developmental in rats at target exposure levels of 100, 500, or 1500 ppm (500,
neurotoxicity test with Aromatol as the test substance. As in 2500, or 7500 mg/m3) (McKee et al. 1990). The study was
the study by Ungvary and co-workers (Ungvary et al. 1983, originally intended to have been a two-generation study, but
Ungvary and Tatrai 1985), pregnant CFY rats were exposed 24 was extended to a third generation to more fully explore the
hours/day on gestational days 7–15. Exposure levels were 600, potential effects of exposure of offspring in the early postnatal
1000, or 2000 mg/m3 (120, 250, or 400 ppm). The dams were period. In the first generation, exposures of rats were initiated
allowed to give birth, and the offspring were held until ter- when the rats were approximately 8 weeks of age. The rats
minal sacrifice at postnatal day 90. There were no significant were exposed for 10 weeks and then mated. The males were
Table 11. Summarized results of developmental toxicity studies of complex C9 aromatic solvents and their constituents.
Test Substance Species Result1 Comment References
C9 aromatic solvent Rat No-effect level  120 ppm Rats were exposed 24 h/d. Maternal toxicity Ungvary et al. (1983),
(Aromatol) (approximately 600 mg/m3) and developmental effects at 200 ppm Ungvary and Tatrai
(1000 mg/m3) (1985)
C9 aromatic solvent Mouse No-effect level  100 ppm Maternal toxicity and developmental delays McKee et al. (1990)
(approximately 500 mg/m3) at 500 ppm (2500 mg/m3)
1,3,5- Rat No-effect level  300 ppm Maternal toxicity and developmental delays Saillenfait et al. (2005)
trimethylbenzene (approximately 1500 mg/m3) at 600 ppm (approximately 3000 mg/m3)
1,2,4- Rat No-effect level  300 ppm Maternal toxicity and developmental delays Saillenfait et al. (2005)
trimethylbenzene (approximately 1500 mg/m3) at 600 ppm (approximately 3000 mg/m3)
Isopropyl benzene Rat No-effect level  1200 ppm Differences in maternal weight gain were Darmer et al. (1997)
(∼ 6000 mg/m3), the highest noted during the study, but there were no
concentration tested. differences in terminal body weights.
Isopropyl benzene Rabbit No-effect level  2300 ppm (∼11500 Two of the does from the high-dose group Darmer et al. (1997)
mg/m3), the highest concentration died during exposure. Body weights and
tested. food consumption were reduced and liver
weights were increased at all levels.
1The result given is the no-effect level for fetal effects. No-effect levels for maternal effects were at the same or in some cases lower levels, as discussed
in the Comment section.
removed but exposure of the females continued until gestational alveolar/bronchiolar adenoma and carcinoma in the mice and
day 20 when they were removed from the exposure chambers nasal tumors in the rats.29 Based on similar responses with
and allowed to give birth. There were no effects on any of the other structurally related materials, Cruzan et al. (2009) pro-
parameters related to the production of the first litter prior to posed a cytotoxicity-based mechanism related to the capacity
culling on postnatal day 8. From that point on, the weights of of mice to convert cumene to cytotoxic metabolites in their
offspring in the high exposure group (1500 ppm or approxi- respiratory systems. The metabolic differences, along with dif-
mately 7500 mg/m3) were significantly below control values. ferences in respiratory architecture, suggest that these effects
When exposures of the F1 offspring was re-initiated, the could be species-specific. An alternative theory is that DNA
animals of the high-dose group had body weights that were damage could contribute to lung cancer (Hong et al. 2008),
below control values, and this continued throughout the expo- suggesting that the results of these rodent studies could be
sure period. There were a few statistically significant findings relevant to humans. With respect to the alternative theory,
in the high-exposure group, but these were more likely to have it should be noted that cumene was not mutagenic in stud-
been due to procedural issues than to exposure. More specifi- ies conducted by the NTP (2009), but one of its metabolites
cally, there were 24 dams for which mating was not confirmed, (a-methylstyrene oxide) has been shown to be mutagenic in
9 of which were in the high-exposure group. Because these vitro (Rosman et al. 1986). However, the separate observation
dams were not known to be pregnant, they were not removed that a-methylstyrene, a direct precursor to a-methylstyrene
from the exposure chamber prior to giving birth. There were oxide, did not induce the mouse lung and rat nasal tumors that
profound effects on the survival and birth weights of the off- are associated with cumene (NTP 2007) seems more compat-
spring that were born in the chambers, affecting the group ible with the hypothesis that cumene acts via a non-genotoxic
mean values for survival and birth weights of offspring in the mode of action. Based on a review of the experimental data
mid- and high-exposure groups. There were essentially no dif- and without direct evidence that cumene causes cancer in
ferences among litters from dams that were removed from the humans, IARC concluded that cumene is possibly carcino-
exposure chambers prior to giving birth. genic to humans (Group 2B) (IARC 2012).
This study was extended to a third generation to investigate
whether the exposures of very young animals had any impact 10. Neurotoxicity
on their reproductive capacity. Accordingly, exposures of F2 10.1 Acute neurological effects
offspring were initiated when the pups were 22 days old.28 In
this part of the experiment, the majority of the offspring from A number of authors have reported observations consistent
the high-exposure group died with the first week of exposure, with acute CNS effects in studies of C9 aromatic solvents and
but, among those that survived, the only apparent effect was their constituents, and acute CNS effects in humans were also
on body weight. suggested by the observations of Battig et al. (1956, 1958).
In summary, this study provided evidence that C9 aromat- These reports led to investigations by Korsak and Rydzynski
ics are toxic to 22-day-old rats exposed at levels of 1500 ppm (1996) and McKee et al. (2010) to characterize the potential
(or approximately 7500 mg/m3), but there were no effects on for acute CNS effects in animals. In the first of these, Korsak
reproductive parameters. It should be noted that, based on and Rydzynski (1996) reported tests of each of the TMB iso-
their body weights, the systemic doses of 22-day-old rats at mers using rotarod performance and pain sensitivity. In the
these exposure levels were approximately 2–3 times those of rotarod study in which the rats were exposed for 4 h, the fail-
adult rats. Thus, it seems reasonable to assume that the appar- ure rates increased with exposure concentrations over a range
ently greater toxicity experienced by young rats was due to of approximately 1200–10000 mg/m3 (240–2000 ppm). Pain
their higher body burdens. Exposures to 500 ppm (approxi- sensitivity was assessed as latency to respond (paw lick) using
mately 2500 mg/m3) had no apparent effects in either 22-day- a hot plate. From the information provided, it appears that dif-
old or adult rats, and exposures to 1500 ppm (approximately ferences were observed at levels  2000 mg/m3 (2000 ppm).
7500 mg/m3) had no effect on reproductive parameters. McKee et al. (2010) assessed the potential for acute CNS
effects using a complex C9 aromatic solvent (200, 1000, or
9. Carcinogenicity 5000 mg/m3 [40, 200, or 1000 ppm], 8 hours/day for 3 con-
secutive days) and one constituent (1,2,4-TMB) (125, 1250, or
There are no reports of carcinogenicity tests of C9 aromatic
5000 mg/m3 [25, 250, or 1000 ppm], 8 hours/day for 3 consec-
solvents or the principal constituents. However, the carcino-
utive days) using a battery of tests for motor activity and func-
genicity of isopropylbenzene (cumene) was assessed by the
tional observations as well as a test of visual discrimination
U.S. National Toxicology Program (NTP 2009). Rats and
performance. In the study of 1,2,4-TMB, the principal finding
mice were exposed to 125 (female mice only), 250, 500, or
in rats exposed to 5000 mg/m3 (1000 ppm) was a significant
1000 ppm (1250, 2500, or 5000 mg/m3) cumene, 6 hours/day,
reduction in short ( 2 s) response latencies and a signifi-
5 days/week, for 105 weeks. At termination, the surviving
cant increase in long ( 6 s) response latencies in the visual
animals were sacrificed for gross and microscopic examina-
tion. The principal findings were dose-related increases in
29Also observed were treatment-related increases in the incidence of
hemangiosarcoma in the spleen of male mice, hepatocellular adenoma
28The first generation offspring were held until all were at least 21 days of the liver of female mice, kidney tumors in male rats, and testicular
old before exposures were reinitiated. Because of differences in time to tumors of male rats. The kidney tumors were considered to have been the
mating, some of the offspring were 28 or more days old before they were consequence of an a2u-globulin-mediated process. The testicular tumors
exposed to test material. are a commonly observed spontaneous tumor in male F-344 rats.
discrimination test. There were no effects in animals exposed auditory brain stem measurements and did not produce patho-
at lower levels. In the study of the complex C9 aromatic solvent logical changes in the central or peripheral nervous system.
the responses were similar but statistically significant at both Changes in motor activity were reported in one study but not
1000 and 5000 mg/m3 (200 and 1000 ppm) with 200 mg/m3 replicated in a second.
(40 ppm) as the no-effect level. When the rats were held for 24 In summary, the neurotoxicity tests in animals revealed that
h without exposure and then tested, no effects were observed. C9 aromatic solvents and some of the C9 isomers tested indi-
These data are consistent with the observations by Douglas vidually could produce acute CNS effects at levels  200 mg/m3
et al. (1993) (discussed in more detail in the following section) (40 ppm), but there was no evidence that repeated treatment
that following repeated exposures, CNS effects, including hot caused pathological changes or neurobehavioral effects in
plate responses, were not observed when the rats were tested conventional assays. There were several studies that suggested
24 h after the last exposure. These results provide evidence that there may have been differences in response to administra-
that these CNS effects are acute and reversible. tion of pain after repeated exposure, but these studies were
Cushman et al. (1995) tested cumene for acute and sub- complicated and the interpretation not straight forward.
chronic neurotoxicity, and reported that acute exposure of
rats to 500 or 1200 ppm (2500 or 6000 mg/m3) resulted in an 11. Observations in humans
increase in motor activity. In order to help define or test the adequacy of occupational
exposure recommendations, there have been several studies
10.2 Subchronic neurological studies of volunteers. Based on their initial observations, Battig et al.
(1958) recommended 35 ppm (175 mg/m3) as a threshold limit
A complex C9 aromatic solvent was tested for neurological
value for an 8-hour day. Nau et al. (1966) recommended 50
effects following repeated exposure (Douglas et al. 1993).
ppm (250 mg/m3) based on a volunteer study, and Gerarde
Male rats were exposed 6 hours/day and 5 days/week, for 13
(1960) made a similar recommendation. Currently recom-
weeks, at target concentrations of 100, 500, or 1500 ppm (500,
mended occupational exposure levels in the US and Europe are
2500, or 7500 mg/m3). Neurobehavioral investigations includ-
in the range of 20–25 ppm (100–125 mg/m3). Caldwell et al.
ing functional observation tests and assessments of motor
(2000) reported that, based on a review of personal breath-
activity were conducted after 5, 9, and 13 weeks; all tests were
ing zone results, the weighted arithmetic mean exposure to
conducted 24 h after the previous exposure period. After 13
trimethylbenzene was 4.1 mg/m3. Jarnberg et al. (1996, 1997)
weeks of exposure, the animals were sacrificed and nervous
reported that there were no signs of irritation or CNS effects

system tissues were examined. The body weights of the rats
among volunteers exposed for 2 h to different TMB isomers at
in the high-exposure group were 12% below and significantly
25 ppm (125 mg/m3). Similarly, Jones et al. (2006) reported
different from control values, but no neurobehavioral effects or
that exposure to 25 ppm (125 mg/m3) to a complex solvent
pathological changes were observed.
composed of TMB isomers for 4 h was well tolerated by vol-
There were three studies of similar design that were con-
unteers. Kostrzewski et al. (1997) conducted a study in which
ducted to characterize the potential for repeated exposure to
volunteers were exposed for periods of 4–8 h to each of the
TMB isomers to cause chronic neurological effects (Grale-
TMB isomers at levels ranging from 5–150 mg/m3 (approxi-
wicz et al. 1997, Gralewicz and Wiaderna 2001, Wiaderna
mately 1–30 ppm) and reported that the volunteers presented
et al. 2002). Rats were exposed to TMB isomers for periods
no abnormalities in routine clinical (internal, laryngological,
ranging from 4 weeks to 3 months at exposure levels rang-
neurological) or hematological investigations.
ing from 25–250 ppm (125–1250 mg/m3). The rats were then
held without exposure for periods ranging from approximately
II. Toxicological properties of C10–C12 Aromatic
2–8 weeks and then evaluated for persistent neurobehavioral solvents (Category 2 solvents)
effects. The tests included radial maze tests to assess short-
term spatial memory, open field tests to assess spontaneous 1. Introduction
motor activity, step-down passive avoidance tests to assess “C10–C12 aromatic solvents” refers to a group of 4 aromatic
long term memory, hot plate tests to assess sensitivity to pain hydrocarbon solvents with complex and variable compositions,
and pain-related stress, and conditioned active avoidance composed of aromatic constituents and boiling in the range
reaction tests to assess learning and memory. There were no of approximately 182–310°C.30 The aromatic constituents of
consistent, treatment-related differences in performance in the these solvents are primarily alkylated one- and two-ring aro-
radial maze tests or open field tests. matics (i.e., alkylated benzenes and alkylated naphthalenes).
The passive avoidance, active avoidance, and hot plate tests The distinguishing characteristics of this group of solvents are
involved the administration of pain. In each of these tests, the carbon number range and the naphthalene content. Most
there were initial trials in which no significant differences were of these substances are listed on the TSCA registry under the
found. The animals were then given foot shocks and re-tested
over a period of several days. Significant differences were
reported in some cases, but not in all tests or with all TMB iso-
30These solvents were identified as “C10–C13 aromatic solvents” in the
mers. Further, there was no clear evidence of dose response.
The toxicological significance of these data is unclear. OECD HPV process and “C10–C12 aromatic solvents” for purposes of
REACH registration. The difference in the category name is because of
Cushman et al. (1995) reported that repeated exposure of differing views about the ways in which the compositions of this group of
rats to cumene vapor at levels up to 1200 ppm (approximately substances should be described by different groups of reviewers, but does
6000 mg/m3) had no effects on functional observations or not reflect compositional differences.
Table 12. Physical and chemical properties of C10–C12 aromatic solvents.

Property C10 Solvents C10–C12 Solvents
Physical state at 20°C and 1013 hPa Colorless to yellow liquid with pungent odor Colorless to yellow liquid with pungent odor
Odor Threshold 0.07 mg/m3 (Ruth 1986) 0.06–0.30 mg/m3 (Ruth 1986)
Melting/Freezing Point  10°C (ASTM D5950)  5°C (ASTM D5950)
Boiling Point 160–230°C (ASTM D86) 200–310°C (ASTM D86)
Relative Density 0.80–1.00 g/cm3 at 15°C (ISO 12185) 0.95–1.05 g/cm3 at 15°C (ISO 12185)
Vapor Pressure 0.006 kPa at 20°C 0.002–0.003 kPa at 20°C
Surface Tension 29–32 mN/m at 15°C 30–37 mN/m at 15°C
Flash Point  62°C (ASTM D93)  94°C (ASTM D93)
Flammability 0.6–7.0% (v/v) 0.6–7.0% (v/v)
Self-Ignition Temperature  400°C at 1atm  400°C at 1atm
Viscosity 0.8–2.0 mm2/s at 20°C (ASTM D7042) 2.7–5.8 mm2/s at 20°C (ASTM D7042)
Maximally Attainable Vapor Concentration Approximately 5000 mg/m3 (1000 ppm) Approximately 5000 mg/m3 (1000 ppm)
Conversion factor (ppm/mg/m3) 1 ppm  ∼ 5.2 mg/m3 1 ppm  ∼ 5.5 mg/m3
Octanol/Water partition coefficient 3.2–4.5 3.2–4.5
Water solubility  0.12 mg/l (at 25°C)  0.12 mg/l (at 25°C)
CAS number 64742-94-5 (although CAS numbers 70693- of rats and mice than humans to the effects of naphthalene
06-0 and 1321-94-4 have also been used) and under the EC and methylnaphthalenes and attributed this to species differ-
numbers 922-153-0 and 919-284-0, depending on the carbon ences in metabolism. PBPK models for naphthalene have been
number range. Benzene levels in these substances are reported published by Sweeney et al. (1996) and Willems et al. (2001).
as  1 ppm (Table 3), and no aromatic molecules with 3 or
more rings are present (OECD 2012b). 4. Acute toxicity
Based primarily on information summarized in the HPV docu-
2. Physical and chemical properties mentation (OECD 2012b), acute oral LD50 values were greater
The physical and chemical properties are summarized in than 2000 mg/kg, acute dermal LD50 values were greater than
Table 12. Of note is that some of the solvents in this group 2000 mg/kg, and acute inhalation LC50 values exceeded the
contain naphthalene at toxicologically relevant levels. Those maximally attainable vapor concentrations (Table 13). Based
solvents are differentiated by the naming convention (Table 2). on other information in the HPV submission (OECD 2012b),
The samples tested as representative of this group of solvents these solvents did not produce skin or eye irritation at toxico-
contained approximately 8–10% naphthalene, making this an logically important levels when tested for these properties in
example of a group for which the characterization of toxico- animals, and did not produce skin irritation or allergic contract
logical hazards of all of the category members was based on a dermatitis in human studies. However, human observations
reasonable worst case approach. indicate that solvent vapors can cause ocular and/or upper
respiratory tract irritation if inhaled at high levels (Nau et al.
3. Toxicokinetic properties 1966, Robins 1951). Sensory irritation studies in mice indicated
The constituents of these solvents are relatively well absorbed respiratory rate decreases with RD50 values of 129 mg/m3 (23
if inhaled or ingested but poorly absorbed dermally. Teshima ppm) and 67 mg/m3 (23 ppm) for the two methylnaphthalene
et al. (1983) reported that at least 80% of an oral dose of naph- isomers (Korsak et al. 1998); however, volunteer studies pro-
thalene was absorbed by guinea pigs within 24 h of adminis- vided evidence that these substances may be less irritating to
tration. In contrast, in dermal administration studies, Riviere humans than they are to rodents (Medeiros et al. 2000).
et al. (1999) reported that only about 1% of the naphthalene
5. Repeated dose toxicity studies
content of jet fuel was absorbed in a porcine skin flap model,
and flux rates for naphthalene and methylnaphthalene isomers 5.1 Repeated inhalation toxicity studies
were in the range of 1–2 mg/cm2/hour (Baynes et al. 2000, Kim
The first systematic investigations of the toxicological proper-
et al. 2006, Singh and Singh 2003), with somewhat higher
ties of complex aromatic solvents were by Nau et al. (1966)
values being reported by Muhammad et al. (2005). These sub-
and Carpenter et al. (1977a), but studies by Carpenter et al.
stances are metabolized and excreted relatively rapidly, with
the majority of the elimination occurring in the first 24 h. The Table 13. Summarized results of acute toxicity tests of C10–C12 aromatic
principal difference among the constituents is that naphthalene solvents.
is primarily metabolized to epoxide intermediates whereas the Test Resulta
alkylated benzenes and alkylated naphthalenes are primarily
Acute Oral Toxicity 2000 mg/kg
metabolized by glucuronidation of the alkyl side chains, as 7000 mg/kg
shown in Figure 2. Based on studies of methylnaphthalene iso- Acute Dermal Toxicity  2000 mg/kg
mers (Grimes and Young 1956, Melancon et al. 1982, Sapota Acute Inhalation Toxicity  4688 mg/m3
et al. 1996, Teshima et al. 1983), alkylated two-ring aromatics ( 890 ppm)
 1073 mg/m3
are primarily metabolized by side chain oxidation and sub- ( 204 ppm)
stantially excreted within 24 h of ingestion (Figure 3). Lin aThe acute toxicity data are mostly unpublished results, summarized in
et al. (2009) took note of the apparently greater susceptibility OECD (2012b).
(1975c) on “70 solvent” are also relevant. Nau et al. (1966) Sprague-Dawley rats (unpublished information summarized
exposed rats for 18 hours/day and 7 days/week to vapors of in OECD 2012b). The test material33 was suspended in
“C11–C12 aromatic fraction”31 at concentrations ranging corn oil and administered by gavage at doses of 300, 600, or
from 50–500 ppm (278–2778 mg/m3). In initial studies, 500 1200 mg/kg (reduced to 1000 mg/kg because of observations
ppm (2778 mg/m3) was acutely toxic, and 50% of the rats died of hypothermia and hypoactivity at the higher dose), 7 days/
within 18 h of the initiation of exposure. When the exposure week for 13 weeks. The corresponding control group received
time was reduced to 8 hours/day and 7 days/week, the rats corn oil only. The majority of the animals were sacrificed at
survived but lost weight, and there were signs of acute CNS the end of the exposure period, but some high dose and con-
depression and upper respiratory tract irritation. When rats trol animals were held without treatment for an additional 4
were exposed to 200 ppm (1111 mg/m3), 8 hours/day and 5 weeks to assess the potential for recovery. There was some
days/week for 90 days, there were effects on body weight gain evidence of mortality in the high-dose group (1 male and 4
and reductions in white blood cell counts. When rats were females died prior to scheduled sacrifice), and terminal body
exposed to 50 ppm (278 mg/m3), 8 hours/day for 90 days, small weights of the high–dose group males were 19% below and
reductions in body weight gain and bone marrow effects were significantly different from control values. In contrast, termi-
reported, but details were not provided. In monkeys exposed nal body weights of high–dose group females were 3% (not
to 200 ppm (1111 mg/m3), 7 hours/day and 5 days/week for
significantly different) below control values. The hemato-

90 days, there was evidence of ocular irritation and bone mar- logical investigation revealed small but statistically significant
row effects, but no details were provided. Similar effects were reductions in hematological values for both male and female
reported in monkeys exposed to 50 ppm (278 mg/m3). rats, with the differences being statistically significant in the
Carpenter et al. (1977a) reported a study of the toxico- 600 and 1000 mg/kg/day groups. There were some small dif-
logical effects of an aromatic solvent containing primarily ferences in clinical values, but all were within normal physi-
C9–C11 constituents.32 There were no effects in either rats or ological limits. There were significant increases in liver and
dogs following exposure for 90 days at levels up to 380 mg/m3 kidney weights in both male and female rats. There were no
(approximately 68 ppm), the highest concentration tested. histological changes in the kidneys. The pathologist described
In a repeated inhalation toxicology study of “70 solvent” the liver changes as hepatocellular hypertrophy, primarily
(approximately 67% aromatic constituents and 33% aliphatic minimal in males and minimal to slight in females, and the
constituents with carbon numbers primarily in the range of levels of liver enzyme markers were not elevated. Increased
C9–C10), the only effects at the highest concentration tested hemosiderosis was reported in male and female rats in the
(1100 mg/m3, approximately 200 ppm) were reductions in 600 and 1000 mg/kg/day groups. Other microscopic changes
body weights and kidney changes in male rats (Carpenter were consistent with adaptive responses. All of these differ-
et al. 1975g). The kidney changes were associated with an a2- ences were reversed in the recovery group animals, i.e., there
globulin-mediated process not relevant to humans. were no significant differences in hematological and/or clini-
Swiercz et al. (2011) reported a 4-week inhalation toxicity cal chemistry values, no differences in organ weights, and no
study in which rats were exposed to vapors of 2-methylnaph- histological changes in animals exposed to the highest dose for
thalene at levels of 2, 10, or 50 mg/m3 (0.4, 2, 10 ppm), 6 hours/ 90 days and then held for 4 weeks without treatment. Although
day and 5 days/week for 4 weeks. At the end of the exposure there were some statistically significant changes in the 600 mg/
period, the animals were sacrificed and samples were taken for kg/day group, the clinical and hematological differences were
clinical and pathological evaluations. All of the rats survived, within normal limits, and the differences in organ weights and
and there were no differences in terminal body weights. There histological findings were more consistent with adaptive than
were small differences in some hematological and clinical val- toxicologically important effects. The majority of the effects
ues, but all were within normal limits. There were some small were reversed in the recovery group animals; however, there
reductions in organ weights, but no pathological changes were was not complete reversibility of the splenic changes. The no
reported other than evidence of irritation in the respiratory observed adverse effect level was reported as 300 mg/kg/day.
tract. These data were consistent with other evidence that the Similar results were reported by Sterner et al. (2005) in a
principal effect of methylnaphthalene isomers in rodents is summary of a repeated oral toxicity study of an aromatic-rich
respiratory tract irritation. material in male B6C3F1 mice and female Sprague-Dawley
rats. The test material was an aromatic fraction derived from
5.2 Repeated oral toxicity studies jet fuel, which, like the C10–C12 aromatic solvents, consisted
The potential for systemic effects following repeated oral principally of alkylated benzenes and naphthalenes.34 The test
administration was assessed in an oral toxicity study in material was suspended in corn oil and administered by gav-
age in doses of 20, 100, or 500 mg/kg/day. Control animals
received corn oil only. Two animals (one mouse and one rat)
31The test substance was described by Nau et al. (1966) as a C11–C12
33The test material was described as a C10–C13 aromatic solvent com-
aromatic solvent with a boiling range of 392–480°F (200–250°C) and
containing 44% alkylbenzenes, 27% indans, 23% naphthalenes, and 6% posed primarily of naphthalene (12%), methylnaphthalene isomers (46%),
paraffins. The distribution of alkylbenzenes was: 7% C9, 10% C10, 24% and dimethylnaphthalene isomers (19%).
34The constituents in the aromatic fraction of jet fuel that were found
C11, 5% C12, and 1% C13.
32The test substance was described by Carpenter et al. (1977g) as con- to be present at levels  1% included ethylbenzene, xylenes, styrene,
taining approximately 99% aromatic constituents of which approximately isopropylbenzene, n-propylbenzene, methylbenzenes, butylbenzenes,
90% were C9–C11 alkylbenzenes. The boiling range was 364–403°F diethylbenzenes, ethylnaphthalenes, dimethylnaphthalenes, biphenyl,
(143–206°C). acenaphthalene, and acenaphthene.
from the high-dose group died during the study, but all other major organs. There were no significant differences in num-
animals survived to scheduled termination. There were no bers of implants, corpora lutea, live fetuses, dead fetuses,
significant differences in body weights. The authors reported resorptions, or sex ratio. Fetal body weights were similar
small reductions in hematological parameters in female rats across groups. There were no differences in frequencies of
from the mid- and high-dose groups, but data were not pro- malformations or variations.
vided. Other reductions in hematological parameters were In summary, the only treatment-related observation was
reported to be small and within normal physiological limits, a reduction in body weight gain in females from the high-
and were not considered to be toxicologically important. Liver dose group, during the dosing period (gestational day 6–15);
and kidney weights were significantly elevated in the rats of however, as the body weights of dams were not different at
the high-dose group; liver weights were elevated but not sig- scheduled termination, the effect on body weights was not
nificantly different in the mice of the high-dose group. How- considered toxicologically important. There were no fetal
ever, there were no histological differences in these or other effects. The overall no adverse effect level was 450 mg/kg/day,
organs, suggesting that the differences in organ weights were the highest dose tested.
due to adaptive rather than pathological processes.
In summary, repeated dose studies of these solvents pro- 8. Reproductive toxicity
vided some evidence for respiratory tract irritation and acute Formal reproductive toxicity tests of complex C10–C12 aro-
CNS effects, but there is no clear evidence for target organ matic solvents have not been conducted, but there are data on
effects aside from adaptive responses at exposure levels of up some constituents. Further, there were no effects on reproduc-
to 380 mg/m3 (68 ppm) in inhalation studies and 300 mg/kg/ tive organs in the repeated dose studies. A two-generation
day in oral studies. reproductive toxicity test was conducted using n-butylbenzene
(a 10-carbon aromatic substance) as the test material (Izumi
6. Genetic toxicity
et al. 2005), administered by gavage to rats at dose levels of
Unpublished information indicates that three independent 0, 30, 100, or 300 mg/kg/day. There were no effects on fertil-
Salmonella studies were conducted following OECD 471 ity. Systemic effects were limited to liver and kidney weight
testing guidelines. Negative results were obtained in all stud- changes. The liver effects were consistent with an adaptive
ies both in the presence and absence of metabolic activation process relating to increased metabolic demands. The principal
(OECD 2102b). histological changes in the kidneys of male rats were described
A bone marrow micronucleus test was conducted in male as increased hyaline droplet formation, providing presumptive
and female CD-1 mice following OECD 474 testing guide- evidence that this was an a2u-globulin-mediated process.
lines. Doses of 250, 500, or 1000 mg/kg were administered by
oral gavage. There was no evidence of micronucleus formation 9. Carcinogenicity
at the highest dose tested (OECD 2012b). Murata et al. conducted carcinogenicity tests of the two meth-
Naphthalene, which has been extensively studied, is con- ylnaphthalene isomers. In the first of these studies (Murata
sidered to be inactive in both in vitro and in vivo mutagenicity et al. 1993), 1-methylnaphthalene was given by dietary admin-
tests (Schreiner 2003). istration to B6C3F1 mice at levels of 0.075% or 0.15% for 81
In summary, the C10–C12 aromatic solvents are not geno- weeks. At terminal sacrifice, the incidence of bronchiolar/
toxic, under either in vitro or in vivo conditions. alveolar adenomas in the lungs of male mice given 0.075%
(26.0% tumor incidence) or 0.15% (24.0%) was elevated by
7. Developmental toxicity
comparison to control mice (4.1%). However, there was no
A developmental toxicity test of a C10–C13 aromatic solvent corresponding increase in frequency of adenocarcinoma in
was conducted in rats in accordance with OECD 414 guide- male mice and no increase in tumors of either type in female
lines (unpublished information summarized in OECD 2012b). mice. In a similar study of 2-methylnaphthalene, the incidence
The test material was suspended in corn oil and administered of bronchiolar/alveolar adenomas was 20.4% in male mice
by gavage at doses of 75, 150, or 450 mg/kg/day in constant given 0.075% in the diet and 12.2% in mice given 0.15% in the
volumes of 5 ml/kg to pregnant Sprague-Dawley rats. A diet (Murata et al. 1997). As in the previous study, there was
control group received corn oil only. The rats were treated no increase in adenocarcinomas in male mice and no increase
from gestational days 6 to 15 and then held without treatment in either type of tumor in female mice. The authors considered
until gestational day 21 when they were sacrificed for uterine that these data provided suggestive but not conclusive evidence
examination. In-life measurements included maternal body that methylnaphthalenes were carcinogenic to mouse lung.
weights and food consumption on days 0, 6, 9, 18 and 21.
On gestational day 21, the rats were weighed and then sacri- 10. Neurotoxicity
ficed. The uteri were removed and weighed, and the uterine In a study to assess the potential for hydrocarbon solvents and
contents were then examined. All fetuses were removed and their constituents to produce acute CNS effects, one of the test
counted. The fetuses were examined for gender and then for substances was a complex C10–C11 aromatic solvent (McKee
developmental effects following standard techniques. et al. 2010). Rats were exposed at levels of 200, 600, or 2000 mg/
Maternal body weight gain was reduced in the dams from m3 (36, 108, or 360 ppm), 6 hours/day for 3 consecutive days,
the high-dose group during the dosing period, but terminal and then examined for deficits in functional observation tests and
body weights of treated animals were similar to control values. in assessments of effects on motor activity and visual discrimina-
There were no differences in uterine weights or in weights of tion performance. The principal effect was a significant increase
in latency response, associated with a significant reduction in aromatic solvents and 25 ppm (132 mg/m3) for C11–C12
short ( 2 s) responses and an increase in long ( 6 s) response aromatic solvents. In similar studies with human volunteers,
times. The largest effects were seen after the first exposure. There Carpenter et al. (1977a) recommended 17 ppm (approximately
were no significant differences in tests conducted 24 h after the 89 mg/m3) for C9–C11 aromatic solvents. The ACGIH® cur-
last exposure, confirming that these effects were acute, not per- rently recommends 10 ppm (approximately 50 mg/m3) as an
sistent. The no-effect level was 600 mg/m3 (108 ppm). 8-hour time-weighted average exposure for naphthalene and
Korsak et al. (1998) investigated the acute of 1- and 2-meth- 0.5 ppm (approximately 2.6 mg/m3) for methylnaphthalene
ylnaphthalene isomers in rats and mice. Rats were exposed for isomers. As discussed in Occupational exposure limits, the
4 h to vapors of methylnaphthalene isomers at levels between hydrocarbon solvent industry recommends that occupational
approximately 150 and 527 mg/m3 (27 and 95 ppm). There were exposure limits for complex hydrocarbon solvents be based
no differences in rotarod performance with either isomer, but on a guidance value of 20 ppm (100 mg/m3) for otherwise
time to paw lick was significantly increased at exposure lev- unspecified C10–C13 aromatics and 10 ppm (50 mg/m3) for
els  229 mg/m3 (41 ppm). Respiratory irritation studies in mice naphthalene. It is also recommended that the occupational
indicated respiratory rate decreases with RD50 values of 129 mg/ limits for methylnaphthalene isomers be respected. Information
m3 (23 ppm) and 67 mg/m3 (12 ppm) for the two isomers. on occupational exposures to these substances has not been
reported, but occupational exposures to naphthalene have been

11. Human observations reported to be  approximately 1 mg/m3 (ATSDR 2005).
11.1 Ocular and respiratory irritation Appendix B. Aliphatic/aromatic hydrocarbon

The earliest reports of effects of exposure of volunteers to solvents [C9–C20 Aliphatic hydrocarbon solvents
C10–C12 aromatic solvents by inhalation were those from
(2–25% Aromatics)]
Nau (1966) who stated that the “minimum concentration I. Toxicological Properties of C9-C14 Aliphatic
which produced lacrimation and/or irritation of the eyes, skin Hydrocarbon Solvents (2-25% Aromatics) (Category 3
and mucous surfaces of humans was found to be 25 ppm for solvents)
C9–C10 and 19 ppm for C11–C12.” Carpenter et al. (1977)
reported that “[O]nly one of six volunteers reported discom- 1. Background
fort while inhaling a concentration of 0.1 mg/l (17 ppm) for The Category 3 hydrocarbon solvents are aliphatic solvents
15 min. This subject experienced mild irritation only from the with carbon numbers ranging from C9–C14 and aromatic
6th through the 15th minute of inhalation.” Robbins (1951) contents ranging from 2–25%. As described previously, the
reported that there were signs of ocular irritation in workers prototypic solvent of this type is “Stoddard solvent”, a des-
exposed to naphthalene at levels above 15 ppm (approximately ulfurized hydrocarbon solvent fractionated from crude oil by
75 mg/m3). As noted previously, respiratory irritation studies distillation, boiling in the range of 149–200°C, and containing
in mice indicated respiratory rate decreases with RD50 values approximately 15–20% aromatic constituents (primarily iso-
of 129 mg/m3 (23 ppm) and 67 mg/m3 (12 ppm) for the two mers of trimethylbenzene and ethyltoluene), depending on the
isomers (Korsak et al. 1998). However, tests of nasal and ocular characteristics of the crude oil from which it was derived. This
sensory detection levels in volunteers indicated that mice are type of solvent is now commonly referred to in the US as a
much more sensitive than humans to the irritant properties of Type 2 mineral spirit and in Europe as “white spirit”, although
naphthalene and methylnaphthalene (Medeiros et al. 2000). the dry cleaning industry continues to use the term Stoddard
solvent. Stoddard solvent was entered into the TSCA inventory
11.2 Studies to assess the potential for dermal irritation under CAS number 8052-41-335; however, other CAS numbers
and allergic contact dermatitis can be used for solvents of this type, as shown in Table 1. Five
In studies to assess the potential for dermal effects, aliquots of solvents of this type were registered under REACH (Table 2).
two commercial C10–C12 aromatic hydrocarbon solvents were
administered to human volunteers. In some cases, the treated 2. Physical and chemical properties
sites were exposed to UVB radiation to assess the potential for The physical and chemical properties of C9–C14 aliphatic
phototoxic effects. There was no indication of more than mild (2–25% aromatic) hydrocarbon solvents are summarized in
irritation at the application sites, indicating that these solvents Table 14. Note that in order to indicate the boundary condi-
are not primary irritants and that they have no phototoxic prop- tions, physical and chemical properties are only described for
erties. Additionally, there were no reactions when additional the category members with constituents composed of the low-
material was administered at remote sites, providing evidence est and highest carbon numbers. Benzene levels for solvents
that these substances do not cause allergic contact dermatitis of this type are generally less than 100 ppm (Table 3). These
(unpublished information summarized in OECD 2012b). solvents are manufactured from petroleum streams that are
11.3 Occupational exposures and recommended 35For purposes of this document, Stoddard solvent, white spirit, and
occupational exposure limits mineral spirit are all common identifiers for substances that fall within
Gerarde (1960) recommended an occupational exposure level the range of Category 3 solvents and can be used interchangeably. To
avoid confusion, in discussions of specific data sets, the term used by the
of 25 ppm (approximately 130 mg/m3) for naphthalene. Based original author will also be used in this document. In those situations in
on results from volunteer studies, Nau et al. (1966) recom- which authors use these terms incorrectly, for example using white spirit
mended 50 ppm (approximately 260 mg/m3) for C9–C10 as a general descriptor for hydrocarbon solvent, it will be noted.
Table 14. Physical and Chemical Properties of C9–C14 aliphatic However, based on studies of some marker constituents
hydrocarbon solvents (2–25% aromatics). (Baynes et al. 2000, Kim et al. 2006, Muhammad et al. 2005,
Hydrocarbons, Hydrocarbons Singh and Singh 2003), percutaneous absorption of these
C8–C12; (2–25% C11–C14; (2–25% constituents is low, and it is not expected that dermal contact
Property aromatics) aromatics)
would result in a substantial contribution to systemic dose
Physical state at 20°C Clear, colorless liquid Clear, colorless under most circumstances.
and 1013 hPa with pungent odor liquid with pungent
odor
Once absorbed, the constituents of C9–C14 aliphatic (2–25%
Odor Threshold 5–158 mg/m3 (Ruth 5–158 mg/m3 (Ruth aromatic) hydrocarbon solvents preferentially distribute to the
1986) 1986) adipose tissues, liver, and the CNS. As shown by Hissink et al.
Melting/Freezing Point   20°C (ASTM   20°C (ASTM (2007), at exposure levels in the range of 600 mg/m3 (approxi-
D5950) D5950)
Boiling Point 110–180°C (ASTM 178–270°C (ASTM mately 110 ppm), steady-state concentrations in the brain are
D86) D86) achieved very quickly, but the levels also decline quickly when
Relative Density 0.70–0.80 g/cm3 at 0.76–0.87 g/cm3 at exposures stop (see Figures 2 and 3 in Hissink et al. 2007).
15°C (ISO 12185) 15°C (ISO 12185) The initial half-lives for C9–C14 aliphatic hydrocarbon solvent
Vapor Pressure 0.5 kPa at 20°C 0.02 kPa at 20°C
Surface Tension 22–25 mN/m at 15°C 25–28 mN/m at 15°C constituents in the CNS are approximately 2 h. In humans,
Flash Point  23°C (ASTM D93)  65°C (ASTM levels of white spirit constituents were below detection levels
D93) within 8 h of exposure. Data from studies in rats, summarized
Flammability 0.6–0.7% (v/v) 0.6–0.7% (v/v) in the publication by Pederson and Cohr (1984a), are similar to
Self-Ignition  200°C at 1atm  200°C at 1atm
Temperature those in Hissink et al. (2007), but Pederson and Cohr (1984a)
Viscosity 0.7–1.5 mm2/s at 20°C 1.3–3.5 mm2/s also indicate that white spirit components are more slowly
(ASTM D7042) at 20°C (ASTM eliminated from depot fat than from other tissues.
D7042)
Maximally Attainable Approximately 10000 Approximately 1500 As discussed in more detail in Metabolism and excretion of
Vapor Concentration mg/m3 (2000 ppm)a mg/m3 (300 ppm)b aliphatic hydrocarbon solvent constituents, the aliphatic con-
Conversion Factors 1 ppm  ∼ 5.25 mg/m3 1 ppm  ∼ 5.5 mg/m3 stituents of these solvents, primarily C9–C11 normal, iso- and
aBased on decane. cycloparaffins, are converted to the corresponding alcohols
bBased on dodecane. and primarily excreted in the urine. The aromatic constituents,
primarily trimethylbenzene isomers, are primarily excreted
in the urine as the dimethylbenzoic and/or dimethylhippu-

derived from crude oil by distillation and commonly known as ric derivatives of the parent molecules (Fukaya et al. 1994,
“straight-run” kerosenes which are complex combinations of Ichiba et al. 1992, Jarnberg et al. 1996, 1997a, b, Jarnberg and
aliphatic and aromatic hydrocarbons in the range of approxi- Johanson 1998, Jones et al. 2006, Kostrzewski et al. 1997,
mately C9–C16 and with boiling ranges of approximately Kostrzeweski and Wiaderna-Brycht 1995).
150–290°C. These substances contain approximately 20% Inhaled material is eliminated primarily by exhalation of
aromatics, primarily alkylated one- and two-ring molecules. volatile parent compounds (Janasik et al. 2008) and urinary
Commercial and military jet fuels (jet A, JP-8) have proper- excretion of metabolites (Fukaya et al. 1994, Ichiba et al.
ties similar to those of kerosene but with additional specifica- 1992, Jarnberg et al. 1997, Jarnberg et al. 1997b, Jarnberg
tions to meet their specific technical properties. Accordingly, et al. 1998, Kostrzeweski and Wiaderna-Brycht 1995). The
toxicology data for “straight-run”36 kerosenes and jet fuels are initial blood clearance of trimethylbenzenes in man is 0.6–1 l/
relevant to aromatic-containing C9–C20 aliphatic hydrocarbon hr/kg (Jarnberg et al. 1996); terminal half-lives are estimated
solvents as the carbon number and distillation ranges overlap to be 78–120 h because of partitioning to adipose tissue. The
both categories of hydrocarbon solvents and the aromatic con- initial half-life for urinary elimination of trimethylbenzenes
tents are similar. For convenience, the relevant kerosene and (the aromatic constituents of these solvents) is 0.45–0.88 h,
jet fuel data are summarized in this section in which data rel- and the terminal half-life is 6.7–19.2 h (Janasik et al. 2008).
evant to C9–C14 aliphatic (2–25% aromatic) hydrocarbon sol- A comparison of the elimination curves of 1,2,4-TMB and
vents are described, and referenced in the following section on n-decane (Figure 4 in Hissink et al. 2007) indicates that the
C14–C20 aliphatic (2–30% aromatic) hydrocarbon solvents. time course for elimination of aliphatic constituents is similar
to that for aromatics.
3. Toxicokinetics
As discussed above, the aliphatic and aromatic constituents 4. Acute toxicity tests
of C9–C14 aliphatic (2–25% aromatic) hydrocarbon solvents
have common pharmacokinetic properties because of the A number of acute toxicity tests have been conducted. Not all
structural similarities of the constituents. of the data have been published but summary information was
If inhaled, C9–C14 aliphatic (2–25% aromatic) hydrocar- published by Amoruso et al. (2008) and is also available in
bon solvent constituents are relatively well absorbed through OECD (2012c). A summary of the information is provided
the lungs (Astrand et al. 1975). If ingested, it is estimated that below (Table 15). Representative solvents were not lethal
approximately 60–80% of the constituents would be absorbed. at the highest administered doses. There have been reports
of ocular and respiratory irritation in volunteer studies and
evidence of respiratory irritation and acute CNS effects in
36“Straight
Run” means that the substances are produced from crude oil toxicology studies in rodents (Carpenter et al. 1975c). These
by atmospheric distillation. hydrocarbon solvents can also produce chemical pneumonitis
Table 15. Summarized results of acute toxicity tests of C9–C14 aliphatic much more profoundly affected than all of the other animals
solvents with 2–25% aromatics. with mortalities, at exposure levels as low as 363 mg/m3
Test Result References (approximately 66 ppm) and a no-effect level of 238 mg/m3
Acute Oral  5000 mg/kg API (1986) (approximately 43 ppm).
Toxicity Due to the striking differences in response between the
 8000 mg/kg Unpublished data summarized in guinea pigs and animals of other species, additional studies
the OECD (2012c)
Acute Dermal  3000 mg/kg API (1986) were conducted in which the animals were exposed 8 hours/
Toxicity day and 5 days/week for 10 weeks at exposure levels ranging
 4000 mg/kg Unpublished data summarized in from 593–1353 mg/m3 (approximately 108 to 246 ppm) (Rec-
OECD (2012c)
Unpublished data summarized in
tor et al. 1966). In this study, all animals, including guinea
 4000 mg/kg
OECD (2012c) pigs, survived to scheduled termination. There was some evi-
 3160 mg/kg Unpublished data summarized in dence of lung irritation in the guinea pigs, but there were no
OECD (2012c) histological changes in other animals.
Acute Inhalation  5500 mg/m3 API (1987)
Toxicity ( 1000 ppm) A later experiment was conducted by the same group to bet-
 8200 mg/m3 Carpenter et al. (1975c) ter understand the species differences. It was hypothesized that
( 1490 ppm) the effects in the guinea pigs had been exacerbated by a defi-
 14000 mg/m3 Unpublished data summarized in ciency in dietary levels of vitamin C. It was noted that when
( 2500 ppm) OECD (2012c)
 2000 mg/m3 Unpublished data summarized in the guinea pigs were maintained on a diet with high levels of
( 2180 ppm) OECD (2012c) ascorbic acid, they were much less susceptible to the effects of
mineral spirit (Jenkins et al. 1971).
In 1975 Carpenter et al. (1975c) reported a study in which
rats were exposed to Stoddard solvent 6 hours/day and 5 days/
if they enter the lungs in a liquid state. There is one report of week for 13 weeks. Exposure levels were approximately 500,
histological changes in the upper respiratory system suggesting 1100, or 1900 mg/m3 (approximately 91, 200, or 345 ppm).
irritation, following exposure of rats to a commercial grade of All animals survived without outward signs of distress. There
white spirit, 4 hours/day for 4 consecutive days to an average were no remarkable findings in the clinical or hematological
concentration of 214 mg/m3 (approximately 39 ppm) (Riley evaluations. There were no pathological findings in the dogs,
et al. 1984). The authors hypothesized that exposures at higher
but marked tubular regeneration was noted in the kidneys

levels for longer periods of time could produce more serious of male rats exposed at the 1100 and 1900 mg/m3 (200 and
upper respiratory effects. However, several longer-term stud- 345 ppm) levels. Aside from the kidney changes in the male
ies in rats have been conducted at higher levels (summarized rats, the overall no-effect level for both rats and dogs was
in Hydrocarbon solvent constituents with unusual toxicologi- 1900 mg/m3 (345 ppm).
cal properties), and none of these has resulted in severe upper A repeated inhalation toxicity study was conducted at higher
respiratory effects. Further, these findings were not substanti- exposure levels in an attempt to better define the no adverse
ated by human evidence (Appendix B,I,9.1.2) effect concentration (Carrillo et al. 2014). Rats were exposed
6 hours/day and 5 days/week for 13 weeks at exposure levels
5. Repeated dose toxicity of 2000, 4000, or 7500 mg/m3 (approximately 364, 727, or
1364 ppm). At termination, the animals were sacrificed and
5.1 Repeated inhalation toxicity studies of complex
examined for gross and pathological changes, and clinical
C9–C14 aliphatic (2–25% Aromatics) solvents
and hematological parameters were evaluated. Statistically
There have been at least 5 repeated inhalation toxicity studies significant effects, only observed in the high-exposure groups,
with protocols similar to OECD 413 (Carpenter et al. 1975a, included a significant reduction in terminal body weights in
Carrillo et al. 2014, Jenkins 1971, Rector et al. 1966, and one female rats and renal changes in the kidneys of male rats. Set-
additional unpublished study summarized in OECD 2012c). ting aside the male rat kidney effects, the low-effect level was
In the first of these (Rector et al. 1966), animals of several 7500 mg/m3 (1364 ppm), and the no observed effect concen-
species (rat, guinea pig, rabbit, dog, monkey) were exposed tration was 4000 mg/m3 (727 ppm).
continuously (i.e., approximately 24 hours/day and 7 days/ In another unpublished study, summarized in Carrillo et al.
week for 90 days) to graded concentrations ranging from 114 (2014), the toxicological significance of the renal changes in
to 1271 mg/m3 (approximately 21 to 231 ppm). The majority male rats was assessed following exposure at levels similar
of the animals survived the dosing period, and, at termina- to those used by Carpenter et al. (1975c). This study better
tion, had body weights similar to control values. The gross characterized the kidney changes and otherwise corroborated
pathological investigation revealed lung congestion in the the conclusion of Carpenter et al. (1975c), that is, that there
high-exposure group animals of all species. There were histo- were no other toxicologically important effects associated with
logical changes in the livers of some animals as well as some repeated exposure to solvents of this type, but the unpublished
small changes in hematological parameters, but the authors study was focused on renal effects and other organs were not
did not consider these effects to have been treatment-related. examined in detail. As described in more detail in a review
The overall conclusion was that for most of the species tested, article by Amoruso et al. (2008), the kidney changes were later
and other than the local effects in the lungs, the overall no- shown to be the consequence of an a2u-globulin-mediated
effect level was 1271 mg/m3 (approximately 231 ppm), the process and not relevant to humans (US EPA 1991, Swenberg
highest concentration tested. However, the guinea pigs were and Leeman-McKeeman 1997). There is also a report in which
male rats were exposed to white spirit for 9 months at a con- vivo conditions in tests for micronucleus induction in mouse
centration of 6500 mg/m3 (approximately 1182 ppm) (Viau bone marrow (McKee et al. 1994).
et al. 1984). The study by Viau et al. (1984) was specifically
an assessment of the potential for renal effects in male rats; 7. Developmental toxicity
results of clinical observations were similar to those of other There have been at least three unpublished developmental
studies, but pathological findings were not reported. toxicity studies of white spirit in which the test material was
In summary, these studies provide evidence that repeated administered by inhalation. These studies provided evidence
inhalation exposure to C9–C14 aliphatic (2–25% aromatic) that at levels high enough to cause maternal effects, there was
hydrocarbon solvents is not associated with target organ effects evidence of delayed fetal development, but no fetal mortality
(other than a2u-globulin-mediated changes in the kidneys of or malformations. It should be noted that this section contains
male rats) and that the overall no observed adverse effect con- only reviews of developmental toxicity studies of conventional
centration is 4000 mg/m3 (727 ppm). design. However, there are also data relevant to developmental
Additional supporting information is provided by repeated toxicity in the following section in which reproductive toxicity
administration studies of two similar substances; specifically, studies are summarized.
repeated inhalation and oral toxicity studies of jet fuel (Jet A) In the first of the developmental toxicity studies (American
and dermal toxicity studies of hydrodesulfurized kerosene. Petroleum Institute 1977), pregnant rats were exposed 6
In the repeated inhalation toxicity study of jet fuel, rats were hours/day, on gestational days 6–15, to either 100 or 400 ppm
exposed continuously to jet fuel for 90 days at levels up to (approximately 550 or 2200 mg/m3) Stoddard solvent. There
1000 mg/m3 (approximately 1667 ppm). The only reported were no differences in frequencies of malformations and no
effect was a2u-globulin-mediated renal nephropathy in male differences in litter size or fetal weight. There was an increased
rats (Mattie et al. 1991). In a repeated oral study in male rats incidence of fetuses with skeletal variations, but this was not
at levels up to 3000 mg/kg/day, there was some evidence of dose-related. In a similar study (unpublished information sum-
gastritis, most likely secondary to the repeated oral admin- marized in OECD (2012c)), pregnant female Sprague-Dawley
istration of large bolus doses of hydrocarbons. Aside from rats were exposed 6 hours/day on days 6–15 of gestation at
kidney changes in male rats, there was minimal evidence of concentrations of either 100 or 300 ppm (approximately 550
systemic effects (Mattie et al. 1995). In the dermal admin- or 1650 mg/m3). The rats were sacrificed on gestational day
istration toxicity studies of hydrodesulfurized kerosene, the 20, and the uterine contents were examined. There were no
test substance was administered repeatedly for 90 days at maternal or fetal effects in either of the experimental groups.
doses up to approximately 500 mg/kg/day, a level considered The no observed adverse effect concentration was 300 ppm
to be the highest that could be repeatedly applied without (approximately 1650 mg/m3).
causing severe dermal irritation and stress. In one repeated A third investigation involved two studies (Jakobsen et al.
dose study, the potential for repeated treatment to cause tar- 1986). In the first, pregnant rats were exposed to white spirit at
get organ and/or systemic effects was assessed (Breglia et al. concentrations of 273, 483, or 953 ppm (approximately 1500,
2014). The other evaluated the potential for effects on fertil- 2656, or 5242 mg/m3), 6 hours/day on days 6–15 of gestation,
ity and on the nervous system (Schreiner et al. 1997). There and in the second study, pregnant rats were exposed to 950
was no evidence of systemic, reproductive or neurological ppm (approximately 5225 mg/m3), 6 hours/day on gestational
effects. In some ways, the absence of effects is not surprising days 3–20. There was maternal toxicity in the highest exposure
considering the limited potential of these substances to be groups of both studies, but there were no effects on fetal sur-
percutaneously absorbed, but they do provide empirical evi- vival and no evidence of an increased frequency of malforma-
dence that repeated dermal exposure would not be associated tion. Fetal weights were reduced in the high-exposure groups,
with systemic toxicity. and there was an increased incidence of delayed ossification.
In summary, the results of the supporting studies are con- It was concluded that the fetal effects were secondary to the
sistent with the results of the inhalation toxicity studies of maternal toxicity.
C9–C14 aliphatic (2–25% aromatic) hydrocarbon solvents, Cooper and Mattie (1996) tested JP-8 jet fuel in a devel-
that is, that repeated exposure does not produce target organ opmental toxicity study, administering the test material by
effects in rats. gavage in doses of 500, 1000, 1500, or 2000 mg/kg on gesta-
tional days 6–15. They reported that maternal weight gain was
6. Genetic toxicity reduced in animals given doses of 1000 mg/kg and that fetal
White spirit was reported as inactive in a battery of genetic weights were reduced in the 1500 and 2000 mg/kg groups.
toxicity tests including Salmonella and an in vivo assessment However, there were no effects on fetal survival, and the fre-
of micronucleus induction in mouse bone marrow cells. White quency of malformations was not increased, indicating that jet
spirit was also inactive in an in vitro assay for sister chroma- fuel does not cause selective developmental effects.
tid exchange using human lymphocytes (Gochet et al. 1984).
8. Reproductive toxicity
Summarized results of other unpublished results provide fur-
ther support for the conclusion that white spirit and similar Although there have not been any formal reproductive toxicity
substances are not genotoxic (Amoruso et al. 2008, IPCS studies of the C9–C14 aliphatic (2–25% aromatic) hydrocar-
1996). bon solvents, there have been reproductive toxicity studies of
In other studies, jet fuel and kerosene have been reported kerosene and jet fuel and also some studies of C9–C14 hydro-
to be inactive when tested in vitro in Salmonella and under in carbon solvent constituents that provide relevant information.
Additionally, there have been several repeated dose studies, 9. Neurotoxicity studies
described in a previous section, in which reproductive organs
were examined. These studies provided no evidence that the 9.1 Studies of acute CNS effects
reproductive system is a target for C9–C14 aliphatic (2–25% 9.1.1 Studies of acute CNS effects in animals
aromatic) hydrocarbon solvents.
The acute CNS effects of white spirit were characterized in
As summarized by Mattie and Sterner (2011), the poten-
rats with a battery of tests that included functional observa-
tial for jet fuel to affect reproductive parameters was assessed
tions and measures of motor activity. The potential for effects
in two studies in rats, one focused on reproductive effects in
on learned visual discrimination was also assessed (Lammers
males and the other on females. In the first of these studies,
et al. 2007). Rats were exposed at levels of 600, 2400, or 4800
male rats were given oral doses at levels up to 3000 mg/kg,
mg/m3 (approximately 109, 436, or 873 ppm), 8 hours/day for
starting 70 days prior to mating. Although male rats in the
3 consecutive days. Assessments were made prior to exposure,
high-dose group had significantly lower body weights than
immediately after exposure on each of the three days, and
controls, there were no effects on pregnancy rate, gestation
24 h after the final exposure, to assess recovery. Ataxia was
length, or sperm parameters. In the second study, female rats
observed in some animals after the first day of exposure, but
were given daily oral doses at levels up to 1500 mg/kg/day,
the magnitude of the differences was not statistically signifi-
starting 90 days prior to mating and continuing to the end of

cant, and effects were not observed on subsequent days. In the
lactation (postnatal day 21). Maternal effects included lower
motor activity tests, there was a small but statistically signifi-
body weights and target organ effects. However, there were
cant reduction in total distance run in the high-exposure group
no statistically significant changes in pregnancy rate, gestation
on the third day of exposure. In the visual discrimination stud-
length, or numbers of offspring per litter. Weights of offspring
ies, there was a significant effect on time to respond among the
in the high-dose group were significantly below control values
animals in the high-exposure group. No significant differences
but by postnatal day 90, there were no significant differences
were observed when the animals were re-tested 24 h after the
in body weights.
last exposure, confirming that the effects were reversible. The
Schreiner et al. (1997) reported a reproductive/develop-
no-effect level in this study was judged to be 600 mg/m3 (109
mental toxicity screening test (OECD 422) in which hydrodes-
ppm). These data were consistent with the results of a previous
ulfurized kerosene37 was administered by dermal administra-
study Kulig (1990) in which it was reported that 1200 mg/
tion in doses up to 495 mg/kg/day, starting 7 weeks prior to
m3 (218 ppm) was a low-effect level for acute CNS effects in
mating and continuing to gestational day 19. There were no

rats.
indications of toxic responses in the parental animals and no
statistically significant changes in any of the reproductive or
9.1.2 Observations of acute CNS effects and upper respiratory
developmental parameters. These data indicated that dermally
tract irritation in humans
applied hydrodesulfurized kerosene had no effects on repro-
ductive or developmental parameters at doses up to 495 mg/ Numerous studies have been conducted to assess the poten-
kg bw/day. tial for Stoddard solvent to produce acute CNS effects and/or
There have also been studies of some C9–C14 hydrocarbon respiratory irritation in humans. In the first of these (Nelson
solvent constituents, as well as a reproductive toxicity study of et al. 1943), volunteers were exposed to a range of concentra-
a complex C9 aromatic substance (McKee et al. 1990, summa- tions for periods of 3–5 min and then asked to classify the
rized in the section on C9 aromatic solvents, Appendix A.1). irritant properties of the vapor on eye, nose, and throat. Each
The C9 aromatics are similar to the constituents that comprise subject was also asked if he believed that he could work under
the aromatic fraction of white spirit. Additional data from a those conditions for 8 hours/day. There were no marked effects
one-generation reproductive toxicity study of decane (unpub- at levels up to 400 ppm (approximately 2500 mg/m3), but the
lished) and a reproductive toxicity screening test equivalent subjects considered that this level was too high for an entire
to OECD 422 on undecane (Ministry of Health and Welfare day.
1996) are summarized in the section on C9–C14 aliphatic This work was extended in a similar study by Carpenter
( 2% aromatics) hydrocarbon solvents (Appendix D). None et al. (1975c) in which volunteers were exposed for 15-minute
of these substances had any effects on reproductive param- periods at levels of 140, 850, or 2700 mg/m3 (approximately
eters. 25, 154, or 490 ppm) and then interviewed to assess the sever-
In summary, although no formal reproductive toxicity tests ity of the perceived effects. At the highest level, all of the vol-
have been conducted on C9–C14 aliphatic (2–25% aromatics) unteers reported eye irritation and 2/6 reported slight dizziness
hydrocarbon solvents, the available data suggest that these which cleared within 15 min of leaving the chamber. The vol-
substances are unlikely to be reproductive toxicants. unteers exposed to 850 mg/m3 (154 ppm) experienced slight
and transitory eye irritation, but no effects were associated
with exposure at the 140 mg/m3 level (25 ppm). The authors
37Hydrodesulfurized kerosene is similar to JP-8 jet fuel and can, in some considered that their data were in line with occupational expo-
ways, be considered as the feed stock for C9–C14 aliphatic (2–25% aro- sure recommendations.38 Hastings et al. (1982) conducted a
matics) hydrocarbon solvent production. As reported by Schreiner et al.
(1997), kerosenes are complex aliphatic substances with approximately
20% aromatics. The carbon number range is approximately C9–C16
and the distillation range is approximately 150–290°C. The aromatic 38As noted previously, the ACGIH TLV® was being revised from 200
constituents are predominantly alkylated one- and two-ring aromatics ppm to 100 ppm (525 mg/m3) during the period when the Carpenter stud-
with  0.01% aromatic constituents with 3 or more aromatic rings. ies were being conducted.
similar study in which volunteers were exposed for 30 min The authors considered that their results suggested “a mild
to 600 mg/m3 (109 ppm) and then interviewed to assess the irritative effect of the eyes… [as]…measured by symptoms
level of irritation. The investigators also measured eye blink, ratings…However…the magnitude of the ratings was not sup-
frequency of swallowing, and rate of respiration to provide ported by any of the objective measures.”
objective measures of effect and used the Purdue Pegboard There have been at least 6 other volunteer studies in which
Test to assess effects on psychomotor function. The authors the potential for irritation and/or acute CNS effects were
concluded that at 600 mg/m3 (109 ppm) Stoddard solvent was, assessed in studies involving longer periods of time. Cohr
at most, only mildly unpleasant or irritating to humans and et al. (1984) compared responses of students (not previously
that psychomotor performance was unaffected. Gamberale exposed) to painters in experiments involving exposure peri-
et al. (1975) exposed volunteers to Stoddard solvent for 30 min ods of 7 h and exposure levels up to 400 ppm (2200 mg/m3).
at levels up to 2500 mg/m3 (454 ppm) in one set of trials and at The authors reported that among the painters, there were
levels up to 4000 mg/m3 (727 ppm) in a second. They reported complaints of eye, nose, and/or respiratory irritation which
prolonged reaction times and also considered that there was were significant at levels of 200 ppm (1100 mg/m3) or higher
evidence for impaired short term memory in subjects exposed and at 100 ppm (550 mg/m3) or higher among the students.
to 4000 mg/m3 (727 ppm). Their overall conclusions were The degree of irritation was not reported. In the assessment of
that exposure to levels  2500 mg/m3 (454 ppm) was associ-
acute CNS effects, an increased incidence of headache, tired-

ated with acute CNS effects, but there were no reported CNS ness, and giddiness was reported at 100 ppm (550 mg/m3) by
effects at exposure levels up to 1850 mg/m3 (336 ppm). The the painters but only at higher levels among the students.
study by Gamberale et al. (1975) was an extension of observa- Pedersen and Cohr (1984a) carried out a volunteer study
tions by Astrand et al. (1975) in which it was reported that in to compare the pharmacokinetic behavior of “white spirit” to
initial studies, the subjects had reported discomfort described aliphatic solvents with low levels of aromatics (i.e., Category
as nausea, vertigo, and a feeling of being severely affected at 8 solvents, as indicated previously). The volunteers were
an exposure to 5000 mg/m3 (909 ppm) and that some symp- exposed for 6 h to each of the solvents at concentrations up to
toms were reported following exposure to 2500 mg/m3 (454 approximately 200 ppm (1100 mg/m3). The authors reported
ppm). Pedersen and Cohr (1984a) exposed volunteers for 6 h that the subjects did not report any symptoms related to either
to white spirit39 at 610 mg/m3 (102 ppm), and followed this respiratory irritation or acute CNS effects.
with examinations of blood and urine biochemistry, lung func- Jarnberg et al. (1997a) compared the pharmacokinetic
tion, odor threshold, and a number of both objective and sub- behavior of white spirit to an aromatic constituent, 1,2,4-trim-
jective tests to assess the potential for acute CNS effects and ethylbenzene. Subjects were exposed for 2 h to levels of 300
upper respiratory irritation. There were no differences in the mg/m3 (54 ppm). The authors reported that “[N]o significant
biochemical assessments, and there were no reports of either irritation or CNS effects were detected at these conditions. All
irritation or CNS effects. mean values of ratings (except those of smell) corresponded
More recently, studies were reported in which the effects of verbally to something from “no effect” to “hardly any effect
short-term exposure to either “regular white spirits” or “de- or discomfort.”
aromatized white spirits”40 were compared (Ernstgard et al. Lammers et al. (2007) compared effects in volunteers
2009a, 2009b). In these studies, volunteers were exposed in exposed for 4 h to regular white spirit at levels of 57 or 570
10-minute steps to doses that were increased from 0.5–600 mg/ mg/m3 (10 or 100 ppm). Irritation was assessed via question-
m3 (0.1 to 109 ppm). Effect levels were based on the reports naires that the volunteers completed prior to and immediately
of the volunteers in a range-finding study (Ernstgard et al. after the exposure period. The potential for acute CNS effects
2009a), with more objective measures used in a follow-up was assessed via a series of tests that was administered 90 min
study in which the volunteers were exposed for 4 h (Ernstgard prior to exposure, 45 and 165 min after the start of exposure,
et al. 2009b). The authors reported that in the studies involving and 90 min after the exposure. A comparison of the responses
short exposure periods, the median levels of irritation were on the questionnaire did not reveal any differences that could
given as “somewhat” for “regular white spirits” and “hardly be attributed to exposure. There were some small but statisti-
at all” for the dearomatized white spirit, and ratings of acute cally significant differences in performance among which the
CNS effects were “hardly at all”. In the follow-up study, vol- outcome most plausibly related to exposure was a 5% reduc-
unteers were exposed for 4 h to regular white spirit at levels tion in simple reaction time.
of 100 mg/m3 or 300 mg/m3 (18 or 54 ppm) (Ernstgard et al. Juran et al. (2014) assessed the potential for acute CNS
2009b). Irritation was assessed via a questionnaire, but addi- effects in volunteers exposed for 4 h to either “regular” or
tional objective measures of ocular and respiratory irritation “dearomatized” white spirit at levels of 100 and 300 mg/m3
were carried out. Acute CNS effects were not investigated. (18 and 54 ppm). They did not identify any consistent effects
associated with exposure to either solvent at either exposure
level.
39The test sample was a commercial solvent identified by the trade In summary, investigations of the acute effects of Stoddard
name “Varnolene” and described as containing 57% alkanes (primarily solvent in humans indicate that it can produce irritation and
C9–C11), 25% cycloparaffins (primarily C8–C11), and 18% alkylben-
zenes (C8–C11). acute CNS effects, but the reported effect levels depend on the
40The de-aromatized white spirits are similar to the complex aliphatic sensitivity of the methods and whether the outcomes are char-
solvents with  2% aromatics. Following the naming convention, those acterized in qualitative (e.g., acceptable versus unacceptable)
solvents are included in Category 8, C9–C14 aliphatic hydrocarbon sol- or quantitative (e.g., statistical differences from control) terms.
vents ( 2% aromatics). In general, the volunteers regarded 100 ppm (approximately
525 mg/m3) as an acceptable level for occupational exposure, during painting (Arlien-Soborg et al. 1979, Hane et al. 1977,
and if there were statistically significant performance deficits Lindstrom and Wickstrom 1983, Lundberg et al. 1995, Mik-
at that level they were too subtle to be noticed by the subjects. kelsen et al. 1988, Seppalainen and Lindstrom 1982). Because
In particular, it should be noted that the exposure levels that it was the solvent most commonly used to formulate construc-
were experienced during construction painting41 may have tion paints, Mikkelsen et al. (1988) concluded that white spirit
caused irritation but are unlikely to have caused noticeable was the causative agent, and this conclusion was carried for-
CNS effects. ward into a subsequent review (IPCS 1996). However, there
are other, more plausible explanations for these findings that
9.2 Studies to assess the potential for persistent CNS do not appear to have been seriously considered. As reported
effects of C9–C14 aliphatic hydrocarbon solvents (2–25% by Riala et al., (1984), construction painters also used epoxy
aromatics) paints which contained toluene, xylene, aliphatic alcohols,
9.2.1 Studies to assess the potential for persistent CNS effects and ketones as solvents. The epoxy paints were used for more
in animals specialized applications and made a smaller contribution to
overall exposure than did white spirit. On the other hand, the
Kulig (1989) conducted a study in which rats were exposed use of epoxy paints was reported to be the cause of episodes
by inhalation to levels of 1200, 2400, or 4800 mg/m3 (218,
of overexposure which were relatively common during that

436, or 873 ppm). Exposures were 8 hours/day and 5 days/ period. Had the differences in the neurobehavioral tests been
week for 26 weeks. In neurobehavioral tests conducted at least the result of repeated overexposure, then the use of epoxy
10 h after cessation of exposure, there were no effects on dis- paints becomes a more likely cause than white spirit. This
criminant performance or in motor activity tests. Further, at alternative hypothesis is supported by evidence that toluene
study termination, a pathological examination did not reveal abuse is associated with persistent CNS effects (Grandjean
any treatment-related changes in the central or peripheral ner- 1985). Another plausible explanation for the data is that the
vous systems. The authors concluded that exposure to white effects were due to exposures to solvents of higher intensity,
spirit could produce acute reversible effects, but there was no experienced in other painting occupations. As documented
evidence of persistent effects. In a recent review, Nielsen et al. by Mikkelsen et al. (1988), many of the construction paint-
(2006) found no evidence for adverse pathological changes ers had also worked in other painting occupations including
in the central or peripheral nervous systems of rats follow- spray painting, and in those jobs, they experienced much more
ing exposure to white spirit. There was also no evidence for
intense solvent exposures than they did during construction

neurobehavioral changes in animals exposed to regular white painting. Further, the solvents used in formulating paints for
spirit. spraying were aromatics, ketones, and chlorinated solvents
There have been a number of reports of neurochemical (Elofsson et al. 1980). Of particular note is that spray paint-
changes in the CNS following repeated exposure to white spirit ers also exhibited neurobehavioral deficits that were at least
(Bondy et al. 1995, Lam et al. 1992, 1995, 2000, Ostergaard as pronounced as those in the construction painting industry,
et al. 1993, Savolainen and Pfaffli 1982, Steensgaard et al. but some of the spray painters had no documented exposure to
1996), but in the absence of any functional or pathological white spirit (Elofsson et al. 1980). Thus, it is possible that the
changes, the interpretation is unclear. At present, there is no neurobehavioral deficits found in construction painters were
animal model for persistent CNS effects of solvents (Bruckner actually due either to higher-intensity exposures to solvents
and Warren 2001). in general or to exposure to solvents other than white spirit
while working in other painting occupations. It should be
9.2.2 Studies to assess the potential for persistent noted that there is a report in which the degree of neurobehav-
CNS effects in humans ioral changes was statistically associated with higher-intensity
As discussed previously in this document, there are reports exposures (Hooisma et al. 1994).
asserting a relationship between long-term exposure to white In summary, there are reports associating long-term occu-
spirit, and neurological deficits. These reports rely primarily pational exposures to white spirit with persistent CNS effects.
on studies of painters who worked in the construction painting However, a critical review of the information suggests that
industry during the period 1960–1980. It was during that time these differences were more likely due to exposure to other,
that formulation of paints for the construction industry evolved more volatile solvents in other types of painting occupations.
from linseed oil and turpentine to hydrocarbon solvents, and
then further, to water-based paints. It was estimated that during 9.2.3 Occupational exposures and recommended occupational
that period, construction painters were exposed to, on average, exposure limits
approximately 40 ppm (approximately 220 mg/m3) of white
spirit with maximal exposures in the range of 200–300 ppm In the United States, there was an ACGIH TLV® of 500 ppm
(1100–1650 mg/m3) (Riala et al. 1984). Neurobehavioral test- (2625 mg/m3) from 1948 to 1969. This value was reduced to
ing of construction painters revealed statistically significant 200 ppm (1050 mg/m3) in 1970 and to 100 ppm (525 mg/m3),
deficits in certain tests that were attributed to solvent exposure the present value, in 1976. The European experience seems
to have been similar. As reported by Lundberg et al. (1995)
the occupational exposure limit (presumably in Sweden) was
41As reported above, the average white spirit exposures were estimated as
1200 mg/m3 prior to 1973, reduced to 600 mg/m3 in 1974, and
40 ppm (approximately 220 mg/m3) with estimated maximal exposures in to 500 mg/m3 in 1979. Anecdotal information in both Mik-
the range of 250–300 ppm (1375–1500 mg/m3). kelsen et al. (1988) and Lundberg et al. (1995) suggests that
at least during the 1960s, the exposure levels in the painting Table 16. Physical/chemical properties of C14–C20 aliphatic solvents
industry were close to the occupational exposure levels, and (2–30%) aromatics.
this is consistent with information on cumulative exposure Hydrocarbons
shown in Figure 3 of Lundberg et al. (1995), and more spe- Hydrocarbons C14-C18, C16-C20, (2-30%
Property (2-30% aromatics) aromatics)
cifically, most of the total exposure took place prior to 1973.
By 1980, it was estimated that exposures to white spirit in
Physical state at Colorless to yellow liquid Colorless to yellow
the construction industry averaged approximately 40 ppm 20°C and 1013 hPa liquid
(estimated as approximately 232 mg/m3) on an 8-hour time- Melting/Freezing  10°C (ASTM D 5950)  0°C (ASTM D
weighted basis (Riala et al. 1984). Exposures to these solvents Point 5950)
in the dry cleaning industry were at similar levels (Oberg Boiling Range 230–320°C (ASTM D 86) 250–350°C (ASTM
D 86)
1969). Caldwell et al. (2000) estimated that the weighted aver- Relative Density 0.8–0.9 g/cm3 at 15°C 0.805–0.920 g/cm3 at
age exposure to white spirit/Stoddard solvent was 466 mg/m3, (ISO 12185) 15°C (ISO 12185)
based on information published between 1961 and 1998. For Vapor Pressure 0.001 kPa at oC  0.001 kPa at oC
comparative purposes, the lowest effect levels for acute effects Flash Point  100°C (ASTM D93)  110°C (ASTM
D93)
in volunteer studies are in the range of 500 mg/m3 (Ernstgard Flammability 0.6–7.0% v/v 0.6–7.0% v/v
et al. 2009a), and no-effect levels in studies of occupationally Self-Ignition  200°C  200°C
exposed painters were estimated to have been between 230 Temperature
Surface Tension 28–30 mN/m at 28°C 28–31 mN/m at 28°C
mg/m3–540 mg/m3 (SCOEL 2007, using information from
(Wilhelmy plate test) (Wilhelmy plate test)
Lundberg et al., 1995). Viscosity 3.5–10.0 mm2/s at 20°C 4.0–10.0 mm2/s at
(ASTM D 7042). 20°C (ASTM D
II. Toxicological Properties of C14-C20 Aliphatic 7042)
Hydrocarbon Solvents (2–30% Aromatics) (Category Water-Solubility  1 mg/kg  1 mg/kg
Limit
4 solvents) Odor Threshold Essentially odorless Essentially odorless
Maximally  200 mg/m3 at 25°C  60 mg/m3 at 25°C
1. Introduction Attainable Vapor
Category 4 solvents, “C14–C20 aliphatic solvents (2–30%) Concentration
(ECETOC, 1997)
aromatics”, are similar to the Category 3 solvents but with
higher molecular weight constituents. More specifically, the
Category 4 solvents contain C14–C20 aliphatic solvents with absorbed, metabolized, and excreted. Albro and Fishbein (1970)
aromatic constituents at levels ranging from 2–30% and with published an empirical relationship in which the percentage of
distillation ranges of approximately 230–350°C. These sol- ingested n-alkanes that were retained was estimated as (per-
vents are derived from crude oil by distillation and may be centage retained)  115.9–3.94  (number of carbon atoms).
further refined by hydrogenation to reduce levels of sulfur and Based on this relationship, the fractional absorption of C14–
other undesirable constituents. As discussed in the previous C20 hydrocarbon solvent constituents would be in the range
section, these solvents are derived from the petroleum streams of approximately 37–60%. The prediction is consistent with
used to manufacture kerosene, jet fuel, and diesel fuel, so the experimental data from Le Bon et al. (1988) who reported
toxicity data for these petroleum substances is relevant to the that following per os administration of radiolabeled pristane
overall toxicological hazard characterization of C14–C20 ali- (2,6,10,14-tetramethylpentadecane), approximately 66% was
phatic (2–30% aromatic) hydrocarbon solvents. recovered in the feces and 14% was excreted in the urine as
pristane metabolites and tritiated water. However, the potential
2. Physical and chemical properties for systemic absorption via non-oral routes is limited. Based
on in vitro studies using porcine skin, Singh and Singh (2003)
There are two types of solvents in this category, defined by
estimated the percutaneous absorption rate for hexadecane as
carbon number range. They are, specifically:
7  10 3 nmol/cm2/hr (approximately 0.002 mg/cm2/hr).
Hydrocarbons C14–C18; normal paraffins, isoparaffinic
The aromatic constituents of the Category 4 solvents are
paraffins, and naphthenes, and containing 2–30% aromatics,
similar to those of the C10–C12 aromatic solvents. As summa-
and
rized in the section for Category 3 solvents (Appendix B.II.3),
Hydrocarbons C16–C20; normal paraffins, isoparaffinic
one- and two-ring aromatic constituents are relatively well
paraffins, and naphthenes, and containing 2–30% aromatics.
absorbed if inhaled or ingested, but percutaneous absorption
Their physical and chemical properties are summarized in
rates are very low. More specifically, the percutaneous absorp-
Table 16.
tion rate for the aromatic contents of C14–C20 is believed to
be similar to or lower than the reported range of 0.59–0.67
3. Toxicokinetics mg/cm2/hr for dimethylnaphthalene (Muhammad et al. 2005,
3.1 Absorption McDougal et al. 2000).
Because they have low vapor pressures, substantial exposure

3.2 Metabolism
to C14–C20 aliphatic (2–30% aromatic) hydrocarbon solvents
by inhalation is not expected, and it is not anticipated that It is believed that regardless of the route of administration,
inhalation would contribute substantially to systemic dose. If C14–C20 aliphatic hydrocarbon solvent constituents would
ingested, the constituents of these solvents are relatively well be metabolized and excreted by similar pathways. McCa-
rthy (1964) showed that long chain alkanes are converted to molecular weights and low vapor pressures, the maximally
fatty acids by oxidation of the terminal carbon. The aromatic attainable vapor concentration for n-tetradecane at 25°C is 202
constituents of the C14–C20 aliphatic (2–30% aromatic) mg/m3 (25 ppm). Thus, the potential for vapor exposure with
hydrocarbon solvents are similar to those of the C10–C12 these solvents is very limited. As described below, Carpenter
aromatic solvents. Once absorbed, the aromatic constituents et al. (1976c) conducted repeated inhalation toxicity studies
are metabolized and excreted relatively rapidly, primarily in with deodorized kerosene, but even under saturated vapor con-
the urine. Naphthalene is metabolized to epoxide intermedi- ditions, the highest concentration that could be stably main-
ates (Figure 2) whereas the alkylated benzenes and alkylated tained was 400 mg/m3. In contrast, Mattie et al. (1991) were
naphthalenes are primarily metabolized by glucuronidation of able to generate atmospheres of 1000 mg/m3 in jet fuel studies.
the alkyl side chains (Figure 3). It should also be noted that as more volatile constituents are
over-represented in the vapor phase, the hydrocarbons to which
3.3 Excretion the animals were exposed would have been primarily lower
The majority of the constituents of C9–C14 aliphatic (2–30% molecular weight constituents, probably more closely related
aromatic) hydrocarbon solvents are excreted within 24 h of to the constituents found in C9–C14 rather than C14–C20 ali-
exposure. phatic hydrocarbon solvents.
One often contentious issue of experimental design is the

4. Acute toxicity route of exposure that should be used in toxicology studies
of low volatility liquids. When studies are done for hazard
As summarized in Table 17, C14–C20 aliphatic hydrocarbons characterization purposes, in particular for purposes of regu-
(2–30% aromatics) were not acutely toxic when tested in stud- latory compliance, it is often required that they be conducted
ies of conventional design for acute oral (LD50  4150 mg/kg, by oral administration, often gavage, as a means of adminis-
or mortality noted), acute dermal (LC50  1700 mg/kg/day, no tering relatively large doses to maximize systemic absorption.
mortality noted) tests. Acute inhalation toxicity tests were not In contrast, studies for risk assessment often utilize inhalation
conducted because of the limited potential for vapor formation; or dermal routes of administration as these are the most com-
but the LC50 (4-hour) value for straight run kerosene is  5280 mon routes of human exposure. Oral administration is not
mg/m3 (American Petroleum Institute 1985), and that for jet a route of administration that is consistent with the recom-
fuel is  3420 mg/m3 (Mattie and Sterner 2011). However, it mended uses of hydrocarbon solvents, and care must be taken
should be noted that these solvents can cause chemical pneu-
to avoid introducing these substances into the lungs in the

monitis if taken into the lungs in a liquid state. liquid state. There may be other complications. For example,
Unpublished data provide evidence that C14–C20 aliphatic in one oral study of JP-8 jet fuel, the investigators adminis-
(2–30% aromatic) hydrocarbon solvents are not irritating to tered large doses by gavage for 90 days (Mattie et al. 1995).
the skin or eyes, and based on results of tests of kerosene Although most of the rats survived the dosing period, jet fuel
(American Petroleum Institute 1985), are unlikely to cause was irritating to the gastrointestinal lining when given repeat-
allergic contact dermatitis. edly by bolus administration. The development of gastritis and
perianal dermatitis secondary to the irritant properties of jet
5. Repeated dose toxicity fuel complicated the interpretation of the results.
Toxicological hazard characterization for solvents of this type In inhalation studies, the vapor levels that can be achieved
is challenging because of their physical and chemical prop- are limited by the vapor pressures of the test materials, and
erties. They are liquids, but because of their relatively high care must be taken to avoid creating aerosols. For example,
Carpenter et al. (1976c) exposed rats and dogs to vapors of
Table 17. Summarized results of acute toxicity tests of C14–C20 aliphatic deodorized kerosene for 13 weeks, at concentrations of 100,
solvents with 2–30% aromatics. 200, or 400 mg/m3 (approximately 20, 40, or 80 ppm), with
Test Result References the highest concentration representing a saturated atmo-
Acute Oral Unpublished study
sphere. The rats and dogs exposed to the highest concentra-
 4150 mg/kg
Toxicity summarized in the REACH tion survived the treatment period with minimal evidence of
registration dossier treatment-related effects.
 5000 mg/kg Mattie and Sterner (2011) In the jet fuel studies, in which animals were exposed con-
(based on jet fuel)
tinuously for 90 days, the principal finding in the inhalation
Acute Dermal  1700 mg/kg Unpublished study studies was that of reduced body weights in rats from the high-
Toxicity summarized in the REACH exposure group, as well as evidence of a2u-globulin neph-
registration dossier ropathy in male rat kidneys (Mattie et al. 1991). In the gavage
 2000 mg/kg API (1985)
(based on kerosene) study in which jet fuel was administered at doses of 750, 1500,
 2000 mg/kg Mattie and Sterner (2011) or 3000 mg/kg/day for 90 days, aside from evidence of gastro-
(based on jet fuel) intestinal irritation, the principal effects again were reductions
Acute Inhalation API (1985)
in body weights of male rats and the induction of a2u-globulin
 5280 mg/m3
Toxicity ( 960 ppm) (based nephropathy in the kidneys of male rats (Mattie et al. 1995).
on kerosene) Dermal administration was used in repeated dose studies
 3420 mg/m3 Mattie and Sterner (2011) of hydrodesulfurized kerosene (Breglia et al. 2014, Schreiner
( 620 ppm) (based
et al. 1997). These tests have the advantage of directly assess-
on jet fuel)
ing the most likely route of human exposure, but the use of
dermal administration has its own limitations; the amount of specifically; however, repeated dose studies of these solvents
material that can be applied is limited both by surface area and and their aliphatic and aromatic constituents provided no evi-
by the potential to cause skin irritation and/or injury, and the dence that the reproductive system was a target for these sub-
internal doses are difficult to assess. In the repeated dermal stances. Further, there were no effects in reproductive toxicity
studies, there were no target organ effects in rats following studies of orally administered jet fuel (Mattie et al. 2000) or
daily administration of 495 mg/kg/day, a dose chosen to avoid dermally administered hydrodesulfurized kerosene (Schreiner
excessive dermal irritation. et al. 1997). Using the same logic as in the previous paragraph,
In summary, the repeated effects of exposure to substances since corresponding petroleum products do not produce repro-
related to C14–C20 aliphatic (2–30% aromatic) hydrocarbon ductive effects, it is unlikely that C14–C20 aliphatic (2–30%
solvents were studied in rats, mice and dogs using oral, der- aromatic) hydrocarbon solvents would have any influence on
mal and inhalation exposures, at the highest possible levels at fertility.
which these tests could be carried out taking into consider-
ation both physical/chemical and animal welfare issues. There 9. Carcinogenicity
was no evidence of treatment-related effects aside from the It has been shown that repeated dermal application of straight
induction of kidney changes in the male rats, an effect that is run kerosene and jet fuel can cause cutaneous tumors in mice
species-specific and not relevant to humans, and gastritis and (Nessel et al. 1998, 1999). However, when the test materials
perianal dermatitis secondary to irritant properties. were applied in a way that minimized dermal irritation, the
animals did not develop tumors. The authors concluded that
6. Genetic toxicity
these tumors were the consequence of a promotional process
A representative C14–C10 aliphatic (2–30% aromatic) hydro- secondary to dermal injury. That these substances were not
carbon solvent was inactive in a Salmonella test conducted genotoxic carcinogens was supported by evidence demonstrat-
both in the presence and absence of metabolic activation ing that they were not mutagenic (Nessel et al. 1998, 1999).
(unpublished information). Kerosene is not mutagenic when
tested under in vitro and in vivo conditions (American Petro- 10. Neurotoxicity
leum Institute 1984a, 1984b, Blackburn et al. 1986, McKee There were no changes in functional observations or motor activ-
et al. 1994, Vijayalaxmi et al. 2006), and similarly, jet fuels ity and no pathological changes in nervous system tissue in a
were reported as inactive as shown in summarized informa- study involving repeated dermal application of hydrodesulfurized
tion published by Mattie and Sterner (2011) and McKee et al. kerosene at levels up to 495 mg/kg/day (Breglia et al. 2014).
(1994).
Appendix C. Volatile aliphatic solvents
7. Developmental toxicity
I. Toxicological Properties of C5 Aliphatic Solvents
No developmental toxicity tests of C14–C20 aliphatic (2–30% (Pentanes) (Category 5 solvents)
aromatic) hydrocarbon solvents specifically have been carried
out. However, as discussed elsewhere, neither kerosene nor 1. Introduction
jet fuel caused maternal or fetal effects in a study in which “C5 aliphatic solvents” include n-pentane, iso-pentane, and/
pregnant rats were exposed by inhalation to 100 or 400 ppm or cyclopentane in various combinations. These substances
(approximately 20 or 80 ppm) of kerosene on gestational days are listed on the Toxic Substance Control Act (TSCA)
6–15 (American Petroleum Institute 1979a, 1979b). Similarly, inventory under CAS registry numbers, in Europe under EC
in a study in which jet fuel was administered orally at doses numbers, and for REACH purposes were described under the
between 500–2000 mg/kg on days 6–15 of gestation, there naming convention, as shown in Table 2.
was no evidence of fetal death or malformations; however,
there were developmental delays at levels that also produced
maternal toxicity (Cooper and Mattie 1996). In a reproductive/ 2. Physical and chemical properties
developmental toxicity screening study in which kerosene was Pentane solvents are manufactured by distillation from crude
dermally applied during gestation, there was no reduction in oil followed by hydrogenation to remove sulfur. Their physical
numbers of live-born offspring, birth weights, or survival to and chemical properties are listed in Table 18.
scheduled termination (Schreiner et al. 1997). The aliphatic
constituents of C14–C20 aliphatic (2–30% aromatic) hydro- 3. Toxicokinetics
carbon solvents have carbon numbers that are similar to those Because of their relatively low boiling points, pentane isomers
in C14–C20 aliphatic ( 2% aromatic) hydrocarbon solvents, are volatile and rapidly evaporate from the skin, so inhala-
and the aromatic constituents are similar to those in C10–C12 tion is the most common route of exposure. However, inhaled
aromatic solvents. By analogy, as it was shown that the C10– pentane isomers are poorly absorbed (Dahl et al. 1988) and
C12 aromatic solvents were not developmental toxicants, the are rapidly eliminated. Filser et al. (1983) reported an elimi-
possibility of developmental effects with C14–C20 aliphatic nation half-life for n-pentane of 0.13 h. A similar value was
(2–30% aromatic) hydrocarbon solvents is also unlikely. obtained in an unpublished pharmacokinetic study of cyclo-
pentane (summarized in Galvin and Marashi 1998c). Tissue
analysis indicated that liver, small intestine, and kidney were
No reproductive toxicity tests have been conducted on the principal sites of pentane distribution (Daugherty et al.
C14–C20 aliphatic (2–30% aromatic) hydrocarbon solvents 1988). If pentanes are absorbed, they are converted to the
Table 18. Physical and chemical properties of C5 hydrocarbon solvents and their constituents.
Property N-Pentane Isopentane Cyclopentane
Molecular Weight 72 72 72
Physical State at 20°C Liquid Liquid Liquid
and 1013 hPA
Pour Point −20°C −20°C −94°C
Boiling Point 36.1°C (ASTM D5950) 31–45°C (ASTM D5950) 48–50°C (ASTM D5950)
Relative Density 0.60–0.65 g/cm3 (ISO 12185) 0.61–0.65 g/cm3 (ISO 12185) 0.75 g/cm3 (ISO 12185)
Vapor Pressure 45–79 kPa at 20°C 74–79 kPa at 20°C 38–45 kPa at 20°C
Surface Tension 13–17 mN/m at 25°C (Wilhelmy 13.7–16 mN/m at 25°C (Wilhelmy 21–22 mN/m at 25°C (Wilhelmy
plate methodology) plate methodology) plate methodology)
Water Solubility 38.5 mg/l at 20°C 48.5 mg/l at 20°C 156 mg/l at 20°C
Octanol/Water 3.45 4.0 3.0–3.45
Partition Coefficient
Flash Point  20°C (DIN 51755)  0°C (DIN 51755)  20°C (DIN 51755)
Flammability 1.3–7.8% (v/v) in air 1.3–7.8% (v/v) in air 1.1-8.7% (v/v) in air
Self-Ignition  200°C (ASTM E 659)  250°C (ASTM E 659)  360°C (ASTM E 659)
Temperature
Viscosity 0.20–0.52 mm2/s (ASTM D7042) 0.21–0.52 mm2/s (ASTM D7042) 0.40–0.59 mm2/s (ASTM D7042)
Odor Threshold 6.6–3000 mg/m3 (2.2–1017 ppm) 6.6–3000 mg/m3 (2.2–1017 ppm) 6.6–3000 mg/m3 (2.2–1017 ppm)
(Ruth 1986) (Ruth 1986) (Ruth 1986)
Maximally Attainable Approximately 500,000 ppm Approximately 500,000 ppm Approximately 500,000 ppm
Vapor Concentration
Conversion Factor 1 ppm  2.95 mg/m3 1 ppm  2.95 mg/m3 1 ppm  2.95 mg/m3
(ppm/mg/m3)
corresponding alcohols and eliminated in the urine. Frommer on survival or body weights, no differences in organ weights,
et al. (1970) reported that 2-pentanol was the principal urinary and no pathological changes. The only statistically significant
metabolite of n-pentane. Ingested pentanes are also reasonably differences were small changes in calcium and phosphorus
well absorbed, but percutaneous absorption of pentanes is not levels. These differences were within normal physiological
limits and reversed during the recovery period. The overall no
expected because they quickly evaporate from the skin.

adverse effect concentration was 10000 ppm (29412 mg/m3),
4. Acute toxicity the highest concentration tested.
In the first of the 90-day inhalation toxicity studies (McKee
At least in part because pentanes are poorly absorbed and rap- et al. 1998), rats were exposed by inhalation at levels of 5000,
idly eliminated, they produce acute effects only at extremely 10000 or 20000 mg/m3 (approximately 1700, 3400, or 6800
high levels. For example, cyclopentane was evaluated as a ppm), 6 hours/day and 5 days/week for 90 days. There were
potential anesthetic agent with the determination that in mice, no significant effects on survival or body weights; no changes
exposures at levels of approximately 8% induced sleep in in organ weights, hematology parameters, or serum chemistry
most animals, and exposures at levels of approximately 12% values that were considered to be treatment-related, and no
resulted in respiratory arrest and death (Virtue 1948). In dose- gross or microscopic findings. The overall no adverse effect
response investigations in mice, it was reported that there level was the analytically determined concentration of 20483
were no apparent acute effects in animals exposed to 16000 mg/m3 (approximately 7000 ppm), the highest concentration
ppm (approximately 470000 mg/m3), no anesthesia among tested. These results were confirmed in a subsequent study (Kim
animals exposed to 32000 ppm (approximately 94000 mg/ et al. 2012) in which similar exposure levels were tested and
m3), and deep anesthesia among animals exposed to 128000 similar results were obtained with an overall no adverse effect
ppm (approximately 375000 mg/m3) (Swann et al. 1974). As concentration of 20300 mg/m3 (approximately 6900 ppm).
summarized below (Table 19), pentane isomers are not acutely The potential for repeated dose effects of cyclopentane
toxic in oral and dermal studies conducted in accordance with was assessed in a 90-day inhalation toxicity study at target
current regulatory testing guidelines; they are minimally irri- concentrations of 5000, 10000, or 30000 mg/m3 (1750, 3500,
tating to the skin and eyes; and they do not produce allergic or 10500 ppm). The test was conducted in accordance with
contact dermatitis (OECD 2008). regulatory guidelines and was similar to the 90-day study of
n-pentane described above. No substance-related effects were
5. Repeated dose toxicity reported; the overall no observed adverse effect concentration
The potential for n-pentane to cause repeated dose toxic- was 30000 mg/m3 (10500 ppm), the highest concentration
ity following exposure by inhalation was assessed in 14-day tested (unpublished laboratory report summarized in Galvin
(Stadler et al. 2001) and 90-day (Kim et al. 2012, McKee et al. and Marashi 1999c).
1998) toxicity studies. In the 14-day study, rats were exposed In addition to the above, there have also been repeated
by inhalation 6 hours/day and 5 days/week for two weeks at dose studies that were intended specifically to investigate the
levels of 1000, 3000, or 10000 ppm (2941, 8824, or 29412 mg/ potential for pentane isomers and related substances to cause
m3). Half of the rats were sacrificed at the end of the exposure renal effects in male rats. Halder et al. (1985) investigated
period; the remaining rats were held for an additional 14 days the potential for n-pentane and 2-methylbutane, among other
without exposure before termination. There were no effects substances, to cause effects following oral administration, and
Table 19. Summarized results of acute and short-term toxicity tests of C5 notable pathological change was an increase in kidney effects
hydrocarbon solvents and their constituents. in male rats. The authors considered that the reduction in body
Substance Endpoint Value References weight gain was a consequence of reduced food intake; how-
n-Pentane Acute Oral  2000 mg/kg McKee et al. (1998) ever, the use of corn oil as a dosing vehicle may have also been
Toxicity a contributing factor.
Acute Dermal Not tested
Toxicity
Acute Stadler et al. (2001)
6. Genetic toxicity
 29,500 mg/
Inhalation m3  6106 Galvin and Marashi Salmonella tests of n- and iso-pentane, using a vapor phase
Toxicity ppm (1999a)
Ocular Minimally McKee et al. (1998) exposure system, produced negative results (Kirwin et al.
Irritation irritating 1980). In subsequent investigations, McKee et al. (1998)
Skin Irritation Minimally McKee et al. (1998) reported equivocal results from an in vitro chromosome aber-
irritating ration study of n-pentane and negative results from an in vivo
Dermal Not sensitizing McKee et al. (1998)
Sensitization micronucleus test in mouse bone marrow.
The results of a series of genetic toxicity tests of cyclopen-
2-Methylbutane Acute Oral Not tested tane that have not been formally published were summarized
Toxicity
Acute Dermal Not Tested by Galvin and Marashi (1998c). Cyclopentane was inactive
Toxicity in Salmonella tests in the presence and absence of S9 acti-
Acute 434,587 ppm Unpublished data vation; it was inactive in a mouse lymphoma (L5178Y) gene
Inhalation (1,282,203 summarized mutation assay in the presence and absence of S9 activation
Toxicity mg/m3) in Galvin and
Marashi (1999b.) at concentrations up to 200 mg/ml, a frankly toxic level; and
Ocular Not Tested it was inactive in an in vitro chromosome aberration assay in
Irritation human lymphocytes and in an in vivo micronucleus test in
Dermal Not Testeda
Irritation mouse bone marrow.
Dermal Not tested
Sensitization 7. Developmental toxicity
Cyclopentane Acute Oral  5000 mg/kg Galvin and Marashi A developmental toxicity test of n-pentane was reported by
Toxicity (1999c) McKee et al. (1998). Female rats were given n-pentane by
Acute Dermal Not Tested

oral administration on gestational days 6–15 in doses of 100,
Toxicity
Acute  11,260 ppm Galvin and Marashi 500, or 1000 mg/kg. The rats were sacrificed on gestational
Inhalation (approximately (1999c) day 21, and the uterine contents were examined. The fetuses
Toxicity 33,000 mg/m3) were weighed, examined externally, and then divided into two
Ocular Minimally Galvin and Marashi
Irritation Irritating (1999c)
groups for visceral and skeletal examination. As there were no
Dermal Minimally Galvin and Marashi maternal or fetal effects, the overall no adverse effect level was
Irritation Irritatinga (1999c) 1000 mg/kg, the highest dose tested.
Dermal Not Tested A preliminary developmental study using inhalation as the
Sensitization
route of exposure has also been reported (Hurtt and Kennedy
a2-Methylbutane and cyclopentane have been reported to be skin irritants 1999). Pregnant rats were exposed at levels of 1000, 3000, or
in humans and to cause burning and blistering of the skin (ACGIH, cit-
ing NIOSH, 1997). It should be noted that under normal circumstances, 10000 ppm, 6 hours/day, on gestational days 6–15. There were
pentane isomers rapidly evaporate from the skin. However, if evaporation no maternal effects and no statistically significant differences
is impeded, exposure can lead to skin irritation. Thus, in studies of this in live fetuses/litter or in fetal weight, and there were no exter-
type, it is always important to assess the conditions of the test. nal malformations. As there were no apparent effects at levels
up to 10000 ppm (approximately 29400 mg/m3), further, more
Aranyi et al. (1986) tested approximately 50:50 mixtures of comprehensive studies, were not conducted.
n-pentane:n-butane and iso-pentane:iso-butane in studies
in which the rats were exposed by inhalation. Halder et al.
(1985) reported that treatment with isoparaffinic hydrocar- The reproductive toxicity information relevant to pentanes
bons with carbon numbers ranging from C6 to C9 resulted in includes a one generation reproductive toxicity test of isopen-
renal effects, but that the results of studies of C5 constituents tane (2-methylbutane, Yu et al. 2011) along with supporting
(n-pentane, 2-methylbutane) were similar to control values. information from complex substances containing pentanes as
Aranyi et al. (1986) reported that the exposures to mixtures principal constituents (Bui et al. 1998, McKee et al. 2000).
containing normal- and iso-paraffin caused transient effects, There have also been repeated dose studies in which it has
but there were no apparent differences at terminal sacrifice. been demonstrated through gross and pathological examina-
In a publication summarizing data from a one-generation tions that repeated exposure to pentanes has no effects on
reproductive toxicity study, Yu et al. (2011) reported a sig- reproductive organs (Galvin and Marashi 1999c, Kim et al.
nificant reduction in body weight gain in male but not female 2012, McKee et al. 1998, Stadler et al. 2001).
rats. There were significant increases in relative weights In the one-generation reproductive toxicity test (Yu et al.
of brain, liver, kidneys and testes in the males administered 2011), Sprague-Dawley rats were given isopentane by oral
1000 mg/kg, and significant increases in absolute and relative gavage in doses of 100, 300, or 1000 mg/kg/day. Dosing of
adrenal weights in the high-dose group females. The only males was initiated 10 weeks prior to mating and continued
to the end of the mating period. Dosing of females was initi- 11. Human observations
ated 2 weeks prior to mating and continued to postnatal day
Patty and Yant (1929) reported that n-pentane had no effects
21. Parental parameters assessed included observations of
on human volunteers exposed for 10 min at levels up to 0.5%
male and female rats prior to mating, and, particularly for
in air.
female rats, through the gestational and lactational periods.
Normal- and iso-pentane were tested for skin irritation
Observations of offspring included number of live births as
potential in humans in unpublished studies involving 30 vol-
well as survival and weight gain during the lactational period.
unteers. The overall irritation scores were reported as 0.3 (of
There were no differences in mating frequency, precoital time,
8.0) in both studies, and the substances were judged as mildly
productive pregnancies, or duration of gestation; neither the
irritating to the skin.
number of stillborn offspring nor the number of offspring with
external anomalies was increased; and there were no differ- II. Toxicological Properties of C6 Aliphatic
ences in offspring viability to terminal sacrifice at postnatal Hydrocarbon Solvents (Hexanes) (Category 6
day 21. The overall no adverse effect level for reproductive solvents)
toxicity was 1000 mg/kg/day, the highest dose tested.
In related studies McKee et al. (2000) reported a two- 1. Introduction
generation reproductive toxicity study of vapor recovery unit C6 aliphatic solvents are complex and variable combinations of
gasoline, a complex substance composed primarily of C4–C6 aliphatic constituents, primarily n-hexane, isohexane isomers,
aliphatic constituents including 37% pentane isomers. Rats cyclohexane, and methylcyclopentane in varying amounts,
were exposed by inhalation at target concentrations of 5000, and boiling in the range of approximately 55–85°C. These
10000, or 20000 mg/m3 (approximately 1700, 3400, or 6800 solvents are manufactured from crude oil by atmospheric
ppm) with the highest concentration being approximately half distillation, fractionated by distillation, and further refined,
of the lower explosive limit. There were no effects in repro- usually by hydrogenation, to reduce levels of sulfur- and
ductive parameters and no changes in sperm counts or time to nitrogen-containing constituents. These substances are listed
onset of developmental landmarks. The only systemic effects under CAS registry numbers for TSCA purposes in the US,
were changes in the kidneys of male rats consistent with a2u- under EINECS numbers in Europe for compliance with the
globulin-mediated effects. The overall no adverse effect con- Dangerous Substance Directive, and, for REACH purposes,
centration was 20000 mg/m3, the highest concentration tested. were registered under the general heading “Hydrocarbons, C6”
in accordance with the naming convention introduced by the

9. Carcinogenicity European Hydrocarbon Solvent Producers Association (HSPA
Because pentane solvents are not mutagenic and do not cause 2011, Table 2). One feature of these solvents is that they may
target organ toxicity, they have not been priority substances for or may not contain n-hexane at toxicologically relevant levels.
carcinogenicity tests. Because the toxicological properties of n-hexane differ from
those of other C6 aliphatic hydrocarbons, substances that con-
10. Neurological studies tain n-hexane at concentrations  5% are differentiated from
As noted previously, early studies in mice demonstrated that those with  5% n-hexane by the naming convention.
n-pentane and cyclopentane could cause anesthetic effects at
2. Physical and chemical properties
very high exposure levels (Swann et al. 1974, Virtue 1948). In
more recent studies of acute CNS effects of n-pentane, isopen- The C6 aliphatic solvents are colorless liquids at room temper-
tane isomers, and cyclopentane (Lammers et al. 2011, McKee ature, with slightly disagreeable odors. The benzene and sulfur
et al. 2011), there were no effects at levels up to 20,000 mg/m3 contents of hydrocarbon solvents in the C6 aliphatic hydrocar-
(approximately 6000 ppm), the highest concentrations tested. bon category are low, with benzene levels typically  1 ppm
Because of the structural similarities between pentane (Table 3). The substances in this category have similar physi-
isomers and n-hexane, studies were conducted to determine cal and chemical properties, as summarized in Table 20.
whether C5 aliphatic molecules could cause peripheral neu-
ropathy. Takeuchi et al. (1980) reported that in studies in which 3. Toxicokinetics
rats were exposed to n-pentane, 12 hours/day for 16 weeks at 3.1 Commercial hexane
3000 ppm (approximately 9000 mg/m3), there were no effects
on nerve conduction velocity or distal latency and no patho- The toxicokinetic properties of hexane isomers were evaluated
logical changes in the peripheral nerves. Frontali et al. (1981) in a study in rats in which commercial hexane42 was adminis-
assessed the neurotoxic potential of n-pentane in a study in tered to rats by intravenous injection, inhalation, and dermal
which the rats were exposed 9 hours/day, 5 days/week for up administration (unpublished information). Inhaled hexane was
to 30 weeks at 3000 ppm (approximately 9000 mg/m3). There well absorbed and widely distributed to various tissues but
were no pathological changes in nervous system tissue. This without evidence of accumulation in any specific tissue.
was in contrast to observations on rats exposed to n-hexane,
in which the characteristic histological changes associated
with axonal neuropathy were observed. Urinary metabolite
42Commercial hexane is a complex C6 aliphatic solvent. The specific
measurements confirmed that n-pentane is metabolized to and
excreted as the corresponding alcohol and does not form the sample tested in this study contained 52% n-hexane, 16% methylcyclo-
pentane, and 3% cyclohexane, with the remaining material consisting of
neurotoxic metabolite 2,5-hexanedione. iso-hexane isomers.
Table 20. Physical and chemical properties of C6 aliphatic solvents.

Property n-Hexane Isohexane Cyclohexanea Methyl-Cyclopentane
Molecular Weight 85 85 84 84
Physical State at 20°C and Liquid, bright and clear Liquid, bright and clear Liquid, bright Liquid, bright and
1013 hPA and clear clear
Pour Point  20°C  20°C  20°C  20°C
Boiling Point 69°C (ASTM D5950) 55–85°C (ASTM D5950) 81°C 72°C (ASTM D5950)
Relative Density 0.66 g/cm3 (ISO 12185) 0.65–0.70 g/cm3 (ISO 0.77 g/cm3 0.75 g/cm3
12185)
Vapor Pressure 20 kPa at 20°C 19–50 kPa at 20°C 12.4 kPa at 24°C Approximately 20 kPa
at 20°C
Surface Tension 18–20 mN/m at 25°C 18–20 mN/m at 25°C 25 mN/m at 25°C 13 mN/m at 25°C
(Wilhelmy plate (Wilhelmy plate (Wilhelmy (Wilhelmy plate
methodology) methodology) plate methodology)
methodology)
Water Solubility 9.5 mg/l at 20°C 9–14 mg/l at 20°C 52 mg/l at 20°C 42 mg/l at 20°C
Octanol/Water Partition 3.6 3.6–4.0 3.4 3.4
Coefficient
Flash Point -20°C (DIN 51755) 0°C (DIN 51755)  20°C  23°C
Flammability 1.1–7.5% (v/v) in air 1.2–8.3% (v/v) in air 1.3–8.4% (v/v) 1.0–8.7% (v/v) in air
in air
Self-Ignition Temperature  225°C (ASTM E 659)  200°C (ASTM E 659) 260°C 258°C
Viscosity 0.4–0.7 mm2/s (ASTM 0.4–0.7 mm2/s (ASTM Not reported Not reported
D7042) D7042)
Odor Threshold ∼ 60 ppm (210 mg/m3) Not reported Not reported Not reported
(IPCS 1991)
Maximally Attainable Vapor Approximately 100,000 ppm Approximately 100,000 ppm Approximately Approximately
Concentration 100,000 ppm 100,000 ppm
Conversion Factor 1 ppm  3.5 mg/m3 1 ppm  3.5 mg/m3 1 ppm  3.5 mg/ 1 ppm  3.5 mg/m3
(ppm/mg/m3) m3
aData taken from US Environmental Protection Agency (1994).
The constituents of commercial hexane were metabolized conducted 6, 24, and 72 h after oral administration indicated
and eliminated, primarily by exhalation and urinary excretion. that the most of the material remaining in the carcass was in
The half-life for hexane in blood was approximately 9–10 h. adipose tissue. The estimated half-lives for plasma and tissue
There were no obvious differences in toxicokinetic properties concentrations were on the order of 10–15 h.
of commercial hexane in acute as compared to repeat exposure Absorption of cyclohexane by male F-344 rats after dermal
studies, suggesting that commercial hexane constituents do exposure was estimated as 35–60% (60–100 mg/cm2/h) after a
not induce their own metabolism. Studies of humans exposed dermal dose of 1 mg/cm2, and 4% (0.65 mg/cm2/h) after admin-
occupationally indicated that n-hexane is metabolized to istration of 100 mg/cm2. However, it should be noted that the
2-hexanol and further to hexanedione, as shown in Figure 4. test materials were applied under an occluded patch, which pre-
In contrast, in rats exposed by inhalation, the principal urinary vented evaporation and exaggerated percutaneous absorption.
metabolite is 2-hexanol. Following dermal contact, it is more
likely that hexane would evaporate than be percutaneously 3.2.2 Toxicokinetic studies of cyclohexane in humans
absorbed. In in vitro studies with rat skin, rates of absorption
Volunteer studies indicated that inhaled cyclohexane is well
for aliphatic C6–C8 constituents were reported to be  1 mg/
absorbed, metabolized to cyclohexanol, and excreted in the
cm2/hr (Tsuruta 1982).
urine, primarily as glucuronide metabolites. The estimated
elimination half-lives were 14–18 h (Mraz et al. 1998). In
3.2 Cyclohexane
a study of lung uptake of cyclohexane, Mutti et al. (1981)
3.2.1 Toxicokinetic studies of cyclohexane in male F344 rats estimated alveolar retention to be approximately 34% of the
In an unpublished study to assess the fate of cyclohexane in inhaled dose.
male F-344 rats following intravenous administration, 54% of
4. Acute toxicity
the administered dose was excreted in the breath after 1 hour,
80% in 24 h, and 83% in 72 h, with 14% being excreted in the Data indicate that C6 aliphatic solvents and their constituents
urine (summarized in the existing substances risk assessment are not acutely toxic when assessed in tests following current
for cyclohexane, European Chemicals Bureau 2004). Virtually guidelines for these properties (Table 21); however, these
all of the exhaled cyclohexane was eliminated as parent com- solvents can produce acute CNS depression if inhaled at high
pound whereas material in the urine was eliminated as polar levels and chemical pneumonitis if taken into the lungs in the
metabolites. liquid state. Similarly, initial reports indicated that hexane sol-
In oral studies in male F-344 rats, approximately 90% of vents were not ocular or dermal irritants and did not induce
the administered material was absorbed. Depending on the allergic contact dermatitis (Hine and Zuidema 1970). Based
dose administered, approximately 60–80% of the cyclohexane on more recent dermal irritation toxicity tests (Jacobs et al.
was eliminated by exhalation and the remaining material was 1987), however, hexane solvents are classified as dermal irri-
eliminated in the urine. Investigations of tissue distribution tants under the provisions of the European adaptation of GHS
Table 21. Summarized results of acute and short-term toxicity tests of C6 hydrocarbon solvents and their constituents.
Substance Endpoint Value Reference
n-hexane Acute Oral Toxicity  16750 mg/kg Hine and Zuidema (1970)
Acute Dermal Toxicity  3350 mg/kg Hine and Zuidema (1970)
Acute Inhalation Toxicity ∼73,000 ppm (255,500 mg/m3) Hine and Zuidema (1970)
Ocular Irritation Practically not irritating Hine and Zuidema (1970)
Skin Irritation Mild irritation Hine and Zuidema (1970)
Dermal Sensitization Not sensitizing, based on animal data Basketter et al. (2000), Fedorowicz et al.
(2004)
Cyclohexane Acute Oral Toxicity  2000 mg/kg Unpublished dataa
Acute Dermal Toxicity  2000 mg/kg Unpublished dataa
Acute Inhalation Toxicity  32880 mg/m3 Unpublished dataa
( 9394 ppm)
Ocular Irritation Practically not irritating Unpublished dataa
Dermal Irritation Practically not irritating Jacobs et al. (1987)
Dermal Sensitization Not sensitizing, based on animal data Unpublished dataa
aAcute toxicity and irritation data are summarized in the existing substances risk assessment for cyclohexane (European Chemicals Bureau 2004).
(European Commission 2008). Studies to assess the potential 4, 6, and 8 weeks of treatment. There was little evidence of
for allergic contact dermatitis using the local lymph node these effects in rats treated with 2-methylpentane, 3-methyl-
assay provided evidence that hexane solvents do not cause skin pentane, and methylcyclopentane (Ono et al. 1981) or complex
sensitization (Basketter et al. 2000, Fedorowicz et al. 2004). n-hexane-free C6 isomeric solvents (Egan et al. 1980).
5. Repeated dose toxicity 5.1.2 Repeated inhalation toxicity studies

5.1 Repeated dose toxicity studies of n-hexane In a 14-week study, male F-344/N rats were exposed by
inhalation to 0 or 2000 ppm (approximately 7000 mg/m3)
5.1.1 Repeated oral toxicity studies
of n-hexane, for 14 hours/day, 7 days/week. There were no
Rats and mice were given n-hexane in doses of 568, 1135, or clinical signs of neuropathy or histopathological changes in
3973 mg/kg/day, by oral administration on a daily basis for peripheral nerves. However, n-hexane exposure resulted in
90 days. During treatment, the rats were examined for body statistically significant reductions in motor activity, fore- and
weight, clinical signs, mortality and neurological effects and hind-limb grip strength, and startle response. The authors
were sacrificed when they were observed to exhibit hind noted an improvement in functional tests over a 6-week recov-
limb paralysis or at scheduled termination at the end of the ery period (Pryor et al. 1983).
dosing period. Histological examination of the nervous sys- Cavender et al. (1982, 1984) exposed rats to 0, 4000, 6500,
tem tissue revealed that n-hexane and its metabolites caused or 10000 ppm (14000, 23000, or 35000 mg/m3) n-hexane via
morphological changes indicative of “giant axonal” neuropa- inhalation, 6 hours/day and 5 days/week for 90 days. There
thy including multifocal axonal swellings, adaxonal myelin were no clinical signs of treatment-related changes, and neuro-
in-folding, and paranodal myelin retraction. The authors also logical function tests were normal. Para-nodal axonal swelling
reported evidence of testicular atrophy, but a detailed char- was however observed in 1/5 and 4/5 male rats in the 6500 ppn
acterization of the testicular effects was outside the scope of (23000 mg/m3) and 10000 ppm (35000 mg/m3) groups.
the study (Krasavage et al. 1980). The study also provided Dunnick et al. (1989) exposed B6C3F1 mice to n-hexane at
evidence that n-hexane-induced “giant axonal” neuropathy levels of 500, 1000, 4000, or 10000 ppm (1500, 3000, 12000,
was dependent on the production of a g-diketone metabolite or 30000 mg/m3), 6 hours/day and 5 days/week for 13 weeks,
which is unique to n-hexane. For example, the authors showed or to 1000 ppm (3000 mg/m3) 22 hours/day and 5 days/week
that the severity of hind limb paralysis in rats exposed to for 13 weeks. Histological findings included mild inflamma-
equimolar concentrations of n-hexane and its metabolites, is tory, erosive, and regenerative lesions in the olfactory and
directly proportional to its step on the metabolic chain (i.e., respiratory epithelium in the high- and continuous-exposure
in order of decreasing potency; 2,5-hexanedione  5-hydroxy- groups. Functional investigations revealed a decrease in loco-
2-hexanone  2,5-hexanediol  2-hexanone  2-hexanol  motor activity in female mice from the 10000 ppm and 1000
n-hexane). Krasavage and colleagues also noted that severity of ppm continuous-exposure groups. Pathological investigations
hind limb paralysis correlated with peak serum concentrations revealed axonal swellings in mice from the 10000 ppm and
of 2,5-hexanedione (threshold  50 mg/ml). Only rats treated 1000 ppm continuous-exposure groups. The authors consid-
with concentrations of n-hexane high enough to achieve the ered that the severity of the peripheral nerve lesion was mild.
serum threshold level of 50 mg of 2,5-hexanedione showed
any signs of peripheral nerve damage or hind limb paralysis 5.2 Repeated dose toxicity studies of commercial hexane
(Krasavage et al. 1980).
5.2.1 Repeated toxicity studies of commercial hexane by
Male rats (5–7/group) were given n-hexane by gavage in
inhalation
olive oil daily for 8 weeks. The rats were given 800 mg/kg/day
for the first 4 weeks, 1047 mg/kg/day for the next 2 weeks, and Rats and mice were repeatedly exposed by inhalation, 6 hours/
2022 mg/kg/day for the last 2 weeks. There were small (5–10%) day and 5 days/week for 13 weeks at target concentrations of
but statistically significant reductions in motor nerve conduc- 900, 3000, or 9000 ppm (2700, 9000, or 27000 mg/m3) (Duffy
tion velocities (MCV) in treated rats compared to controls after et al. 1991). There were no treatment-related effects on survival
or terminal body weights. Liver weights were increased in mice (McCarroll et al. 1981a, 1981b) using bacterial assays. These
and male rats in the high-exposure group; the histopathologi- tests all produced negative results, and n-hexane was not con-
cal examination revealed slight hemorrhage in the livers of 3 sidered to have mutagenic potential.
high-dose male rats, with inflammation in two. The authors
also reported an increased incidence of male rat kidney effects 6.2 Genetic toxicity studies of other C6 aliphatic solvents
consistent with an a2u-globulin-mediated process. The no Commercial hexane was found to be inactive in in vitro tests
observed adverse effect concentration was 3000 ppm (9000 for gene mutation using both bacterial (Salmonella) and mam-
mg/m3), based on liver and kidney effects in the male rats. malian cell (CHO/HGPRT) mutagenesis tests (Kirwin et al.
1991). In chromosomal aberration tests, commercial hexane
5.3 Repeated dose toxicity studies of cyclohexane was not active either under in vitro conditions in assays using
The potential for repeated exposure to cyclohexane to cause CHO cells or in vivo in rat bone marrow following 5 con-
systemic effects was investigated in a repeated inhalation toxic- secutive days of inhalation exposure at levels up to 9000 ppm
ity study in which rats and mice were exposed 6 hours/day and (27000 mg/m3) (Daughtrey et al. 1994b).
5 days/week for 14 weeks at target concentrations of 500, 2000,
or 7000 ppm (1500, 6000, or 21000 mg/m3) (Malley et al. 6.3 Genetic toxicity studies of cyclohexane
2000). One group of animals was exposed at the high level and A summary of published and unpublished information pro-
then held for 4 weeks without treatment, to assess recovery. The vided no indication that cyclohexane was mutagenic under
animals survived the exposure period with no effects on termi- in vitro or in vivo conditions (US Environmental Protection
nal body weights. In the rats, the only effect was an increase Agency 2003).
in relative liver weights with an increased incidence of hepatic
centrilobular hypertrophy at the end of the exposure period. 7. Developmental toxicity
The liver changes had reversed by the end of the recovery
period. In the mice there were small increases in hematological 7.1 Developmental toxicity studies of n-hexane
parameters in both males and females from the high-exposure Several developmental toxicity studies of n-hexane (Bus, et
group as well as increased liver weights. The liver weights were al., 1979; Marks, 1980; Mast, 1987; Mast et al., 1988a) by
not significantly different at the end of the recovery period. As oral and inhalation routes of administration provided results
the changes in liver weight appeared to be evidence of adaptive similar to those in the commercial hexane studies (Appendix
rather than adverse processes and the hematological changes section 7.2). There was some evidence of developmental
were within the normal range, the no adverse effect concentra- delays in some studies but no reports of fetal death and or
tion for both rats and mice was judged to be 7000 ppm (21000 malformation.
mg/m3), the highest concentration tested.
7.2 Developmental Toxicity Studies of Commercial Hexane
5.4 Repeated dose toxicity studies of The potential for developmental toxicity of commercial
methylcyclopentane hexane was assessed in rats and mice (Keenan et al. 1991,
Yang et al. (2014) reported a study in which rats were exposed Daughtrey et al. 1994a). Animals were exposed 6 hours/day
by inhalation, 6 hours/day and 5 days/week for 13 weeks, at to vapors of commercial hexane at concentrations of 900,
levels of 290, 1300, or 5870 ppm (880, 3900, or 18000 mg/ 3000, or 9000 ppm (880, 3900, or 18000 mg/m3) on ges-
m3). Following the exposure period, the surviving animals tational days 6–15. The animals were examined on a daily
were sacrificed and examined at the gross and microscopic lev- basis during the gestational period for clinical signs, body
els. There were no unscheduled mortalities during the study. weight changes, and food and water consumption. The ani-
Terminal body weights of treated animals were not different mals were sacrificed on gestational day 20, and the uterine
by statistical criteria. There were no statistically significant contents were examined. The rats from the high-exposure
hematological changes, and the only significant change in group exhibited reduced body weight gain and reduced food
clinical chemistry values was a decrease in Cl levels; however, consumption, and both rats and mice from the high-exposure
the differences were small and within the normal physiological group had discoloration in the lungs. There were no differ-
range. There were no differences in absolute organ weights, but ences in gestational parameters, including the number of
there were significant increases in relative liver (both genders) viable and non-viable fetuses. There were no differences in
and kidney (females only) weights. There were no gross abnor- fetal body weights and no treatment-related differences in the
malities and no pathological findings in any of the organs. In frequencies of external, visceral, or skeletal malformations.
summary, repeated exposure of rats to methylcyclopentane The frequencies of variations were not increased in the rats
resulted in small increases in liver and kidney weights in the at any treatment level; in the mice there was a statistically
high-exposure group (5870 ppm, 18000 mg/m3), but these significant increase in the frequencies of two skeletal varia-
were not associated with clinical or pathological changes. tions in the 9000 ppm (27000 mg/m3) group. The overall
conclusions were that there were no developmental effects in
6. Genetic toxicity rats at exposure levels up to 9000 ppm (27000 mg/m3), and
that in mice, there were no serious developmental effects but
6.1 Genetic toxicity studies of n-hexane
there was slight evidence of developmental delays at 9000
The genotoxic potential of n-hexane was tested for mutation ppm (27000 mg/m3), a level that also produced some slight
(Mortelmans et al. 1986) as well as DNA damage and repair evidence of maternal effects.
7.3 Developmental toxicity studies of cyclohexane re-initiated on postnatal day 4 and continued through weaning
(postnatal day 21). At that point the surviving animals in the
The potential for cyclohexane to cause developmental effects
parental generation were sacrificed, and the F1 offspring were
was assessed by Kreckman et al. (2000). Rats and rabbits were
exposed on a similar schedule through a second generation.
exposed to cyclohexane by inhalation at target concentrations
The F2 offspring were examined and sacrificed on postnatal
of 500, 2000, or 7000 ppm (1500, 6000, or 21000 mg/m3). The
day 28. There were no statistically significant differences in
rats were exposed on gestational days 6–15, and rabbits were
any of the reproductive parameters. The only exposure-related
exposed on gestational days 6–18. The animals were sacrificed
effects observed were reductions in body weight gain in off-
near the end of gestation (gestational day 21 for rats, gestational
spring exposed to 9000 ppm (27000 mg/m3) and evidence
day 29 for rabbits), and the uterine contents were examined.
of hyaline droplet nephropathy in the kidneys of the parental
In the rat study, there were no unscheduled deaths. Maternal
and F1 males exposed to 9000 ppm (27000 mg/m3). The no
weight gain was significantly reduced in the 7000 ppm (21000
observed adverse effect concentration for reproductive effects
mg/m3) group during gestation, but there were no differences
was 9000 ppm (27000 mg/m3), and the no observed effect
in terminal body weights. There was a small but statistically
level for all effects was 3000 ppm (9000 mg/m3).
significant decrease in the number of implantations in the 7000
ppm (21000 mg/m3) group, but there were no significant dif-
8.3 Reproductive toxicity studies of cyclohexane

ferences in numbers of live fetuses, fetal weights, frequency of
malformations, or variations. In the rabbit study, there were no The potential for cyclohexane to cause reproductive effects
significant differences in survival or body weight gain. There was assessed by Kreckmann et al. (2000) in a two-generation
was a significant reduction in corpora lutea in the 7000 ppm reproductive toxicity test in rats at exposure levels of 500, 2000,
(21000 mg/m3) group, but no differences in implantations, live- or 7000 ppm (1500, 6000, or 21000 mg/m3). Exposures were
born fetuses, or fetal weights, and the numbers of fetuses with initiated 10 weeks prior to mating, and continued through the
malformations and/or variations was not increased. Accord- mating period to gestational day 20. Exposure of dams was
ingly, based on studies in both rats and rabbits, the no observed suspended from gestational day 20 to lactational day 4 and then
adverse effect concentration for developmental effects was re-initiated and continued to the end of lactation. Exposure of
7000 ppm (21000 mg/m3), the highest level tested. offspring was initiated after the lactational period and continued
for a second cycle to the end of the lactational period for the F1
8. Reproductive toxicity females. Exposures were for 6 hours/day, 5 days/week during
the pre-mating periods and on a daily basis through the mat-

8.1 Reproductive toxicity studies of n-hexane
ing, gestational, and lactational periods (aside from the suspen-
O’Donoghue et al. (1978) reported that exposure of rats to 2,5- sion at the end of gestation and start of lactation, as described
hexanedione resulted in testicular atrophy. A subsequent publi- above). There were small differences in body weight gain in
cation from the same group (Krasavage et al. 1980) confirmed the rats exposed to 7000 ppm (21000 mg/m3). However, there
that these effects could be produced by n-hexane and other were no differences in reproductive indices (mating, fertility, or
n-hexane metabolites, albeit at higher treatment levels than were gestation indices, implantation efficiency, or gestation length)
required with 2,5-hexane dione. In a series of publications in in either generation. The only significant difference was a small
which 2,5-hexanedione was used as the test substance, Boekel- but statistically significant reduction in the percentage of live-
heide and colleagues (Boekelheide et al. 1987, Boekelheide born offspring in the first generation, but this was not considered
et al. 1988, Boekelheide and Eveleth 1988) showed that the to have been toxicologically important as the result was higher
underlying process leading to testicular atrophy involved inhi- than the mean of the historical incidence of live-born offspring
bition of microtubule assembly leading to compromised Sertoli in the testing laboratory, and no differences were found in the
cell function. This hypothesis was supported by evidence that second generation. As weight gain in offspring from the 7000
the testicular atrophy was not associated with changes in levels ppm (21000 mg/m3) group was reduced in comparison to con-
of testosterone or gonadotropins (Chapin et al. 1982). trol values, the authors concluded that the no observed adverse
Studies of the potential for reproductive effects of n-hexane effect concentration was 2000 ppm (6000 mg/m3).
were similar to the results of the studies of commercial hexane.
There were no effects on fertility (Marks et al. 1980, Mast et al. 9. Carcinogenicity
1988c) or sperm morphology (DeMartino et al. 1987, Linder Daughtrey et al. (1999) assessed the potential for commercial
et al. 1992, Mast et al. 1988b). hexane to cause cancer in rats or mice. The animals were
exposed to hexane vapor, 6 hours/day and 5 days/week for
8.2 Reproductive toxicity studies of commercial hexane two years, at levels of 900, 3000, or 9000 ppm (2700, 9000,
The potential for commercial hexane to cause reproductive or 27000 mg/m3). At termination, the surviving animals
effects was assessed in a two-generation study in Sprague- were sacrificed and grossly examined. Tissues were taken for
Dawley rats (Daughtrey et al. 1994a). The animals were microscopic examination. There were small but statistically
exposed for 6 hours/day to commercial hexane vapor at levels significant differences in terminal body weights in the mid-
of 900, 3000, or 9000 ppm (2700, 9000, or 27000 mg/m3). Prior and high- exposure group rats and in female mice from the
to mating, the animals were exposed 6 hours/day and 5 days/ high-exposure group. In the rats, the only histological finding
week for 10 weeks. During mating and gestation, the rats were of note was epithelial cell hyperplasia in the nasoturbinates
exposed daily. Exposures were suspended on gestational day and larynx of the exposed animals, considered to be indicative
20, to allow the dams to give birth. Exposure of the dams was of upper respiratory tract irritation. There was no evidence of
tumor induction in the rats. In the mice, there was a significant In summary, these studies demonstrated that cyclohexane
decrease in the severity of cystic endometrial hyperplasia. can produce subtle CNS effects in rats at levels  7000 ppm
There was a statistically significant increase in the combined (21000 mg/m3) but also confirmed that the effects were acute
incidence of hepatocellular adenomas and carcinomas in the and reversible within 24 h of exposure.
female mice from the high-exposure group. There was also an
increase in the incidence of pheochromocytomas in the female 10.1.2 Acute neurological effects in humans
mice, but this was considered to have been a statistical effect
The potential for cyclohexane to cause acute CNS effects in
associated with an unusually low incidence in the control
humans was investigated by Lammers et al. (2009). Volunteers
group and not treatment-related. There were no tumors in the
were exposed to cyclohexane vapors at 86 mg/m3 or 860 mg/
male mice. The authors concluded that chronic exposure to
m3 (28 or 280 ppm) for 4 h. Neurobehavioral testing was con-
commercial hexane vapor was not carcinogenic to F-344 rats
ducted 90 min before exposure, 45 min and 165 min after the
or to male B6C3F1 mice but that it did result in an increase in
start of exposure, and 90 min after exposure, using a comput-
liver tumors in female mice.
erized neurobehavioral test battery. Based on self-reporting,
some subjective measures including headache and dry throat
10. Neurological studies
were increased, but there was no apparent eye or respiratory
The primary concern about n-hexane relates to the associa- tract irritation. Further, there were no differences in any of the
tion between repeated exposure of humans to n-hexane at high objective tests that were considered treatment-related. In sum-
levels and the development of peripheral neuropathy. As summary, exposures of volunteers for 4 h to cyclohexane at levels
marized by the IPCS (1991), the first reports of peripheral up to 860 mg/m3 (280 ppm) produced no objective evidence
neuropathy appeared in the Japanese literature in the 1960s for acute CNS effects.
and were quickly followed by publications in Italian journals.
The characteristic neurological effects were reproduced in rats 10.2 Persistent neurological effects
in studies involving exposure by inhalation (Altenkirch et al.
10.2.1 Persistent neurological effects in animals
1982, Frontali et al. 1981, Takeuchi et al. 1980) and oral admin-
istration (Krasavage et al. 1980). The neurological effects are Repeated exposure to n-hexane can cause peripheral neuro-
mediated through 2,5-hexanedione, which is derived from logical effects in rodents (Altenkirch et al. 1982, Frontali et al.
n-hexane but not from other structurally similar hydrocarbons. 1981, Krasavage et al. 1980, Ono et al. 1981, Takeuchi et al.
Studies in rats have shown that although n-hexane is neuro- 1980). The underlying mechanism involves metabolism of
toxic, similar substances including n-pentane, iso-hexane iso- n-hexane to 2,5-hexane dione (Figure 4) which inhibits micro-
mers, cyclohexane, and n-heptane do not produce peripheral tubule assembly (Boekelheide 1987), ultimately interfering
neuropathy (Egan et al. 1980, Frontali et al. 1981, Ono et al. with axoplasmic flow. Other alkanes with similar structures,
1981, Takeuchi et al. 1980). including n-pentane, iso-hexane, cyclohexane, and n-heptane
are not metabolized to 2,5-hexane dione at toxicologically rel-
10.1 Acute neurological studies evant levels and do not cause similar peripheral nervous system
10.1.1 Acute neurological effects in animals effects (Egan et al. 1980, Frontali et al. 1980, Malley et al. 2000,
Ono et al. 1981, Soiefer et al. 1991, Takeuchi et al. 1980).
The potential for cyclohexane to cause acute CNS effects
in animals was studied in separate investigations. In a study
10.2.1.1 Hypothesized mode of action for n-hexane-induced
by Christoph et al. (2000), rats were trained to press levers
peripheral neuropathy
on a fixed ratio-fixed interval schedule of food presentation
and then exposed to cyclohexane at levels of 500, 2000, or The mode of action of n-hexane-induced peripheral neuro-
7000 ppm (1500, 6000, or 21000 mg/m3) for 6 h. When tested toxicity has been extensively researched in several animal
shortly after exposure (the session began 30 min after expo- species (ATSDR 1999, Couri and Milks 1982, USEPA 2005)
sure), the fixed ratio response rate was decreased by 11% in the and several required and measurable steps (or key events)
7000 ppm (21000 mg/m3) group, but there were no effects on leading up to the peripheral nerve damage response have been
pause duration or performance pattern. There were no effects described (Table 22). The first step in the mode of action for
in the groups exposed to either 500 ppm or 7000 ppm (1500 n-hexane neurotoxicity involves the metabolism of n-hexane to
or 21000 mg/m3) immediately after exposure, and no effects 2,5-hexanedione, which preferentially takes place in the liver
in rats from any of the treatment groups in sessions conducted (Couri and Milks 1982). This is followed by the formation of
24 or more hours after exposure. pyrrole adducts with the free epsilon-amino groups of lysine
In a study reported by Lammers et al. (2009), rats were residues in neurofilament proteins, which are then modified to
exposed to cyclohexane at levels of 1400, 8000, or 28000 mg/ covalent crosslinks by oxidation (Lapadula et al. 1986, Sanz
m3 (462, 2640, or 9240 ppm), 8 hours/day for 3 consecutive et al. 1995). Pyrrole adducts have been consistently found in
days, and then evaluated for effects on performance using the urine of rats exposed to n-hexane or its metabolites (Kessler
a visual discrimination task in a functional observation test et al. 1990, Mateus et al. 2002). Additionally, rats exposed to
battery, and for effects on motor activity. The only significant a 2,5-hexanedione analog with increased capacity for pyrrole
effect was an increase in errors and in long ( 6 s) response formation induce more severe forms of hind limb paralysis and
times. The differences were significantly different only in the at faster rates (Anthony et al. 1983a, Anthony et al. 1983b,
28000 mg/m3 (9240 ppm) group and were not observed when Graham et al. 1982b). Oxidation of pyrrole adducts then leads
the animals were tested 24 h after exposure. to the formation of covalent cross links. The direct effects of
Table 22. Hypothesized key events, with animal evidence, for n-hexane-induced peripheral neuropathy.
# Key Events Evidence in Animals Key References
1 Metabolism to the reactive 2,5- YES. (Abou-Donia et al. 1982, Couri and Milks 1982,
hexanedione • Metabolism of n-hexane is a requirement for Krasavage et al. 1980, Schaumburg and Spencer
end effects. Hind limb paralysis following 1978)
oral exposure in rats, in order of increasing
severity, correlates with step on metabolic
chain (2,5-HD most potent, n-hexane least
potent).
• Severity of hind limb effects in rats
correlates with peak serum levels of 2,5-
hexanedione.
• 1-hexanol, which is not metabolized to
2,5-hexanedione, does not cause peripheral
neuropathy.
• Methyl-n-butyl ketone, which is also
metabolized to 2,5-HD, also causes
peripheral neuropathy.
• Chronic (4-month) exposure to n-hexane-
free mixture of hexane isomers in rats does
not induce neuropathy.
2 Pyrrole adduct formation YES (Anthony et al. 1983a, Anthony et al. 1983b,
• 2,5-hexanedione induces pyrrole formation DeCaprio et al. 1988, Graham et al. 1982b,
with neurofilament proteins in spinal cord Lapadula et al. 1986a, Mateus
and peripheral nerves both in vivo and in et al., 2002, Sanz et al. 1995)
vitro.
• 3,4-dimethyl-2,5-hexanedione, a 2,5-
hexanedione analog with increased capacity
for pyrrole formation, induces more severe
limb paralysis, at a lower dose and at a faster
rate.
• 2,5-hexanedione analogs with slower

pyrrole-forming capacity and competitive
inhibition with metal chelation induce
milder symptoms of hind limb paralysis at
equivalent doses.
3 Oxidation of pyrrole adducts to YES (Genter et al. 1988)
form protein crosslinks • Diketones which form stable pyrrole adducts
that cannot be oxidized to form protein
crosslinks do not induce typical signs of
peripheral neuropathy.
4 Axonal swelling and degeneration YES (Genter et al. 1988, Politis et al. 1980,
• Axonal swelling, due to condensation Schaumburg and Spencer 1978)
of neurofilament protein crosslinks, and
axonal degeneration, are the earliest
histopathological indicator of peripheral
neuropathy.
• Axonal neurofilament accumulation is seen
with direct 2,5-hexanedione exposure to
nerve fibers.
• No axonal swelling is seen with exposure to
diketones which are incapable of forming
protein crosslinks. Animals in these studies
show no signs of peripheral nerve damage.
5 Peripheral nerve damage YES (Abou-Donia et al. 1982, Dunnick et al. 1989,
• Multiple species Krasavage et al. 1980, Schaumburg and Spencer
1978)
the covalently-linked neurofilaments include neurofilament methyl ethyl ketone (which by itself does not produce neurolog-
aggregation, axonal swelling, and degeneration, all of which ical effects) potentiates the neuropathic response with n-hexane
ultimately lead to the disruption of axoplasmic flow and even- exposure (Altenkirch et al. 1978). In addition, n-heptane, which
tually neuronal death (Politis et al. 1980). is also metabolized to a g-diketone (2,5-heptanedione) but
The study by Krasavage and colleagues (1980) showed that much less efficiently than n-hexane, does not cause peripheral
the production of the g-diketone metabolite is a requirement for neuropathy (Bahima et al. 1984, Takeuchi et al. 1980).
the histopathological changes in proximal and distal nerves that When pyrrole-forming capacity is blocked, either by metal
are the hallmark of n-hexane-mediated neuropathy. This is also chelation or with 2,5-hexanedione analogs with reduced pyr-
bolstered by evidence showing that stimulation of liver CYP2E1 role-forming capacity, effects on the peripheral nervous system
(the liver enzyme responsible for n-hexane metabolism) by neurotoxicity can be prevented (DeCaprio et al. 1988, Mateus
et al. 2002). To demonstrate the essential requirement for pyr- metabolism of n-hexane is qualitatively similar across several
role formation (note that pyrrole formation is a necessary step, animal species and humans. 2,5-Hexanedione, the key metabo-
but is not sufficient in and of itself without protein crosslink- lite driving axonal neuropathy, has been detected in human urine.
ing) in the mode of action for n-hexane-induced neuropathy, Mutti et al. (1984) evaluated the urine samples of shoe factory
rats were treated with equimolar concentrations of 2,5-hexane- workers and showed that urinary 2,5-hexanedione was the
dione and 3,4-dimethyl-2,5-hexanedione every 8 h (Graham best biomarker for occupational n-hexane exposure. Although
et al. 1982a). Rats treated with 3,4-dimethyl-2,5-hexanedione, other elements of the mode of action of n-hexane-induced dis-
but not 2,5-hexanedione, developed severe hind limb paralysis tal axonopathy (see 10.2.1.1), they have not been evaluated in
within 3 days. In all cases, hind limb paralysis was consistently humans in great detail compared to rodents; pyrrole-like deriva-
preceded by neurofilament aggregation and para-nodal axonal tives have been found in the urine of human volunteers exposed
swelling. To prove that pyrrole oxidation was a critical step to 146 ppm (442 mg/m3) n-hexane for 3 h (Kessler et al. 1990).
in the formation of covalent crosslinks, rats were treated with Furthermore, there is an abundance of evidence for axonal swell-
similar concentrations of 2,5-hexanedione and 3-acetyl-2,5- ing, disruption of axonal transport, and distal axonopathy in
hexanedione, a substance with pyrrole derivatives that cannot humans occupationally exposed at high levels to n-hexane and/
be oxidized to form covalent crosslinks (Genter et al. 1988). or methyl n-butyl ketone (ATSDR 1999). In addition, a consid-
Rats treated with 3-acetyl-2,5-hexanedione showed no signs erable amount of data comparing evidence for distal neuropathy
of neurofilament aggregation, axonal swelling, or hind limb following exposure to various hexacarbons in humans has been
paralysis compared to 2,5-hexanedione-treated rats, although published (ATSDR 1999, US Environmental Protection Agency
similar amounts of stable pyrrole adducts were formed. These 2005). These data provide evidence that n-hexane exposure
data identify crosslinking of the neurofilaments as a key event affects the human peripheral nervous system in a manner that is
in the proposed mode of action. consistent with that observed in rodent, avian, and other mam-
Overall, there is strong evidence that the ability of n-hexane malian species. More importantly, there is evidence showing
to induce peripheral neuropathy is entirely dependent on the that while 2-hexanol is the major n-hexane metabolite in rats,
generation of 2,5-hexanedione at levels above critical serum 2,5-hexanedione is the major metabolite in humans, suggesting
thresholds for effect. Other C6 isomers are not metabolized to that humans may be more susceptible than rats to the adverse
2,5-hexanedione and do not cause peripheral neuropathy. For effects of n-hexane exposure (ATSDR 1999).
example, a comparative study of the neurotoxic effects of C6 Limited data exist for the evaluation of quantitative dif-
isomers43 and n-hexane was conducted in male rats exposed ferences in the hypothesized mode of action in humans and
via inhalation to 0, 125 ppm of (378 mg/m3) n-hexane, 125 rodents. Mortensen and Nilsen (1998) compared the n-hexane
ppm of n-hexane plus 125 ppm of (378 mg/m3) C6 isomers, metabolic capacity of human liver S9 fractions to mice, rats,
125 ppm of n-hexane plus 375 ppm of (1136 mg/m3) C6 iso- and guinea pigs. Vmax, Km and intrinsic clearance values were
mers, 125 ppm of n-hexane plus 1375 ppm of (4166 mg/m3) similar across species, suggesting that humans most likely
C6 isomers, or 500 (1500 mg/m3) ppm of n-hexane, 22 hours/ produce n-hexane metabolites at a rate similar to that observed
day and 7 days/week for 6 months. Rats in the 500 ppm (1500 in rodents and other species.
mg/m3) n-hexane group developed atrophy of the sciatic and
anterior tibial nerves, skeletal muscle atrophy, and abnormal III. Toxicological properties of C7–C9 aliphatic
gait, which increased in severity with prolonged exposure. hydrocarbon solvents (Category 7 solvents)
Similar effects were not seen in any other treatment group 1. Introduction
(American Petroleum Institute 1983a, b).
The category of “C7–C9 aliphatic solvents” includes both
11. Human observations mono-constituent (n-heptane, n-octane, n-nonane) and com-
plex hydrocarbon solvents with aliphatic constituents pre-
The neurotoxicity of n-hexane was first observed in the shoe
dominantly in the C7–C9 range.
industries of Japan and Italy in the 1960s and early 1970s.
Among the first reports were Buiatti et al. (1978), Herskow- 2. Physical and chemical properties
itz et al. (1971), Inoue et al. (1970), Paulson and Waylonis
(1976) and Yamada (1964, 1967). Summarized information The C7–C9 aliphatic solvents are colorless liquids at room
can be found in reports by the Agency for Toxic Substances temperature. The physical and chemical properties are sum-
and Disease Registry (ATSDR 1999), the US Environmental marized in Table 24.
Protection Agency (EPA 2005) and IPCS (1991). Once the
hazards of n-hexane exposure were recognized, risk manage- 3. Toxicokinetics
ment measures including reductions in occupational exposure 3.1 Absorption of C7–C9 aliphatic hydrocarbons
limits and the introduction of substance classification were
instituted in the United States and Europe. Dahl et al. (1988) reported that inhaled C7–C9 aliphatic
The hypothesized mode of action for n-hexane-induced hydrocarbons were relatively well absorbed. The efficiency of
peripheral neuropathy in rodents, as described earlier (10.2.1.1), absorption increased with increasing molecular weight, and
appears relevant to humans (Table 23). For example, the normal paraffins were better absorbed than branched paraffins.
In a comparison of the properties of n-octane to iso-octane
43C6 isomers were an isomeric hexane mixture that did not contain (2,2,4-trimethylpentane), the uptake rates for octane were
n-hexane. Major constituents were methylcyclopentane, 3-methylpentane reported as 6.1 and 3.4 nmol/kg/min/ppm for “low” and “high”
and 2-methylpentane. exposure concentrations, whereas the corresponding rates for
Table 23. Hypothesized key event interspecies concordance for n-hexane-induced peripheral neuropathy.
# Key Events Rat Mice Human Other
1 Metabolism to the reactive 2,5- Yes, in vivo data with Yes. Pathway Yes. Pathway similar 2,5-hexandione found in the
hexanedione 2-hexanol metabolism similar in mice in humans. Urinary urine of monkeys, rabbits,
and ultimately distal metabolites have been cats and chickens.
axonopathy potentiated found.
by liver enzyme
inducers
2 Pyrrole adduct formation Yes, in vivo and in vitro. No data? Evidence of pyrrole-like No data
substances in urine
3 Oxidation of pyrrole adducts to Yes, based on Yes, based on Yes, based on Yes, based on
form protein cross-links histopathological data histopathological histopathological data histopathological data
data
4 Axonal swelling and Yes Yes Yes Yes – cats, chickens
degeneration
5 Peripheral nerve damage/ Yes Yes Yes Yes
axonopathy
“low” and “high” isooctane were 3.4 and 2.2 nmol/kg/min/ metabolized to the corresponding alcohols and carboxylic
ppm (Dahl 1989). acids. Olson et al. (1986) reported that the principal urinary
These solvents are reasonably well absorbed following oral metabolites of octane were 2-octanol, 3-octanol, 5-oxo-
administration. The fraction absorbed is estimated to be in hexanoic acid, and 6-oxohexanoic acid. Charbonneau et al.
the range of 80%–90%, based on the empirical relationship (1987) reported that 30% of ingested trimethylpentane can be
defined by Albro and Fishbein (1970). However, percutaneous recovered as exhaled material within the first 8 h of dosing.
absorption is not anticipated to make a large contribution to The majority of the remaining material is eliminated in the
systemic dose unless evaporation is inhibited. Tsuruta (1982) urine within 24 h of administration. Dahl (1989) reported that
reported that the percutaneous absorption rate for heptane half of the octane retained following exposure by inhalation
was 0.1ug/cm2/hr and that for octane was 0.0005 mg/cm2/hr. had been eliminated 5–10 h after exposure, and essentially all
McDougal et al. (2000) reported that the percutaneous absorp- of the absorbed material had been eliminated within 70 h.
tion rate for nonane was 0.38 mg/cm2/hr, but the value reported
by Muhammad et al. (2005), 0.03 mg/cm2/hr, was an order of 3.4 Pharmacokinetic modeling
magnitude lower. Singh et al. (2002) estimated that 0.14% of
a dermal dose of heptane would be systemically absorbed. In Robinson (2000) developed a physiologically-based pharma-
short, these solvents are more likely to evaporate than to be cokinetic model for nonane which was then validated against
absorbed following dermal contact. blood concentration data developed in their own laboratory.
The data fit the model reasonably well, at exposure levels of
3.2 Distribution of C7–C9 aliphatic hydrocarbons 100–500 ppm (526–2632 mg/m3) n-nonane, but did not pro-
vide a good fit for late time-points following exposure to 1000
Charbonneau et al. (1987) reported that 24 h after oral
ppm (5263 mg/m3). The simulation predicted that, following
administration of 2,2,4-trimethylpentane, the material remaining
exposure by inhalation, blood concentrations would quickly
in the body was primarily in the liver and kidneys, with smaller
reach steady state, and that the concentrations would decline
amounts in the plasma and even smaller quantities in the fat.
quickly once exposures were terminated.
3.3 Metabolism and elimination of C7–C9 aliphatic
hydrocarbons 4. Acute toxicological effects
Inhaled octane is primarily metabolized to CO2 and elimi- The acute toxicity data indicated that the C7–C9 aliphatic
nated by exhalation (Dahl 1989). Ingested octane is primarily hydrocarbon solvents and their constituents have a low order
Table 24. Physical and chemical properties of C7–C9 aliphatic solvents.

Property n-Heptane n-Octane n-Nonane Complex C7–C9 Solvent
Molecular Weight 100 114 128 100–128
Physical State at 20°C and 1013 hPA Colorless liquid with Colorless liquid with Colorless liquid with Colorless liquid with slight odor
slight odor slight odor slight odor
Pour Point  20°C  20°C  20°C  20°C
Boiling Point 99°C 125°C 151°C 90–165°C
Relative Density 0.68 g/cm3 0.70 g/cm3 0.72 g/cm3 0.71 to 0.78 g/cm3
Vapor Pressure 5.3 kPa at 20°C 1.5 kPa at 20°C 1.3 kPa at 20°C 2 kPa at 20°C
Surface Tension 21 mN/m at 25°C 20 to 24 mN/m at 25°C.
Water Solubility 3.4 mg/l 0.66 mg/l 0.22 mg/l Insoluble in water
Octanol/Water Partition Coefficient 4.7 5.2 5.3 Not reported
Flash Point Not reported Not reported Not reported  10°C at 1 atm.
Flammability Not reported Not reported Not reported 0.9–6.7%
Self-Ignition Temperature Not reported Not reported Not reported 200°C
Viscosity Not reported Not reported Not reported 0.5–1.5 mm2/s at 20°C.
Conversion Factor (ppm/mg/m3) 1 ppm  4.2 mg/m3 1 ppm  4.7 mg/m3 1 ppm  5.2 mg/m3 1 ppm  ∼ 5.0 mg/m3
of acute oral, dermal, and inhalation toxicity when tested in of repeated inhalation studies, the earliest of which was an
accordance with current guidelines for those effects. These assessment of “varnish makers’ and painters’ naphtha”
solvents can cause acute CNS effects if inhaled at high vapor (VM&P naphtha) 45 (Carpenter et al. 1975b–d). In this study,
concentrations (Lammers et al. 2011, McKee et al. 2011) and rats and dogs were exposed 6 hours/day and 5 days/week for
can cause chemical pneumonitis if they enter the lungs in a 13 weeks to vapors at concentrations of 1300, 2800, or 5800
liquid state. mg/m3 (280, 600, or 1200 ppm). At scheduled termination,
the animals were sacrificed and examined. Two rats died of
4.1 Acute inhalation toxicity studies lung abscesses and pneumonia prior to the scheduled termi-
Carpenter et al. (1975d) reported that the 4-hour LC50 value nation, but as these were in the lowest exposure group, they
in rats for rubber solvent44 was 61000 mg/m3 (15000 ppm). were not considered to have been treatment-related. There
There were no obvious effects in rats exposed for 4 h to 11000 were no significant differences in terminal body weights or
mg/m3 (2000 ppm) but at intermediate levels (21000 mg/m3, clinical chemistry values. The authors commented that the
39000 mg/m3), there were observations of eye irritation and erythrocyte count was reduced in the high exposure group,
loss of coordination, although all of the animals survived to but the difference was not statistically significant. There were
scheduled termination, 14 days after exposure. The 4-hour no significant differences in organ weights. The pathologist
LC50 value for n-nonane in rats was reported as 17000 mg/m3 reported tubular regeneration, but did not consider these
(or 3200 ppm) by the same group (Carpenter et al. 1978), but changes to have been treatment-related. Among the dogs, the
in a later study (Nilsen et al. 1988), the 8-hour LC50 value in authors reported a few statistically significant findings, but
rats was reported as 4467 ppm (23500 mg/m3). the only effect that was regarded as treatment-related was an
Kinkead et al. (1985) summarized older studies (Fuehner increase in alkaline phosphatase levels. The overall conclu-
1921, Lazarew 1929, Lehman and Flury 1943, Treon et al. sion was that there may have been slight effects at the 5800
1943) in which it was reported that in acute inhalation stud- mg/m3 level.
ies of various species, exposures to heptane and/or methyl- Carpenter et al. (1975d) also investigated the potential
cyclohexane at levels up to at least 10,000 ppm (41000 mg/ effects of “rubber solvent”, a commercial hydrocarbon sol-
m3) produced narcosis but were not lethal. The apparent vent composed primarily of C6 and C7 constituents.46 Rats
decrease in LC50 values with increasing carbon numbers and dogs were exposed 6 hours/day, 5 days per week for 13
seems consistent with the evidence of an increasing tendency weeks at exposure levels of 1900, 3700, or 7900 mg/m3 (480,
of aliphatic constituents to distribute to the CNS across the 930, or 2000 ppm). There were no pre-termination mortali-
C6–C10 carbon number range (Zahlsen et al. 1992, 1993). ties that were considered treatment related and no significant
However, aliphatic hydrocarbons with carbon numbers differences in body weights or in hematological parameters at
 C10 do not partition as easily into the CNS as the C6–C10 terminal sacrifice. There were statistically significant eleva-
aliphatic hydrocarbon constituents, and alkanes with carbon tions in alkaline phosphatase levels, but as the differences
numbers  C10 do not produce acute CNS effects (Nilsen were small and within normal physiological limits and not
et al. 1988). dose-responsive, they were not considered to have been treat-
ment-related. Notably, levels of AST and ALT (identified in
the paper as SGOT and SGPT) were not elevated. There were
4.2 Acute oral and dermal toxicity
no pathological findings in the rats. There was an increased
Based on results of the studies summarized in the HPV dos- incidence of foreign body granulation nodules in the lungs
sier (OECD 2010), the oral LD50 values were greater than the of dogs, but this finding was considered to have more likely
highest levels tested (5000–15000 mg/kg), the dermal LD50 been the consequence of a surgical procedure that had been
values were  3000 mg/kg, and the inhalation LC50 values performed on the dogs than a consequence of treatment. An
were  23,000 mg/m3 (approximately 4600 ppm). overall conclusion was that the highest exposure level tested
(7900 mg/m3 or 2000 ppm) had no effects on the rats and most
4.3 Dermal and ocular irritant effects likely had no effects on the dogs.
More recently, Schreiner et al. (1998) assessed the potential
Based on summarized information (Johnson et al. 2012,
for effects following repeated exposure of rats to vapors of
OECD 2010), C7–C9 aliphatic hydrocarbon solvents are slight
light alkylate naphtha distillate.47 Rats were exposed to vapors
to moderate dermal irritants, but they are not ocular irritants
6 hours/day and 5 days/week for 13 weeks and then sacrificed
and do not cause allergic contact dermatitis.
for hematological, clinical, and pathological evaluation. The
5. Repeated dose studies

5.1 Repeated inhalation studies of C7–C9 aliphatic 45The sample of VM&P naphtha was described by Carpenter et al. (1975b)
hydrocarbon solvents as containing 90% aliphatic constituents; 13% heptanes, 41% octanes,
29% nonanes, and 13% higher molecular weight aliphatic constituents.
The potential for exposure to C7–C9 aliphatic hydrocarbon 46“Rubber solvent” contained 41% hexanes and 54% heptanes. The
solvents and their constituents has been assessed in a number remaining material was primarily toluene (Carpenter et al. 1975d).
47“Light alkylate distillate” is a gasoline blending stream manufactured
by reacting olefins, resulting in process streams of almost pure ( 99%)

44The “rubber solvent” tested in this study was a commercial product sup- branched paraffins. The test sample contained primarily C5–C8 isoparaf-
plied in approximately 1966. As described by Carpenter et al. (1975d), finic components, essentially covering the aliphatic constituents of the
this material was composed primarily of C6–C7 aliphatic constituents. Category 5, 6 and 7 hydrocarbon solvents.
exposure levels were 2400, 8100, or 24300 mg/m3 (668, 2200, in these studies were reductions in body weight gain in male
or 6646 ppm). Some of the animals from the highest exposure rats and male hamsters. There were some small hematologi-
group were held for an additional 28 days to assess recovery. cal and/or clinical changes, but the results were within normal
The highest concentration tested was 75% of the lower expo- biological limits and not considered toxicologically important.
sure limit. All of the animals survived the treatment period The only notable pathological change was an increase in renal
with little evidence of treatment-related effects. There were tubular dilation at exposure termination and progression of
small (∼ 5%) reductions in some hematological parameters renal pathology to papillary hyperplasia and medullary min-
in the male rats from the high-exposure group, but these dif- eralization in the kidneys of rats from the 4000 ppm (16393
ferences were not considered toxicologically important as mg/m3) group. This change was referred to by the authors as
they were within normal physiological ranges. ALT and AST “light hydrocarbon nephropathy” but is now be understood to
levels were significantly reduced in female rats from the high- be evidence of an a2u-globulin-mediated effect. The authors
exposure group; elevated levels of these enzyme markers are also reported that there was no evidence for a carcinogenic
considered to be indicative of the potential for liver effects, response. The authors considered that their data supported the
but the biological significance of reductions in levels of these TLV® of 400 ppm (1639 mg/m3).
enzymes is unknown. There was evidence of increased kidney Sung et al. (2010) summarized the results of a repeated
weights in male rats and elevations in levels of hyaline drop- inhalation study of n-octane. Rats were exposed to target con-
lets in the proximal convoluted tubules. These changes were centrations of 930, 2620, or 7480 mg/m3 (195, 550, or 1571
considered the consequence of an a-2u-globulin-mediated ppm), 6 hours/day and 5 days/week for 13 weeks. At the end of
process (identified by the authors as “light hydrocarbon neph- the exposure period, the animals were sacrificed and examined
ropathy”). There were also increases in liver weights in both for hematological, clinical, and/or pathological effects. There
male and female rats from the high-exposure group, but as were no significant differences in terminal body weights or
there were no pathological changes and liver enzyme markers in hematological or clinical chemistry parameters, and there
were not elevated, the liver weight changes were considered were no pathological findings. The no observed adverse effect
to be evidence of an adaptive response and not toxicologically level was 7480 mg/m3 (1571 ppm), the highest concentration
important. The overall no adverse effect concentration in this tested.
study was 24,300 mg/m3 (6646 ppm), the highest concentra- Carrillo et al. (2013) summarized the results of a repeated
tion tested. inhalation study of a C8 isoparaffinic solvent. Rats were
exposed 6 hours/day and 5 days/week to vapors at concentra-

5.2 Repeated dose studies of C7–C9 Aliphatic tions of 400 or 1200 ppm (approximately 1800 or 5500 mg/
hydrocarbon solvent constituents m3). At termination, the animals were sacrificed and examined;
In a repeated dose study of n-heptane, rats were exposed by however, as this study had been conducted primarily to assess
inhalation 6 hours/day and 5 days/week for 26 weeks at exposure potential effects on the kidneys, the clinical examination was
levels of 398 or 2970 ppm (1631 or 12172 mg/m3) (American restricted to urinary parameters, and the pathological evalua-
Petroleum Institute 1980b). There were no treatment-related tion included only the major organs. The only reported find-
mortalities and no differences in terminal body weights. There ings were increased kidney weights and histological changes
were no differences in hematological parameters, and the only in the kidneys of high exposure male rats. Setting aside the
clinical finding was a small but statistically significant eleva- kidney changes, the overall no-effect concentration was 5500
tion in alkaline phosphatase. There were no gross findings at mg/m3 (1200 ppm).
necropsy. There were no apparent toxicologically important Carpenter et al. (1978) reported a repeated inhalation toxic-
effects at the highest exposure level (approximately 2790 ppm ity study of n-nonane using a protocol similar to the rubber
or 12171 mg/m3). In this respect, this study is similar to those solvent study summarized above. The rats and dogs were
by Takeuchi et al. (1980) and Frontali et al. (1981), in which exposed at levels of 1900, 3100, or 8400 mg/m3 (360, 590,
no apparent effects were observed following repeated expo- or 1600 ppm). Two rats from the highest exposure group died
sure at levels of 1500 or 3000 ppm (6148 or 12295 mg/m3). on the first day of exposure; other unscheduled animal deaths
However, as the experimental objective of the Takeuchi and were considered to have been unrelated to treatment. The body
Frontali studies was to assess the potential for neurological weights of rats in the high-exposure group were significantly
effects, the investigation of other systemic effects was limited. below control values, but there were no hematological, clini-
The API study confirmed that there were no systemic effects cal, or pathological findings that were considered to have been
at exposure levels up to approximately 3000 ppm (12295 mg/ treatment-related. The authors considered that the no-effect
m3). concentration was 3100 mg/m3 (590 ppm).
Kinkead et al. (1985) assessed the potential for methylcy-
6. Genetic toxicity
clohexane to cause systemic effects in repeated dose studies.
Mice, rats, dogs, and hamsters were exposed to vapors, 6 A comprehensive in vitro evaluation of the mutagenic poten-
hours/day and 5 days/week for 12 months, at concentrations tial of n-heptane was published by Brooks et al. (1988). More
of either 400 or 4000 ppm (1639 or 16393 mg/m3). At the specifically, n-heptane was tested for mutation in Salmonella
end of the exposure period, some of the animals were sacri- and E. coli, gene conversion in Saccharomyces cerevisiae, and
ficed for hematological, clinical, and pathological evaluation, for chromosomal aberrations in rat liver cells (RL4) in culture.
whereas others were held for either one (rodents) or 5 (dogs) n-Heptane did not show any evidence of genotoxic potential in
years for post-exposure observations. The principal findings any of these assays.
Richardson et al. (1986) investigated the genotoxic potential It should also be noted that there were no pathological
of 2,2,4-trimethylpentane (TMP, isooctane) as part of a broader changes in reproductive organs in any of the repeated dose
investigation into the underlying mechanisms of male rat kid- toxicity studies in which these were examined.
ney tumors in chronic studies of gasoline. TMP did not increase
the frequency of mutation or sister chromatid exchange in TK6 9. Carcinogenicity
cells (derived from a human lymphoblastoid cell line). Similarly, As the C7–C9 aliphatic hydrocarbon solvents are not genotoxic
Loury et al. (1986) reported that TMP did not induce unsched- and do not cause cumulative toxicity in repeated dose studies,
uled DNA synthesis in human or rodent hepatocytes in studies it is unlikely that these substances would be carcinogenic.
involving both in vivo (rodents) and in vitro (primary cultures
of rodent and human liver cells) test material administration. 10. Neurotoxicity
The authors found no evidence that TMP was genotoxic.
Additional supporting evidence that the C7–C9 aliphatic 10.1 Studies of acute neurological effects
hydrocarbon solvents are not genotoxic can be found in John- The acute CNS effects of C7–C9 aliphatic hydrocarbon sol-
son et al. (2012) and in the HPV documentation for this cat- vents and their constituents have been the subject of numerous
egory of solvents (OECD 2010). investigations. Patty and Yant (1929) and later Carpenter et al.
(1975d) reported observing behavior suggestive of acute CNS

7. Developmental toxicity effects in volunteers exposed at levels  1000 ppm. Bowen
In an unpublished developmental toxicity study (summarized and Balster (1998) reported that exposure to complex isoparaf-
in Johnson et al. 2012), a C8 isoparaffinic solvent was tested finic solvents resulted in statistically significant reductions in
at levels of 400 or 1200 ppm (1900 or 5700 mg/m3). Mated motor activity in mice at exposure levels  4000 ppm. Lam-
female rats were exposed 6 hours/day on gestational days 6–15 mers et al. (2011) reported that exposure to n-octane at levels
and were then sacrificed on gestational day 21 for uterine and up to 14,000 mg/m3 (2940 ppm) did not produce acute CNS
fetal examinations. There were no treatment-related effects on effects in mice, but Sung et al. (2010) reported acute CNS
uterine implantations, fetal size, or sex distribution, or in fre- effects in rats exposed for 4 h to n-octane at 23360 mg/m3
quencies of malformations. There was an increase in skeletal (4900 ppm). McKee et al. (2011) did not observe CNS effects
variations in the high-exposure group, but the types and fre- in rats exposed to isooctane at levels  14000 mg/m3 (2940
quencies of variations were similar to those in the unexposed ppm). There were some small but statistically significant CNS
control group, and therefore, were most likely evidence of effects at 14000 mg/m3 (2940 ppm) in a study of a C6/C7
developmental delays. A positive control group (acetylsalicylic cycloparaffinic solvent; the no-effect level was 4200 mg/m3
acid) caused developmental effects in this study, as expected. (882 ppm) (McKee et al. 2011). These observational data are
A second study (summarized in Mullin et al. 1990) investi- consistent with reports from Zahlsen et al. (1992, 1993) that
gated the potential for a C9–C11 isoparaffinic solvent to cause levels of normal and isoparaffinic hydrocarbons in the CNS
developmental effects using a study design similar to that in increase with increasing number over the range of C6–C10.
the C8 isoparaffinic solvent discussed in the paragraph above Levels of cycloparaffins are higher than those of the corre-
but at exposure levels of 300 or 900 ppm (1429 or 4286 mg/ sponding alkanes and are higher at carbon numbers ranging
m3). There were no developmental effects under the conditions from C6–C10, but lower at C10.
of this study.
10.2 Studies of persistent neurological effects
The potential for n-heptane to cause peripheral neurological
Bui et al. (1998) assessed the potential for a complex C5–C8 effects was assessed by experimental studies in rats (Frontali
isoparaffinic hydrocarbon substance to affect fertility and devel- et al. 1981, Takeuchi et al. 1980). In the Frontali study, rats were
opment in rats, in a study following the guidelines of OECD exposed to 1500 ppm heptane (approximately 6000 mg/m3), 9
422 in which male and female rats were exposed beginning hours/day and 5 days/week for up to 30 weeks. There were no
2 weeks prior to mating and continuing through the mating differences in terminal body weights and no apparent signs of
period. Exposure of mated females was then continued to ges- neurological impairment. Similarly, there were no pathological
tational day 19. The dams were allowed to give birth, and then findings in rats sacrificed after 7, 14, or 30 weeks of exposure.
held with their litters to postnatal day 4, at which time all sur- The authors concluded that exposure to n-heptane did not cause
viving dams and offspring were sacrificed. The animals were alterations in the peripheral nervous system, although such
exposed 6 hours/day at levels of 5000, 12500, or 25000 mg/m3 effects were shown to be the consequence of n-hexane exposure
(1650, 4040, or 8000 ppm). All parental animals survived to in the same set of studies. In similar studies, Takeuchi et al.
scheduled termination without evidence of clinical findings and (1980) exposed rats to n-heptane, 12 hours/day for 16 weeks
with no differences in body weights. There were no differences at levels of 3000 ppm (12295 mg/m3). The authors reported
in fertility index, number of implantation sites, average number that exposure to n-heptane had no effects on nerve conduction
of offspring/litter, fraction of offspring born live, or fraction of velocity and did not produce any pathological changes in the
offspring surviving to postnatal day 4, and no differences in peripheral nervous system, although such changes were caused
average body weights of offspring on days 1 or 4. The authors by exposure to n-hexane under the same circumstances.
concluded that the no observed adverse effect level for repro- Schreiner et al. (1998) assessed the potential for light alky-
ductive effects was 24,700 mg/m3 (8000 ppm), the analytically- late naphtha distillate, containing primarily C5–C8 isopar-
determined concentration for the most highly exposed group. affins, to produce neurological changes in rats exposed by
inhalation for 13 weeks at levels up to 6646 ppm (24300 mg/ and boiling in the range of approximately 110–280°C. The sol-
m3). There were no differences in functional observations or vents in this category can be either mono-constituent (primar-
motor activity in rats tested after 5, 9, or 14 weeks of exposure. ily normal paraffins) or complex substances. There are several
Further, there were no pathological changes in nervous system manufacturing processes that can be used to make solvents of
tissue. The authors considered that the highest level tested this type, including hydrogenation of C9–C14 aliphatic solvents,
(6646 ppm or 24300 mg/m3) was a no-effect concentration for 2–25% aromatics, to reduce aromatic contents, resulting in
neurological effects. complex aliphatic solvents. However, solvents in this category
can be manufactured by other processes including an alkylation
11. Occupational exposures and recommended process in which olefins react to form complex isoparaffin-rich
occupational exposure limits streams that can be separated by distillation to produce solvents
Carpenter et al. (1975b, 1975d) assessed the effects of VM&P with the desired distillation ranges. The Category 8 hydrocar-
naphtha and rubber solvent exposure in humans. Volunteers bon solvents are listed in the US EPA toxic substances Control
were exposed to these solvents in 15 minute periods. In the study Act (TSCA) registry under a number of CAS registry numbers
of VM&P naphtha, the test concentrations were 660, 1400, including 64742-47-8 and 64742-48-9 which are the most com-
2100, or 4100 mg/m3 (140, 300, 450, or 880 ppm). The authors monly used CAS registration numbers for the complex solvents,
reported ocular and upper respiratory irritation in volunteers as well as separate CAS numbers for the mono-constituent
exposed to 4100 mg/m3 (880 ppm) and concluded that approxi- solvents. CAS registry numbers and EINECS numbers for the
mately 2000 mg/m3 (430 ppm) was appropriate as an occupa- solvents in this category are shown in Table 2. For REACH reg-
tional exposure level. In the rubber solvent study, the volunteers istration purposes these solvents were grouped into Category
were exposed at levels of 1700, 3100, 6700, and 8100 mg/m3 8 with the generic descriptor “Hydrocarbons, C9-C14,  2%
(430, 780, 1700, and 2000 ppm). There was some evidence of aromatics” and the various substances were registered in accor-
reversible CNS effects in one subject exposed to 8100 mg/m3 dance with the naming convention introduced by the Hydrocar-
(2000 ppm), and some evidence of eye and/or respiratory irrita- bon Solvent Producers Association (HSPA 2011). Names of the
tion in volunteers exposed to either 3100 or 6700 mg/m3 (780 or individual solvents are listed in Table 2.
1700 ppm). There were no effects among individuals exposed
2. Physical and chemical properties
to 1700 mg/m3 (430 ppm). These observations are consistent
with those previously reported by Patty and Yant (1929) who The C9–C14 aliphatic solvents are colorless and essentially
had observed evidence of acute CNS effects in humans exposed odorless liquids at room temperature. The physical and chemi-
to heptane at levels of 1000 ppm (4100 mg/m3). cal properties of several examples of C9–C14 aliphatic ( 2%
Current occupational exposure recommendations for these aromatic) hydrocarbon solvents covering the range of carbon
substances in the United States are summarized in Table 6. numbers in the category are summarized in Table 25.
According to Caldwell et al. (2000), the weighted average expo-
sures for C5–C9 aliphatic hydrocarbon solvent constituents, 3. Toxicokinetics
between 1961 and 1998 were: pentane (9.5 mg/m3); n-hexane 3.1 Absorption
(93 mg/m3); other hexanes (0.7 mg/m3); cyclohexane (160 mg/
m3); heptane (37 mg/m3); octane (0.02 mg/m3); and nonane Inhaled C9–C14 aliphatic ( 2% aromatic) hydrocarbon sol-
(467 mg/m3); however, some of these values were based on vents are systemically absorbed, distributed to various tissues,
very small numbers of samples. They also reported values for metabolized, and eliminated. For aliphatic hydrocarbons in
exposure to rubber solvent (263 mg/m3) and VM&P naphtha general, the potential for distribution to the CNS is of par-
(48 mg/m3). The value for total volatile hydrocarbons was 67 ticular interest. As shown by Zahlsen and associates (Nilsen
mg/m3. With respect to rubber solvent and VM&P naphtha, et al. 1988, Zahlsen et al. 1992, 1993) partitioning of aliphatic
it should be noted that it is not clear how these historical for- hydrocarbons to the CNS increases with increasing carbon
mulations relate to products currently in the market place. In number (and increasing octanol/water partition coefficients)
particular, Carpenter et al. (1975d) reported that the rubber over the range of approximately C6–C10. However, lower lev-
solvent (which was obtained as a commercial solvent in the els are found in studies of aliphatic hydrocarbons with carbon
1960s) contained 1.5% benzene. As noted in the discussion on numbers  C10 and cycloparaffins with carbon numbers 
Benzene, steps were being taken in the late 1970s to reduce C9 (Zahlsen et al. 1992, 1993). In part, this is related to the
benzene levels. Thus, the historical information on exposure to more limited vapor concentrations that can be achieved with
formulated products may not reflect current usage. these constituents due to their relatively low vapor pressures,
but there may also be blood/brain effects leading to inhibition
of uptake into the CNS (Nilsen et al. 1988). Ingested aliphatic
Appendix D. Non-volatile aliphatic hydrocarbons are also well absorbed. Based on the empirical
hydrocarbon solvents relationship of Albro and Fishbein (1970), systemic absorp-
I. Toxicological Properties of C9-C14 Aliphatic tion of 60–80% of the constituents of these solvents would be
Hydrocarbon Solvents (<2% aromatics) (Category expected. However, percutaneous absorption of these solvents
8 solvents) and their constituents is much more limited (Baynes et al.
2000, Kim et al. 2006, Muhammad et al. 2005, Singh and
1. Introduction
Singh 2003). Based on an in vitro study with human skin, Kim
“C9–C14 aliphatic solvents” are aliphatic hydrocarbon solvents et al. (2006) reported percutaneous absorption values in the
with carbon numbers predominantly in the range of C9–C14 range of 1–2 mg/kg/day for decane, undecane, and dodecane.
Table 25. Physical and chemical properties of some representative C9–C14 aliphatic solvents,  2% aromatics.
Complex C9–C10 Complex C10–C13 Complex C11–C14 n-Decane (C10
Property Aliphatic Solvent Aliphatic Solvent Aliphatic Solvent aliphatic substance)
Molecular Weight Approximately 135 Approximately 163 Approximately 177 142.28
Physical State at 20°C Colorless liquid with slight Colorless liquid with Colorless liquid with slight Liquid
and 1013 hPA odor slight odor odor
Pour Point  20°C  20°C  0°C  20°C
Boiling Point 110–190°C 160–245°C 210–280°C 160–190°C
Relative Density 0.73–0.80 g/cm3 0.75–0.85 g/cm3 0.79–0.85 g/cm3 0.733–0.738 g/cm3
Vapor Pressure 0.5 kPa at 20°C 0.05 kPa at 20°C 0.003 kPa at 20°C 0.135 kPa at 20°C
Surface Tension 22–26 mN/m at 25°C 23–28 mN/m at 25°C 25–29 mN/m at 25°C ∼ 23 mN/m at 25°C
Water Solubility 0.08–0.3 mg/l at 25°C 0.001–0.08 mg/l at 25°C 0.0003–0.02 mg/l at 25°C 0.0826 mg/l at 25°C
Flash Point  23°C  61°C  90°C 57.4°C
Flammability 0.6–7.0% v/v 0.6–7.0% v/v 0.6–7.0% v/v 0.6–7.0% v/v
Self-Ignition  200°C  200°C  200°C 206°C
Temperature
Viscosity 0.8–1.8 mm2/sec at 20°C 1.3–2.5 mm2/sec at 20°C 2.2–5.0 mm2/sec at 20°C 1.10–1.20 mm2/s at 20°C
Odor Threshold Virtually odorless Virtually odorless Virtually odorless Virtually odorless
Maximally  31  103 mg/m3 ( 5300 ppm)  10  103 mg/m3  3.5  103 mg/m3 10  103 mg/m3
Attainable Vapor ( 1800 ppm) ( 636 ppm) (1800 ppm)
Concentration
Conversion Factor 1 ppm  5.2  5.8 mg/m3 1 ppm  5.8  7.5 mg/m3 1 ppm  6.3  8.1 mg/m3 1 ppm  5.832 mg/m3
(ppm/mg/m3)
3.2 Distribution materials for pharmacokinetic modeling. In their initial modeling

which focused primarily on the fat compartment, Pedersen et al.
An investigation of the systemic distribution of representative
(1984) showed that with repeated exposure, levels of white spirit
normal, isoparaffinic, and naphthenic hydrocarbons in rats fol-
increased in the fat and then slowly declined following cessation
lowing exposure by inhalation was reported by Zahlsen et al.
of exposure. The authors also summarized the results of studies
(1992, 1993). In tissues taken from animals sacrificed at the
in rats (data provided by Edelfors and Ravn-Jonsen). The rat
end of the exposure period, the highest hydrocarbon constitu-
data indicated that concentrations of white spirit in blood and

ent levels were found in fat, with lesser amounts in brain, liver,
brain rapidly declined at exposure termination but that the con-
kidney, and blood. When the rats were held without treatment
centrations in perirenal fat declined more slowly; in this respect,
for 12 h, hydrocarbon levels in fat remained relatively high.
the data were similar to those reported later by Zahlsen et al.
In contrast, the levels were low in other tissues, and in some
(1992, 1993). Based on the rat data, a pharmacokinetic model
cases, below detection levels.
was constructed, with which it was shown that concentrations
in blood and brain declined very rapidly following exposure
3.3 Metabolism
termination (see Figure 6 in Pedersen et al. 1987) but that con-
As discussed in Metabolism and excretion of aliphatic centrations in fat gradually increased with repeated exposures
hydrocarbon solvent constituents and shown in Figure 1, (Figure 5 in Pedersen et al. 1987).
alkanes are metabolized by w-oxidation and either excreted or Hissink et al. (2007) reported a study similar to that of
re-incorporated into other macromolecules, depending on the Edelfors and Ravn-Jonsen, in which rats were exposed to
length of the carbon chain. white spirit for periods of up to 8 h at levels ranging from
600–4800 mg/m3 (approximately 109–873 ppm), and samples
3.4 Excretion were collected for use in developing a physiologically-based
The kinetics of C9–C12 aliphatic solvents48 were investigated in pharmacokinetic model in which 1,2,4-trimethylbenzene was
volunteers (Pedersen and Cohr 1984a, Pedersen et al. 1984b). In selected to represent the aromatic constituents and n-decane
the first study (Pedersen and Cohr 1984a), the participants were was selected to represent the aliphatic constituents. The model
exposed to the solvent by inhalation for 6 h at levels of 304 mg/ predictions were then compared to blood and air samples from
m3 (51 ppm), 611 mg/m3 (102 ppm), or 1228 mg/m3 (200 ppm). volunteers exposed for 4 h at 570 mg/m3 (104 ppm). The data
Urine samples were collected 1 hour prior to exposure and 3 and and modeled results were similar to those previously reported,
6 h after the start of the exposure. Blood samples were collected and, of particular relevance to this report, confirmed that levels
after 6 h of exposure. In the second study (Pedersen and Cohr of n-decane rapidly declined in blood and brain at termination
1984b), the volunteers were exposed to an isoparaffinic solvent, of exposure. Based on inspection of the figures (Figures 2 and
6 hours/day for 5 days at 616 mg/m3 (103 ppm). In this study, 3 in Hissink et al. 2007), the initial half-life for n-decane in the
blood, alveolar air, and fat were sampled to obtain biological CNS was in the order of 2 h. Decane levels were below detec-
tion levels in blood within about 4 h of exposure cessation and
were below detection limits in expired air within about 8 h
48The test sample used in the single exposure study by Pedersen and of exposure termination. These data confirmed that C9–C14
Cohr (1984a) contained primarily paraffinic (52%) and cycloparaffinic aliphatic constituents were rapidly eliminated from physi-
(48%) constituents with carbon numbers ranging from C8–C12. The test
sample used in the repeated exposure study (Pedersen and Cohr 1984b) ological compartments other than depot fat once exposure was
contained primarily (99%) isoparaffins with carbon numbers ranging terminated. PBPK models for decane have been published by
from C10–C12. Hissink et al. (2007) and Perleberg et al. (2004).
4. Acute effects have been conducted have focused on the most volatile of the
solvents in the category. Nau et al. (1966) reported a study in
4.1 Acute toxicity which rats were exposed to 540 ppm (3200 mg/m3) n-decane,
Much of the acute toxicity information on C9–C14 aliphatic 18 hours/day and 7 days/week for 90 days.49 At termination,
solvents is unpublished. However, based on summaries of the the exposed rats had body weights that were greater than those
available information (Amoruso et al. 2008, Johnson et al. of the corresponding control animals. The only other observa-
2012, OECD 2012d), these substances are not acutely toxic tion was that after 57 days of exposure the treated animals had
when tested at limit doses by the oral and dermal routes. In one a reduced number of white blood cells in comparison to the
published study, Johannsen and Levinskas (1987) reported control values. However, at terminal sacrifice, the white blood
that the acute oral LD50 of a mixture of tetramethylcyclo- cell counts in the treated rats were above control values.
hexanes (C10 cycloparaffins) was greater than 15800 mg/kg, Carpenter et al. (1978) exposed rats to n-nonane by inhala-
the highest dose administered to the rats. Acute dermal toxic- tion at levels of 360, 590, or 1600 ppm (1900, 3100, or 8400
ity values were greater than 2000 mg/kg, the limit doses for mg/m3), 6 hours/day and 5 days/week for 13 weeks. Two of the
tests of this type (OECD 2012d). As reported by Nilsen et al. rats in the high-exposure group died on the first exposure day.
(1988), the acute inhalation LC50 (8 h) for n-nonane was 4467 The authors also reported that on the first 4 days of exposure,
rats in the 1600 ppm (8400 mg/m3) group exhibited saliva-
ppm (23500 mg/m3), similar to the value reported by Car-

penter et al. (1978) of 14000–21000 mg/m3 (approximately tion, mild coordination loss, and fine tremors (presumably
2700–4000 ppm). However, there were no deaths or adverse evidence of upper respiratory irritation and CNS effects). On
behavioral effects reported in studies of rats exposed for 8 h to subsequent exposure days, salivation and lacrimation contin-
nC10–nC13 at the maximally attainable vapor concentrations ued to be observed, but there did not appear to have been any
(1369, 442, 142, and 41 ppm; 8000, 2700, 1000, and 315 mg/ evidence of CNS effects. The terminal body weights of the rats
m3 respectively) (Nilsen et al. 1988). in the high exposure group were lower than (but apparently
In summary, C9–C14 aliphatic ( 2% aromatic) hydro- not statistically different from) those of the control animals.
carbon solvents and their constituents do not produce acutely There were no statistically significant differences in clinical or
toxic effects except at very high levels and/or under unusual hematological findings at termination, and no notable patho-
circumstances. The acute oral toxicity data indicate that in logical observations. The authors considered that the no-effect
single dose studies, these solvents do not produce lethality concentration was 590 ppm (3100 mg/m3).
even when tested at levels of 5000–15000 mg/kg. No lethal- Studies of C9–C11 and C10–C12 isoparaffinic hydrocarbon
ity was observed in acute dermal studies conducted at limit solvents were carried out to further investigate the toxicologi-
doses of 2000 mg/kg. In acute inhalation toxicity studies, cal consequences of the kidney findings in male rats that had
LC50 values  2700 ppm ( 14000 mg/m3) were reported for been reported in some of the studies by Carpenter. In these
n-nonane, but it was not possible to produce mortality or acute studies, rats were exposed to the solvents by inhalation at tar-
CNS effects at maximally attainable vapor concentrations of get concentrations of 300 or 900 ppm (approximately 1800
nC10–nC14 aliphatic hydrocarbon constituents. It should be or 5300 mg/m3) (Phillips and Egan 1984a, 1984b, Phillips
noted, however, that these solvents can cause chemical pneu- and Cockerell 1984). Exposures were 6 hours/day and 5 days/
monitis if aspirated into the lungs as liquids. week for 8–12 weeks. In one of the studies, the animals were
exposed for 12 weeks with intermediate sacrifices after 4 or 8
4.2 Ocular and dermal irritation, sensitization weeks of exposure. In the second study, the rats were exposed
for 8 weeks, and a group of rats that had been exposed at the
Data summarized in the OECD HPV assessments on hydro- highest concentration was held for an additional 4 weeks to
carbon solvents (OECD 2014d) indicated that these solvents assess recovery. Because these investigations were focused
do not produce ocular or dermal irritation at classifiable levels on the potential for these solvents to produce kidney effects,
when tested in animals, nor do they produce allergic contact the assessment of more general systemic parameters was more
dermatitis. Kristiansen and Nielsen (1988) assessed the poten- limited. Nevertheless, the only notable effects of repeated
tial for C7–C11 n-alkanes to produce sensory irritation in exposures to isoparaffinic hydrocarbons were changes in male
mice using the method originally described by Alarie (1966). rat kidneys, consistent with a2u-globulin-mediated processes
They were unable to achieve 50% reductions in respiratory rate (summaries of these data may be found in Amoruso et al. 2008,
(RD50) for alkanes with carbon numbers  C7, and, accord- Carrillo et al. 2013, Johnson et al. 2012, OECD 2012d).
ingly, were unable to calculate RD50 values. More recently, Carrillo et al. (2013) reported a study of a
Studies with human volunteers indicated that these solvents C10–C12 isoparaffinic solvent50 in which rats were exposed
were not primary dermal irritants and that they did not pro- by inhalation to test material at levels of 2600 mg/m3, 5200
duce allergic contact dermatitis (Amoruso et al. 2008, Johnson mg/m3, or 10400 mg/m3 (416, 832, or 1664 ppm). Exposures
et al. 2012, OECD 2014d).
5. Repeated dose studies 49In tabulated information (Table 2 of Nau et al. 1966), the maximum
number of days of exposure is given as 90; however, in the text there is a
5.1 Repeated inhalation toxicity studies discussion of observations after 123 days of exposure. Based on informa-
tion in Table 3 (Nau et al. 1966), it seems most likely that exposure was
Because of the relatively low vapor pressures of these sol- terminated after 90 days.
vents, vapor exposure levels that can be assessed in inhala- 50The solvent contained 99% C9–C12 isoparaffins and 1% C10 cyclopar-
tion toxicity tests are limited, and the inhalation studies that affins.
were for 6 hours/day and 5 days/week for 13 weeks. All of the repeated oral toxicity studies. It should be noted that the use of
rats survived the treatment period, and there were no signifi- oral administration does have some complications. In particu-
cant differences in terminal body weights. There were small lar, care must be taken to avoid introducing test material into
but statistically significant differences in some of the clinical the lungs during dosing, and it should be noted that repeated
and hematological values at study termination, but these were bolus dosing may cause gastro-intestinal irritation which may
all within normal physiological limits and not considered to be complicate the data analysis. Additionally, oral administration
toxicologically important. Liver and kidney weights were sig- is not a common route of human exposure, so data from gavage
nificantly increased, but the only pathological findings were administration studies may have limited use for risk assess-
changes in the kidneys of male rats, consistent with induction ment. Those issues notwithstanding, there have been repeated
of an a2u-globulin-mediated process that was not considered oral studies of all 4 types of solvents in this category; normal
to be relevant to humans. As the liver weight increases were paraffins, iso-paraffins, cycloparaffins, and complex aliphatic
not associated with pathological changes and liver enzyme hydrocarbon solvents. Results of representative studies of
markers were not elevated, the increased liver weights were C9–C14 aliphatic ( 2% aromatic) hydrocarbon solvents and
considered to be adaptive rather than adverse effects. The their constituents are summarized below.
overall no-effect concentration was the highest level tested in
the study, approximately 10400 mg/m3 (1664 ppm).
5.2.1 Results of repeated oral administration studies of normal

There have been at least 3 other unpublished repeated dose paraffins
studies in which rats were exposed by inhalation for periods of There have been two repeated oral toxicity studies of normal
approximately 90 days to vapors of isoparaffinic hydrocarbons paraffins, specifically nC10 and nC11. These studies, which
at levels up to 1800 ppm (12,540 mg/m3). The results of these were actually assessments of both repeated dose toxicity
studies are similar to those above with the principal findings and reproductive/developmental toxicity, were conducted
being male rat kidney effects consistent with a2u-globulin- according to OECD 422 guidelines. In the study of n-decane,
mediated processes (summarized in OECD 2012d). There is in male and female Sprague-Dawley rats were given doses of
addition another unpublished study in which Rhesus monkeys 25, 150 or 1000 mg/kg/day52 by oral gavage in constant
were exposed to a C10–C13 complex hydrocarbon solvent, for volumes of 10 ml/kg. Dosing of males and females was initi-
6 hours/day and 3 days/week for 4 weeks at 4200 mg/m3 (615 ated 14 days prior to mating and continued through a 14-day
ppm). There were no toxicologically important findings, and mating period when the males were sacrificed. The females
the no observed adverse effect level was the highest concen- continued to be treated throughout the gestational period to
tration tested, 4200 mg/m3 (615 ppm) (results summarized in postnatal day 4 when all surviving parental animals and off-
OECD 2012d). spring were sacrificed. There were no clinical, hematologi-
The U.S. National Toxicology Program (NTP 2005) assessed cal or pathological findings. The overall no adverse effect
the potential for decalin to cause systemic effects following level was 1000 mg/kg/day (unpublished study summarized
repeated exposure by inhalation. Male and female F-344 rats in OECD 2012d).
and B6C3F1 mice were exposed for 6 hours/day and 5 days/ The study of undecane followed a similar protocol with
week for 3 months to decalin vapors at levels ranging from doses of 100, 300, or 1000 mg/kg/day. As in the n-decane
25–400 ppm (139–2200 mg/m3). In the rats, the only effects study, there was no evidence of repeated dose effects, and the
reported were increases in absolute and/or relative liver and overall no adverse effect level was 1000 mg/kg/day (Ministry
kidney weights in the male rats. There was also evidence of of Health and Welfare 1996).
histological changes in the kidneys of the male rats consistent
with an a2u-globulin-mediated process. In the mice, increased 5.2.2 Results of repeated oral toxicity studies of isoparaffinic
liver weights were observed in the 400 ppm (2200 mg/m3) hydrocarbons
group along with evidence of centrilobular cytomegaly.
Carpenter et al. (1977b) assessed the effects of a “high In a previously unpublished oral toxicity study of a C11–C15
naphthenic solvent”51 in a repeated inhalation toxicity study isoparaffinic solvent,53 Sprague-Dawley rats were given test
in male rats at exposure levels of 610, 2100, and 5500 mg/ material at levels of 100, 500, or 1000 mg/kg/day. The test
m3 (110, 380, or 1000 ppm). The principal effect was slight material was administered daily, by gavage, in constant doses
tubular regeneration in the kidneys of animals from the high- of 2 ml/kg. The control animals were given vehicle (corn oil)
exposure group. Assuming the kidney effects were due to an only. The majority of the animals were treated daily (7 days/
a2u-globulin-mediated process and not relevant to humans, week) for 13 weeks and then sacrificed for clinical and sys-
the overall NOAEC was 5500 mg/m3 (1000 ppm), the highest temic observations. There was one additional group that was
concentration tested. treated at the high dose and then held for 4 weeks without
treatment to assess recovery. All of the animals survived to
5.2 Repeated oral toxicity studies scheduled termination, and there were no differences in body
weights. There were some statistically significant clinical and
In order to maximize the potential for systemic dose, some of hematological changes but these were small and within normal
the higher molecular weight aliphatic solvents were tested in
51The 52For studies of this type, 1000 mg/kg/day is the “limit” dose.
“high naphthenic solvent” had a distillation range of 157–183°C
53This solvent contained aliphatic hydrocarbons with carbon numbers
and contained approximately 69% naphthenes (25% cyclopentanes, 26%
cyclohexanes and 19% dicyclonaphthenes). The remaining constituents ranging from C12–C15. Approximately 2/3 of the constituents were
were primarily normal- (19%) and iso- (10%) paraffins. isoparaffins, with the remainder being cycloparaffins.
physiological limits. There was a significant increase in liver 5.3 Repeated dermal toxicity studies
weights in the 500 and 1000 mg/kg bw/day dose groups, but
There is one unpublished study (summarized in OECD
there were no pathological changes, and liver enzyme markers
2012d) in which a C12–C14 normal paraffinic solvent was
were not elevated. The only pathological findings were kidney
administered to rabbits in daily dermal applications equiva-
changes consistent with an a2u-globulin-mediated process.
lent to 100, 500 or 2000 mg/kg. The doses were applied under
Aside from the kidney effects, these changes were all reversed
occlusive patches, to prevent test material evaporation. Under
in the recovery study. Considering the liver weight increases
these test conditions, the solvent was so irritating to the skin
to have been adaptive rather than adverse effects, the overall
that all of the rabbits in the 2000 mg/kg/day group either died
no adverse effect level was 1000 mg/kg/day, the highest dose
or were euthanized prior to scheduled termination. The rab-
tested (summarized data in Amoruso et al. 2008, Carrillo et al.
bits in the 500 mg/kg/day group survived to scheduled termi-
2013, Johnson et al. 2012, OECD 2014d).
nation, and there were no significant differences in terminal
body weights. There was evidence of severe skin irritation
5.2.3 Results of repeated dose studies of cycloparaffinic
in the 500 mg/kg/day group but no significant differences
hydrocarbons
in hematological or clinical parameters. There were no sig-
Johannsen and Levinskas (1987) reported a study in which nificant organ weight differences, and the only pathological
male and female Sprague-Dawley rats were fed diets con- changes of note were associated with dermal irritation at the
taining trimethylcyclohexane (a C10 cycloparaffin) at levels sites of test material application. In summary, setting aside
of 3000, 10000, or 30000 ppm (corresponding to dietary effects at the site of administration, the no adverse effect level
levels of approximately 300, 1000, or 3000 mg/kg/day) for for systemic effects following repeated dermal administration
90 days. There were no effects on survival, no differences in was 500 mg/kg/day.
terminal body weights, and no significant changes in clini-
cal or hematological parameters. The only treatment-related 6. Genetic toxicity
effects were increased kidney weights in male rats, along A number of genetic toxicity tests have been conducted
with pathological changes consistent with an a2u-globulin- including bacterial mutagenesis tests in Salmonella, in vitro
mediated process. The overall no adverse effect level was mammalian cell mutagenesis tests (V79 cells), in vitro sister
3000 mg/kg. chromatid exchange assays, as well as dominant lethal tests
As part of the same study, beagle dogs were also exposed and micronucleus tests under in vivo conditions. The majority
to tetramethylcyclohexane by dietary administration at levels of these studies are unpublished but are summarized in Amo-
equivalent to approximately 2.5, 7.5, or 25 mg/kg/day for 90 ruso et al. (2008), Johnson et al. (2012), and OECD (2012d).
days. All of the animals survived to scheduled termination, Moreover, it has also been reported that a complex C9–C14
and there were no notable treatment-related effects, but the aliphatic solvent was inactive in Salmonella and in a micro-
treatment levels tested were very low by comparison to the rat nucleus test in mice (NTP 2004). Decalin was reported as not
studies. mutagenic in Salmonella (NTP 2005). In micronucleus tests
in mice, exposure to decalin was reported to have resulted in
5.2.4 Results of repeated dose studies of complex C9–C14 a small but statistically significant increase in micronucleus
aliphatic( 2% aromatic) hydrocarbon solvents frequency in male, but not in female mice (NTP 2005).
There have been two repeated oral toxicity studies of repre-
sentative complex C9–C14 aliphatic (2% aromatic) hydro- 7. Developmental and reproductive toxicity
carbon solvents. In one study, a C10–C13 complex aliphatic There have been two prenatal developmental toxicity studies
solvent was administered by gavage to rats in daily doses of on substances in this category, along with two repeated dose/
500, 2500, or 5000 mg/kg bw/day for 90 days. The principal reproductive/developmental toxicity screening tests. As dis-
findings in these studies were liver enlargement with centrilob- cussed previously, the data on the C9–C14 aliphatic (2–25%
ular hypertrophy and renal lesions in male rats, consistent with aromatic) hydrocarbon solvents and related materials are rel-
an a2u-globulin-mediated response (Adenuga et al. 2014b). evant on a “worst case” basis and will be briefly summarized
The clinical chemistry investigation revealed elevated levels of in this section but not discussed in detail.
alanine aminotransferase (2500 and 5000 mg/kg/day in male Pregnant female Sprague-Dawley rats were exposed by
rats only), and gamma glutamyl transferase and total bilirubin, inhalation to vapors of either a complex C9–C11 aliphatic
which were elevated only in the 5000 mg/kg/day males. A ( 2% aromatic) hydrocarbon solvent or a C10–C12 isoparaf-
similar dose-response trend in ALT, GGT and bilirubin was finic ( 2% aromatic) hydrocarbon solvent at target concentra-
noted in females, but these differences were not statistically tions of 300 or 900 ppm (1800 or 5220 mg/m3). Exposures
significant compared to control rats. A benchmark dose analy- were for 6 hours/day on gestational days 6–15. All surviving
sis, based on the increase in ALT levels in male rats, provided females were sacrificed on gestational day 21, and the uterine
an estimated point of departure (expressed as the 95% lower contents were examined. It was noted that rats in the high-
confidence limit of the benchmark dose) of 1857 mg/kg/day. exposure groups of both studies exhibited slight increases in
This result was consistent with the data from a repeated oral lacrimation, but there were no other obvious treatment-related
toxicity study of a complex C11–C14 aliphatic hydrocarbon effects during the exposure period. There were also no effects
solvent (summarized in Adenuga et al. 2014b) in which there on body weight gain and no remarkable postmortem changes.
were no toxicologically important findings in rats given daily All data on fetal survival, size, and gender obtained from
doses ranging from 100–1000 mg/kg/day for 13 weeks. the treated groups were comparable to the negative controls.
There was also no evidence of increased frequencies of mal- 8. Carcinogenicity studies

formations or variations and no evidence of developmental
A carcinogenicity study of Stoddard solvent IIC (CAS registry
delays. A positive control substance (acetylsalicylic acid)
number 64742-88-7), corresponding to a complex C9–C14
produced developmental effects as expected. The overall no
aliphatic ( 2% aromatic) hydrocarbon solvent, was con-
adverse effect levels were the highest concentrations tested in
ducted by the NTP (2004). Male and female F-344 rats and
the two studies, that is, 900 ppm or approximately 5220 mg/
B6C3F1 mice were exposed by inhalation, 6 hours/day and 5
m3 (unpublished data summarized in Amoruso et al. 2008,
days/week for 105 weeks. The rats were exposed at concentra-
Johnson et al. 2012, OECD 2012d).
tions of 138 (males only), 550, 1100 and 2200 (females only)
There have also been repeated dose/reproductive toxicity
mg/m3.54 Mice were exposed to 550, 1100, or 2200 mg/m3. At
screening tests of n-decane and n-undecane. In these studies,
termination, the survivors were sacrificed and evaluated for
which followed the OECD 422 guidelines, Sprague-Dawley
the presence of gross and pathological lesions.
rats were given test material in daily oral doses. In the n-de-
In the male rats, there was an increase in benign and malig-
cane study (unpublished, summarized in Amoruso et al. 2008,
nant adrenal gland tumors (pheochromocytomas), with the
Johnson et al. 2012, OECD 2012d), rats were given test mate-
tumor frequencies in the 550 and 1100 mg/m3 (100 and 200
rial in daily doses of 25, 150, or 1000 mg/kg bw/day. Dosing
ppm) groups being significantly elevated with respect to control
of males was initiated 14 days prior to mating and continued

values. There was also a slightly increased incidence of renal
to the end of the mating period. Dosing of females was initi-
tubule adenoma in the male rats. There were no tumors that
ated 14 days prior to mating and continued to postnatal day 4,
were significantly elevated in the female rats. As shorter-term
at which time all surviving females and their offspring were
studies had provided evidence of renal effects in male rats, addi-
examined and then sacrificed. There were no treatment-related
tional studies were conducted to provide mechanistic informa-
effects in any parameter; the rats survived, grew and mated
tion relative to the kidney changes. Among other things, it was
normally. There were no differences in gestational length, litter
shown that the mean numbers of labeled cells and the labeling
size, or number of surviving offspring, and there were no dif-
indices (evidence of increased cell replication) were increased
ferences in offspring survival or body weight to lactational day
in the kidneys of male rats exposed to 550 and 1100 mg/m3
4. Finally, there was no evidence of external malformations.
(100 and 200 ppm) groups. There was evidence that levels of
The no observed adverse effect level was 1000 mg/kg bw/day.
a2u-globulin were increased in male rat kidneys, along with
The study of n-undecane was similar but with a somewhat dif-
histological findings including evidence of granular casts and
ference dose range (100, 300, or 1000 mg/kg/day). However,

cortical tubule degeneration and regeneration, and severity of
as in the n-decane study, there were no differences in any of
hyaline droplets. None of these changes were observed in the
the parameters evaluated, and the no observed adverse effect
kidneys of female rats. Kidney tumors in male rats that are
level was 1000 mg/kg bw/day (Ministry of Health and Welfare
associated with the induction of a2u-globulin are not consid-
1996).
ered to be relevant to humans (US EPA 1991, Swenberg and
Reproductive tissues of male and female rats and mice
Lehman-McKeeman 1998). The toxicological significance
exposed to decalin (0, 25, 50, 100, 200, or 400 ppm) 6 hours/
of the increased incidence of adrenal gland tumors in male
day and 5 days/week for 14 weeks were also examined for
mice is questionable. As noted by Nyska et al. (1999) and later
potential reproductive organ effects (NTP 2005). Samples
by Greim et al. (2009), hyperplasia and neoplasia in the rat
were collected for the evaluation of sperm count, motility, and
adrenal gland have been commonly observed to occur both
vaginal cytology evaluations on rats and mice exposed to 0,
spontaneously and in association with some chemicals. At one
100, 200 or 400 ppm. Parameters evaluated included sperma-
time, the spontaneous incidence in male F-344 rats was nearly
tid counts per testis, per gram testis, per cauda and per gram
40%, but it has been in the range of 12%–16% since a dietary
cauda, and epididymal spermatozoal motility. The left cauda,
change in 2000. In contrast, pheochromocytomas are very rare
left epididymis, and left testis were also weighed. There were
in humans, occurring at a frequency of 1/100,000 (Greim et al.
no effects on sperm motility, sperm counts, or vaginal cytology,
2009). It has also been observed that the adrenal gland tumors
including time spent in various estrous cycle stages and estrous
in rats are associated with kidney changes of certain types,
cycle length, in rats or mice. The only statistically significant
leading Greim et al. to include Stoddard solvent IIC among
finding in mice was a decrease in spermatid head counts per
those substances for which the pheochromocytomas may have
testis (107) in the 400 ppm (2200 mg/m3) dose group. This
been secondary to nephrotoxic effects.
finding did not appear to be of toxicological relevance as there
There was no evidence of carcinogenic activity of Stod-
were no changes in spermatid counts on a per gram testis, per
dard solvent IIC in male mice. In female mice, there was an
gram cauda epididymis, or per cauda epididymis when com-
increase in hepatocellular carcinoma that was associated with
pared to chamber controls. In conclusion, the authors consid-
increased body weight. According to the report, the NTP had
ered that the lack of consistent evidence by other spermatid
developed a statistical model (Haseman et al. 1997) that esti-
and sperm measurements indicated a lack of biological signifi-
mated liver tumor occurrence based on 52-week body weights
cance for the spermatid head counts (NTP 2005).
and age at death. Based on this model, the slight increase in
Additionally, as previously discussed, there were no devel-
incidence of liver neoplasms of female mice was primarily due
opmental effects in 3 studies of C9–C14 aliphatic (2–25% aro-
to the increased incidence of body weights.
matic) hydrocarbon solvents (OECD 2012c), in a developmen-
tal toxicity study of jet fuel (Cooper and Mattie 1996), or in a
repeated dose/reproductive toxicity test of hydrodesulfurized 54These concentrations are equivalent to approximately 28 (males only),
kerosene (Schreiner et al. 1997). 100, 200, and 400 (females only) ppm.
In summary, a C9–C14 aliphatic ( 2% aromatic) hydro- robehavioral effects and others were held for 65 days and
carbon solvent was tested for carcinogenic properties by the then evaluated for persistent effects. The acute studies pro-
NTP. There was evidence of increased frequencies of kidney vided little evidence of effects. In the longer-term studies,
tumors in male rats (most likely the consequence of an a2u- the authors reported that repeated exposure to 1000 mg/m3
globulin-mediated process), adrenal gland tumors in male rats induced significant deficits in acquisition or performance of
(plausibly a secondary consequence of the renal effects), and moderately difficult or difficult tasks, but not simple learn-
liver tumors in female mice (most likely secondary to increased ing tasks as compared to those animals exposed to 500 mg/
body weights in these animals). m3. The toxicological relevance of these data is unclear as
In the older literature, there are reports that aliphatic hydro- the animals in the intermediate exposure group (500 mg/m3)
carbons with carbon numbers ranging from approximately performed better than, but were not significantly different
C10–C16 could potentially promote skin tumors (Horton from, the air-exposed control animals, and the performance
et al. 1957, Sice 1966). It was noted that the peak in promot- of animals in the high exposure group was not significantly
ing potential approximated the peak in irritancy in a study that different from that of the air-exposed control animals, but,
examined hydrocarbons with carbon numbers ranging from rather, was different from that of the animals in the interme-
C6–C24 (Brown and Box 1970). Nessel et al. (1999) assessed diate exposure group.
the extent to which repeated dermal irritation could explain

the promoting effects of these hydrocarbon constituents using 10. Observations in humans
a C10–C14 normal paraffinic solvent. The solvent was diluted 10.1 Studies of acute central nervous system effects and
in a non-irritating mineral oil and then tested for promotional upper respiratory irritation
properties under conditions in which the solvent was applied in
constant weekly volumes but at differing frequencies, to control The earliest reports of the effects in humans with exposure
the level of skin irritation. The results of this study indicated to C9–C14 aliphatic solvents with low aromatic content were
that tumor promotion could only be observed when there was from Pedersen and Cohr (1984a, 1984b). In the first of these
also evidence of skin irritation, providing evidence that the reports, volunteers were exposed for 6 h to dearomatized ali-
promotional effects were a secondary consequence of repeated phatic solvents at levels of 304 mg/m3 (51 ppm), 611 mg/m3
skin irritation and not an intrinsic property of the hydrocarbons (102 ppm), or 1228 mg/m3 (205 ppm). The participants were
themselves. evaluated in a toxicological screening program including bio-
chemistry of blood and urine, lung function, odor threshold,

Romberg test, nystagmus, ECG, blood pressure, pulse, eye
9. Neurotoxicity studies examination and mucociliary function. The volunteers were
There have been several studies in which the potential for the also given questionnaires on subjective symptoms and given
C9–C14 aliphatic ( 2% aromatic) hydrocarbon solvents to stress tests. The authors reported that the subjects did not
produce acute CNS effects have been investigated in animals. experience dryness of the mucous membranes, loss of appe-
Carpenter et al. (1977) and later Nilsen et al. (1988) reported tite, nausea, vomiting, diarrhea, or fatigue. Sleeping patterns
evidence of acute CNS effects in studies in which rats were were not disturbed. There were no reports of effects sugges-
exposed by inhalation to vapors of n-nonane. However, Nilsen tive of acute CNS effects; specifically, headache, dizziness,
et al. (1988) reported that there was no evidence of acute CNS feeling of inebriation, visual disturbances, tremor, feeling of
effects in inhalation studies of aliphatic hydrocarbons with muscle weakness, impairment of coordination or paresthesia
10 or more carbons. Bowen and Balster (1998) reported that of skin. The results of the clinical evaluation revealed small
it was not possible to achieve stable vapor concentrations of but statistically significant reductions in serum alpha amylase
C10–C12 isoparaffinic hydrocarbons that were sufficiently high and urate levels. These differences were statistically signifi-
to produce acute CNS effects in mice. Lammers et al. (2011) cant 48 h after exposure and only in the group exposed to
and McKee et al. (2011) reported subtle CNS effects in rats 205 ppm.
exposed to 5000 mg/m3 (850 ppm) n-decane and to C9–C11 In a second study, Pedersen and Cohr (1984b) exposed vol-
isoparaffinic and cycloparaffinic solvents. Lower concentra- unteers to a C10–C11 isoparaffinic solvent, 6 hours/day for 5
tions were without effect, and there was no evidence of acute consecutive days at a concentration of 600 mg/m3 (100 ppm).
CNS effects in rats tested 24 h after the last exposure period. There were no changes in levels of IgG, IgA, IgM, or oroso-
There are no reported studies in which the neurological mucoid, providing evidence that exposure at this level does
effects of repeated exposures to C9–C14 aliphatic solvents not cause upper respiratory tract irritation or inflammatory
have been assessed in laboratory animals. However, repeated responses.55
exposure to a C9–C14 aliphatic solvent with approximately
20% aromatics did not cause any persistent CNS effects
(Kulig 1990), and no effects were reported in a repeated dose 55The investigation also included an assessement of serum chemistry
neurotoxicity study of hydrodesulfurized kerosene in which changes. These are provided for completeness but not discussed in the
the test material was administered by dermal application in text as they are unrelated to either CNS effects or upper respiratory tract
rats at doses up to 495 mg/kg/day (Breglia et al. 2014). irritation. The principal changes in serum chemistry were an increase
Ritchie et al. (2001) investigated the effects of inhaled in serum levels of creatinine kinase and an 11% reduction in follicle
stimulating hormone levels. The increase in creatinine kinase levels was
JP-8 jet fuel. Rats were exposed 6 hours/day and 5 days/ plausibly associated with higher levels of physical activity in the exposed
week for 6 weeks to jet fuel vapors at levels of 500 or 1000 subjects in comparison to the controls. The reasons for the differences in
mg/m3. Some rats were tested immediately for acute neu- FSH levels are unknown.
Ernstgard et al. (2009a, 2009b) exposed volunteers to varying plausibly associated with higher levels of physical activity in
levels of two solvents, “regular” white spirit56 and a “dearomatized the exposed subjects in comparison to the controls. The rea-
white spirit”57, to compare the potential of these two substances sons for the differences in FSH levels are not known.
to cause ocular and/or respiratory irritation and acute central
nervous system depression. In initial studies, Ernstgard et al. con- 10.3 Occupational exposures and recommended
ducted tests at levels up to approximately 600 mg/m3 (109 ppm); occupational exposure limits
there was little evidence that either irritation or central nervous The recommended occupational exposure limit for nonane in
system effects were associated with exposure to the dearomatized the United States is 200 ppm (approximately 1050 mg/m3).
white spirit (a C9–C14 aliphatic ( 2% aromatic) hydrocarbon There are no specific recommendations for higher molecular
solvent) (Ernstgard et al. 2009a). By comparison, higher levels weight, lower volatility hydrocarbon solvent constituents; how-
of respiratory irritation were associated with exposure to regular ever, a guidance value of 1200 mg/m3 for C9–C15 aliphatic
white spirit, but the differences were only statistically significant hydrocarbons is recommended for use in calculating occu-
at levels of 500–600 mg/m3 (91–109 ppm). In more detailed stud- pational exposure limits for complex hydrocarbon solvents
ies in which volunteers were exposed to these solvents at levels of (ACGIH 2009, Appendix H). As noted in Section Occupa-
either 100 or 300 mg/m3 (18–55 ppm) for 4 h, the only statistically tional exposure limits, it is also recommended that exposures
significant difference was an increase in eye irritation in subjects to aerosols of these substances be limited to 5 mg/m3.
exposed to regular white spirit at 300 mg/m3 (55 ppm). No statis- Caldwell et al. (2000) reported that between 1961 and 1998,
tically significant differences in measures of irritation were found the weighted average exposure to C10–C12 hydrocarbons was
in subjects exposed to the dearomatized white spirit. In addition 1.8 mg/m3 (n  96).
to the assessment of irritation which was primarily subjective, the
authors reported that there were no significant changes in more
objective indicators of upper respiratory irritation tests includ- II. Toxicological properties of C14–C20 aliphatic
ing pulmonary function, nasal swelling, nasal airway resistance, ( 2% aromatic) hydrocarbon solvents
breathing frequency, blinking frequency, and plasma inflamma- (Category 9 solvents)
tory markers (C-reactive protein, interleukin-6). Ernstgard et al.
1. Introduction
did not find any consistent evidence that exposure to either of the
solvents at either of the exposure levels resulted in neurological Category 9 solvents are hydrocarbon solvents composed of ali-
impairment (Ernstgard et al. 2009a, 2009b). phatic hydrocarbon solvent constituents ( 2% aromatics) with
Kjaergaard et al. (1989) investigated the potential for n-decane carbon numbers in the range of C14–C20 and boiling in the range
to cause ocular and/or respiratory irritation in a study in which 63 of approximately 200–350°C. There are three general types of
volunteers were exposed for 6 h to decane at levels given as “0, solvents in this category, namely complex aliphatic solvents com-
10, 35, or 100 ml/l pure n-decane (1 ml/L or approximately 5.82 posed of normal-, iso-, and cycloparaffins in varying amounts,
mg/m3)”, making the exposure levels 58, 204, and 582 mg/m3. complex isoparaffinic/cycloparaffinic solvents, and mono-constit-
The principal indicators of effect in this study, mucus membrane uent solvents, primarily normal paraffins. For REACH purposes,
irritation threshold and skin irritation, may have been associated these solvents were registered generically as “Hydrocarbons,
with increasing exposure, but the authors could not be certain C14–C20 aliphatics,  2% aromatics” in accordance with the
that the effects were due to exposure and not a consequence of naming convention introduced by the European Hydrocarbon
bias, as odor would have provided some indication of the levels Solvent Producers Association (HSPA 2011, Table 2).
to which the subjects were being exposed. In summary, some There have been few repeated exposure studies with the C14–
irritation may have been experienced by the subjects, but this C20 aliphatic ( 2% aromatic) hydrocarbon solvents specifically,
seemed to have been mostly discomfort and hard to differentiate in part for technical reasons discussed in previous sections, but
from simply a reaction to odor of the solvent. the potential for toxicological effects of hydrocarbon substances
with similar physical and chemical properties has been addressed
10.2 Investigations involving other physiological and/or in studies of petroleum substances with either lower (e.g., stud-
clinical parameters ies of kerosene and jet fuel) or higher (mineral oils) molecular
weight petroleum substances. As discussed in more detail in
Pedersen and Cohr (1984b) exposed volunteers to a C10–C11
previous sections, “kerosene” is a generic descriptor used in the
isoparaffinic solvent, 6 hours/day for 5 consecutive days at a
petroleum industry for complex hydrocarbon fuels with boiling
concentration of 600 mg/m3 (100 ppm). The clinical chemistry
ranges of approximately 150–290°C and carbon numbers in the
investigation revealed an increase in serum levels of creatinine
range of C9–C16, and containing approximately 80% aliphatic
kinase and an 11% reduction in follicle stimulating hormone
and 20% aromatic constituents (American Petroleum Institute
(FSH) levels. The increase in creatinine kinase levels was
2010). The kerosene-range material that is derived from crude
56The test sample was a commercial solvent with a boiling range of oil by distillation is commonly referred to as “straight-run”. This
142–200°C, containing 80% aliphatic constituents (primarily C10–C12, material can be refined by treatment with hydrogen to remove
of which 23% were normal paraffins, 28% were isoparaffins, and 29% sulfur- and nitrogen-containing compounds. The majority of this
were naphthenes), and 20% aromatic constituents, primarily C9–C11 material is used for aviation fuels (jet fuels) although there are
alkyl benzenes.
57The test sample was a commercial solvent with a boiling range of other uses. Straight-run kerosene can also be used as a feedstock
145–200°C, containing  99% aliphatic constituents, primarily C9–12, for the production of hydrocarbon solvents. Because straight-run
of which 22% were normal paraffins, 23% were isoparaffins, and 55% kerosene and jet fuels contain approximately 20% aromatics, they
were naphthenes. are more closely aligned with C9–C14 aliphatic solvents (2–25%
aromatic), and C14–C20 aliphatic (2–30% aromatic) hydro- sure would not normally result in substantial systemic absorp-
carbon solvents, but can also be considered as “worst case” in tion. The low vapor pressures of the constituents of these solvents
comparison to the category of C9–C14 aliphatic solvents, ( 2% limit the potential for exposure by inhalation under normal cir-
aromatic) and C14–C20 aliphatic ( 2% aromatic) hydrocarbon cumstances. For example, the saturated vapor concentration for
solvents because of their higher levels of aromatic constituents. n-tetradecane at 25°C is 202 mg/m3, and that for iso-octadecane
For purposes of this review, the data from studies of kerosene and is 11 mg/m3 (ECETOC 1997). However, if aerosol formation is
jet fuel are used to characterize the hazards of the higher molecu- a possibility, it would be reasonable to consider these as mineral
lar weight solvents in this group. oil mists for purposes of occupational control measures.
Mineral oils are complex petroleum substances with carbon
numbers broadly in the range of C16–  C35. They are com- 3.1 Absorption
monly manufactured from the residuum of atmospheric distil- The potential for percutaneous absorption of jet fuel constituents
lation of crude oil and separated under vacuum into streams of was measured using perfused porcine skin flaps (Riviere et al.
varying viscosity for use in the manufacture of lubricating oils. 1999); the fraction absorbed was reported as 0.18%. In studies
Crude oils often contain polycyclic aromatic hydrocarbons with human skin, the percutaneous absorption rate for hexa-
which predominantly segregate to the atmospheric residuum decane was reported as 7  10 3 nmol/cm2/hr (approximately
because of their boiling points. The polycyclic aromatic hydro- 0.0002 mgm/cm2/hr) (Singh et al. 2002). In short, the constitu-
carbons are removed from the lubricating oil feed-stocks during ents of these solvents are not well absorbed dermally; rather,
the refining process, most commonly by a process called solvent the exposure route most likely to result in systemic doses is oral
extraction which selectively removes aromatics with 2 or more administration. Based on the empirical relationship of Albro and
rings. The products of vacuum distillation, commonly referred Fishbein (1970), the absorption efficiency drops from approxi-
to as base oils, are primarily long-chain aliphatic molecules. In mately 60% to 30% over the C14–C20 carbon number range.
some cases, these oils are further refined either by more exten-
sive hydrogenation or acid extraction to reduce the aromatic 3.2 Metabolism, distribution, and excretion
constituents to very low levels, to meet regulatory requirements
Le Bon et al. (1988), studied the metabolic fate of pristane
for food contact applications and other sensitive end uses.
(2,6,10,14-tetramethylpentadecane) in rats. Radiolabeled
These more highly refined base oils are commonly referred
test material was administered as a single gavage dose. Most
to as “mineral oils”. For purposes of this report, mineral oils,
of the administered material (66%) was excreted in the feces,
which are complex aliphatic hydrocarbon substances with low

primarily as non-metabolized material. The remaining radio-
aromatic content, are similar to the C14–C20 aliphatic ( 2%
activity was either excreted in the urine, as pristane metabo-
aromatic) hydrocarbon solvents. In particular data obtained
lites and tritiated water or remained in the carcass, primarily
with a low viscosity mineral oil with an average carbon number
in liver or adipose tissue, as pristane metabolites and tritiated
of C20 are directly relevant to the upper range of the C14–C20
water. The principal identified metabolites were pristan-1-ol,
aliphatic ( 2% aromatic) hydrocarbon solvents.
pristan-2-ol, pristanic acid, and 4,8,12-trimethyldecanoic
2. Physical and chemical properties acids. The metabolic pattern indicates that pristane is metab-
olized by subterminal hydroxylation or terminal oxidation
The C14–C20 aliphatic ( 2% aromatic) hydrocarbon solvents followed by the classic ß-oxidation process. As described
are colorless, odorless liquids with low vapor pressures. The previously, this is typical for aliphatic hydrocarbon solvent
physical and chemical properties are summarized in Table 26. constituents, particularly those with longer carbon chains.
In summary, constituents of C14–C20 aliphatic ( 2%
3. Toxicokinetic properties of C14–C20 aliphatic aromatic) hydrocarbon solvents are absorbed to some extent
( 2% aromatic) hydrocarbon solvents
if ingested, but percutaneous absorption is limited, and
Because of the physical and chemical properties of the C14–C20 exposure by inhalation is unlikely except under conditions
aliphatic ( 2% aromatic) hydrocarbon solvents, external expo- of aerosol formation. Once absorbed, these constituents are
Table 26. Physical and chemical properties of Category 9 aliphatic solvents.

Property Complex Aliphatic Solvents (C14–C20) Complex Isoparaffinic/Cycloparaffinic Solvents (C14–C19)
Molecular Weight Range 198–282 198–282
Physical State at 20°C and 1013 hPA Clear, colorless liquid Clear, colorless liquid
Pour Point  0°C  0°C
Boiling Point 230–370°C 229–355°C
Relative Density 0.79–0.95 g/cm3 0.75–0.90 g/cm3
Vapor Pressure  0.001kPa  0.001kPa
Surface Tension 25–30 mN/m using (Wilhelmy plate method) 25–30 mN/m (Wilhelmy plate method)
Water Solubility Essentially insoluble Essentially insoluble
Flash Point  97°C  93°C
Flammability 0.5–7.0% (v/v) 0.6–7.0% (v/v)
Self-Ignition Temperature  200°C  200°C
Viscosity 3.3–20 mm2/second at 20°C 5.0–25 mm2/second at 20°C
Odor Threshold Odorless Odorless
Maximally Attainable Vapor  200 mg/m3 ( 25 ppm)  200 mg/m3 ( 25 ppm)
Concentration
Conversion Factor (ppm/mg/m3) 1 ppm   8 mg/m3 1 ppm   8 mg/m3
metabolized by ß-oxidation and either excreted in the urine, levels of up to 1000 mg/m3, there was little evidence of tox-
primarily in the first 24 h, or reincorporated into other mac- icity in rats or mice. The principal effects were decreased
romolecules. body weight in male rats along with evidence of a2u-globulin
nephropathy, a characteristic effect in male rat kidneys that is
4. Acute toxicity not considered relevant to humans. In oral studies, jet fuel was
The constituents of C14–C20 aliphatic ( 2% aromatic) hydro- administered to rats by gavage in daily doses of 750, 1500,
carbon solvents have limited potential to produce acute toxic- or 3000 mg/kg/day for 90 days. As in the inhalation studies,
ity. Unpublished data from acute oral and dermal toxicity tests there was a reduction in terminal body weights and evidence
indicate that the LD50 values are greater than the highest levels of a2u-globulin induction in male rats. There was also evi-
tested. There was no evidence of acute inhalation toxicity, but the dence of gastritis and perianal dermatitis which were probably
ability to test was limited by the low vapor pressures of the sol- related to repeated irritation of the gastrointestinal tract, sec-
vent constituents. Vapor concentrations high enough to produce ondary to repeated administration of large bolus doses of liq-
acute CNS effects cannot be achieved under experimental con- uid hydrocarbons. The authors also reported increased levels
ditions for aliphatic hydrocarbons with carbon numbers greater of the liver enzyme markers ALT and AST, but there were no
than approximately C10 (Bowen and Balster 1998, Nilsen pathological changes in the livers themselves. In repeated der-
et al. 1988). The acute toxicity data are summarized in Table mal toxicity studies, there were no effects at treatment levels
27. These solvents produced mild skin irritation when tested in up to 495 mg/kg/day, considered to be the highest level that
rabbits and in volunteers, essentially no ocular irritation when could be applied without causing excessive dermal irritation.
tested in rabbits, and are not classified for ocular or dermal irri- In a study of higher molecular weight aliphatic constitu-
tant properties. These solvents did not produce allergic contact ents, female F-344 rats were fed diets containing 0.02%, 0.2%,
dermatitis when tested in guinea pigs or in volunteers. or 2% of a white mineral oil (equivalent to approximately 20,
200, or 2000 mg/kg/day) with an average carbon number of
5. Repeated dose studies C20, on a daily basis for 90 days (McKee et al. 2012). At
the end of the scheduled treatment period, the rats were sac-
Skyberg et al. (1990) reported an inhalation study in which rificed and examined for evidence of toxicologically relevant
rats were exposed 7 hours/day and 5 days/week for 2 weeks findings. All of the rats survived the treatment period with-
at levels of either 70 or 700 mg/m3 to petroleum-derived min- out evidence of clinical signs or differences in terminal body
eral oils or synthetic lubricants produced from alkylbenzenes
weights. There were a few differences in clinical chemistry

or polybutene. The substances of most direct relevance to this values but as the differences were small and within normal
report were two synthetic oils, oil A which was composed physiological ranges, they were considered to be unimport-
of C14–C18 alkylbenzenes, and oil C which was composed ant. The only statistically significant difference in the hema-
of C15–C19 alkylbenzenes. As indicated by the authors, the tological investigation was an increase in neutrophils in the
exposures were primarily to aerosols. Body weight gain was high-dose group. Kidney and liver weights were significantly
significantly reduced in animals exposed to oil A, and to the increased in the high-dose group. The histological findings
low level of oil C, but rats exposed to the high level of oil C were limited to evidence of microgranuloma formation in the
lost weight. The principal effects were increased lung weights, livers and mesenteric lymph nodes in the animals of the high-
most likely associated with oil deposition, but there were no dose group. The principal objective of this study was to exam-
pathological changes or systemic effects. The authors did not ine the potential for mineral oils to accumulate in the liver
consider that these oils produced toxic effects in the lung. and mesenteric lymph nodes, an effect previously reported to
As noted above, there have been few repeated dose studies occur in F-344 rats but not in rats of other strains or animals
on the C14–C20 aliphatic hydrocarbon solvents specifically, of other species (Baldwin et al. 1992, Bird et al. 1990, Firriolo
but the potential for effects has been addressed through stud- et al. 1995, Smith et al. 1995, 1996, Scotter et al. 2003). A
ies of petroleum substances with either lower (e.g., studies principal finding was that the hydrocarbons that preferentially
of kerosene and jet fuel) or higher (mineral oils) molecular accumulated in the liver, leading to granuloma formation,
weight petroleum substances. As discussed in more detail had carbon numbers  C20. In contrast, the mineral oil con-
in summaries of studies of other solvents, Carpenter et al. stituents with carbon numbers in the range of C15–C20, that
(1976c) reported minimal effects in rats and beagle dogs is, corresponding approximately to the C14–C20 aliphatic
exposed for 90 days to vapors of deodorized kerosene. The ( 2% aromatic) hydrocarbon solvents, were metabolized and
highest concentration tested was 400 mg/m3, indicated by excreted. Based on a pathological investigation of human liver,
the authors as the maximally attainable vapor concentration. Barp et al. (2014) confirmed that aliphatic hydrocarbons with
Mattie et al. (1991) reported that even under conditions of carbon numbers  C20 did not accumulate in liver, spleen,
continuous exposure (i.e., approximately 24 hours/day) to lung, heart, or kidney. Additionally, Barp et al. (2014) found
no evidence that aromatic hydrocarbons accumulate in these
Table 27. Summarized results of acute toxicity studies of Category 9 tissues. In short, these studies provided evidence that the only
solvents.
systemic change in rats that could be associated with the inges-
Endpoint Complex Aliphatic Solvents (C14–C20) tion of mineral oils, was the development of granulomas in the
Acute Oral Toxicity  5000 mg/kg liver and mesenteric lymph nodes; that this change was not
Acute Dermal Toxicity  2000 mg/kg associated with aliphatic hydrocarbons with carbon numbers
Acute Inhalation Toxicity  5800 mg/m3 (nominal) less than C20; and that aliphatic hydrocarbons with carbon
( 725 ppm)
numbers less than C20 did not accumulate in human liver.
6. Genetic toxicity section, they are not developmental toxins. Accordingly, it is

anticipated that these substances would not cause reproductive
Results of unpublished in vitro tests for gene mutation and chro-
toxicity. That hypothesis is further supported by evidence that
mosomal aberration indicate that C14–C20 aliphatic ( 2%
jet fuel did not affect reproductive processes, in an oral study
aromatic) hydrocarbon solvents are not genotoxic under in vitro
at doses  1500 mg/kg/day (Mattie et al. 2000). Similarly,
conditions. These studies are supported by data from studies of
dermally-administered hydrodesulfurized kerosene did not lead
kerosene (American Petroleum Institute 1984a, 1987, Black-
to reproductive or developmental effects in a study involving
burn et al. 1987), jet fuel (Mattie and Sterner 2011, McKee
dermal administration at levels up to 495 mg/kg/day, the highest
et al. 1994), and diesel fuel (McKee et al. 1994). No in vivo
doses applied (Schreiner 1997). An unpublished one-generation
studies of the genotoxic potential of C14–C20 aliphatic ( 2%
reproductive toxicity study of a C16–C30 mineral oil (summa-
aromatic) hydrocarbon solvents were identified, but there have
rized in Dalbey et al. 2014) was conducted to evaluate the repro-
been both chromosomal aberration studies and micronucleus
ductive performance (gonadal function, all 4 stages of the estrus
tests of kerosene (American Petroleum Institute 1984b), jet
cycle, fertilization, implantation, length of gestation, and parturi-
fuel (American Petroleum Institute 1980a, Mattie and Sterner
tion) in Sprague-Dawley rats. The animals were exposed to test
2011) and diesel fuel (McKee et al. 1994). As all of these tests
material by dermal administration at levels of 125, 500, or 2000
produced negative results, C14–C20 aliphatic ( 2% aromatic)
mg/kg bw/day. The treatment began 10 weeks prior to mating

hydrocarbon solvents are considered to be non-genotoxic.
and continued through a 3-week mating period and continued to
gestational day 20, at which point dosing was suspended, and the
7. Developmental toxicity animals were held without further treatment to postnatal day 21,
In a developmental toxicity study summarized in the REACH when all surviving dams and offspring were sacrificed. There
registration dossier (ECHA 2010b), a complex C16–C20 ali- were no clinical signs of systemic toxicity and no effects on
phatic hydrocarbon solvent was given by gavage to Sprague- maternal body weight or food consumption. As described in the
Dawley rats at doses of 400, 800, or 1000 mg/kg/day on days previous section, there were no effects on survival or develop-
6–15 of gestation. In the gross examination, there were no ment of offspring, no evidence of malformation, and there were
signs of maternal toxicity. In the uterine examination, there no pathological findings. The overall no-effect level in this study
were no differences in fetal survival or weight. Similarly the was  2000 mg/kg bw/day, the highest dose tested.
external, visceral, and skeletal fetal examinations provided no
evidence of malformation or developmental delays. Accord- 9. Carcinogenicity

ingly, the no observed adverse effect concentration was 1000 As C14–C20 aliphatic ( 2% aromatic) hydrocarbon solvents
mg/kg/day, the highest dose tested. are non-genotoxic and do not produce evidence of cumula-
There is an unpublished developmental toxicity study tive toxicity in repeated dose studies, they are considered to be
that assessed the potential effects of a C16–C30 mineral most likely non-carcinogenic.
oil (60% paraffins, 40% cycloparaffins) in Sprague-Dawley rats
(summarized in Dalbey et al. 2014). The study compared the
10. Neurotoxicity
effects following exposure on gestational days 6–19 by oral
(5000 mg/kg/day), dermal (2000 mg/kg/day), or inhalation No specific neurotoxicity studies of C14–C20 aliphatic ( 2%
(1000 mg/m3 as aerosol, 6 hours/day). The dams were sacrificed aromatic) hydrocarbon solvents have been identified. Based on
on gestational day 20, and the uterine contents were examined. data from Nilsen et al. (1988), acute CNS effects are unlikely
There was no evidence of maternal toxicity or fetal effects. for aliphatic hydrocarbons with carbon numbers  C10, in
Accordingly, the no-effect levels/concentrations were the high- part because of difficulties in maintaining stable vapor con-
est doses/concentrations used in this study. In separate studies centrations at sufficiently high levels. The blood/air and brain/
(also summarized in Dalbey et al. 2014), the developmental air ratios in the same paper suggest that in addition to the low
toxicity of a C16–C30 mineral oil (60% paraffins, 40% vapor levels, there are additional factors that limit the hydro-
cycloparaffins) was evaluated in a one generation reproductive/ carbon concentrations that can be achieved in the CNS. Hau
developmental toxicity study in which rats were exposed to test et al. (2001) suggested that uptake of hydrocarbons with car-
material by dermal administration in daily doses of 125, 500, bon numbers  C10 might be impeded by blood/brain barrier
or 2000 mg/kg bw. There were no differences in mean numbers effects. Thus, the association of exposure to high molecular
of implantation sites, live pups/litter, live-birth indices (total weight aliphatic hydrocarbons and chronic CNS effects seems
number offspring born), and survival of offspring on postnatal unlikely due to the physical/chemical properties of these mol-
days 4 and 21. There were no differences in the body weights ecules, and this is supported by empirical evidence.
of offspring, and there were no pathological differences in the
organs of offspring evaluated at postnatal day 21. The NOAEL 11. Observations in humans
for mineral oil was  2000 mg/kg bw/day for prenatal effects.
The potential for dermal irritancy was assessed using a panel
of human volunteers. As described in the study report, a
C14–C19 aliphatic hydrocarbon solvent was diluted in mineral
There have been no formal reproductive toxicity tests of C14–C20 oil administered to 30 volunteers at a 50% concentration under
aliphatic ( 2% aromatic) hydrocarbon solvents. However, there a semi-occlusive patch in a volume of 0.3 grams. None of the
is evidence from repeated dose studies that these substances do subjects developed irritant responses, and the substance was
not affect reproductive organs, and, as indicated in the previous judged to be non-irritating to human skin.
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tatives who reviewed the manuscript for technical accuracy. rat toxicology studies. Food Chem Toxicol, 36, 831–839.
American Petroleum Institute (1977). Teratology study in rats. Stoddard
The authors would like to specifically acknowledge Pierre- Solvent. API Medical Research Publication 26–60026. API,
Yves Guyomar and Niels Vaeck for assistance with analytical Washington, DC.
information and otherwise reviewing this paper before it was American Petroleum Institute (1979a). Teratology study in rats: Kerosene.
API Medical Research Publication 27–32175. American Petroleum
submitted for publication, Andrea Holliday for conducting
Institute, Washington, D.C.

the literature search, and Lynn Bennett for administrative American Petroleum Institute (1979b). Inhalation/Teratology study
assistance in manuscript preparation. Funding for manuscript in rats: Jet Fuel A. API Medical Research Publication 27-32173.
preparation was provided to RHM and MDA by the Hydro- American Petroleum Institute, Washington, DC.
American Petroleum Institute (1980a). Mutagenicity evaluation of Jet Fuel
carbon Solvents Panel of the American Chemistry Council. A in the mouse dominant lethal assay. API Medical Research Publication
The corporate members of the Hydrocarbon Solvents Panel 28-31345. American Petroleum Institute Washington, D. C.
are Chevron Phillips Chemical Company, CITGO Petroleum American Petroleum Institute (1980b). Project No. 78-7233. A 26 week
Corporation, ExxonMobil Chemical Company, and Sasol inhalation toxicity study of heptane in the rat. API Medical Research
Publication 28-31209. American Petroleum Institute, Washington, D.C.
North America, Inc. American Petroleum Institute (1983a). Six-month continuous inhalation
exposures of rats to hexane mixtures – phase 1. API Medical Research
Declaration of Interest American Petroleum Institute (1983b). Six-month continuous inhalation
exposures of rats to hexane mixtures – phase II. API Medical Research
The authors are employed by companies that manufacture

hydrocarbon solvents. The company affiliations of the authors American Petroleum Institute (1984a). Mutagenicity evaluation of
are indicated on the cover page. The authors had sole respon- API#81-07 in the mouse lymphoma forward mutation assay. Medical
Research Publication 32-30240. American Petroleum Institute,
sibility for the writing and content of the paper, although some
Washington, DC.
financial support was provided by the Hydrocarbon Solvents American Petroleum Institute (1984b). Mutagenicity evaluation studies in
Panel. One of the authors (RHM) has been an expert witness the rat bone marrow cytogenetic assay. Hydrodesulfurized Kerosene,
in cases involving hydrocarbon solvent composition and/or API sample 81-07. API Medical Research Publication 32-30240.
American Petroleum Institute, Washington, DC.
toxicity but not within the past 5 years. American Petroleum Institute (1985). Acute oral toxicity study in rat,
acute dermal toxicity study in rabbits, primary dermal irritation study
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Crit Rev Toxicol, 2015; 45(4): 273–365

Characterization of the toxicological hazards of hydrocarbon solvents

irritation at exposure levels exceeding occupational recommendations. Otherwise, there are

Abbreviations: ACGIH, American College of Governmental Industrial Hygienists; AIHA, American

Address for correspondence: Richard H. McKee, ExxonMobil Biomedical

Table of Contents Exposure to hydrocarbon solvents and occupational

and other solvents ����������������������������������������������������������������������������������� 287

Hydrocarbons, 64742-94-5 C10-–C13  99% aromatics  1% naphthalene

Hydrocarbons, 64742-81-0 C10-–C13 10-–25% aromatics  2% aromatics

cyclics,  2% aromatics

Hexadecane 544-76-3 C16 monoconstituent  2% aromatics

as C10-–C13 aromatics in the OECD HPV program.

Absorption (C10 cycloparaffin), 53–67% of the administered material was

studies by Tulliez and Bories (1978), in which it was shown that

to be limited because of their low vapor pressures, and may be

found in urine as sulfated and glucuronidated conjugates.

(Tulliez and Bories 1979). The authors concluded as follows:

of hydrocarbon solvents guinea pigs (hippuric conjugate of p-butylbenzoic acid) were

only 15–20% of 2-methylnaphthalene metabolism occurs via

naphthalenes through ring oxidation decreases with increasing

ficients for alkanes with carbon numbers  C10 suggest blood/

which summarized an investigation of neurological effects hazards of these specific substances.

analytical information to confirm that the solvent used was

effects at exposure levels above the thresholds for irritation

C14–C20 Aliphatics, Yes No No None

mental delays, but these have only been observed at levels

Occupational exposure limits In which Fa is the mass fraction of constituent a, Fb is the

for C9–C15 aromatics. Concentrations of cumene, indene, (excluding use:

ppm, 176 mg/m3; naphthalene  10 ppm, 52 mg/m3. The

II. Toxicological properties of C14–C20 Aliphatic

5.1 Repeated inhalation toxicity studies .. . .. . .. . .. . .. . .. . .. . .. . .. 311

3.3 Metabolism and elimination of C7–C9 aliphatic

Appendix D. Non-Volatile aliphatic hydrocarbon solvents 1. Introduction

3.3 Metabolism and excretion

sistent or significant peripheral blood changes, weight gains,

weeks. All rats survived to scheduled termination, and there

administered by gavage at levels of 50, 200, or 600 mg/kg/ 7. Developmental toxicity

(i.e., 24 hours/day) from days 7–15 of gestation to 120, 200, or

27Aromatol is the brand name for a commercial solvent containing 48%

As summarized in Table 11, repeated exposure to C9

reported that there were no signs of irritation or CNS effects

Table 12. Physical and chemical properties of C10–C12 aromatic solvents.

significantly different) below control values. The hemato-

reported, but occupational exposures to naphthalene have been

11.1 Ocular and respiratory irritation Appendix B. Aliphatic/aromatic hydrocarbon

in the urine as the dimethylbenzoic and/or dimethylhippu-

but marked tubular regeneration was noted in the kidneys

starting 90 days prior to mating and continuing to the end of

mating and continuing to gestational day 19. There were no

acute CNS effects, an increased incidence of headache, tired-

of overexposure which were relatively common during that

intense solvent exposures than they did during construction

Because they have low vapor pressures, substantial exposure

One often contentious issue of experimental design is the

to avoid introducing these substances into the lungs in the

expected because they quickly evaporate from the skin.

Acute Dermal Not Tested

in accordance with the naming convention introduced by the

Table 20. Physical and chemical properties of C6 aliphatic solvents.

5. Repeated dose toxicity 5.1.2 Repeated inhalation toxicity studies

8.3 Reproductive toxicity studies of cyclohexane

the pre-mating periods and on a daily basis through the mat-

and other solvents �� 287