Endocrine Disruption Potentials of Bisphenol A Alternatives - Are Bisphenol A Alternatives Safe from Endocrine Disruption?

Abstract

Objectives: Although a great body of knowledge is available on the toxicity of bisphenol A (BPA), little is known about that of BPA alternatives, such as bisphenol analogues (BPs) or Tritan TM copolyesters. This review provides a summary of the available information on the toxicity of BPs and three components of Tritan TM , with a special focus on endocrine disruption. Methods: We collected from the literature a battery of in vitro and in vivo assay data developed to assess endocrine disruption of four BPs (bisphenol AF, B, F, and S) and three major components of Tritan TM ((di-methylterephthalate (DMT), 1,4-cyclohexanedimethanol (CHDM), and 2,2,4,4-tetramethyl-1,3-cyclobutanediol (TMCD)). Results: Several alternative compounds were identified as possessing comparable or even greater endocrine-disrupting effects than BPA in in vitro and in vivo studies. Conclusions: Potential endocrine disruption of BPA alternatives requires further studies on health consequences in experimental animals and in humans following longer term exposure.

Full text

한국환경보건학회지
,
제
39
권제
1
호
(2013)
J Environ Health Sci, 2013: 39(1): 1-18
1
http://dx.doi.org/10.5668/JEHS.2013.39.1.1
Endocrine Disruption Potentials of Bisphenol A Alternatives
- Are Bisphenol A Alternatives Safe from Endocrine Disruption?
Kyunghee Ji* and Kyungho Choi
School of Public Health, Seoul National University, Korea
ABSTRACT
Objectives: Although a great body of knowledge is available on the toxicity of bisphenol A (BPA), little is
known about that of BPA alternatives, such as bisphenol analogues (BPs) or TritanTM copolyesters. This review
provides a summary of the available information on the toxicity of BPs and three components of TritanTM, with
a special focus on endocrine disruption.
Methods: We collected from the literature a battery of in vitro and in vivo assay data developed to assess endocrine
disruption of four BPs (bisphenol AF, B, F, and S) and three major components of TritanTM ((di-methylterephthalate
(DMT), 1,4-cyclohexanedimethanol (CHDM), and 2,2,4,4-tetramethyl-1,3-cyclobutanediol (TMCD)).
Results: Several alternative compounds were identified as possessing comparable or even greater endocrine-
disrupting effects than BPA in in vitro and in vivo studies.
Conclusions: Potential endocrine disruption of BPA alternatives requires further studies on health consequences
in experimental animals and in humans following longer term exposure.
Keywords: bisphenols, endocrine disruption, Tritan
I. Introduction
Endocrine disruptions due to exposure to chemicals
in various consumer products, e.g., plastics have
received great attention.1) Among them, bisphenol A
(BPA; 2,2-bis(4-hydroxydiphenyl)propane, which has
been produced over eight billion pounds each year
worldwide, is frequently used as a monomer in the
manufacture of polycarbonates and epoxy resins.2)
As BPA can disrupt steroidogenesis and act as a
weak estrogen receptor agonist, concerns on adverse
health outcomes, especially on reproduction and
development, are increasing.3,4) A large number of
biomonitoring studies indicate widespread exposure
to BPA in adults, adolescents, and children from
several different countries,5) while the results from
toxicokinetic studies that determined the disposition
of BPA in humans after oral administration of BPA
are at odds with them.6,7) In 2011, the European
Commission has applied the precautionary principle
on BPA and restricted its use in plastic infant feeding
bottles.8) In response to this restriction, a number of
alternative compounds, such as bisphenol AF (BPAF;
2,2-bis(4-hydroxyphenyl)hexafluoropropane), bisphenol
B (BPB; 2,2-bis(4-hydroxyphenyl)butane), bisphenol
F (BPF; bis(4-hydroxydiphenyl)methane), and bisphenol S
(BPS; bis(4-hydroxyphenyl)sulfone), began to be
often used increasingly as component of plastic
substitutes.2) In addition, a novel plastic is also
manufactured by Eastman Chemical Company
(Kingsport, TN, USA) utilizing three monomers, di-
methylterephthalate (DMT), 1,4-cyclohexanedimethanol
(CHDM), and 2,2,4,4-tetramethyl-1,3-cyclobutanediol
(TMCD) in various ratios, marketed under a trade
name of TritanTM.9)
The production and consumption of bisphenol
analogues (BPs; Table 1) that are structurally similar
to BPA with two hydroxyphenyl functionalities have
[특집]
†Corresponding author: School of Public Health, Seoul National University, Gwanak, Seoul, 151-742, Republic of Korea,
Tel: 82-2-880-2795, Fax: 82-2-745-9104 E-mail: jkh526@snu.ac.kr
Received: 7 February 2013, Revised: 13 February 2013, Accepted: 22 February 2013

2Kyunghee Ji and Kyungho Choi
J Environ Health Sci 2013: 39(1): 1-18 http://www.kseh.org/
increased recently.10) BPAF, a fluorinated derivative
of BPA, is widely used in polycarbonate copolymers
in high-temperature composites, electronic materials,
gas-permeable membranes, and specialty polymer
applications.11-14 ) Approximately 10,000-500,000 pounds
of BPAF are produced annually in the United
States.15) BPB, a BPA analogue having a butyl chain
instead of a propyl chain between the two phenol
moieties, is utilized in the manufacture of resins and
plastics.16) BPF, which differs from BPA only by the
lack of two methyl groups on the central carbons,
has a broad range of industrial applications such as
lacquers, varnishes, liners, adhesives plastics, food
packaging, dental sealants, and water pipes.17) BPS,
whose two phenolic rings are joined together with
sulfur, has excellent stability against high temperature
and resistance to sunlight.18) BPS has been
introduced to the market as a component of plastic
substitutes for the production of babybottles19) or
used as a developer in dyes for thermal paper.20)
Tritan copolyester is used in packaging of
beverages, edible oil, and foods, as well as for food
contact films and foils including microwave
packaging.21) Three important co-monomers of
Tritan, namely CHDM, DMT, and TMCD, were
used for production of polyethylene terephthalate
(PET) bottle in Lock & Lock® company. DMT is
nominated as a high production volume chemical,
both in the United States22) and Organization for
Economic Co-operation and Development.23)
Recent studies have reported the occurrence of
BPA alternatives in environmental samples,
consumer products, food, and human specimens
(Table 2). BPAF has been found in 76% of the 41
Table 1 . Chemical structures of bisphenol A and its alternatives which are commonly used in consumer products
Abbreviation Systematic name CAS Number Structure
BPA 2,2-bis(4-hydroxydiphenyl)propane 80-05-7
BPAF 2,2-bis(4-hydroxyphenyl)hexafluoropropane 1478-61-1
BPB 2,2-bis(4-hydroxyphenyl)butane 77-40-7
BPF Bis(4-hydroxydiphenyl)methane 87139-40-0
BPS Bis(4-hydroxyphenyl)sulfone 80-09-1
CHDM 1,4-cyclohexanedimethanol 105-08-8
(mixture of cis and trans)
DMT Dimethyl terephthalate 120-61-6
TMCD 2,2,4,4-tetramethyl-1,3-cyclobutanediol 3010-96-6
(mixture)

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Tab l e 2 . Concentrations of bisphenol A alternatives in environment, consumer products, and biota
Compounds Location Sample N Median Range LOQ References
BPAF USA Sediment 82 ND ND 0.25 ng/g dw Liao et al., 2012c
Japan Sediment 56 ND ND 0.25 ng/g dw Liao et al., 2012c
Korea Sediment 34 ND <LOQ ~ 4.23 ng/g dw 0.25 ng/g dw Liao et al., 2012c
USA Indoor dust 38 ND ND 0.5 ng/g Liao et al., 2012d
China Indoor dust 55 ND ND 0.5 ng/g Liao et al., 2012d
Japan Indoor dust 22 ND ND 0.5 ng/g Liao et al., 2012d
Korea Indoor dust 41 4.8 ng/g <LOQ ~ 91 ng/g 0.5 ng/g Liao et al., 2012d
BPB Italy Peeled canned tomatoes in canes with
epoxyphenolic lining 6 33.4 µg/kga27.1 ~ 85.7 µg/kg 2.3 µg/kg Grumetto et al., 2008
Italy Peeled canned tomatoes in canes with
low BADGE coating 3 37.7 µg/kga31.3 ~ 45.5 µg/kg 2.3 µg/kg Grumetto et al., 2008
Italy Serum 69 5.15 ng/mLa0.88 ~ 11.94 ng/mL 0.18 ng/mL Cobellis et al., 2009
Portugal Serumb20 0.68 ng/mL <LOQ ~ 1.15 ng/mL 0.05 ng/mL Cunha and Fernandes,
2010
Spain Glass beverage soft-drink cola 1 ND ND 167 ng/L Gallart-Ayala et al., 2011
Spain Glass beverage soft-drink soda 5 ND ND 167 ng/L Gallart-Ayala et al., 2011
Spain Glass beverage soft-drink tonic 1 ND ND 167 ng/L Gallart-Ayala et al., 2011
USA Sediment 82 ND ND 0.5 ng/g dw Liao et al., 2012c
Japan Sediment 56 ND ND 0.5 ng/g dw Liao et al., 2012c
Korea Sediment 34 ND <LOQ ~ 10.6 ng/g dw 0.5 ng/g dw Liao et al., 2012c
USA Indoor dust 38 ND ND 1.0 ng/g Liao et al., 2012d
China Indoor dust 55 ND ND 1.0 ng/g Liao et al., 2012d
Japan Indoor dust 22 ND ND 1.0 ng/g Liao et al., 2012d
Korea Indoor dust 41 ND ND 1.0 ng/g Liao et al., 2012d
BPF Germany Surface-water 30 - <LOQ ~ 180 ng/L 2 ng/L Fromme et al., 2002
Germany Sewage water 25 - 22 ~ 123 ng/L 2 ng/L Fromme et al., 2002
Germany Sediment 7 - 1,200 ~ 7,300 ng/kg
dw 5 ng/kg Fromme et al., 2002
Spain Glass beverage soft-drink cola 1 ND ND 132 ng/L Gallart-Ayala et al., 2011
Spain Glass beverage soft-drink orange soda 1 218 ng/L 218 ng/L 132 ng/L Gallart-Ayala et al., 2011
Spain Glass beverage soft-drink lemon soda 1 141 ng/L 141 ng/L 132 ng/L Gallart-Ayala et al., 2011

4Kyunghee Ji and Kyungho Choi
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Tab l e 2 . Continued
Compounds Location Sample N Median Range LOQ References
BPF Spain Glass beverage soft-drink tonic 1 ND ND 132 ng/L Gallart-Ayala et al., 2011
USA Sediment 82 1.44 ng/g dw <LOQ ~ 27.5 ng/g dw 1.0 ng/g dw Liao et al., 2012c
Japan Sediment 56 3.57 ng/g dw <LOQ ~ 9.11 ng/g dw 1.0 ng/g dw Liao et al., 2012c
Korea Sedim ent 34 ND <LOQ ~ 9,650 ng/g dw 1.0 ng/ g dw Liao et al., 2012c
USA Indoor dust 38 49 ng/g <LOQ ~ 240 ng/g 2.0 ng/g Liao et al., 2012d
China Indoor dust 55 38 ng/g <LOQ ~ 1,890 ng/g 2.0 ng/g Liao et al., 2012d
Japan Indoor dust 22 57 ng/g <LOQ ~ 2,780 ng/g 2.0 ng/g Liao et al., 2012d
Korea Indoor dust 41 450 ng/g <LOQ ~ 1,070 ng/g 2.0 ng/g Liao et al., 2012d
BPS SpaincPeas and carrots (supernatant) 3 175 ng/mLa- 0.0083 ng/mL Viñas et al., 2010
SpaincPeas and carrots (food) 3 36.1 ng/ga- 0.073 ng/g Viñas et al., 2010
SpaincPeas (supernatant) 3 16.7 ng/mLa- 0.0083 ng/mL Viñas et al., 2010
SpaincNatural peas (supernatant) 3 30.9 ng/mLa- 0.0083 ng/mL Viñas et al., 2010
SpaincArtichoke (supernatant) 3 34.3 ng/mLa- 0.0083 ng/mL Viñas et al., 2010
SpaincMushroom (supernatant) 3 11.5 ng/mLa- 0.0083 ng/mL Viñas et al., 2010
SpaincBean shoot (supernatant) 3 14.0 ng/mLa- 0.0083 ng/mL Viñas et al., 2010
SpaincMixed vegetables (supernatant) 3 70.1 ng/mLa- 0.0083 ng/mL Viñas et al., 2010
Spainc
Natural peas, sweet corn, artichoke,
mushroom, bean shoot, and mixed
vegetables (food)
3 ND ND 0.073 ng/g Viñas et al., 2010
Spain Glass beverage soft-drink cola 1 ND ND 167 ng/L Gallart-Ayala et al., 2011
Spain Glass beverage soft-drink soda 5 ND ND 167 ng/L Gallart-Ayala et al., 2011
Spain Glass beverage soft-drink tonic 1 ND ND 167 ng/L Gallart-Ayala et al., 2011
USA(Albany) Thermal receipt paper 81 7,440 µg/g 0.0138 ~ 22,000 µg/g 0.0001 µg/g Liao et al., 2012a
Japan Thermal receipt paper 6 5,500 µg/g 0.546 ~ 6,130 µg/g 0.0001 µg/g Liao et al., 2012a
Korea Thermal receipt paper 11 0.8 µg/g 0.0896 ~ 11 µg/g 0.0001 µg/g Liao et al., 2012a
Vietnam Thermal receipt paper 3 0.3 µg/g 0.105 ~ 0.554 µg/g 0.0001 µg/g Liao et al., 2012a
USA(Albany) Several paper products 157 8.5 µg/g <LOQ ~ 8,380 µg/g 0.0001 µg/g Liao et al., 2012a
USA Urineb31 0.263 ng/mL <LOQ ~ 21.0 ng/mL 0.02 ng/mL Liao et al., 2012b
China Urineb89 0.297 ng/mL <LOQ ~ 3.16 ng/mL 0.02 ng/mL Liao et al., 2012b
India Urineb38 0.055 ng/mL <LOQ ~ 0.881 ng/mL 0.02 ng/mL Liao et al., 2012b

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Tab l e 2 . Continued
Compounds Location Sample N Median Range LOQ References
BPS Japan Urineb36 1.040 ng/mL 0.147 ~ 9.57 ng/mL 0.02 ng/mL Liao et al., 2012b
Korea Urineb33 0.014 ng/mL <LOQ ~ 1.98 ng/mL 0.02 ng/mL Liao et al., 2012b
Kuwait Urineb30 0.371 ng/mL <LOQ ~ 12.1 ng/mL 0.02 ng/mL Liao et al., 2012b
Malaysia Urineb29 0.084 ng/mL <LOQ ~ 0.922 ng/mL 0.02 ng/mL Liao et al., 2012b
Vietnam Urineb29 0.157 ng/mL 0.037 ~ 0.932 ng/mL 0.02 ng/mL Liao et al., 2012b
USA Sediment 82 ND <LOQ ~ 4.65 ng/g dw 0.25 ng/g dw Liao et al., 2012c
Japan Sediment 56 ND <LOQ ~ 4.46 ng/g dw 0.25 ng/g dw Liao et al., 2012c
Korea Sediment 34 ND <LOQ ~ 1,970 ng/g dw 0.25 ng/g dw Liao et al., 2012c
USA Indoor dust 38 630 ng/g 5.6 ~ 25,500 ng/g 0.5 ng/g Liao et al., 2012d
China Indoor dust 55 170 ng/g 0.83 ~ 12,600 ng/g 0.5 ng/g Liao et al., 2012d
Japan Indoor dust 22 810 ng/g 250 ~ 2,550 ng/g 0.5 ng/g Liao et al., 2012d
Korea Indoor dust 41 360 ng/g 90 ~ 26,600 ng/g 0.5 ng/g Liao et al., 2012d
aMean value.
bValues indicate creatinine-unadjusted concentration.
cValues are analyte concentrations with derivatization procedures using bis-(trimethylsilyl)trifluoroacetamide (BSTFA).
LOQ: limit of quantification, ND: non-detected, -: not available.

6Kyunghee Ji and Kyungho Choi
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indoor dust samples collected in South Korea
(median 4.8 ng/g).24) BPB has been found in human
serum from Italy (mean 5.15 ng/mL)25) and Portugal
(mean 0.68 ng/mL),26) and in canned foods (mean
42.3 ng/g).27) BPF has been reported in surface
water, sewage, and sediments at concentrations
ranging from below the limit of quantification
(LOQ) to 0.180 µg/L, 0.022 to 0.123 µg/L, and 1.2
to 7.3 µg/kg, respectively.17) BPF was reported to
occur in soft drinks at concentrations ranging from
below LOQ to 0.22 µg/L.28) The highest median
concentration of BPF (450 ng/g) was found in dust
from South Korea, which was ten folds higher than
that detected in samples of USA, China, and
Japan.24) BPS has been found in thermal receipt
papers at concentrations comparable to those of BPA
(several tens of milligrams per gram)29,30) and in
sediment samples collected from various
countries.31) Widespread exposure of the general
population in various countries to BPS has been
demonstrated through biomonitoring studies.2) BPS
has been found in canned foodstuffs at
concentrations on the order of several tens of
nanograms per gram.27,32)
Since the discharges into the environment of BPs
and Tritan are estimated to increase rapidly,9,33)
environmental and health risk potentials of BPA
alternatives are of growing concern. Unlike BPA of
which endocrine toxicity and various health
consequences have received thorough investigations,
very limited attention has been paid to the toxicity
of BPA alternatives until now. This review focuses
on endocrine disruption and presents what we know
about the endocrine disruption potentials of BPs and
the three monomers of Tritan to understand the
current status of knowledge and to identify areas of
future research.
II. Methods
In this review, we provides a summary of the
available information on the estrogenicity and
androgenicity of four BPs (BPAF, BPB, BPF, and
BPS) and three monomers of Tritan copolyesters
(CHDM, TMCD, and DMT) which have been
frequently used as BPA alternatives. Only toxicity
data that measured estrogenicity/anti-estrogenicity
and androgenicity/anti-androgenicity in in vitro cell-
based and in in vivo assay in rat were summarized.
Specifically, the following studies were summarized:
•In vitro estrogen receptor binding assays (alpha
and beta isoforms)
•In vitro androgen receptor binding assays
•In vitro estrogen and androgen receptor
transactivation assays (mammalian cells and
yeast)
•In vivo estrogenicity assays (uterotrophic assay
and steroidogenic assay)
•In vivo androgenicity assays (Hershberger assay)
III. Results and Discussion
A. Estrogenic activities of BPs
Several studies have been published confirming
the estrogenic and anti-androgenic activity of BPA
alternatives in diverse in vitro and in vivo assay
which are summarized in Tables 3-4. Analysis of the
structure-activity relationship of BPA and its related
compounds implied that key structural requirement
for estrogenic and anti-androgenic activity of BPs is
the phenolic hydroxyl group (Fig. 1).34) In addition,
4-hydroxyl group on the A-phenyl ring and a
hydrophobic group of the propane moiety are
suggested to regulate estrogenic and anti-androgenic
activities (Fig. 1).34) For example, the increase of E2
activity by BPAF and BPB could be explained by
hydrophobic substituents in place of the 1-methyl
group of the propane moiety. Unhindered hydroxyl
group on an aryl ring and a hydrophobic group
attached para to the hydroxyl group are also
important factors for estrogen receptor (ER) ligand
activity.35) It was predicted that BPF and BPS, which
have a para hydroxyl group on each of the phenol
rings, may have modulating effects toward ER
binding potency.18,36)
1. Estrogenic activities of BPAF
BPAF may possess greater toxicological
implication than BPA because trifluoromethyl (CF3)
group which is substituted for methyl (CH3) group
of BPA is much more electronegative and therefore
potentially more reactive. This type of substitution
has been reported to increase estrogenic activity in
vivo and in vitro.12,13,34,37) An ER-luciferase reporter
assay using MCF-7 cell line demonstrated that the
estrogen activity of BPAF was about one order of
magnitude greater than that of BPA.34) Daily
subcutaneous injections of 100 mg/kg BPAF to

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Tab l e 3 . Estrogenicity/anti-estrogenicity and androgenicity/anti-androgenicity of bisphenol A alternatives in in vitro studies
Compounds Test type Endpoint Toxicity data Reference
BPAF Recombinant gene assay in yeast EC50, Estrogenic activity 7.44E-7 M Zhang et al., 2009
ER transactivation assay in human T47D-KBluc cell EC50, Estrogenic activity 2.248E-8 M Bermudez et al., 2010
ER transactivation assay in human Ishikawa cell LOEC, ERα luciferase activity 1E-9 M Li et al., 2012
ER transactivation assay in human Ishikawa cell NOEC, ERα luciferase activity <1E-9 M Li et al., 2012
ER transactivation assay in human Ishikawa cell LOEC, ERβ luciferase activity 1E-6 M Li et al., 2012
ER transactivation assay in human Ishikawa cell NOEC, ERβ luciferase activity 1E-7 M Li et al., 2012
ER transactivation assay in human HeLa cell LOEC, ERα luciferase activity 1E-7 M Li et al., 2012
ER transactivation assay in human HeLa cell NOEC, ERα luciferase activity 1E-8 M Li et al., 2012
ER transactivation assay in human HeLa cell EC50, ERα luciferase activity 5.87E-8 M Matsushima et al., 2010
ER transactivation assay in human HeLa cell LOEC, ERβ luciferase activity 1E-7 M Li et al., 2012
ER transactivation assay in human HeLa cell LOEC, ERβ luciferase activity 1E-8 M Li et al., 2012
ER transactivation assay in human HepG2 cell LOEC, ERα luciferase activity 1E-8 M Li et al., 2012
ER transactivation assay in human HepG2 cell NOEC, ERα luciferase activity 1E-9 M Li et al., 2012
ER transactivation assay in human HepG2 cell LOEC, ERβ luciferase activity 1E-8 M Li et al., 2012
ER transactivation assay in human HepG2 cell LOEC, ERβ luciferase activity 1E-9 M Li et al., 2012
ER binding assay to human ERá Log relative ER binding affinity -0.11 Akahori et al., 2008
E-screen (cell proliferation) assay in human MCF-7 cell Proliferative effect over control a5.5 (E2: 6.7, BPA:6.0) Perez et al., 1998
E-screen (cell proliferation) assay in human MCF-7 cell Proliferative effect over control a5.5 Rivas et al., 2002
E-screen (cell proliferation) assay in human MCF-7 cell Relative proliferative effect b78.94 Rivas et al., 2002
E-screen (cell proliferation) assay in human MCF-7 cell LOEC, Estrogenic activity 1E-7 M Rivas et al., 2002
E-screen (cell proliferation) assay in human MCF-7 cell Relative proliferative potency c0.01 Rivas et al., 2002
ERE-luciferase reporter assay in human MCF-7 cell EC50, Estrogenic activity 5E-8 M Kitamura et al., 2005
ERE-luciferase reporter assay in human MCF-7 cell LOEC, Estrogenic activity 1E-7 M Kitamura et al., 2005
ERE-luciferase reporter assay in human MCF-7 cell NOEC, Anti-estrogenic activity
at 1E-11 M E2 1E-5 M Kitamura et al., 2005
Radioligand binding assay for saturation binding of ER IC50, inhibit ability to [3H]17β-estradiol
binding in ERα ligand 5.34E-8 M Matsushima et al., 2010
Radioligand binding assay for saturation binding of ER IC50, inhibit ability to [3H]17β-estradiol
binding in ERβ ligand 1.89E-8 M Matsushima et al., 2010

8Kyunghee Ji and Kyungho Choi
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Tab l e 3 . Continued
Compounds Test type Endpoint Toxicity data Reference
BPAF Radioligand binding assay for saturation binding of ER IC50, inhibit ability to [3H]17β-estradiol
binding in ERRγ ligand 3.58E-7 M Matsushima et al., 2010
ARE-luciferase reporter assay in mouse NIH3T3 cell NOEC, Androgenic activity 1E-4 M Kitamura et al., 2005
ARE-luciferase reporter assay in mouse NIH3T3 cell IC50, Anti-androgenic activity at 1E-10 M
dihydrotestosterone 1.3E-6 M Kitamura et al., 2005
BPB Two-hybrid system in yeast 10% REC, β-galactosidase activity Greater than bisphenol A Chen et al., 2002
Two-hybrid system in yeast without S9 mix LOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-5 M Hashimoto et al., 2001
Two-hybrid system in yeast without S9 mix NOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-6 M Hashimoto et al., 2001
Two-hybrid system in yeast with S9 mix LOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-5 M Hashimoto et al., 2001
Two-hybrid system in yeast with S9 mix NOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-6 M Hashimoto et al., 2001
Fluorescence polarization system LOEC, Estrogenic activity 1E-7 M Hashimoto et al., 2001
Fluorescence polarization system NOEC, Estrogenic activity <1E-7 M Hashimoto et al., 2001
ER transactivation assay in human HeLa cell PC10, ERα luciferase activity 4.09E-8 M Yamasaki et al., 2002
ER transactivation assay in human HeLa cell PC50, ERα luciferase activity 6.63E-7 M Yamasaki et al., 2002
ER transactivation assay in human HeLa cell EC50, ERα luciferase activity 1.67E-7 M Yamasaki et al., 2002
E-screen (cell proliferation) assay in human MCF-7 cell LOEC, Estrogenic activity 1E-9 M Hashimoto et al., 2001
E-screen (cell proliferation) assay in human MCF-7 cell NOEC, Estrogenic activity <1E-9 M Hashimoto et al., 2001
E-screen (cell proliferation) assay in human MCF-7 cell Proliferative effect over control a5.9 Rivas et al., 2002
E-screen (cell proliferation) assay in human MCF-7 cell Relative proliferative effect b85.96 Rivas et al., 2002
E-screen (cell proliferation) assay in human MCF-7 cell LOEC, Estrogenic activity 1E-7 M Rivas et al., 2002
E-screen (cell proliferation) assay in human MCF-7 cell Relative proliferative potency c0.01 Rivas et al., 2002
ERE-luciferase reporter assay in human MCF-7 cell EC50, Estrogenic activity 7E-8 M Kitamura et al., 2005
ERE-luciferase reporter assay in human MCF-7 cell LOEC, Estrogenic activity 1E-7 M Kitamura et al., 2005
ERE-luciferase reporter assay in human MCF-7 cell NOEC, Estrogenic activity 1E-8 M Kitamura et al., 2005
ERE-luciferase reporter assay in human MCF-7 cell LOEC, Anti-estrogenic activity at 1E-11 M E2 1E-6 M Kitamura et al., 2005
ERE-luciferase reporter assay in human MCF-7 cell NOEC, Anti-estrogenic activity at 1E-11 M E2 1E-7 M Kitamura et al., 2005

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Tab l e 3 . Continued
Compounds Test type Endpoint Toxicity data Reference
BPB ER competitive-binding assay in Sprague-Dawley rat uterine cytosol IC50, Inhibition of [3H]-E2 binding 1.05E-6 M Blair et al., 2000
ER competitive-binding assay in Sprague-Dawley rat uterine cytosol Log relative ER binding affinity -1.07 Blair et al., 2000
ARE-luciferase reporter assay in mouse NIH3T3 cell NOEC, Androgenic activity 1E-4 M Kitamura et al., 2005
ARE-luciferase reporter assay in mouse NIH3T3 cell LOEC, Anti-androgenic activity at 1E-10
M dihydrotestosterone 1E-5 M Kitamura et al., 2005
ARE-luciferase reporter assay in mouse NIH3T3 cell NOEC, Anti-androgenic activity at 1E-10
M dihydrotestosterone 1E-6 M Kitamura et al., 2005
ARE-luciferase reporter assay in mouse NIH3T3 cell IC50, Anti-androgenic activity at 1E-10 M
dihydrotestosterone 1.7E-6 M Kitamura et al., 2005
BPF Two-hybrid system in yeast 10% REC, β-galactosidase activity Similar to bisphenol A Chen et al., 2002
Two-hybrid system in yeast without S9 mix LOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-4 M Hashimoto and Nakamura,
2000; Hashimoto et al., 2001
Two-hybrid system in yeast without S9 mix NOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-5 M Hashimoto and Nakamura,
2000; Hashimoto et al., 2001
Two-hybrid system in yeast with S9 mix LOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-5 M Hashimoto et al., 2001
Two-hybrid system in yeast with S9 mix NOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-6 M Hashimoto et al., 2001
Recombinant gene assay in yeast EC50, Estrogenic activity 7.52E-6 M Zhang et al., 2009
Fluorescence polarization system LOEC, Estrogenic activity 1E-4 M Hashimoto and Nakamura,
2000; Hashimoto et al., 2001
Fluorescence polarization system NOEC, Estrogenic activity 1E-5 M Hashimoto and Nakamura,
2000; Hashimoto et al., 2001
E-screen (cell proliferation) assay in human MCF-7 cell Proliferative effect over control a7.1 (E2: 6.7, BPA:6.0) Perez et al., 1998
E-screen (cell proliferation) assay in human MCF-7 cell LOEC, Estrogenic activity <1E-7 M Hashimoto and Nakamura,
2000
E-screen (cell proliferation) assay in human MCF-7 cell LOEC, Estrogenic activity 1E-8 M Hashimoto et al., 2001
E-screen (cell proliferation) assay in human MCF-7 cell NOEC, Estrogenic activity 1E-9 M Hashimoto et al., 2001
E-screen (cell proliferation) assay in human MCF-7 cell EC50, Estrogenic activity 8.48E-8 M Stroheker et al., 2004
ERE-luciferase reporter assay in human MCF-7 cell EC50, Estrogenic activity 1E-6 M Kitamura et al., 2005
ERE-luciferase reporter assay in human MCF-7 cell NOEC, Anti-estrogenic activity at 1E-11 M E2 1E-5 M Kitamura et al., 2005

10 Kyunghee Ji and Kyungho Choi
J Environ Health Sci 2013: 39(1): 1-18 http://www.kseh.org/
Tab l e 3 . Continued
Compounds Test type Endpoint Toxicity data Reference
BPF ER transactivation assay in human HeLa cell PC10, ERα luciferase activity 2.84E-6 M Yamasaki et al., 2002
ER transactivation assay in human HepG2 cell LOEC, ERα transcriptional activity 1E-7 M Cabaton et al., 2009
ER transactivation assay in human HepG2 cell EC50, ERα transcriptional activity 2.39E-6 M Cabaton et al., 2009
ER transactivation assay in human HepG2 cell LOEC, ERβ transcriptional activity 1E-6 M Cabaton et al., 2009
ER transactivation assay in human HepG2 cell EC50, ERβ transcriptional activity 6.04E-6 M Cabaton et al., 2009
ER competitive-binding assay in Sprague-Dawley rat uterine cytosol IC50, Inhibition of [3H]-E2 binding 9.50E-5 M Blair et al., 2000
ER competitive-binding assay in Sprague-Dawley rat uterine cytosol Log relative ER binding affinity -3.02 Blair et al., 2000
AR-binding assay in hamster CHO-K1 cell IC50, AR binding activity 9.0E-6 M Satoh et al., 2004
ARE-luciferase reporter assay in hamster CHO-K1 cell NOEC, Androgenic activity 1E-4 M Satoh et al., 2004
ARE-luciferase reporter assay in hamster CHO-K1 cell NOEC, Anti-androgenic activity 4.8E-6 M Satoh et al., 2004
ARE-luciferase reporter assay in mouse NIH3T3 cell NOEC, Androgenic activity 1E-4 M Kitamura et al., 2005
ARE-luciferase reporter assay in mouse NIH3T3 cell LOEC, Anti-androgenic activity at 1E-10
M dihydrotestosterone 1E-5 M Kitamura et al., 2005
ARE-luciferase reporter assay in mouse NIH3T3 cell NOEC, Anti-androgenic activity at 1E-10
M dihydrotestosterone 1E-6 M Kitamura et al., 2005
ARE-luciferase reporter assay in mouse NIH3T3 cell IC50, Anti-androgenic activity at 1E-10 M
dihydrotestosterone 1.2E-5 M Kitamura et al., 2005
ARE-luciferase reporter assay in human MDA0MB453 cell LOEC, Anti-androgenic activity at 4E-10
M dihydrotestosterone 1E-10 M Stroheker et al., 2004
AR transactivation assay in human MDA-kb2 cell LOEC, AR transcriptional activity 1E-5 M Cabaton et al., 2009
BPS Two-hybrid system in yeast 10% REC, β-galactosidase activity Lesser than BPA Chen et al., 2002
Yeast two-hybrid system without S9 mix LOEC, Estrogenic activity based on
relative β-galactosidase activity >1E-3 M Hashimoto and Nakamura,
2000; Hashimoto et al., 2001
Yeast two-hybrid system without S9 mix NOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-3 M Hashimoto and Nakamura,
2000; Hashimoto et al., 2001
Yeast two-hybrid system with S9 mix LOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-3 M Hashimoto et al., 2001
Yeast two-hybrid system with S9 mix NOEC, Estrogenic activity based on
relative β-galactosidase activity 1E-4 M Hashimoto et al., 2001
Fluorescence polarization system LOEC, Estrogenic activity 1E-3 M Hashimoto and Nakamura,
2000; Hashimoto et al., 2001

Endocrine Disruption Potentials of Bisphenol A Alternatives - Are Bisphenol A Alternatives Safe from Endocrine Disruption? 11
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Tab l e 3 . Continued
Compounds Test type Endpoint Toxicity data Reference
BPS Fluorescence polarization system NOEC, Estrogenic activity 1E-4 M Hashimoto and Nakamura,
2000; Hashimoto et al., 2001
ERE-luciferase reporter assay in human BG1Luc4E2 cell EC50, Estrogenic activity 4.93E-6 M Grignard et al., 2012
E-screen (cell proliferation) assay in human MCF-7 cell LOEC, Estrogenic activity <1E-7 M Hashimoto and Nakamura,
2000
E-screen (cell proliferation) assay in human MCF-7 cell LOEC, Estrogenic activity 1E-7 M Hashimoto et al., 2001
E-screen (cell proliferation) assay in human MCF-7 cell NOEC, Estrogenic activity 1E-8 M Hashimoto et al., 2001
ERE-luciferase reporter assay in human MELN cell EC50, Estrogenic activity 4.24E-6 M Grignard et al., 2012
ERE-luciferase reporter assay in human MCF-7 cell EC50, Estrogenic activity 1.75E-6 M Kuruto-Niwa et al., 2005
ERE-luciferase reporter assay in human MCF-7 cell EC50, Estrogenic activity 1.1E-6 M Kitamura et al., 2005
ERE-luciferase reporter assay in human MCF-7 cell NOEC, Anti-estrogenic activity at 1E-11
M E2 1E-5 M Kitamura et al., 2005
ER competitive-binding assay in Sprague-Dawley rat uterine cytosol IC50, Inhibition of [3H]-E2 binding 1.05E-4 M Blair et al., 2000
ER competitive-binding assay in Sprague-Dawley rat uterine cytosol Log relative ER binding affinity -3.07 Blair et al., 2000
ARE-luciferase reporter assay in mouse NIH3T3 cell NOEC, Androgenic activity 1E-4 M Kitamura et al., 2005
ARE-luciferase reporter assay in mouse NIH3T3 cell IC50, Anti-androgenic activity at 1E-10 M
dihydrotestosterone 1.7E-5 M Kitamura et al., 2005
CHDM E-screen (cell proliferation) assay in human MCF-7 cell Estrogenic activity Yes Yang et al., 2011
ER binding assay to human ERá and ERâ NOEC, binding to ERα and ERβ1E-3 M Osimitz et al., 2012
ER transactivation assay in yeast NOEC, Estrogenic activity 1E-3 M Osimitz et al., 2012
ER transactivation assay in human T47D-KBluc cell NOEC, Estrogenic activity 1E-3 M Osimitz et al., 2012
ER transactivation assay in human T47D-KBluc cell NOEC, Anti-estrogenic activity 1E-3 M Osimitz et al., 2012
ER transactivation assay in yeast NOEC, Estrogenic activity 1E-3 M Osimitz et al., 2012
AR binding assay NOEC, binding to AR 1E-3 M Osimitz et al., 2012
AR transactivation assay in human MDA-kb2 cell NOEC, Androgenic activity 1E-3 M Osimitz et al., 2012
AR transactivation assay in human MDA-kb2 cell NOEC, Anti-androgenic activity 1E-3 M Osimitz et al., 2012
AR transactivation assay in yeast NOEC, Androgenic activity 1E-3 M Osimitz et al., 2012
DMT E-screen (cell proliferation) assay in human MCF-7 cell Estrogenic activity Yes Yang et al., 2011
ER binding assay to human ERá and ERâ NOEC, binding to ERα and ERβ1E-3 M Osimitz et al., 2012
ER transactivation assay in yeast NOEC, Estrogenic activity 1E-3 M Osimitz et al., 2012

12 Kyunghee Ji and Kyungho Choi
J Environ Health Sci 2013: 39(1): 1-18 http://www.kseh.org/
Tab l e 3 . Continued
Compounds Test type Endpoint Toxicity data Reference
DMT ER transactivation assay in human T47D-KBluc cell NOEC, Estrogenic activity 1E-3 M Osimitz et al., 2012
ER transactivation assay in human T47D-KBluc cell NOEC, Anti-estrogenic activity 1E-3 M Osimitz et al., 2012
ER transactivation assay in yeast NOEC, Estrogenic activity 1E-3 M Osimitz et al., 2012
AR binding assay NOEC, binding to AR 1E-3 M Osimitz et al., 2012
AR transactivation assay in human MDA-kb2 cell NOEC, Androgenic activity 1E-3 M Osimitz et al., 2012
AR transactivation assay in human MDA-kb2 cell NOEC, Anti-androgenic activity 1E-3 M Osimitz et al., 2012
AR transactivation assay in yeast NOEC, Androgenic activity 1E-4 M Osimitz et al., 2012
TMCD ER binding assay to human ERá and ERâ NOEC, binding to ERα and ERβ1E-3 M Osimitz et al., 2012
ER transactivation assay in yeast NOEC, Estrogenic activity 1E-3 M Osimitz et al., 2012
ER transactivation assay in human T47D-KBluc cell NOEC, Estrogenic activity 1E-3 M Osimitz et al., 2012
ER transactivation assay in human T47D-KBluc cell NOEC, Anti-estrogenic activity 1E-3 M Osimitz et al., 2012
ER transactivation assay in yeast NOEC, Estrogenic activity 1E-4 M Osimitz et al., 2012
AR binding assay NOEC, binding to AR 1E-3 M Osimitz et al., 2012
AR transactivation assay in human MDA-kb2 cell NOEC, Androgenic activity 1E-3 M Osimitz et al., 2012
AR transactivation assay in human MDA-kb2 cell NOEC, Anti-androgenic activity 1E-3 M Osimitz et al., 2012
AR transactivation assay in yeast NOEC, Androgenic activity 1E-3 M Osimitz et al., 2012
AR: Androgen receptor, EC50: Median effective concentration, ER: Estrogen receptor, IC50: Median inhibition concentration, LOEC: lowest observed effective concentration,
NOEC: no observed effective concentration, PC10: concentrations estimated to show 10% of the transcriptional activity of 1 nM E2, PC50: concentrations estimated to show 50%
of the transcriptional activity of 1 nM E2, REC: Relative effective concentration.
a Proliferative effect over control = maximal cell count of test compounds/cell count of control.
b Relative proliferative effect = (proliferative effect of test compounds-1/proliferative effect of E2-1)×100.
c Relative proliferative potency = ratio between E2 and test compounds doses to produce maximal yield ×100.

Endocrine Disruption Potentials of Bisphenol A Alternatives - Are Bisphenol A Alternatives Safe from Endocrine Disruption? 13
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Tab l e 4 . Summary of in vivo studies published on the estrogenic and androgenic activity of bisphenol A alternatives
Compounds Test type Test organisms Exposure duration Endpoint Toxicity data Reference
BPAF Steroidogenic assay Adult male rat 14 d NOED, T levels in serum 50 mg/kg/d Feng et al., 2012
Steroidogenic assay Adult male rat 14 d LOED, T levels in serum 200 mg/kg/d Feng et al., 2012
Steroidogenic assay Adult male rat 14 d NOED, LH levels in serum 10 mg/kg/d Feng et al., 2012
Steroidogenic assay Adult male rat 14 d LOED, LH levels in serum 50 mg/kg/d Feng et al., 2012
Steroidogenic assay Adult male rat 14 d NOED, FSH levels in serum 2 mg/kg/d Feng et al., 2012
Steroidogenic assay Adult male rat 14 d LOED, FSH levels in serum 10 mg/kg/d Feng et al., 2012
Steroidogenic assay Adult male rat 14 d NOED, transcription in genes in
steroidogenesis 50 mg/kg/d Feng et al., 2012
Steroidogenic assay Adult male rat 14 d LOED, transcription in genes in
steroidogenesis 200 mg/kg/d Feng et al., 2012
Uterotrophic assay Immature female rat 1 d Log LOED, estrogenic effects 1.08 ìmol/kg/d Akahori et al., 2008
Uterotrophic assay Immature female rat 3 d LOED, estrogenic effects 8 mg/kg/d Yamasaki et al., 2003
Hershberger assay Adult male rat 10 d LOED, anti-androgenic effects 200 mg/kg/d Yamasaki et al., 2003
BPB Uterotrophic assay Immature female rat 3 d LOED, estrogenic effects 200 mg/kg/d Yamasaki et al., 2002
Hershberger assay Adult male rat 10 d LOED, anti-androgenic effects 600 mg/kg/d Yamasaki et al., 2003
BPF Uterotrophic assay Immature female rat 3 d LOED, estrogenic effects 200 mg/kg/d Yamasaki et al., 2002
Hershberger assay Adult male rat 10 d LOED, anti-androgenic effects 1,000 mg/kg/d Yamasaki et al., 2003
CHDM Uterotrophic assay Female rat 3 d NOED, estrogenic effects 10 mg/kg/d Osimitz et al., 2012
Hershberger assay Male rat 10 d NOED, androgenic effects 10 mg/kg/d Osimitz et al., 2012
DMT Uterotrophic assay Female rat 3 d NOED, estrogenic effects 10 mg/kg/d Osimitz et al., 2012
Hershberger assay Male rat 10 d NOED, androgenic effects 10 mg/kg/d Osimitz et al., 2012
TMCD Uterotrophic assay Female rat 3 d NOED, estrogenic effects 10 mg/kg/d Osimitz et al., 2012
Hershberger assay Male rat 10 d NOED, androgenic effects 10 mg/kg/d Osimitz et al., 2012
NOED: no observed effective dose, LOED: lowest observed effective dose, T: testosterone, LH: luteinizing hormone, FSH: follicle-stimulating hormone.

14 Kyunghee Ji and Kyungho Choi
J Environ Health Sci 2013: 39(1): 1-18 http://www.kseh.org/
immature female rats for three days led to a 337%
increase in uterus size, compared to only a 197%
increase when exposed to 200 mg/kg BPA.15)
BPAF has been shown to induce estrogen-
dependent responses via binding to ERα and
ERβ.11,12,13,37) The binding affinity of BPAF was
approximately 20 times stronger and 48 times
stronger than that of BPA as a ligand for ERα and
ERβ, respectively.13) High binding activity of BPAF
for ERβ suggests that the binding pocket of ERβ
possesses specific structural elements that interact
much more favorably with the CF3 groups of BPAF
than with the CH3 groups of BPA. In Ishikawa and
HepG2 cells, the agonistic effects of BPAF for ERα
were stronger than that of BPA (10 nM BPAF vs.
100 nM BPA).12) However binding to estrogen-related
receptor gamma (ERRγ) was weaker than BPA,
suggesting less favorable interaction of ERRγ-ligand
binding domain (LBD) with the CF3 groups.13)
In uterotrophic assay employing immature rat,
uterine weight increased significantly in rats given
8, 40, and 100 mg/kg BPAF, suggesting estrogen
agonist.38) The results of Hershberger assay showed
that BPAF decreased body weight-gain and
spontaneous locomotion in the rats treated with 200
and 600 mg/kg BPAF, suggesting anti-androgen
activity.38)
BPAF may impair pituitary-gonadal function at
different levels by increasing LH and FSH
concentrations and decreasing testosterone levels in
serum.39) It appears that the inhibition of androgen
biosynthesis was not a result of altered regulatory
function of the LH-dependent signaling pathway but
was more likely a direct result of BPAF’s ability to
reduce the expression of steroidogenic genes such
as SR-B1, StAR, P450scc, and 17βHSD. Sharp
decrease in testosterone concentration and reduction
in gene transcriptions and protein levels involved in
steroidogenesis suggested that the testes may be a
primary target organ of BPAF exposure.
2. Estrogenic activities of BPB
Estrogenicity of BPB is suspected in part because
of the substitution of the propane bridge of BPA to
butane, which is related to its estrogenic activity. It
was reported that higher estrogenic responses were
obtained from longer alkyl substituent at the
bridging carbon in MCF-7 cells.14) Recently Chen et
al. ranked diphenylalkanes without any modifications
by their estrogenic potency, and reported the order
of BPB (C4) > BPA (C3) bisphenol E (1,1-bis(4-
hydroxyphenyl)ethane; BPE) (C2) BPF (C1).33)
BPB has been shown to possess estrogenic
properties in various in vitro and in vivo
experiments. BPB showed considerably higher
estrogenic activity than BPA in the yeast two-hybrid
assay.33) This compound also exhibited significant
increase of MCF-7 cell growth in the E-screen
test40,41) as well as greater luciferase activity in MCF-
7 cells.34) Moderate binding affinity to ER was also
reported in Sprague-Dawley rat uterine cytosol.35)
Yamasaki et al. found that BPB has estrogenic
activity both in ER transactivation assay in HeLa
cell and uterotrophic assay in female rat, however,
the estrogenic potencies obtained in in vitro assay
do not completely correspond to the uterotrophic
potency in in vivo test.42)
3. Estrogenic activities of BPF
BPF has been shown to induce estrogenic activity
in vivo and in vitro. In in vivo, BPF exhibited
estrogen agonistic properties in the uterotrophic
assay.42) In in vitro assay using a yeast two-hybrid
system BPF was identified as the most estrogenic
compound among the tested chemicals that were
present in food packaging material or used in
dentistry.40,43) BPF has also exhibited estrogenic
activity in yeast recombinant gene assay.44) In human
cells, the proliferative response of MCF-7 cells (E-
Screen assay) increased in a concentration dependent
manner.14,40,43,45) The latter authors showed that,
according to the respective median effective
concentration (EC50) values for proliferation of
MCF-7 cell, BPF was more pronounced than BPA.
Fig. 1. Structural characteristics of bisphenol-relate
d
compounds which might possess endocrine-
disrupting activity (modified from Kitamura et al.
(2005)34)

Endocrine Disruption Potentials of Bisphenol A Alternatives - Are Bisphenol A Alternatives Safe from Endocrine Disruption? 15
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Moderate binding affinities of BPF to ER in MCF-
7 cell,34) HeLa cell,42) HepG2 cell,46) and Sprague-
Dawley rat uterine cytosol35) were reported.
Some of effects of BPF exposure are mediated by
binding to nuclear steroid receptors (ERα and ERβ)
and inducing estrogenic signals, which may
subsequently modify estrogen-responsive gene
expression in HepG2 cells.46) These data are in
agreement with Kitamura et al. who used an ERE-
luciferase reporter assay in MCF-7 cells.34)
Anti-androgenic activities of BPF were reported in
in vitro and in vivo systems. BPF can compete with
5-α dihydro-testosteron (5-α DHT) for binding with
AR and exhibits a significant anti-androgenic
activity in MDA-kb2 cells.46) These results are in
agreement with Satoh et al. (2004) who observed a
decrease of 5-α DHT-induced luciferase at 10-6 M
in CHO-K1 cells using the AR-EcoScreen assay.47)
BPF decreased luciferase induction by DHT in
mouse NIH3T3 cells30) as well as human
MDA0MB453 cells.45)
4. Estrogenic activities of BPS
Estrogenic activity of BPS is rather controversial.
It was first reported that BPS had no estrogenic
activity using the yeast two-hybrid system.33) Several
authors, however, reported that BPS possessed
estrogenic activity in MCF-7 cell40,43) as well as
weak estrogenic transcriptional activities in human
MELN cells derived from MCF-7 cells.36) Since the
basic structural features which have been linked to ER
binding,48) in particular the presence of a para
hydroxyl group on each of the phenol rings, are shared
by both BPA and BPS, BPS also may have
modulatory effects toward ER binding potency.18,34,35,36)
BPS is also reported to have anti-androgenic activity
in mouse NIH3T3 cells.34)
B. Estrogenic activities of Tritan copolyesters
Three monomers of Tritan exhibited no evidence
of interaction with either the AR or the alpha or
beta ER receptors.9) Similarly, the AR and ER
transactivation assays, conducted with human cells
and yeast reporters were negative as well. The lack
of an estrogenic effect in in vitro assays was in good
agreement with the in vivo uterotrophic assay in
which none of the monomers demonstrated
biological activities consistent with agonism of
natural estrogens when administrated orally to
ovariectomized female rats using a very wide range
of dose levels. Similarly, the in vivo Hershberger
assay shows no evidence of androgenic or anti-
androgenic effects.
These results, however, are contrary to those
reported by Yang et al.49) They reported estrogenic
effects of CHDM and DMT using the MCF-7 cell.
Although details of the test results are not given, the
authors report both compounds to be “estrogenic
active”. Authorities in the US and Europe have
reviewed Tritan copolyesters for safety for food
contact use,50,51) however, further study of polymer
as well as monomer on endocrine disruption appear
to be warranted.
IV. Conclusion
According to the investigations that employed
several estrogenicity and androgenic assays, most
BPs used as alternatives of BPA appear to have
estrogenic and anti-androgenic activity as a common
property. The modification of phenolic rings and
bridging carbon, or the longer length of the alkyl
substituents seems to influence the estrogen and
anti-androgen activity, although the apparent
relationship between their structure and estrogenic
activity was not clarified.33) For components of
Tritan, no evidence of estrogen- or androgen-related
effects was reported in one study, but a report
suggesting otherwise is available. More studies with
thorough study design, e.g., with long-term exposure
period are warranted.
If current trends continue, production and subsequent
environmental release of BPA alternatives are
expected to increase. As some BPA substitutes such
as BPF and BPS could be more persistent in
environments compared to BPA,52 toxicological
consequences in ecosystem should receive more
attention. Further toxicological information of BPA
alternatives is required to understand the
environmental health implications of these
alternatives and to develop proper management
plans.
References
1. Diamanti-Kandarakis E, Bourguignon JP, Giudice
LC, Hauser R, Prins GS, Soto AM, et al. Endo-
crine-disrupting chemicals: an Endocrine society

16 Kyunghee Ji and Kyungho Choi
J Environ Health Sci 2013: 39(1): 1-18 http://www.kseh.org/
scientific statement. Endocrine Reviews. 2009; 30:
293-342.
2. Liao C, Liu F, Alomirah H, Loi VD, Mohd MA,
Moon HB, et al. Bisphenol S in urine from the
United States and seven Asian countries: occur-
rence and human exposures. Environ Sci Technol.
2012b; 46: 6860-6866.
3. Goodman JE, Witorsch RJ, McConnell EE, Sipes
IG, Slayton TM, Yu CJ, et al. Weight-of-evidence
evaluation of reproductive and developmental
effects of low doses of bisphenol A. Crit Rev Tox-
icol. 2009; 39: 1-75.
4. Zhang X, Chang H, Wiseman S, He Y, Higley E,
Jones P, et al. Bisphenol A disrupts steroidogene-
sis in human H295R cells. Toxicol Sci. 2011; 121:
320-327.
5. Vandenberg LN, Chahoud I, Heindel JJ, Pad-
manabhan V, Paumgartten FJR, Schoenfelder G.
Urinary, circulating, and tissue biomonitoring stud-
ies indicate widespread exposure to bisphenol A.
Environ Health Perspect. 2010; 118: 1055-1070.
6. Volkel W, Colnot T, Csanady GA, Filsner JG,
Dekant W. Metabolism and kinetics of bisphenol
A in humans at low doses following oral admin-
istration. Chem Res Toxicol 2002; 15: 1281-1287.
7. Volkel W, Bittner N, Dekant W. Quantitation of
bisphenol A and bisphenol A glucuronide in bio-
logical samples by high performance liquid chro-
matography-tandem mass spectrometry. Drug
Metab Dispos 2005; 33: 1748-1757.
8. European Commission. 2011. Commission direc-
tive 2011/8/EU of 28 January 2011 on the restriction
of use of Bisphenol A in plastic infant feeding bot-
tles. Available: http://eur-lex.europa.eu/LexUriServ/
LexUriServ.do?uri=OJ:L:2011:026:0011:0014:EN:PDF
[accessed 4 February 2013].
9. Osimitz TG, Eldridge ML, Sloter E, Welsh W, Ai
N, Sayler GS, et al. Lack of androgenicity and
estrogenicity of the three monomers used in East-
man’s TritanTM copolyesters. Food Chem Toxicol.
2012; 50: 2196-2205.
10. Danzl E, Sei K, Soda S, Ike M, Fujita M. Biodeg-
radation of bisphenol A, bisphenol F and bisphe-
nol S in seawater. Int J Environ Res Public Health.
2009; 6: 1472-1484.
11. Akahori Y, Nakai M, Yamasaki K, Takatsuki M,
Shimohigashi Y, Ohtaki M. Relationship between
the results of in vitro receptor binding assay to
human estrogen receptor alpha and in vivo
uterotrophic assay: comparative study with 65
selected chemicals. Toxicol In Vitro. 2008; 22:
225-231.
12. Li Y, Burns KA, Arao Y, Luh CJ, Korach KS. Dif-
ferential estrogenic actions of endocrine-disrupt-
ing chemicals bisphenol A, bisphenol AF, and
zearalenone through estrogen receptor α and β in
vitro. Environ Health Perspect. 2012; 120: 1029-
1035.
13. Matsushima A, Liu X, Okada H, Shimohigashi M,
Shimohigashi Y. Bisphenol AF is a full agonist for
the estrogen receptor ERα but a highly specific
antagonist for ERâ. Environ Health Perspect. 2010;
118: 1267-1272.
14. Perez P, Pulgar R, Olea-Serrano F, Villalobos M,
Rivas A, Metzler M, et al. The estrogenicity of
bisphenol A-related diphenylalkanes with various
substituents at the central carbon and the hydroxyl
groups. Environ Health Perspect. 1998; 106: 167-
174.
15. Stout MD. 2008. NTP research concept: Bisphe-
nol AF. Available: http://ntp.niehs.nih.gov/files/
BPAF_Concept_final-100608_508.pdf [accessed 4
February 2013].
16. Yoshihara S, Mizutare T, Makishima M, Suzuki N,
Fujimoto N, Igarashi K, et al. Potent estrogenic
metabolites of bisphenol A and bisphenol B
formed by rat liver S9 fraction: their structures and
estrogenic potency. Toxicol Sci. 2004; 78: 50-59.
17. Fromme H, Küchler T, Otto T, Pilz K, Müller J,
Wenzel A. Occurrence of phthalates and bisphe-
nol A and F in the environment. Water Res. 2002;
36: 1429-1438.
18. Kuruto-Niwa R, Nozawa R, Miyakoshi T, Shiozawa
T, Terao Y. Estrogenic activity of alkylphenols,
bisphenol S, and their chlorinated derivatives using
a GFP expression system. Environ Toxicol Phar-
macol. 2005; 19: 121-130.
19. Simoneau C, Valzacchi S, Morkunas V, Van den
Eede L. Comparison of migration from polyether-
sulphone and polycarbonate baby bottles. Food
Addit Contam Part A Chem Anal Control Expo
Risk Assess. 2011; 28: 1763-1768.
20. Watanabe M, Fukazawa H, Shiraishi F, Shiraishi
H, Shiozawa T, Terao T. Analysis and estrogenic
activity of bisphenol A and other chemicals
released from waste paper by pulping. J Environ
Chem. 2004; 14: 65.
21. Ball GL, McLellan CJ, Bhat VS. Toxicological
review and oral risk assessment of terephthalic
acid (TPA) and its esters: a category approach. Crit
Rev Toxicol. 2012; 42: 28-67.
22. US EPA. 2010. Office of Pollution Prevention and
Toxics. ChemRTK 1990 HPV challenge program
chemicals list. Available: http://www.epa.gov/hpv/
pubs/update/hpv_1990.pdf [accessed 4 February
2013].

Endocrine Disruption Potentials of Bisphenol A Alternatives - Are Bisphenol A Alternatives Safe from Endocrine Disruption? 17
http://www.kseh.org/ J Environ Health Sci 2013: 39(1): 1-18
23. OECD. 2009. The 2007 OECD list of high pro-
duction volume chemicals. ENV/JM/MONO(2009)40.
OECD. Paris, France 23 October 2009. Available:
http://search.oecd.org/officialdocuments/display-
documentpdf/?cote=env/jm/mono(2009)40&doc-
language=en [accessed 4 February 2013].
24. Liao C, Liu F, Guo Y, Moon HB, Nakata H, Wu
Q, et al. Occurrence of eight bisphenol analogues
in indoor dust from the United States and several
Asian countries: Implications for human exposure.
Environ Sci Technol. 2012d; 46: 9138-9145.
25. Cobellis L, Colacurci N, Trabucco E, Carpentiero
C, Grumetto L. Measurement of bisphenol A and
bisphenol B levels in human blood sera from
healthy and endometriotic women. Biomed Chro-
matogr. 2009; 23: 1186-1190.
26. Cunha SC, Fernandes JO. Quantification of free
and total bisphenol A and bisphenol B in human
urine by dispersive liquid-liquid microextraction
(DLLME) and heart-cutting multidimensional gas
chromatography-mass spectrometry (MD-GC/MS).
Talanta. 2010; 83: 117-125.
27. Grumetto L, Montesano D, Seccia S, Albrizio S,
Barbato F. Determination of bisphenol A and
bisphenol B residues in canned peeled tomatoes by
reversed-phase liquid chromatography. J Agric
Food Chem. 2008; 56: 10633-10637.
28. Gallart-Ayala H, Moyano E, Galceran M.T. Anal-
ysis of bisphenols in soft drinks by on-line solid
phase extraction fast liquid chromatography-tan-
dem mass spectrometry. Anal Chim Acta. 2011;
683: 227-233.
29. Liao C, Kannan K. Widespread occurrence of
bisphenol A in paper and paper products: Implica-
tions for human exposure. Environ Sci Technol.
2011; 45: 9372-9379.
30. Liao C, Liu F, Kannan K. Bisphenol S, a new
bisphenol analogue, in paper products and cur-
rency and its association with bisphenol a resi-
dues. Environ Sci Technol. 2012a; 46: 6515-6522.
31. Liao C, Liu F, Moon HB, Yamashita N, Yun S,
Kannan K. Bisphenol analogues in sediments from
industrialized areas in the United States, Japan,
and Korea: spatial and temporal distributions.
Environ Sci Technol. 2012c; 46: 11558-11565.
32. Viñas P, Campillo N, Martínez-Castillo N, Hernán-
dez-Córdoba M. Comparison of two derivatization-
based methods for solid-phase microextraction-gas
chromatography-mass spectrometric determination
of bisphenol A, bisphenol S and bisphenol
migrated from food cans. Anal Bioanal Chem.
2010; 397: 115-125.
33. Chen MY, Ike M, Fujita M. Acute toxicity,
mutagenicity, and estrogenicity of bisphenol-A and
other bisphenols. Environ Toxicol. 2002; 17: 80-
86.
34. Kitamura S, Suzuki T, Sanoh S, Kohta R, Jinno N,
Sugihara K, et al. Comparative study of the endo-
crine-disrupting activity of bisphenol A and 19
related compounds. Toxicol Sci. 2005; 84: 249-
259.
35. Blair RM, Fang H, Branham WS, Hass BS, Dial
SL, Moland CL, Tong W, Shi L, Perkins R, Shee-
han D.M. The estrogen receptor relative binding
affinities of 188 natural and xenochemicals: struc-
tural diversity of ligands. Toxicol Sci. 2000; 54:
138-153.
36. Grignard E, Lapenna S, Bremer S. Weak estrog-
nic transcriptional activities of bisphenol A and
bisphenol S. Toxicol In Vitro. 2012; 26: 727-731.
37. Bermudez DS, Gray Jr LE, Wilson VS. Modeling
the interaction of binary and ternary mixtures of
estradiol with bisphenol A and bisphenol AF in an
in vitro estrogen-mediated transcriptional activa-
tion assay (T47D-KBluc). Toxicol Sci. 2010; 116:
477-487.
38. Yamasaki K, Takeyoshi M, Sawaki M, Imatanaka
N, Shinoda K, Takatsuki M. Immature rat
uterotrophic assay of 18 chemicals and Hersh-
berger assay of 30 chemicals. Toxicology. 2003;
183: 93-115.
39. Feng Y, Yin J, Jiao Z, Shi J, Li M, Shao B.
Bisphenol AF may cause testosterone reduction by
directly affecting testis function in adult male rats.
Toxicol Lett. 2012; 211: 201-209.
40. Hashimoto Y, Moriguchi Y, Oshima H, Kawagu-
chi M, Miyazaki K, Nakamura M. Measurement of
estrogenic activity of chemicals for the develop-
ment of new dental polymers. Toxicol In Vitro.
2001; 15: 421.
41. Rivas A, Lacroix M, Olea-Serrano F, Laïos I,
Leclercq G, Olea N. Estrogenic effect of a series
of bisphenol analogues on gene and protein
expression in MCF-7 breast cancer cells. J Steroid
Biochem Mol Biol. 2002; 82: 45-53.
42. Yamasaki K, Takeyoshi M, Yakabe Y, Sawaki M,
Imatanaka N, Takatsuki M. Comparison of reporter
gene assay and immature rat uterotrophic assay of
twenty-three chemicals. Toxicology. 2002; 170:
21-30.
43. Hashimoto Y, Nakamura M. Estrogenic activity of
dental materials and bisphenol-A related chemi-
cals in vitro. Dent Mater J. 2000; 19: 245.
44. Zhang HC, Chen LY, Liu SS, Yin DQ. Jointed
estrogenic activities of bisphenol A and three of its
analogs. Huan Jing Ke Xue. 2009; 30: 260-265.

18 Kyunghee Ji and Kyungho Choi
J Environ Health Sci 2013: 39(1): 1-18 http://www.kseh.org/
45. Stroheker T, Picard K, Lhuguenot JC, Canivenc-
Lavier MC, Chagnon MC. Steroid activities com-
parison of natural and food wrap compounds in
human breast cancer cell lines. Food Chem Toxi-
col. 2004; 42: 887-897.
46. Cabaton N, Dumont C, Severin I, Perdu E, Zalko
D, Cherkaoui-Malki M, et al. Genotoxic and endo-
crine activities of bis(hydroxyphenyl)methane
(bisphenol F) and its derivatives in the HepG2 cell
line. Toxicology 2009; 255: 15-24.
47.Satoh K, Ohyama K, Aoki N, Iida M, Nagai F.
Study on anti-androgenic effects of bisphenol A
diglycidyl ether (BADGE), bisphenol F diglycidyl
ether (BFDGE) and their derivatives using stably
transfected with human androgen receptor, AR-
EcoScreen. Food Chem Toxicol. 2004; 42: 983-
993.
48. Gallegos Saliner A, Amat L, Carbo-Dorca R,
Schultz TW, Cronin MT. Molecular quantum sim-
ilarity analysis of estrogenic activity. J Chem Inf
Comput Sci. 2003; 43: 1166-1176.
49. Yang C, Yaniger S, Jordan VC, Klein D, Bittner G.
Most plastic products release estrogenic chemi-
cals: a potential health problem that can be solved.
Environ Health Perspect. 2011; 119: 989-996.
50. US FDA. 2011. Inventory of Effective Food Con-
tact Substance (FCS) Notifications: FCN No.
1041. Available: http://www.accessdata.fda.gov/
scripts/fcn/fcnDetailNavigation.cfm?rpt=fcsListing
&id=1041 [accessed 4 February 2013].
51. European Commission. 2011. Commission Regula-
tion (EU) No. 10/2011 of 14 January 2011 on
Plastic Materials and Articles Intended to Come
into Contact with Food Text with EEA Relevance.
Available: http://eurlex.europa.eu/LexUriServ/Lex-
UriServ.do?uri=OJ:L:2011:012:0001:01:EN:HTML
[accessed 4 February 2013].
52. Ike M, Chen MY, Danzl E, Sei K, Fujita M. Bio-
degradation of a variety of bisphenols under aero-
bic and anaerobic conditions. Water Sci Technol.
2006; 53: 153-159.