OFFICE OF ENVIRONMENTAL HEALTH HAZARD ASSESSMENT

Proposition 65

Evidence on the Female
Reproductive Toxicity ofBisphenol S
October 2023

Reproductive and Cancer Hazard Assessment Branch
Office of Environmental Health Hazard Assessment
California Environmental Protection Agency

CONTRIBUTORSThe Office of Environmental Health Hazard Assessment’s (OEHHA) Reproductive
Toxicology and Epidemiology Section within the Reproductive and Cancer Hazard
Assessment Branch was responsible for the preparation of this document.
Authors (listed alphabetically by last name)
Faye V. Andrews, Ph.D., M.P.H.
Research Scientist III

Farla Kaufman, Ph.D.
Staff Toxicologist

Erin Delker, Ph.D., M.P.H.
Research Scientist III

Francisco Moran, Ph.D.
Senior Toxicologist

Poorni Iyer, DVM, Ph.D., DABT
Staff Toxicologist

Yassaman Niknam, Ph.D.
Staff Toxicologist

Acknowledgments
The valuable support of the following OEHHA staff is also acknowledged: Nancy
Firchow, MLS for conducting the literature search.
Internal OEHHA Reviewers
Martha S. Sandy, Ph.D., M.P.H.
Chief, Reproductive and Cancer Hazard Assessment Branch
Elaine Khan, Ph.D.
Chief, Pesticide and Environmental Toxicology Branch
Vince Cogliano, Ph.D.
Deputy Director, Division of Scientific Programs
David Edwards, Ph.D.
Chief Deputy Director
Director
Lauren Zeise, Ph.D.

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PREFACE
Proposition 65 1 requires the publication of a list of chemicals “known to the state” tocause cancer or reproductive toxicity. The Office of Environmental Health Hazard
Assessment (OEHHA) of the California Environmental Protection Agency maintains thislist in its role as lead agency for implementing Proposition 65. The Developmental andReproductive Toxicant Identification Committee (DARTIC) advises and assists OEHHA,
and adds chemicals to the Proposition 65 list of chemicals that cause reproductive
toxicity, as required by Health and Safety Code section 25249.8. The DARTIC serves
as the state’s qualified experts for determining whether a chemical has been clearlyshown to cause reproductive toxicity.
The Committee also provides advice and consultation regarding which chemicals
should receive their review. At their meeting in December 2020, the DARTIC
recommended that bisphenol S be placed in a ‘high’ priority group for future listing
consideration. OEHHA selected bisphenol S for consideration for listing by the DARTIC,
and in March 2022, OEHHA solicited from the public information relevant to theassessment of the evidence on the reproductive toxicity of this chemical. In response tothis solicitation, several studies were submitted by the BASF Corporation. The
submitted information was considered in the development of this document.
This document presents evidence relevant to the evaluation of the female reproductive
toxicity of this chemical. On December 12, 2023, the DARTIC is scheduled to deliberateon the female reproductive toxicity of bisphenol S. The Committee will be convened in
2024 to consider the male reproductive toxicity and developmental toxicity of thischemical.
OEHHA developed this document as part of the hazard identification materials that areprovided to the DARTIC to assist it in its deliberations on whether or not bisphenol S
should be listed under Proposition 65. The original papers and reports discussed in the
document are provided to the DARTIC.
OEHHA is holding a public comment period on this hazard identification document.
Comments on this document will be included in the hazard identification materials thatare provided to the DARTIC members prior to the meeting.

1
The Safe Drinking Water and Toxic Enforcement Act of 1986 (California Health and Safety Code section
25249.5 et seq.).

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TABLE OF CONTENTSPREFACE ....................................................................................................................... ii
LIST OF ABBREVIATIONS ............................................................................................ vii
SUMMARY .................................................................................................................... xiii
Introduction ................................................................................................................ xiii
Uses, Occurrence and Exposure ............................................................................... xiii
Systematic Literature Review Approach .................................................................... xiii
Pharmacokinetics of BPS .......................................................................................... xiii
Studies in Humans .................................................................................................... xiv
Studies in Animals ......................................................................................................xv
Mechanistic Considerations ....................................................................................... xix
1. INTRODUCTION ......................................................................................................... 1
1.1 Identity of bisphenol S (BPS) ................................................................................. 1
1.2 Uses, occurrence, and exposure ........................................................................... 1
Use and production .............................................................................................. 1
Occurrence .......................................................................................................... 2
Exposure .............................................................................................................. 3
Considerations for exposure assessment in epidemiologic studies ..................... 6
1.3 Reviews by other health agencies ......................................................................... 7
2. Overview of Systematic Literature Review Approach .................................................. 8
2.1. Primary search process ........................................................................................ 8
2.2. Literature screening process ................................................................................ 9
3. PHARMACOKINETICS ............................................................................................. 10
3.1 Absorption ........................................................................................................... 10
3.2 Distribution ........................................................................................................... 11
Maternal-fetal placental transfer......................................................................... 12
Breast milk transfer ............................................................................................ 12
3.3 Metabolism .......................................................................................................... 12
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3.4 Excretion.............................................................................................................. 13
4. FEMALE REPRODUCTIVE TOXICITY ..................................................................... 14
4.1 Studies in humans ............................................................................................... 15
4.1.1 Gestational diabetes mellitus .................................................................... 16
4.1.2 Thyroid hormones during pregnancy ........................................................ 17
4.1.3 Sex steroid hormones and related proteins during pregnancy .................. 17
4.1.4 Sex steroid hormones and related proteins in young girls ........................ 18
4.1.5 Sex steroid hormones in women ............................................................... 18
4.1.6 Polycystic ovary syndrome........................................................................ 18
4.1.7 Other female reproductive outcomes ........................................................ 19
4.2 Studies in animals ............................................................................................... 44
4.2.1 Overview ................................................................................................... 44
4.2.2 Organ weight and histology....................................................................... 44
4.2.3 Endocrine effects ...................................................................................... 51
4.2.4 Puberty onset ............................................................................................ 57
4.2.5 Effects on estrous cycle ............................................................................ 58
4.2.6 Mammary gland development ................................................................... 59
4.2.7 Reproductive performance ........................................................................ 61
4.2.8 Sex ratio .................................................................................................... 63
4.3 Mechanistic considerations and other data relevant to female reproductive
toxicity ................................................................................................................ 80
4.3.1 Effects on ovarian development and maturation of oocytes ...................... 80
4.3.2 Effects in placenta..................................................................................... 83
4.3.3 Effects on the endocrine system ............................................................... 84
4.3.4 Key characteristics of female reproductive toxicants ................................ 88
5. REFERENCES .......................................................................................................... 98
APPENDIX A. LITERATURE SEARCH APPROACH ON THE FEMALEREPRODUCTIVE TOXICITY OF BPS ........................................................ 108
Primary Search Process .......................................................................................... 108

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1. Data Sources ............................................................................................... 108
2. Search term identification ............................................................................ 109
3. Primary search execution ............................................................................ 109
4. Other data sources ...................................................................................... 113
Literature Screening Process .................................................................................. 113
Use of health assessment workspace collaborative......................................... 113
Detailed PubMed Literature Search Strategies – Primary Searches ....................... 115
APPENDIX B ............................................................................................................... 135

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LIST OF FIGURES
Figure 1 Structure of BPS .............................................................................................. 1
Figure 2 BPS and BPA in pregnant women measured in urine samples collected
between 2007 and 2015 in four biomonitoring studies. ..................................... 6
Figure 3 Proposed metabolism of BPS in rodents. ....................................................... 13

LIST OF TABLES
Table 1.1 BPS serum concentrations (µg/L) in recent studies of California
residents ........................................................................................................... 4
Table 1.2 BPS exposure in pregnant women in California during 2007-2014 froma high-familial risk autism spectrum disorder cohort ......................................... 5
Table 4.1.1 BPS: Epidemiologic studies of female reproductive toxicity. ..................... 21
Table 4.2.1 BPS: Evidence on the female reproductive toxicity in animal studies ........ 64
Table 4.3.1 Key characteristics of female reproductive toxicants ................................. 88
Table 4.3.2 Key characteristics of endocrine-disrupting chemicals .............................. 88
Table 4.3.3 Female reproductive toxicity: studies with mechanistic data notalready included in Table 4.2.1 ....................................................................... 91
Table A.1 Human DART studies search structure (PubMed, Embase, Scopus) ........ 110
Table A.2 Animal DART studies search structure (PubMed, Embase, Scopus) ......... 110
Table A.3 ADME studies search structure (PubMed, Embase, Scopus) .................... 111
Table A.4 Human DART studies search structure (SciFindern) .................................. 111
Table A.5 Animal studies search structure (SciFindern) ............................................. 112
Table A.6 DART studies search structure (Google Scholar) ...................................... 112
Table A.7 BPS DART studies search results (totals from all search dates) ............... 112
Table A.8 PubMed search strategy for human DART studies .................................... 115
Table A.9 PubMed search strategy for animal DART studies .................................... 123
Table A.10 PubMed search strategy for ADME .......................................................... 134
Table B.1 BPS: Summary of epidemiologic studies of female reproductive
toxicity. .......................................................................................................... 135

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LIST OF ABBREVIATIONS
Abbreviation
β
∆
µg
µM
5-HT
5meC
5α-R
ADME

Full namebeta estimatechange / delta
micrograms
micromolar
5-hydroxytryptamine (serotonin)
5-methylcytosine
5α-reductase
Absorption, distribution, metabolism, excretion

AGI
AMH
AMHRII
AR
BCS
BKMR
BMI
Bmp15
BPA
BPAF
BPAP
BPB
BPF
BPP
BPS
BPZ
CARE
CAS
CAT
CHDS
CI

anogenital index
anti-müllerian hormone
anti-müllerian hormone receptor
androgen receptor
body condition score
Bayesian kernel machine regression
body mass index
bone morphogenetic protein 15
bisphenol A
bisphenol AF
bisphenol AP
bisphenol B
bisphenol F
bisphenol P
bisphenol S
bisphenol Z
California Regional Exposure Studychemical abstracts service
catalase
Canadian House Dust Study
confidence interval

CL
CMC
COC

corpus luteum / corpora lutea
carboxymethylcellulose
cumulus-oocyte complex

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Abbreviation
CompToxCRH
CXCYP
CYP11A1
CYP19A1
DART
DEG
DHEA
DHT
dL
DMSO
DNA
DOR
DPF
DPP
DSBs
E1
E2
ECHA
EDC
EE

Full namecomputational toxicology
corticotropin-releasing hormone
connexins
cytochrome P450 enzyme
P450 side chain cleavage [P450scc]
P450 aromatasedevelopmental and reproductive toxicity
differentially expressed genes
dehydroepiandrosterone
dihydrotestosterone
deciliters
dimethyl sulfoxide
deoxyribonucleic aciddiminished ovarian reservedays post-fertilization
days post-partum
double strand breaks
estroneestradiol
European Chemicals Agency
endocrine disrupting chemical
ethinyl estradiol

EGF-R
EPA
ER
ERK
ERα
ERβ
Esr1
Esr2
EU
Exp
FAI
FDA
FF

epidermal growth factor 1
Environmental Protection Agency
estrogen receptor
extracellular signal regulated kinase
estrogen receptor alphaestrogen receptor beta
estrogen receptor alphaestrogen receptor beta
European Union
experiment
free androgen index
Food and Drug Administration
follicular fluid

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Abbreviation
fM
FPG
FR
FSH
FT
FT3
FT4
g
GD
Gdf9
GDM
GJIC
GM
GSI
GV
HAWC
HDL
hpf
HPG
HR
HSD
HSD3B1

Full namefemtomolar
fasting plasma glucose
fecundability ratio
follicular stimulating hormone
free testosterone
free triiodothyronine
free thyroxine
gram
gestational day
growth differentiation factor 9
gestational diabetes mellitus
gap junction intercellular communicationgeometric mean
gonadosomatic index
germinal vesicle
Health Assessment Workspace Collaborative
high-density lipoprotein
hours post-fertilization
hypothalamic-pituitary-gonadal
hazard ratiohydroxysteroid dehydrogenases
3-beta-hydroxysteroid dehydrogenase

HSI
ICR
IGF-1
ip
IQR
IVF
KC
kg
L
LD
LH
ln
LOD

hepatic somatic index
Institute of Cancer Research
insulin-like growth factor 1
intraperitoneal
inter quartile range
in vitro fertilization
key characteristic
kilogram
liter
lactation day
luteinizing hormone
natural logarithm
limit of detection

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Abbreviation
LOQ
m
MAPK
MARBLES
MDA
mg
MI
MII
mL
mmol
mRNA
MVH
n
ng
NHANES
nM
nmol
NS
NTP
OEHHA
OR
P

Full namelimit of quantification
meter
mitogen-activated protein kinase
Markers of Autism Risk in Babies - Learning Early Signs
malondialdehyde
milligram
metaphase I
metaphase II
milliliter
millimolemessenger ribonucleic acidmouse vasa homolog
sample size
nanogram
National Health and Nutrition Examination Survey
nanomolar
nanomole
non-significant
National Toxicology Program
Office of Environmental Health Hazard Assessment
odds ratio
progesteronepP450scc
PAS
PBS
PCNA
PCOS
PC-PLC
PCR
pg
PG
PIP
PI-PLC
PKA

p-value
P450 side chain cleavage
periodic acid-schiff
phosphate buffered saline
proliferating cell nuclear antigen
polycystic ovary syndrome
phosphatidyl choline specific phospholipase Cpolymerase chain reaction
picograms
plasma glucose
posterior inclusion probability
phosphatidylinositol-specific phospholipase C
protein kinase A

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Abbreviation
PKC
pM
pmol
PND
PR
PRL
PrlR
PROTECT
Q1
Q2
Q3
Q4
r
RBC
REACH
RF
RoC
sc
Scp3
SD
SERM
SG
SHBG
SIRT-1
SOD
STAR
STAT5
T
T1
T2
T3
T4
TAC
TBARS

Bisphenol S

Full nameprotein kinase C
picomolar
picomoles
postnatal day
progesterone receptor
prolactin
prolactin receptor
Puerto Rico Testsite for Exploring Contamination Threats
quartile 1
quartile 2
quartile 3
quartile 4
correlation coefficient
red blood cell
Registration, Evaluation, Authorization and Restriction ofChemicals
restricted feed
report on carcinogens
subcutaneoussynaptonemal complex protein 3
standard deviation / Sprague-Dawley
selective estrogen receptor modulator
specific gravity
sex hormone binding globulin
silent information regulator 1
superoxide dismutase
steroidogenic acute regulatory protein
signal transducer and activator of transcription 5
testosterone
tertile 1
tertile 2
triiodothyronine / tertile 3
thyroxine
total antioxidant capacity
thiobarbituric acid reactive substances

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Abbreviation
TBBPA
TC
TCBPA
TEB
TM1
TM2
TM3
Tmax
TNF-α
TSH
TT
TT3
TT4
TUNEL
UL
URM
US
VEGF
VTG
WF
WHO

Bisphenol S

Full nametetrabromobisphenol A
total cholesterol
tetrachlorobisphenol A
terminal end buds
trimester 1
trimester 2
trimester 3
time to get to maximum concentration
tumor necrosis factor-α
thyroid stimulating hormone
total testosterone
total triiodothyronine
total thyroxine
terminal deoxynucleotidyl transferase dUTP nick endlabellinguterine leiomyomata
unexplained recurrent miscarriage
United States
vascular endothelial growth factor
vitellogenin
well fed
World Health Organization

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SUMMARY
Introduction
This document presents evidence relevant to the evaluation of the female reproductive
toxicity of bisphenol S (BPS), also identified as 4,4'-sulfonyldiphenol. BPS consists of
two hydroxyphenyl groups connected by a sulfonyl group and is an analogue ofbisphenol A (BPA).
Uses, Occurrence and Exposure
BPS is part of polyethersulfone plastic, which is used to make hard plastic items andsynthetic fibers for clothing and other textiles. BPS is also used as a color developer inthermal paper and is in protective coatings inside some food cans. It has been detectedin cash register receipts, personal care products, foods, baby bottles, and otherproducts. Recent data from Biomonitoring California (https://biomonitoring.ca.gov) found
BPS is detected in approximately two-thirds of Californians tested. Data from other USbased human biomonitoring studies indicate that as BPA has been removed from many
products over the last decade, the levels of BPS detected in human samples has
increased.
Systematic Literature Review Approach
Using a systematic approach, the Office of Environmental Health Hazard Assessment(OEHHA) conducted literature searches on the developmental and reproductive toxicity
of BPS, including primary searches in major biomedical databases, searches in otherdata sources such as reports by other health agencies, and additional focusedsearches. In addition, OEHHA conducted a data call-in from March 4 to April 18, 2022
to solicit relevant information. For purposes of preparing this document, OEHHA has
focused on literature relevant to female reproductive toxicity (last comprehensive
search, July 2023).
Pharmacokinetics of BPS
BPS is rapidly absorbed in humans by the oral and dermal routes and is distributed
throughout the body. BPS has been detected in human blood, umbilical cord blood,amniotic fluid, breast milk, semen, skin, and urine. BPS does not accumulate in tissues
or blood over time. Once absorbed, BPS undergoes metabolism primarily in the liver,

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where it can be conjugated, and then excreted in urine and feces. Estimates of the halflife of BPS in humans exposed by the oral route range between 7 to 9 hours.
Studies in Humans
OEHHA identified 23 epidemiologic studies of effects of BPS on the female reproductive
system. The female reproductive outcomes that were assessed in more than one study
were gestational diabetes, thyroid hormones, sex steroid hormones, and polycystic
ovary syndrome.
BPS was typically not the sole chemical exposure of interest and methodologicalchallenges related to valid measurement of BPS in observational studies should beconsidered. Most studies measured BPS levels in a single urine spot sample, which
may be insufficient to assess exposure considering BPS’ short half-life. Also, the levelsof detection (LOD) for BPS varied widely across studies, from 0.002 to 0.20 ng/mL. The
percentage of samples with BPS detected for each study was similarly varied, ranging
from 14.8% to 97.7%, and the sample median BPS levels were generally low. Imprecise
exposure assessment resulting in non-differential misclassification of exposure would
likely bias the estimate of any association of risk towards the null.
Evidence for associations of BPS exposure and gestational diabetes (GDM) was mixed
across three studies and seemed to vary by population subgroup. Positive associations
between BPS and odds of GDM were observed among pregnant individuals with prepregnancy body mass index over 23 kilograms per square meter (kg/m2), among thosecarrying a female fetus, and among those identifying as non-Asian/Pacific Islanders(data from two different studies). A third study did not find significant associations with
BPS exposure and onset of GDM but did report increases in screening measures of
GDM, including fasting, 1- and 2-hour plasma glucose with higher BPS exposure levels
in pregnancies with a female fetus.
Four prospective cohort studies and one nested case-control study investigated theassociations between urinary BPS levels and thyroid hormone levels and showed mixed
results. There were observations that BPS was associated with higher levels of total
thyroxine (TT4), free triiodothyronine (FT3) or total thyroid stimulating hormone (TSH)and lower levels of total triiodothyronine (TT3) or free thyroxine (FT4) at various
timepoints over pregnancy. However, study findings related to specific thyroid hormones
did not overlap and the studies conducted many statistical tests, varying times of BPS
and thyroid hormone assessment, with no adjustment for multiple comparisons.

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One study of sex steroid hormones during pregnancy reported an association of BPS
with lower corticotropin-releasing hormone levels, while another study reported noassociations with kisspeptin.
BPS exposures were not associated with changes in sex steroid hormones in young
girls within one cohort and two cross-sectional studies. The only significant finding
reported was the association between BPS exposure and a higher total testosterone(TT)/estradiol (E2) ratio in female adolescents. In adult women, one study reported apositive association between BPS and TT, with no differences in levels of luteinizing
hormone, follicle stimulating hormone, E2, or prolactin.
Two case-control studies reported higher odds of polycystic ovary syndrome in womenwith higher BPS levels.
Other female reproductive outcomes were examined in one study each. Findings
suggested BPS exposure was associated with a decrease in gestational weight gain
over pregnancy, decreases in anti-müllerian hormone, higher odds of diminishedovarian reserve, and delayed onset of menstruation. Associations between BPS and
unexplained recurrent miscarriage varied by age, such that BPS was associated withhigher odds of unexplained recurrent miscarriage among women at 30 years and older
and lower odds among women younger than 30 years old. Conflicting evidence wasgiven for uterine fibroids where within the same study higher BPS levels wereassociated with a significant lower risk for developing uterine fibroids, but among
women with existing fibroids, higher levels of BPS were associated with increases infibroid volume growth. There were no associations between BPS exposure andendometriosis, placental angiogenic/hemodynamic function measures, and gestational
hypertension disorders.
BPS was not significantly associated with fecundability, though there was a suggested
association of longer time to pregnancy with higher BPS exposure among women withinadequate folic acid intake. There were no observed effects on infertility, defined as notconceiving after one year of trying.
Studies in Animals
BPS has been evaluated for its effects on the female reproductive system in 43 in vivo
animal studies, including 18 studies in mice, 13 in rats, three in ewes, one in gerbils,
one in hamsters, five in zebrafish, one in chickens, and one in C. elegans. Most
mammals were exposed to BPS orally by gavage, diet or drinking water, chickens were

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exposed by gavage, zebrafish were exposed in tank water, and C. elegans were
exposed directly via the nematode growth medium. Exposures varied by life stage (e.g.,
gestational, pubertal, adult) and duration (ranging from a few hours in studies assessing
the effects of BPS on hormone levels to 90 days in a repeated-dose oral toxicity study).
Ovarian weight and histology
Effects of BPS on ovarian weight and gonadosomatic index were mixed and only
apparent in a few studies. In the studies that reported changes in ovarian weight, thedirection of the effect appeared to be related to the magnitude of the dose administered.
Seven studies in mice, three of four studies in rats, and one study in hamsters reported
that BPS exposure was consistently associated with effects on ovarian follicle
development, including alterations in the timing of germ cell nest breakdown anddecreases in the numbers of primary, secondary, antral follicles, and corpora lutea (CL).
Gestational BPS exposure of mice was observed to accelerate meiotic progression,
defined by a smaller percentage of oocytes at earlier stages of meiosis prophase I anda larger percentage of oocytes at later stages. Neonatal BPS exposure was observed toincrease the number of atretic cystic follicles in rats and pubertal BPS exposure wasassociated with dose-dependent decreases in primary, preantral, and antral follicles inmice. Exposure to adult rats was observed to decrease numbers of antral and atretic
follicles. BPS exposure to adult mice was associated with atrophy in immature follicles
and increased numbers of atretic follicles, an effect also reported in BPS exposed adult
golden hamsters. Exposure to adult chickens with 50 µg/kg-day for three months wasobserved to decrease the number of preovulatory follicles. One study examined follicle
development in ewes and reported no effects.
Across five mouse studies, the other BPS-related effects observed were damaged
oocyte structure, increased incidence of spindle damage and spindle malformations,and increased numbers of abnormal oocytes at doses as low as 0.001 μg/kg-day. In
zebrafish, observed effects of BPS included increased numbers of perinucleolar
oocytes, decreased numbers of cortical alveolar oocytes, and decreased percentages oflate vitellogenic and spawning oocytes at 1, 10, and 100 µg/L for 14 days, increased
numbers of regressed oocytes, perinucleolar oocytes, and yolk vesicle oocytes at 8, 40,
and 200 µg/mL, and fewer oocytes at the primary growth stage and more oocytes at the
full-grown stage at 1 and 100 µg/L. In C. elegans, one study reported that BPS
exposure was associated with a dose-dependent irregular distribution of nuclei along
the distal to proximal axis, nuclear loss, and increased apoptosis.
Disruptions to follicle and oocyte maturation were often reported alongside observationsof downstream effects on ovulation, measured by changes in the number of CL.
Specifically, three rat studies and one golden hamster study reported associations
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between BPS exposure and reductions of CL and one rat study reported no changes in
the number of CL with BPS exposure.
Uterine weight and histology
In rats, BPS was observed to reduce relative uterine weights in two studies with doses
ranging from 5 to 50 mg/kg-day and to increase uterine weight in two studies with doses
ranging from 40 mg/kg-day to 500 mg/kg-day. Four other studies, also in mice and rats,
reported no effects on uterine weight. Across three studies, BPS exposure wasassociated with changes in uterine histology, including narrowing of the uterine cavity,
increased numbers of uterine glands, reduced endometrial area, atrophied lamina
propria myometrium and plasma layer in adult ICR mice, increased numbers of glandsin the endometrium and thickness of the myometrium in adult C57BL/6J mice, andincreased squamous metaplasia in Wistar rats. In estrogen challenge studies, BPS at
20 mg/kg-day was observed to further increase the uterine weight of ethinyl estradiol(EE) treated rats, and in mice, treatment with BPS + EE was observed to increase
uterine epithelial height, whereas treatment with only BPS had no observed effect onuterine tissue organization.
Endocrine effects
BPS exposure was observed to decrease gonadotropin levels [luteinizing hormone (LH)and follicle-stimulating hormone (FSH)] in rats and mice, and down-regulate expressionof the gonadotropin releasing hormone gnrh3 and fshβ genes in zebrafish brain tissue.
BPS exposure to rats, mice, ewes, and zebrafish was associated with changes inprogesterone (P) levels that were mixed in direction depending on the dose, time of
exposure, and time of assessment.
In mice and rats, increases in serum testosterone (T) levels were observed after BPS
exposure during pregnancy or early in life and decreases in T levels were observed
after BPS exposure later in life (postnatal day [PND] 35 or at eight months of age). No
effects of BPS exposure were observed on plasma T in zebrafish.
Three studies in mice, one in rats, and one study in ewes reported that BPS exposure
was associated with decreased E2 levels in various body fluids (e.g., serum, plasma,
oviduct fluid, urine). Also, in mice, BPS exposure was associated with fewer granulosa
cell layers – the cells producing E2. Two studies in mice reported increased serum E2
levels with BPS exposure during gestation, or by subcutaneous injection every threedays from birth to PND 60. In zebrafish, BPS exposure was associated with increases in
E2 (in plasma or whole-body) and the E2/T ratio, decreases in estrogen receptor alphaBisphenol S

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(ERα) (and vitellogenin) mRNA levels, and non-significant changes in ER beta (ERβ)
mRNA levels.
In the mammary gland of female mice exposed to BPS during gestation and via
lactation (through PND 20), an increase in progesterone receptor (PR) expression wasobserved at puberty, no effects on ERα or PR expression were observed at PND 24,
and a decrease in expression of ERα without effects on PR was observed in adulthood.
Increased mammary gland expression of ERα at PND 31, but not PR, was reported inmice exposed to BPS during gestation and for a shorter time after birth (to PND 2). In
the ovary and uterus of mice, non-significant increases in the expression of ERα andERβ (uterus only) were observed at PND 22 after exposure during gestation andlactation.
Puberty onset
BPS exposure was observed to delay puberty onset in mice exposed during gestation at
20 µg/kg-day or on the first 10 days of life, and in rats exposed perinatally at 60 mg/kgday but not at other doses. Puberty onset was observed to occur 1-2 days earlier inmice exposed during gestation at 0.5 μg/kg-day. Four other studies, with similar dosing
strategies, reported no effects on pubertal timing.
Estrous cycle
BPS exposure was associated with estrous cycle irregularity in mice and rats exposed
prenatally, postnatally, and as adults. The most frequently observed effect was
lengthening of the estrous cycle, and specifically, more days spent in diestrus. Only one
study in mice reported no effects on estrus cyclicity.
Mammary gland development
The effects observed with BPS exposure on mammary gland development varied bytiming of outcome assessment. Gestational and lactational exposure to BPS wasobserved to accelerate the development of terminal end buds (TEBs) in pre-pubertal
mice (2 studies), have no effect on TEBs in pubertal mice (2 studies), and be associatedwith retention of TEBs in adult mice (2 studies). Other effects observed with BPS
exposure on the mammary gland were increased numbers of alveolar buds andincidence of intraductal hyperplasia, increased incidence of mixed cell inflammation,
non-neoplastic lesions, and lobuloalveolar hyperplasia at 14 months of age, decreased
ductal area at PND 31, and increased mammary gland development score at multiple
times and doses. In addition to these mammary gland effects, BPS exposure during
pregnancy and lactation was observed to alter dam nursing behaviors.

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Reproductive performance – including fertilization and implantation
Five studies examined BPS-related effects on fertility rats and mice, of which threestudies observed effects. One reported reduced fertility in rats, one reported nonsignificant decreases in SD rats with 300 mg/kg-day in rats, and a third reported
decreased fertilization at 10 μg/kg-day and increased fertilization at 100 μg/kg-day inpubertal mice.
Treatment of adult rats with BPS was observed to decrease the mean number of
implantation sites in one study, while no effects were reported in one rat and one mousestudy, both with shorter durations of exposure.
An increase in the rate of post-implantation loss was observed in one study of mice
exposed to BPS for 14 weeks from premating to lactation. However, no differences in
the rate of post-implantation loss were observed in rats treated with BPS for a shorter
duration (gestational day 6 to 19) or in mice treated with a lower dose (200 μg/kg-day
for 2 weeks prior to mating through gestational day 12.5). Two studies reported
decreased numbers of live pups born, one in mice and the other in rats, while three
separate studies in mice and one in rats reported no effects on litter size.
In zebrafish, BPS exposure was associated with a decrease in the number of eggs
during a 7-day spawning period, a lower hatching rate, an increase in time to hatchingof embryos, and altered spawning behavior, i.e., debilitated female shoaling. In C.
elegans BPS exposure was associated with a dose dependent increase in embryoniclethality and a decrease in brood size.
Sex ratio
In zebrafish, exposure to BPS from day post fertilization 2 to 75 at doses of 10 and 100
μg/L was observed to skew the sex ratio towards female. No effects on the sex ratio
were observed in rodent models.
Mechanistic Considerations
Numerous in vitro studies, as well as multiple in vivo studies, provide mechanistic
evidence on the potential for BPS to cause female reproductive toxicity.
With respect to adverse effects on whole ovaries, BPS exposure was associated withalterations in α-tubulin assembly, decreased numbers of tubulin filaments that was
associated with spindle disorganization, and chromosome misalignment in oocytes.

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BPS interfered with theca cell gap junction intercellular communication in ovarian thecacells from sheep and primary human ovarian theca cells obtained from women at
reproductive age. It is hypothesized that BPS can modulate gap junction intercellularcommunication through activation of kinase and kinase-independent pathways.
Gestational exposure to BPS in mice was observed to alter ovarian differentiation andreduce oocyte quality, while in a human cell line in vitro exposure to BPS resulted inincreased cell proliferation that is thought to be mediated by a member of the mitogenactivated protein kinase family pathway.
BPS exposure was associated with epigenetic effects, including an observed increase
in DNA methylation in oocytes in one study in mice and an observed decrease intrimethyl-histone H3 (Lys4) (H3K4me3) methylation in in vitro fertilized embryos (fromoocytes collected from BPS exposed mice).
Some studies reported that BPS altered the expression of genes related to ovarianfunction. In mature female mice, BPS exposure was associated with changes in hub
genes (Synp, Fam46c, Cpeb1) that have been linked to mammary gland and breast
neoplasms, polycystic ovary syndrome, hypercholesterolemia, ovarian neoplasm,
ovarian and breast carcinoma, endometriosis, and estrogen resistance.
BPS exposure was associated with alterations to the endocrine system, including
effects on the hypothalamus pituitary gonadal (and liver in zebrafish) axis. Decreases in
gonadotropin levels were reported in two studies in rat and one in mice and changes in
gonadal steroid levels (P, T, and E2) were reported in several in vivo and in vitro
studies. Alterations in hormone receptor mediated events, such as changes in estrogen
receptor expression and/or activity were also observed in several cell types.
Key Characteristics of Female Reproductive Toxicants
Exogenous agents that cause female reproductive toxicity frequently exhibit one ormore of the key characteristics of female reproductive toxicants (See Table 5.1 in
Section 5). The available mechanistic evidence suggests that BPS exhibits several ofthese key characteristics (KCs), including KC 1: alters hormone receptor signaling;alters reproductive hormone production, secretion, or metabolism; KC 2: chemical or
metabolite is genotoxic; KC 8: alters direct cell-cell interactions; and KC 10: alters
microtubules and associated structures.

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1. INTRODUCTION
1.1 Identity of bisphenol S (BPS)
Bisphenol S, or “BPS”, consists of two hydroxyphenyl groups connected by a sulfonylgroup and has the molecular formula C12H10O4S. It is an analog of bisphenol A (BPA)
and its IUPAC name is 4-(4-hydroxyphenyl) sulfonylphenol. Other names for BPS
include 4,4'-sulfonyldiphenol, 4-hydroxyphenyl sulfone, and 4,4′-sulfonylbisphenol. The
chemical structure of BPS is shown in Figure 1. The Chemical Abstracts Service (CAS)registration number for BPS is 80-09-1.

Figure 1 Structure of BPS
1.2 Uses, occurrence, and exposure
Use and production
BPS, a BPA analog, is used to make plastics and is part of polyethersulfone (PES)
plastic, which is used to make hard plastic items and synthetic fibers for clothing andother textiles. BPS may also be used to make colors last longer in some fabrics. It is acommon replacement for BPA in some types of paper receipts and is also in protective
coatings inside some food cans. It is used in coatings of some nonstick pans, as well as
in some baby bottles and parts of electronics, like screens for mobile phones(Biomonitoring California 2020).
The US nationally aggregated production volume range for BPS in 2019 was between
1,000,000 and 10,000,000 pounds per year, as reported by the US Environmental
Protection Agency (US EPA) for the entities required to report and appears to be largely
or completely imported (US EPA 2020). The European Chemicals Agency (ECHA)
reported that at least 10,000 million metric tons of BPS are manufactured or imported
annually into the European Economic Area per year (ECHA 2023).
Some increases in BPS use are associated with actions taken to reduce the use of
BPA. In 2011 Governor Brown signed into law a ban on the use of BPA in baby bottles
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and spill-proof cups; the US Food and Drug Administration (FDA) similarly banned thisuse in 2012, and restricted use of BPA-based epoxy resins in packaging materials for
infant formula in 2013 (US FDA 2014). The European Union (EU) instituted greater
restrictions on BPA that became effective in 2020, including an EU-wide ban on its use
in thermal paper. Since the restrictions on BPA use and public concerns regarding its
safety, manufacturers have been gradually replacing BPA with BPS and other analogs
such as bisphenol F (BPF). ECHA projected the use of BPS in thermal paper in the EU
to be 61% in 2022, and for BPA to be mainly replaced by BPS for this use (ECHA2022). Between 2020 and 2030 the online market analysis platform Chem Analyst(2021) projects a possible 3.6-fold increase in market global demand for BPS.
ChemAnalyst (2021) notes that
“More than 42% of the demand is generated by Heat Sensitive Developers thatare used for the manufacture of Thermal Papers. … Polyurethane systems havebeen associated with offering strength, durability, and resilience while dealing
with challenging conditions such as heavy footfall, physical impacts, extreme
temperatures, and corrosive chemicals. These offerings by [polyurethane] resins
are driving the BPS market growth” (ChemAnalyst 2021).
Occurrence
The European Chemicals Agency (ECHA 2023) notes that BPS:
“can be found in products with material based on: “leather (e.g. gloves, shoes,
purses, furniture), paper (e.g. tissues, feminine hygiene products, diapers, books,
magazines, wallpaper), paper used for articles with intense direct dermal (skin)contact during normal use such as printed articles (e.g. newspapers, books,
magazines, printed photographs) and paper used for articles with intense direct
dermal (skin) contact during normal use such as personal hygiene articles (e.g.
diapers, feminine hygiene products, adult incontinence products, tissues, towels,
and toilet paper)” (ECHA 2023)
BPS has also been detected in personal care products (e.g., hair care products,
makeup, lotions, toothpaste), paper products (e.g., currency, tickets, mailing envelopes,
airplane boarding passes) and food (dairy products, meat, vegetables, canned foods).
BPS has been detected in environmental samples such as dust and sediment, andhuman biological samples such as blood and urine (Fan et al. 2021; Pelch K et al.
2017).
Some examples of BPS detections in environmental samples and consumer products
are provided below.
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•

BPS was detected in all house dust samples from 38 families in Northern
California, collected between May 2015 to August 2016 (Shin et al. 2020).
Similarly, BPS was detected in over 75% of house dust samples collected from2007 to 2010 in 13 cities across Canada under the Canadian House Dust Study(CHDS). The median concentration was 0.242 micrograms per gram (µg/g), andthe range was <0.017 to 35.1 (Fan et al. 2021).

•

BPS was detected in 75% of receipts submitted by volunteers from several US
locations, with the majority from southeast Michigan (Ecology Center 2019).

•

Fifty adults in North Carolina collected 776 duplicate-diet solid food samples overa six-week monitoring period in 2009–2011. A 32% detection frequency wasreported for BPS. The researchers calculated a daily dietary exposure of 13,640
nanograms per day (ng/day) and a dietary intake dose of 238 nanograms per
kilogram per day (ng/kg/day) (Morgan and Clifton 2021).

•

Fish packaged with thermal labels on plastic film were found with BPS with amean of 147 ng/g, and a maximum level of 1140 ng/g, exceeding the European
Union specific migration limit (50 ng/g ww) (Xu et al. 2023).

•

In a study in Albany, NY, BPS was detected in 43% of fresh meat and meatproducts (n = 51) with a mean of 0.609 ng/g, and 95th percentile of 0.780 (Liaoand Kannan 2013).

•

Cao et al (2022a, b) reported concentrations of BPS in composite samples ofmeat and meat products in the Canadian Total Diet Study from 2008 to 2020. In
2008, BPS was detected in meat samples with measured concentrations ranging
from 0.070 ng/g in canned luncheon meats to 105 ng/g in fresh pork and 140
ng/g in veal cutlets. In 2020, measured concentrations of BPS in meat samples
ranged from 0.14 ng/g in cured pork to 26 ng/g in ground beef and 118 ng/g inroast beef. The authors noted that, while levels varied across years, there was no
trend, and that:“The lack of trend for BPS over the period of 13 years (2008-2020) does
not support the speculation that BPS is being used to replace BPA in foodpackaging, and sources other than food packaging may be possible andshould be investigated for BPS.” (Cao et al. 2022a, b)

Exposure
Human biomonitoring studies indicate that exposure to BPS is widespread. The tableand figures below present data on serum BPS concentrations from recent biomonitoringstudies. Table 1.1 presents data from the state’s Biomonitoring California program.
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Table 1.1 BPS serum concentrations (µg/L) in recent studies of California
residents
Geometric mean(µg/L)
0.38(95% CL, 0.270.34)

50th
Percentile

90thPercentile

Number ofParticipants

Detection
Frequency

0.342

2.42

60

76.7%

Adults

NC*

0.233

2.25

151

64.90%

Adults

NC

0.288

2.85

90

64.4%

Study
Name1

SampledGroup

CARE-LA

Women

CARE-2
CARE-3

CARE-LA: California Regional Exposure Study (CARE), Los Angeles County. Sample collection in 2018;
CARE-2: Region 2. Sample collection in 2019; CARE-3: Region 3. Sample collection in 2020.
*Geometric mean not calculated (NC) when the chemical was found in <65% of samples; the Limit ofDetection was 0.100 µg/L (wet weight)
CL: confidence interval. Data available at https://biomonitoring.ca.gov/. (Biomonitoring California 2020)
1

In another study conducted in California, urine samples were taken from pregnant
women, and analyzed for several chemicals including BPS. This is the MARBLES
study, a high-familial risk autism spectrum disorder cohort, in which 218 pregnant
women in California were sampled between 2007-2014. The researchers reported
increasing BPS detection in urine across time, from 0% in 2007 to 32% in 2014. The
95th percentile BPS level was 2.8 nanograms per milliliter (ng/mL, equivalent tomicrograms per liter (µg/L) reported in other studies). Because the limit of detection(LOD) was lower than for the recent Biomonitoring California studies, frequencies ofdetections cannot be directly compared across these studies. Also as noted by Kim etal. (2021) the sample sizes are small and therefore the temporal trends and geometricmean concentrations should be interpreted with caution.

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Table 1.2 BPS exposure in pregnant women in California during 2007-2014 from ahigh-familial risk autism spectrum disorder cohort (Adapted from Kim et al. 2021)
Year

No. of Samples

BPS Detection
Frequency(%)

BPA Detection
Frequency
(%)

2007

51

0

67

2008

121

5

63

2009

181

9

58

2010

112

14

59

2011

67

6

55

2012

65

26

62

2013

106

26

62

2014

57

32

42

At the national level, biomonitoring for BPS has been conducted under the US Center
for Disease Control and Prevention’s National Health and Nutrition Examination Survey(NHANES). Urinary BPS concentration levels were reported in summary form for 1808
adults and 868 children for the years 2013- 2014. Participants were selected using arandom selection approach. BPS was detected in 89.4% of urine samples with medianlevels of 0.37µg/L for adults and 0.29 µg/L for children. Urinary BPS levels in adults
varied by race/ethnicity, where non-Hispanic Black and Hispanic adults had significantly
higher BPS levels compared to non-Hispanic White adults. Urinary BPS levels inchildren varied by age (6-11 year olds had higher BPS levels than 12-19 year olds),
gender (females had higher BPS levels than males), and race/ethnicity (Hispanic
children had higher BPS levels than White children) (Lehmler et al. 2018).
Kim et al. 2021 presented data on temporal trends in BPA and BPS urinary levels for
pregnant women in the NHANES and MARBLES studies, as well as two additional
studies: LIEFECODES (pregnant individuals in Massachusetts, 2007-2009), andPROTECT (pregnant individuals in Puerto Rico, 2011- 2016). They also presented
NHANES data for non-pregnant women. For NHANES they selected the ages 20 to 50
stratified by pregnancy status, for the years 2013 – 2015. Figure 2, adapted from Kim etal. (2021), indicates decreasing BPA levels with concomitant increases in BPS levels
measured in urine samples of pregnant and non-pregnant women during the period
covered by these studies, 2007-2015.

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Figure 2 BPS and BPA in pregnant women measured in urine samples collected
between 2007 and 2015 in four biomonitoring studies. (Source: Kim et al. 2021
[adapted])
Finally, of note in an online crowdsourced study, BPS was detected above the medianmethod reporting limit in urine at 81% of 726 US participants with values ranging from
0.084–921 ng/mL, and a median of 0.48 ng/mL. Samples were collected between
February 2017 and October 2018 (Dodson et al. 2020).
Considerations for exposure assessment in epidemiologic studies
Several characteristics for each study related to exposure magnitude and assessment
influence the study’s ability to detect an association if one exists. These include theLOD of BPS in the analysis of the samples taken, the percentage of the samples with
BPS levels above the LOD, and the median and variability in BPS exposure levels. In
the epidemiologic studies presented in this document there was considerable variationin the LODs and, as such, in the ability to detect BPS in the samples. Also, measured
BPS levels in the participants across studies differed substantially. Some of thevariation in BPS exposure across studies may be explained by study year, as BPS
occurrence has changed over time. Other very important considerations are that (i) BPS
is rapidly eliminated in urine, with a short half-life (estimates range between 7 to 9 hours
for oral exposures; see Section 3. Pharmacokinetics of BPS), and (ii) most of theepidemiological studies relied on a single urine sample to classify exposure. There is a
growing consensus that in many circumstances a single urine sample may be
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insufficient to adequately assess exposure to non-persistent chemicals, such as BPS(Fays et al. 2020; Verner et al. 2020).
Imprecise exposure assessment resulting in non-differential misclassification of
exposure would likely bias the estimate of any association of risk towards the null, thatis, toward not detecting an effect even if one were present.
1.3 Reviews by other health agencies
The US EPA, the National Institute for Occupational Safety and Health (NIOSH), the
FDA, and the National Toxicology Program (NTP) have not reached conclusions orclassified BPS as to its potential to cause female reproductive toxicity.
ECHA has identified BPS as a substance meeting the criteria of a reproductive toxicant(toxic for reproduction category 1B, H360FD” 2). ECHA’s description for category 1B is
as follows (ECHA 2022):“Reproductive toxicant category 1B adverse effects on sexual function andfertility or on development or reproductive toxicant category 2 with R60 (Mayimpair fertility) or R61 (May cause harm to the unborn child)”.
In the summary of classification and labelling, the hazard statement listed is “Maydamage fertility. May damage the unborn child.” (See:https://echa.europa.eu/substance-information/-/substanceinfo/100.239.213).
ECHA (2022, pp 12-13), in identifying BPS as a substance of “very high concern
because of its toxic for reproduction”, stated:“Based on all available scientific evidence, it can be concluded that BPS fulfils theWHO/ International Programme on Chemical Safety (2002) definition of an
endocrine disruptor:• It shows clear reproductive adverse effect in rodents and fish. The reproductive
endocrine system is highly conserved not only between mammals, but also
between mammals and other vertebrates like fish.
• It has endocrine modes of action: clear estrogenic mode of action and alteration ofsteroidogenesis.
• The adverse effects, including the recognized EAS-mediated effects (e.g., on
estrous cycle and sex ratio) and effects sensitive, but not diagnostic of EAS (e.g.
fecundity, fertility, implantation sites and number of pups), are a consequence ofthe endocrine modes of action.”
H360FD is the hazard statement code for “May damage fertility. Suspected of damaging the unborn
child”
2

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“The assessment performed demonstrates that there is scientific evidence of
probable serious effects of BPS to the environment and human health due to its
endocrine disrupting properties, which give rise to an equivalent level of concern tothose of other substances listed in points (a) to (e) of Article 57 of the REACH
Regulation” (ECHA 2022)

2. OVERVIEW OF SYSTEMATIC LITERATURE REVIEW APPROACH
Searches of the published scientific literature on the developmental and reproductive
toxicity (DART) of bisphenol S (BPS) were conducted in January 2022 and updates tothe original search were conducted in February, and July 2023. The goal was to identify
peer-reviewed open source and proprietary journal articles, print and digital books,
reports, and gray literature that potentially reported toxicological and epidemiologicalinformation on the DART of this chemical.
The searches were conducted using the following three approaches:•••

Primary searches in major biomedical databases, conducted by OEHHA librarian
Nancy Firchow, MLS
Searches in other data sources, including authoritative reviews and reports, anddatabases or web resources, conducted by OEHHA scientists
Additional focused searches, conducted by OEHHA scientists

In addition to information identified from these searches, OEHHA also considered thefollowing:•

One submission received during the data call-in period (March 4 – April 18, 2022)(https://oehha.ca.gov/proposition-65/comments/comment-submissions-requestrelevant-information-reproductive-toxicity)

2.1. Primary search process
The US EPA Computational Toxicology (CompTox) Chemicals Dashboard(https://comptox.epa.gov/dashboard) was used to identify synonyms for BPS. The
PubMed MeSH database (https://www.ncbi.nlm.nih.gov/mesh/) was used to identify
subject headings and other index terms related to the chemical, reproduction anddevelopment, and adverse effects on reproduction and development.

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Preliminary searches were run and results evaluated to identify additional relevant
search terms. The resulting search strategies were then executed in PubMed where
searches were divided as follows:•••

Human DART studies (including in vitro studies)
Animal DART studies (including in vitro studies)
Absorption, distribution, metabolism, and excretion (ADME) studies

There were no restrictions in the searches on exposure route or duration of exposure, oron publication language. The full DART search strings used in PubMed are included inAppendix A.
The PubMed search strategies were then tailored for use in the additional databases
and data sources listed below, according to the search interface and features unique toeach resource. For instance, MeSH terms were replaced with Emtree terms for the
Embase search strategies.
2.2. Literature screening process
The results of the literature searches were uploaded to EndNote libraries and duplicateswere removed. In addition to the studies identified through this process, other relevant
studies were identified from citations in individual articles, through alert services (e.g.,
ScienceDirect, Google Scholar, etc.), and through February and July 2023 updates tothe searches. A total of 1,400 references were identified in the searchers for human andanimal DART data and a total of 759 references were identified in the searches for
ADME data (see Appendix Table A.7).
The EndNote libraries containing the literature search results (citations) for BPS were
uploaded to HAWC (Health Assessment Workspace Collaborative,
https://hawcproject.org). HAWC is a tool used for multi-level screening of literaturesearch results.
In Level 1 screening, citations were reviewed independently by OEHHA scientists,based solely on study titles and abstracts, using specific inclusion and exclusion criteria
to eliminate studies or articles that did not contain information on DART or other key
related topics (e.g., pharmacokinetics, mechanisms of action). This initial screen (Level1) was intended to identify all studies deemed to have a reasonable possibility ofcontaining information relevant to DART that could be useful for the review process, andto further identify (i.e., tag in HAWC) studies relevant to particular aspects of DART
(e.g., male reproductive toxicity, female reproductive toxicity, developmental toxicity).
For purposes of identifying the available evidence on the female reproductive toxicity of
BPS, citations identified as having a reasonable possibility of containing information
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relevant to female reproductive toxicity underwent Level 2 screening. In the Level 2
screening of this subset of citations, the full text was obtained. These full papers werescreened independently by one OEHHA scientist, using similar inclusion/exclusion
criteria as was used in the Level 1 screening. However, Level 2 reviewers could makemore accurate judgements about the relevance of the articles because they werereviewing the full text in addition to the title and abstract. Following Level 2 screening,
the tagging of articles according to key topics was updated in HAWC. Level 1 and 2
screenings were repeated as search results were updated, and with additional relevant
studies identified from citations in individual articles and alert services (e.g.,
ScienceDirect, Google Scholar) (See Appendix A for additional details).
One hundred and twenty-four references were cited in this document.

3. PHARMACOKINETICS
This section briefly describes the absorption, distribution, metabolism, and excretion of
BPS in humans and laboratory animals. Since humans and laboratory animals sharemany of the reported metabolic pathways for, and metabolites of, BPS, data fromanimal studies are included when human data are unavailable or incomplete. Data fromanimal studies are useful to complement human data and highlight observed route,species, and gender differences.
3.1 Absorption
In general, BPS can be absorbed following oral, inhalation or dermal exposures (Kumar
et al. 2018). BPS is rapidly absorbed in humans, on the order of hours through oral
exposures and seconds through dermal exposures, with higher oral bioavailability of
BPS compared to bisphenol A (BPA) (Khmiri et al. 2020). For example, in humans
serum concentrations of total and unconjugated d4-BPS increased rapidly within one
hour after a single oral administration (Oh et al. 2018). In dermal absorption studies inhumans, exposure to BPS from thermal papers and receipts showed slower dermal
absorption (Liu and Martin 2019; Reale et al. 2021). In humans the average BPS post
exposure BPS measurement in hand wipes was 3.9 µg and the time to get the
maximum concentration in urine (Tmax) ranged from 14 – 26 hours. It was reported that
78% of urine samples had detectable BPS after two days. BPS was not detectable inserum collected over 7.5 hours (Liu and Martin 2019). In vitro absorption in frozen
human skin (800 µm) skin after 24 h of exposure, 73% of BPS remain in skin swab0.4%
of BPS was absorbed at 2h (Reale et al. 2021).
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In one study the half-life of BPS in human serum was 4.06 and 6.81 hours forunconjugated and total BPS, respectively, after a single oral administration (Oh et al.
2018) and in another study the half-life in human plasma was 7.9 and 9.3 hours forunconjugated and conjugated BPS, respectively, after a single oral dose (Khmiri et al.
2020).
Studies in rats and mice administered a single gavage dose of [14C]BPS have also
found that BPS is well absorbed (Waidyanatha et al. 2018; Waidyanatha et al. 2020).
There was a slower absorption in females rats compared to males after a single oral
BPS exposure 110 mg/kg (Waidyanatha et al. 2020). The Tmax was 5.88 hours forfemale and 1.17 hour for male (Waidyanatha et al. 2018; Waidyanatha et al. 2020).
Another study conducted in female Wistar rats reported that BPS was rapidly absorbed
and detected in plasma following a single oral dose of 30 or 300 mg/kg [14C]BPS, with
87% of the lower dose and 96% of the higher dose absorbed within one hour (BASF
2019b).
The plasma half-life of unconjugated BPS following a single oral dose was estimated tobe 2.86–4.21 hours in mice and 5.77–11.9 hours in rats (Waidyanatha et al. 2020).
3.2 Distribution
Once absorbed into the circulation, BPS is distributed throughout the body (Oh et al.
2018; Khmiri et al. 2020). BPS has been detected in human cord blood, amniotic fluid,breast milk, semen, and skin (Niu et al. 2021).
As shown in single dose gavage studies in rats and mice, extensive biliary excretionoccurs within the first two hours of dosing, with significant re-uptake from the small
intestine (i.e., enterohepatic recirculation) (Waidyanatha et al. 2018; Karrer et al. 2018).
Similar findings were reported in another study in rats (BASF 2019b). In the rat BPS has
been detected in gastrointestinal track tissues, liver, kidney, heart, spleen, lung, andmuscle following gavage administration (Mao et al. 2022). In the BASF (2019b) study infemale Wistar rats, the tissue distribution (ordered from highest to lowest levels ofradiolabel without considering the highest levels in the GI tract with its content) after 1
hour in the 300 mg/kg dose group was: brain, plasma, liver, thyroid, pancreas, lung,skin, and carcass, and in the 30 mg/kg dose group the distribution was: liver, kidney,
thyroid, brain, bone, adipose tissue, and muscle. The maximum plasma concentration
measured in this study was at 1 hour in the 300 mg/kg dose group and at 4 hours in the
30 mg/kg dose group (BASF 2019b). BPS did not accumulate in tissues or blood overtime in rats and mice administered BPS as a single dose (Mao et al 2022; BASF 2019b;
Waidyanatha et al. 2018).

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Maternal-fetal placental transfer
As reviewed by Abrantes-Soares et al. 2022, there is maternal-fetal transfer of BPS via
the placenta in humans, with detection in cord blood and amniotic fluid (AbrantesSoares et al. 2022; Liu et al. 2017; Pan et al. 2020), and in animals (e.g., Gingrich et al.
2018; Grandin et al. 2018; Mao et al. 2020).
Breast milk transfer
BPS has been measured in human breast milk in the US and other populations aroundthe world (Iribarne-Durán et al. 2022; Luo et al. 2021).
3.3 Metabolism
BPS undergoes metabolism via sulfation, glucuronidation and hydroxylation to form
BPS-sulfate, BPS-glucuronide, and hydroxylated bisphenol S (Waidyanatha et al.
2018). Figure 3 depicts the main metabolic pathways and metabolites for BPS in
humans, rats and mice (Oh et al. 2018; Sonker et al. 2021; Waidyanatha et al. 2018).
Studies indicate that BPS-glucuronide is the primary metabolite of BPS formed inhumans (Khmiri et al. 2020; Oh et al. 2018), rats, and mice (Mao et al. 2022;
Waidyanatha et al. 2018). In an in vitro study in HepaRG cells, a human hepatic cellline, 85.8% of BPS was metabolized to BPS-glucuronide and 10.5% to BPS-sulfate (Ohet al. 2018). Comparative metabolism studies in male and female rats and miceadministered a single gavage dose of BPS reported that BPS-glucuronide accounted forapproximately 40-50% of the administered dose in both rats and mice, while BPSsulfate accounted for approximately 18-21% of the administered dose in mice, and 4%
in rats (Waidyanatha et al. 2018).
Considerable inter-individual variation in metabolism of BPS was observed in a study
conducted in women, with the percentage of a single administered dose recovered inurine as conjugated BPS during the first 72 hours ranging from 37 to 72% (Khmiri et al.
2020). Another single dose study conducted in adult men and women reported that thepercentage of the dose recovered in urine as conjugated BPS during the first 48 hours
was lower in women (59-77%) compared to men (67-104%) (Oh et al. 2018).

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Figure 3 Proposed metabolism of BPS in rodents. Abbreviations: SULTs
sulfotransferases; UGTs = uridine 5′-diphospho-glucuronosyl-transferase; CYP=
cytochrome P450 enzyme (Adapted from Waidyanatha et al. (2018))
3.4 Excretion
BPS is excreted in the urine, feces and breast milk. In humans, BPS is excreted
primarily via urine. In one study, the total urinary excretion of a single oral dose of BPS
was 92% in men and 70% in women after 48 hours, with an estimated half-life ofelimination of 6.93 hours, representing metabolism and excretion (Oh et al. 2018).
Another study in women estimated a similar half-life of elimination for a single oral doseof BPS of around 7.9-9.3 hours and reported that urinary excretion was almost complete
after 72 hours (Khmiri et al. 2020).
BPS is also primarily excreted via urine in mice, rats, and sheep. No substantial species
or sex differences in urinary BPS excretion were identified in studies of male and female
rats and mice administered a single gavage dose of BPS, with approximately 67-70 %of an administered dose of 150 mg/kg excreted within 72 hours (Waidyanatha et al.
2018). Greater variability was reported for fecal excretion, however, with 22, 16, 12, and17 percent of the administered dose excreted by male rats, female rats, male mice, andfemale mice, respectively, within 72 hours (Waidyanatha et al. 2018).
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In a study conducted in female Wistar rats, 2% of a single oral 30 mg/kg dose of BPS
was reported to be excreted in expired air (BASF 2019b). In rats receiving an oral BPS
dose of 30 mg/kg 51% was excreted in the urine and 43% in the feces, and in rats thatreceived an oral BPS dose of 300 mg/kg 39.02% was excreted in the urine and 55.71%
in the feces (BASF 2019b).
The elimination half-life in rats was 4.11 hours in females and 8.06 hours for males after
a single oral exposure to BPS at 110 mg/kg. The plasma elimination half-life in malemice was 2.86 hours at 34 mg/kg and 4.21 hours 340 mg/kg, and in male rats was 5.77
hours at 34 mg/kg and 11.9 hours at 340 mg/kg after a single gavage administration(Waidyanatha et al. 2020).

4. FEMALE REPRODUCTIVE TOXICITY
Development of the female reproductive system starts in utero when fetal follicle andoocyte formation and development begin. At birth, the life-long reserve of ovarianfollicles is established, and oocytes have started miotic division. Oocytes remainarrested in meiosis prophase I until follicle development is resumed during puberty, aphysiologic stage that brings up a series of coordinated events to prepare the body forreproduction. There are several steps in female reproduction that are possible targets
for environmental or chemical insult. These steps include development of the
reproductive tract (e.g., the ovaries, oviducts, uterus, cervix, and vagina), early oocyte
and follicle development, oocyte and follicle maturation in preparation for ovulation,
oocyte fertilization within the oviduct, transport of the embryo to the uterus, implantation,and formation of the placenta.
Female reproductive biology is under tight control of the endocrine system. In addition
to the intrinsic regulation present in the organism, environmental factors may influence
the reproductive process by altering the ovarian physiology and the endocrine balancenecessary for reproductive success. Endocrine effects can occur through a variety ofmechanisms, including effects on hormone synthesis or metabolism, hormone transportand binding to proteins in the blood, and cell receptor mediated events, such as
receptor transactivation and alterations in intracellular signaling. Endocrine-disrupting
chemicals, such as bisphenols, have been recognized to interfere with some of theseprocesses.
In this section, the available evidence on the effects of BPS on female reproduction ispresented in the following order: evidence from human epidemiologic studies, evidence
from whole animal in vivo mammalian and non-mammalian studies, and evidence fromstudies most often employing in vitro experimental designs, which help inform
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consideration of mechanisms of action. This is followed by an integrated summary of
the evidence on the female reproductive toxicity of BPS. Some studies presented here
included examination of the effects of prenatal BPS exposure through the pregnant
mother on offspring outcomes, so summary of these studies may appear in thedevelopmental toxicity section also.
4.1 Studies in humans
OEHHA identified 23 epidemiologic studies of possible effects of BPS on the female
reproductive system for inclusion. These were cohort, case-control, or cross-sectional
studies that assessed individual-level exposure to BPS and several different female
reproductive outcomes. Included studies evaluated associations between BPS
exposure and female reproductive outcomes utilizing multivariable epidemiologicalmethods, reported original data analyses with details about methods, and were
published in peer-reviewed journals. Two studies were excluded because of the lack ofmultivariable methods. One was a comparative correlation study of women withpolycystic ovary syndrome (PCOS) by (Šimková et al. 2020) and the other was a study
measuring levels of BPS in breast cancer patients and survivors by (Segovia-Mendoza
et al. 2022) that reported only correlations or mean levels without adjustment. Abstracts
from conferences, opinions, and reviews were also excluded.
Female reproductive outcomes that were assessed in more than one study included
gestational diabetes mellitus (GDM), thyroid hormones during pregnancy, sex steroid
hormones during pregnancy, sex steroid hormones in young girls, and PCOS. All
studies reported BPS exposure as nanograms per milliliter (ng/mL) BPS measured inurine (most common) or serum. As discussed in the exposure section, Appendix B
Table B.1 presents the limit of detection (LOD), percent of samples with BPS detected,
and sample matrices for each study.
The studies are ordered by outcome in the following discussion. Key elements of eachstudy are presented in Table 4.1. Statistically significant results (α = 0.05) in all thetables are shown in bold along with p-values and confidence intervals (CIs). In thesummary presented below, all results are statistically significant unless otherwise noted.
Estimation of an unbiased association between BPS and reproductive outcomesrequires adjustment for potential confounding variables. Covariates included in study
adjustment sets are listed in Table 4.1. The most frequently adjusted variables wereage, body mass index (BMI), parity and/or gravidity, exposure to smoking, and markers
of socioeconomic status (e.g., household income, occupation, maternal education).
Most adjustment sets were comprehensive, though bias due to unmeasured or residual
confounding is possible in observational research.
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All but one of the studies included measurement of additional bisphenols, mainly
bisphenol A (BPA) and bisphenol F (BPF). The correlations between BPS and otherbisphenols varied across studies. In general, most studies showed positive correlations
between BPS and other bisphenols in the range of 0.10 to 0.30. Four studies controlled
for exposure to other bisphenols using mutual adjustment in multivariable models. Nine
studies conducted analyses using Bayesian kernel machine regression (BKMR) models,
which can estimate the contributions of BPS to the joint effects of a chemical mixture. It
is important to note that BPS was typically not the sole exposure of interest. This may
suggest that publication bias for BPS is low, as pressure to publish or not publish basedon statistical significance or findings related specifically to BPS are unlikely.
4.1.1 Gestational diabetes mellitus
Evidence for associations of BPS exposure and development of GDM was mixed,
specific to effect modification, and based on three studies. One of the three studies was
a nested case-control study from a prospective birth cohort in Guangxi Province, China(Tang et al. 2021). Overall, pregnant women with BPS levels in the middle tertile of
exposure had 1.77 times higher odds of GDM (odds ratio (OR) 1.77; 95% CI: 1.01,
3.13) compared with women in the lowest tertile of exposure. The OR for the highest
versus lowest tertile was similar but not statistically significant (OR 1.68, 95% CI: 0.95,
2.99). There was effect modification by BMI and fetus sex such that associations werestronger for pregnant women with pre-pregnancy BMI at or above 23 kilograms permeters squared (kg/m2) (OR 3.89; 95% CI: 1.29, 13.20) and for pregnancies with afemale fetus (OR 2.74; 95% CI: 1.07, 7.47) when comparing the highest to lowest
tertiles of BPS exposure (Tang et al. 2021). Within a nested case-control study from acohort in California, pregnant women in the highest tertile of BPS exposure during thefirst trimester had 2.12 times higher odds of GDM (OR 2.12; 95% CI: 1.00, 4.50)
compared to those in the lowest tertile of exposure. Effect modification was seen as
participants identifying as non-Asian/Pacific Islander (race/ethnicities of white, black,
Hispanic or other) who had BPS exposure in the highest tertile during the first trimester
had 4.6 times higher odds of developing GDM (OR 4.60; 95% CI: 1.55, 13.70)
compared to non-Asian/Pacific Islanders in the lowest exposure tertile (Zhu et al. 2022).
No elevation in risk of GDM was observed in participants identifying as Asian or PacificIslander; the study did not report evaluating effect modification by BMI. Zhu et al (2022)
also analyzed BPS exposure as the area-under-the-curve across the first and second
trimesters to incorporate both concentration and timing of exposure into one index andfindings were consistent; among non-Asian/Pacific Islanders, higher GDM risk was
observed with higher cumulative exposure across first and second trimesters (Zhu et al.
2022). The third study in a cohort in Wuhan, China did not report significant
associations with BPS exposure and onset of GDM, although the authors reported
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increases in screening measures of GDM (increased fasting, 1- and 2-hour plasma
glucose) with increased BPS exposure among women with female fetuses (Zhang et al.
2019).
4.1.2 Thyroid hormones during pregnancy
Four prospective cohort studies and one nested case-control study investigated theassociation between urinary BPS levels and thyroid hormone levels, which aresupportive of pregnancy, including thyroid stimulating hormone (TSH), total
triiodothyronine (TT3), free triiodothyronine (FT3), total thyroxine (TT4), and freethyroxine (FT4). Derakhshan et al. (2021) analyzed data collected prior to the 18th weekof pregnancy from a prospective birth cohort in the Netherlands and reported that forevery unit increase in natural-log (ln) BPS, serum TT4 levels increased by 0.97 (95%
CI: 0.03, 1.91) (Derakhshan et al. 2021). Huang et al. (2022) analyzed data collectedduring the first and second trimesters from a prospective birth cohort in Guangxi, China,
and reported that first trimester TT3 levels were lower by percent change (%Δ) of -10.90(95% CI: -18.16, -2.99) for those with BPS measurements in the middle tertile comparedto the lowest tertile (Huang et al. 2022). Analyses of second trimester data found FT3increased by %Δ of 6.07 (95% CI: 1.20, 11.17) for those with BPS measurements in thehighest tertile compared to the lowest tertile, with no significant effect on TT3 (Huang etal. 2022). No associations between urinary BPS and thyroid hormones were observedin prospective birth cohorts in Puerto Rico (Aker et al. 2019) or Sweden (Derakhshan etal. 2019). A nested case-control study of pregnant women in Boston reported that
detectable levels of BPS were associated with changes in FT4 (Aker et al. 2018).
Authors originally recruited participants as a nested case-control study and incorporated
inverse probability weighting in analyses to account for the over selection of preterm
births and to allow the sample to be representative of the original cohort. Analyses
proceeded in a cohort study design. This study had very low detection frequency of BPS(26.2%), thus exposure was modeled as a binary variable of above or below the LOD.
Aker et al. (2018) observed that in women with BPS above the LOD (compared to below
LOD), FT4 was decreased at less than 15 weeks gestational age (%Δ = -9.63, 95% CI: 18.10, -0.33). Additionally, for women with term births (>37 weeks gestation) and BPS
above the LOD, TSH was increased at less than 15 weeks gestational age (%Δ = 11.77,
95% CI: 0.08, 24.8). There were no significant associations observed at later gestational
ages (Aker et al. 2018).
4.1.3 Sex steroid hormones and related proteins during pregnancy
One prospective cohort study in Puerto Rico analyzed data on urinary BPS levels andserum levels of corticotropin-releasing hormone (CRH), estriol, progesterone,
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testosterone (T), and sex hormone binding globulin (SHBG) during pregnancy (Aker etal. (2019). Increased levels of BPS were associated with a decrease in CRH perinterquartile range (%Δ = -11.35, 95% CI: -18.71, -3.33), with the strongest associations
observed at 16 to 20 weeks. There were no statistically significant associations reported
for other outcomes (Aker et al. 2019). One prospective cohort study in China reported
no significant associations between urinary BPS and total kisspeptin levels in pregnant
women, as assessed during the third trimester (Wang et al. 2022). Kisspeptin is a familyof peptides involved in a number of reproductive processes, including the secretion of
gonadotrophins and the regulation of fertility and puberty onset.
4.1.4 Sex steroid hormones and related proteins in young girls
One prospective cohort study in China reported no association between gestational
exposure to BPS (measured in third trimester maternal urine) and levels of kisspeptin ingirls 6 years of age (Wang et al. 2022).Two cross-sectional studies, both using datafrom the same National Health and Nutrition Examination Survey (NHANES) survey
cycles (2013-2014, 2015-2016), investigated associations between urinary levels of
BPS and serum estradiol (E2), total testosterone (TT), TT/E2, SHBG, and free
androgen index (FAI; TT/SHBG) in female participants ages 6-19 years (Hu et al. 2022;
Wang et al. 2021). No statistically significant associations were reported by Hu et al.
(2022), while Wang et al. (2021) reported an increase in TT/E2 between the first andsecond quartiles of BPS exposure in female adolescents. Differences in findings
between the studies may be due to the analysis of BPS exposure as continuous by Hu
et al. (2022) versus as categorical (in quartiles) by Wang et al (2021). When Wang et al(2021) did analyze the data treating BPS as a continuous variable using restricted cubic
spline models, there was evidence to suggest potential non-linear associations between
BPS and two outcomes, FAI and SHBG.
4.1.5 Sex steroid hormones in women
A case-control study in China analyzed associations between BPS and hormones inwomen without PCOS (i.e., women in the study control group) and reported that per unit
increases in creatinine-adjusted BPS were positively associated with T levels (β = 0.07,
95% CI: 0.02, 0.12). There were no differences observed for luteinizing hormone, follicle
stimulating hormone, E2, and prolactin (Zhan et al. 2023).
4.1.6 Polycystic ovary syndrome
Two case-control studies investigated the association between PCOS and BPS levels.
One study in China reported women with higher levels of urinary BPS had higher odds
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of PCOS compared to women with lower levels of BPS, when BPS was analyzed as acontinuous variable and by quartiles (p < 0.001 for trend) (Zhan et al. 2023). Anotherstudy in Poland estimated associations between per unit increases in ln BPS and PCOS
odds within each tertile of BPS. Within the lowest exposure tertile, per unit increases in
ln BPS were associated with higher odds of PCOS (OR 1.12, 95% CI: 1.03, 3.71). Per
unit increases in ln BPS were also associated with higher odds of PCOS in the middle
exposure tertile (OR 1.29, 95% CI: 0.79, 6.89) and in the highest exposure tertile (OR1.33, 95% CI: 0.58, 4.88), though these estimates were not statistically significant(Jurewicz et al. 2020). This study had reporting limitations, namely, they only reported
associations estimated within each tertile of BPS exposure and did not report the
number of PCOS cases within each stratum (Jurewicz et al. 2020).
4.1.7 Other female reproductive outcomes
Weight Gain During Pregnancy
A prospective cohort study from Rotterdam, Netherlands assessed associations
between BPS levels measured during early pregnancy and gestational weight gain overpregnancy and reported that for every unit increase in ln BPS, there was a 261-gram
reduction in gestational weight gain over the total pregnancy (95% CI: -466, -56) (Philips
et al. 2020).
Recurrent Miscarriage
One case-control study of unexplained recurrent (≥ two) miscarriage (URM) in China
reported higher odds of URM with BPS exposure levels in quartile two (OR 1.27, 95%
CI: 1.22, 2.22) and quartile three (OR 1.37, 95% CI: 1.02, 1.85) versus in quartile one(Ao et al. 2022). There were no differences between quartile four and quartile one (OR0.92, 95% CI: 0.69, 1.23). Secondary analyses showed that associations between BPS
and URM may vary by age. Specifically, there were higher odds of URM observed inwomen aged 30 or older in the middle tertile of BPS exposure compared to the lowest
tertile (OR 1.62, 95% CI: 1.13, 2.33) and lower odds of URM observed in women lessthan 30 years old in the highest tertile compared to the lowest tertile (OR 0.65, 95% CI:
0.44, 0.97) (Ao et al. 2022).
Uterine Fibroids
In a case-cohort study of Black women followed over the course of 60 months, higher
urinary BPS levels were associated with a significant lower risk for developing uterine
leiomyomata (fibroids) (Hazard ratio 0.93 (95% CI: 0.87, 0.99) (Wesselink et al. 2021).
Growth of a subset of uterine fibroids detected during the study was assessed via
transvaginal ultrasound at 20 months (n = 67) and 60 months (n = 145), withmeasurements conducted on as many as six fibroids per participant. The authors
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reported that a 50% increase in urinary BPS was associated with a 4.1% increase infibroid volume growth over 18 months (95% CI: 0.3%, 8.0%) (Wesselink et al. 2021).
Fecundability/Fertility
A cross-sectional study in China at an infertility clinic reported associations between
urinary BPS measurements and decreases in anti-müllerian hormone (β = -0.287, 95%
CI: -0.505, -0.070) (Zhang et al. 2023). Zhang et al. (2023) also reported higher odds ofdiminished ovarian reserve among individuals with high versus low levels of urinary
BPS (OR 6.85, 95% CI: 1.24, 37.82). A prospective cohort study in the Netherlands
found no association between first trimester urinary BPS levels and time to pregnancy(fecundability), though there was a suggested association of longer time to pregnancy
with per unit increases in ln BPS exposure among women with inadequate folic acidintake (fecundability ratio 0.94, 95% CI: 0.87, 1.01) (Philips et al. 2018). A crosssectional study of NHANES data by Zhan et al. (2022) found no association between
urinary BPS levels and infertility in women 18 to 45 years of age.
Onset of Menstruation
A prospective birth cohort study in Rotterdam, Netherlands examined associations
between average gestational exposure to BPS and offspring reproductive development
at ages 10 and 13 years. BPS was associated with delayed onset of menstruation by
0.17 years, on average, among female offspring (β = 0.17, 95% CI: 0.02, 0.31)(Blaauwendraad et al. 2022).
Placental Function and Hypertension During Pregnancy
Urinary levels of BPS were not associated with placental angiogenic markers, placental
hemodynamic function, or hypertensive end points, in a prospective cohort in the
Netherlands (Philips et al. 2019).
Endometriosis
No statistically significant associations were reported between urinary BPS levels andendometriosis (Peinado et al. 2020).
Thyroid Levels in Adult WomenUrinary BPS levels from adult women were not associated with changes in thyroid
hormones (TT4, TT3, FT4, FT3, and TSH) in a cross-sectional study from Beijing, China(Yue et al. 2023a).

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Table 4.1.1 BPS: Epidemiologic studies of female reproductive toxicity.
Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Outcomes

Results

Covariates, otherbisphenols analyzed

Comments

Gestational Diabetes Mellitus (GDM)
Tang et al. 2021

Serum samples

Guangxi Providence, China

Limit of Detection (LOD)
0.046

2015-2021
Nested case-control study,
hospital-based
Guangxi Zhuang Birth
Cohort (GZBC) prospective
birth cohort
n = 500; n = 100 cases,
matched 4:1 on age andfetal sex
Mean ages for cases (±Standard Deviation [SD]:30.62 ± 6.46 years
Mean ages for controls(±SD): 30.6 ± 6.41 years
Inclusion criteria:
singleton pregnancy,
Zhuang population (motheror father), enrolled at ≤ 12
weeks gestation

Bisphenol S% < LOD: 17.8
Values below the LOD
were calculated asLOD÷√2
Geometric mean, cases:
0.103
Geometric mean, controls:
0.092
Percentiles:
25th: 0.05
50th: 0.097
75th: 0.107
95th: 0.83
Timing of samples:
Samples taken in firsttrimester

One step GDM
screening via blood:oral glucose tolerance
test at 24-28 weeks
gestation.

Odds of GDM, BPS tertiles

Criteria for GDM
based on meeting oneof three criteria:

Female fetus:
T2 vs T1, OR: 2.34 (95% CI: 0.95, 6.18)
T3 vs T1, OR: 2.74 (95% CI: 1.07, 7.47)
P trend = 0.03

Fasting plasma
glucose (FPG) > 5.1
millimoles per liter(mmol/L);
1 hour plasma glucose(1 h-PG) > 10.0
mmol/L;2 hour plasma glucose(2 h-PG) > 8.5mmol/L.

Tertile 2 (T2) vs Tertile 1 (T1), odds ratio (OR):1.77 (95% Confidence Interval [CI]: 1.01, 3.13)
Tertile 3 (T3) vs T1, OR: 1.68 (95% CI: 0.95, 2.99)
P trend: 0.049

No association with male fetus:
T2 vs T1, OR: 1.61 (95% CI: 0.74, 3.55)
T3 vs T1, OR: 1.45 (95% CI: 0.67, 3.20)
P trend = 0.28
High body mass index (BMI) (≥ 23 kg/m2):
T2 vs T1, OR: 1.72 (95% CI: 0.55, 5.80)
T3 vs T1, OR: 3.89 (95% CI: 1.29, 13.20)
P trend = 0.03

Models adjusted for: area ofresidence, BMI, passive smoking
during pregnancy, parity,
gravidity, regular exercise.
Matched on maternal age andfetal sex.
Effect modification: BMI, fetal
sex
BKMR models and quantilebased g-computation used toestimate effects of BPS as partof a mixture of chemicals.
Other Bisphenols: Bisphenol A(BPA), Bisphenol B (BPB),
Bisphenol F (BPF),
Tetrabromobisphenol A (TBBPA)

Approach used to classify
BPS tertiles was not clearly
described and ranges of BPS
included in each tertile were
not reported.
BPA negatively associatedwith GDM in main analyses
and in BKMR/quantile gcomputation models.
Correlations between BPS
and other bisphenols: TBBPA(r = 0.31), BPF (r = 0.32), BPA(r = -0.27), BPB (r = -0.03).

No association at BMI < 23 kg/m2:
T2 vs T1, OR 1.16 (95% CI: 0.55, 2.42)
T3 vs T1, OR: 0.84 (95% CI: 0.40, 1.74)
P trend = 0.78
BPS consistently showed increased risk of GDM
through Bayesian Kernel Machine Regression(BKMR) and g-computation analysis (Posterior
inclusion probability (PIP) for BPS = 0.99). Showed
an inverted U shape dose-response.

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Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Zhang et al. 2019

Urine spot samples

Wuhan, China

Adjusted with specificgravity (SG)2013-2015
Prospective cohort
n = 1,841
Mean age (±SD): 28.58 ±3.27 years
Inclusion criteria:
singleton pregnancy,
enrolled at < 16 weeks
gestation, no family historyof diabetes, no diabetes
before pregnancy

LOD: 0.2% < LOD: 10
Values below the LOD
were calculated asLOD÷√2
Geometric mean (95% CI),
SG adjusted: 0.36
(0.33,0.38)
Percentiles:
25th: 0.14
50th: 0.31
75th: 0.81
95th: 5.51
Timing of samples:
Samples collected between
weeks 8-18 gestationalage, on average at 13
weeks

Bisphenol S

Outcomes
One step GDM
screening via blood:oral glucose tolerance
test at 24-28 weeks
gestation
Criteria for GDM
based on meeting oneof three criteria:
FPG > 5.1 mmol/L;
1 h-PG > 10.0 mmol/L;
2 h-PG > 8.5 mmol/L
Screening measures:
FPG, 1 h-PG, 2 h-PG,
Sum of PG z-scores

Results
Odds of GDM, BPS tertiles:
No significant association between BPS tertiles and
GDM in full or BMI-stratified analyses (ORs ranged
from 0.83 to 1.66)
GDM screening measures:
For continuous, per unit increase in ln BPS:
FPG [β = 0.03 (95% CI: 0.01, 0.04)] (p < 0.01)
Sum PG z-score [β = 0.07 (95% CI: −0.00, 0.14)] (p= 0.05)
No associations with 1 h-PG or 2 h-PG
Significant interaction with fetal sex for FPG, 1 h
PG, Sum PG z-score (p < 0.05)
Female fetus:
For continuous, per unit increase in ln BPS:
FPG [β = 0.04 (95% CI: 0.02, 0.06)] (p < 0.01)
1 h-PG [β = 0.11 (95% CI: 0.04, 0.17)] (p <0.01)
Sum PG z-score [β = 0.19 (95% CI: 0.08, 0.30)] (p< 0.01)

Covariates, otherbisphenols analyzed
Models adjusted for: maternalage, education level, passive
smoking, pre-pregnancy BMI,
parity, infant sex.
GDM models additionallyadjusted for other bisphenols.
Effect modification: fetal sex
Other Bisphenols: BPA, BPF,
Bisphenol AF (BPAF)

Comments
BPA had a U-shaped
exposure pattern for FPG and
PG z-scores, but noassociation with linear
models.
Ranges of BPS values
included in each tertile were
not reported.
Correlations between BPS
and other bisphenols: BPA (r= 0.14), BPF (r = 0.11), BPAF
(r = 0.20).
BPAF was associated with anincreased risk of GDM inwomen of normal range prepregnancy BMI.

Similar results when exposure was categorized into
tertiles for pregnancies with female fetus
No associations in pregnancies with male fetus

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Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)Zhu et al. 2022

Urine Spot Samples

Northern California, USA

Adjusted with creatinine

2015-2017

Values below the LOD
were calculated asLOD÷√2

Nested case-control study
within the Kaiser
Permanente Northern
California – Pregnancy
Environment and Lifestyle
Study (PETALS) prospective
birth cohort
n = 111 and n = 222 controls
Mean ages (±SD): 31.2 ±4.6
Inclusion criteria:
singleton pregnancy, <11weeks gestation, no
evidence of recognized preexisting cancer, diabetes, or
hepatic diseases

LOD: 0.1% < LOD:
Trimester 1.
Cases: 16.2, controls: 15.8
Trimester 2.
Cases: 11.8, controls: 18.3
Values below the LOD
were calculated asLOD÷√2
Median (Interquartile range[IQR] 25th, 75th):
Trimester 1:cases: 0.6 (0.3,1.3),
controls: 0.4(0.2,0.9)
Trimester 2:cases: 0.5 (0.2,1.2),
controls: 0.4 (0.2,1.1)
Timing of samples(gestational weeks ± SD):

Outcomes
One step GDM
screening 24-28
weeks gestation: oral
glucose tolerance test
or fasting glucose test
≥ 5.1 mmol/L

Results
First trimester BPS tertiles
T2 (IQR: 0.4-0.6 ng/mL) vs T1 (IQR 0.1-0.3 ng/mL),
OR: 1.86 (95% CI: 0.94, 3.67)
T3 (IQR: 1.0-2.7 ng/mL) vs T1, OR: 2.12 (95%
CI:1.00, 4.50)
Significant interaction with race/ethnicity (p < 0.001)
Non-Asian/Pacific Islanders:
T2 vs T1, OR: 1.43 (95% CI: 0.48, 4.27)
T3 vs T1, OR: 4.60 (95% CI: 1.55, 13.70)
No associations among Asian/Pacific Islanders:
BPS area-under-the-curve first and second
trimester:T2 (IQR: 78.8-111.2 ng/mL) vs T1 (IQR: 19.7-51.0
ng/mL), OR: 1.59 (95% CI: 0.67, 3.77)
T3 (IQR: 189.8, 443.7 ng/mL) vs T1, OR: 2.49
(95% CI: 1.01, 6.17)
No significant associations when BPS was
analyzed per IQR increase.

Covariates, otherbisphenols analyzed
Models adjusted for: maternal
age at childbirth, race/ethnicity,
pre-pregnancy BMI, creatinine.
Effect modification: race/ethnicity
Other Bisphenols: BPA, BPF
BKMR models used to estimate
effects of BPS as part of a
mixture of chemicals.

Comments
Non-Asians/Pacific Islander
individuals had higher risk of
GDM with higher levels of
BPA exposure.
Authors stated “…wecalculated the area-under-thecurve as a proxy of overall
exposure to phenols in early
to mid-pregnancy, whichshowed overall consistent
results with time-specificanalysis.”
Correlations between BPS
and other bisphenols notreported.
IQR of each BPS tertile
reported, but specific bounds
were not.

BPS showed an inverted u-shaped association with
GDM risk through cubic splines for both firsttrimester BPS and area-under-the-curve
Contribution to BPS to mixture in BKMR models
was low to moderate (PIPs ranged from 0.24 to
0.57)

Visit 1: 14.0 ± 2.3
Visit 2: 20.5 ± 2.4

Bisphenol S

23

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Outcomes

Results

Pregnancy thyroid
hormones via bloodplasma at gestational
visits 1-4: freethyroxine (FT4), total
thyroxine (TT4), total
triiodothyronine (TT3),and thyroid stimulating
hormone (TSH),
TT3/TT4 ratio.

Analyzed as values above or below the LOD sincevery few were above the LOD:

Covariates, otherbisphenols analyzed

Comments

Thyroid Hormones During Pregnancy
Aker et al. 2018

Urinary spot samples

Boston, MA

Adjusted with SG

2006-2008

LOD: 0.4**

Nested case-control study
within the LIFECODES
Cohort*%< LOD: 73.8**

n = 439, n = 130 preterm
births and n = 352 controls

Geometric mean: <LOD

Mean ages: 31.8 years (27%35+)
Inclusion criteria: singleton,
non-anomalous fetus, deliverat participating hospital, ≥18 years, < 15 weeks, no
pre-existing thyroid
conditions such as thyroid
cancer, Graves’ disease andhyper- or hypothyroidism

Values below the LOD
were calculated as LOD/√2
Timing of samples gestation week (medianand range):
Visit 1: 9.64 (5.43-19.1)
Visit 2: 17.9 (14.9-32.1)
Visit 3: 26.0 (22.9-36.3)
Visit 4: 35.1 (33.1-38.3)

Outcomes analyzed
as percent change(%Δ)

Non-significant increase for TSH
Non-significant decrease for FT4, TT4, TT3,
TT3/TT4 ratio
Stratified by gestational age:
FT4 at <15 weeks gestational age %Δ = –9.63(95% CI: –18.10, –0.33) (p = 0.04)
No associations observed at other gestational
ages.
Among births > 37 weeks
TSH at <15 weeks gestational age %Δ = 11.77(95% CI: 0.08, 24.8) (p = 0.05)
No associations observed at other gestational ages

Models adjusted for: SG, study
visit, BMI at the first study visit,
insurance provider, maternal age
and gestational age at time ofsample collection.
Stratification: gestational age (<15, 15-21, 21-30, > 30 weeks)
Sensitivity analysis limited tobirths > 37 weeks

Benzophenone-3, 2,4dichlorophenol, and 2,5dichlorophenol were also
analyzed, and weakly
correlated with BPS (range ofcorrelation coefficients 0.08 to0.11).
* Authors originally recruitedparticipants as a nested casecontrol study and incorporated
inverse probability weighting
in analyses to account for theover selection of preterm
births and to allow the sample
to be representative of theoriginal cohort. Analyses
proceeded in a cohort study
design.
**Mistakes were noted in themethods section where the
BPS LOD is given as 0.1
ng/mL in text and 0.4 ng/mL in
Table 2 of the study. The
percentage of samples with
BPS above the LOD wasgiven as <25% in text andgiven as 26.2% in Table 2.
Many statistical tests were runwith no adjustment for multiple
comparisons.

Bisphenol S

24

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Derakhshan et al. 2019

Urine spot samples

SwedenAdjusted with creatinine

2007-2010

LOD: 0.03

Prospective cohort study% < LOD: 19.8

Swedish Environmental
Longitudinal, Mother andchild, Asthma and allergy(SELMA) cohort

Reported concentrations
were used for values below
LOD

n = 1,996

5th: 0.03

Mean ages (±SD): 30.9 ±4.9 years
Inclusion criteria: singleton
pregnancy, no pre-existing
thyroid issues

Bisphenol S

Outcomes
Thyroid hormones inserum measured at
first prenatal visit:
TSH, FT4, FT3, TT4,
and TT3, TT4/TT3
ratio, FT4/FT3 ratio

Results
For continuous, per unit increase in ln BPS
No associations of BPS with thyroid hormones
Non-significant increase for TSH, FT4, FT3,
FT4/FT3 ratio, TT4, TT4/TT3 ratio
Non-significant decrease for TT3

Covariates, otherbisphenols analyzed
Models adjusted for: maternalage, thyroid peroxidase
antibodies, thyroglobulin
antibodies, human chorionicgonadotropin, urinary creatinine,smoking status by cotininelevels, BMI, education, ethnicity,and parity.
Other Bisphenols: BPA, BPF

Percentiles:
50th: 0.08
95th: 0.88

Comments
BPA associated with reduced
FT4/FT3 ratio, TT4 (non-linear
relationship) and TT4/TT3
ratio. BPA had significant
interactions with timing of
FT3, FT4/FT3, and TT4/TT3
between weeks 7 and 12 ofgestation. BPF wasassociated with higher FT3.
Correlations between BPS
and other bisphenols: range
0.17 to 0.33 (exact correlation
coefficients not provided).

Timing of samples(gestation week):
median 10
range 6-14

25

OEHHA
October 2023

Reference; study
location, period,
design, sample
Derakhshan et al. 2021
Rotterdam, Netherlands
2002-2006
Prospective Cohort Study
Generation R Studyn = 1,267
Mean ages (±SD): 30.5 ±4.8 years
Inclusion criteria: singleton
pregnancy, ≤ 18 weeks
gestation

Exposure matrix
and concentration(ng/mL)
Urine spot samples(creatinine adjusted)
LOD: 0.05% < LOD: 33.3 earlypregnancy, 70.2 middle
pregnancy, 81.1 latepregnancy

Outcomes

Results

Maternal thyroid
function via serum <18
weeks gestation: TSH(mU/L), FT4 (pmol/L),
TT4 (nmol/L)

For continuous, per unit increase in ln BPS (earlypregnancy):
TT4 [β = 0.97 (95% CI: 0.03 1.91)] (p = 0.04)
Non-significant increase for FT4, non-significant
decrease for TSH
There was a significant interaction (p = 0.001) for
FT4 and ln-TSH (i.e., BPS associated with anattenuation of the association between FT4 and lnTSH)

Values below the LOD
were calculated asLOD÷√2

Covariates, otherbisphenols analyzed
Models adjusted for: week ofgestation, maternal age, thyroid
peroxidase antibodies, humanchorionic gonadotropin, urinary
creatinine, BMI, education,
ethnicity, smoking status, andparity.
Other Bisphenols: BPA, BPF

Comments
No associations between BPA
and outcomes.
Correlations between BPS
and other bisphenols: BPA (r= 0.19), BPF (r = 0.30) at time
1; BPA (r = -0.08) at time 2;
estimates at time 3 (latepregnancy) not reported due
to > 80% below the limit ofdetection.
Percentiles of BPS at time 3(> 25 weeks) not reported.

Percentiles (earlypregnancy, middle
pregnancy):
5th: < LOD, < LOD
50th: 0.34, 0.24
95th: 8.83, 1.69
Timing of samples(gestation week):
Early: <18, middle: 18-25,
late: >25

Huang et al. 2022

Serum samples

Guangxi Providence, China

LOD: 0.046

2015-2018% < LOD: 13.7

Prospective birth cohort

Values below the LOD
were calculated asLOD÷√2

Guangxi Zhuang Birth
Cohort (GZBC) prospective
cohortThyroid hormone
testing for: TT3, TT4,
FT3, FT4, and TSH in
first or second
trimester.
Analyzed as %Δ

First trimester thyroid hormone:T2 vs T1, TT3 %Δ = –10.90 (95% CI: –18.16, –2.99)
T3 vs T1, TT3 %Δ = -4.76 (95% CI: -12.40, 3.54)
No associations with TT4, FT3, FT4, TSH

Models adjusted for: maternalage, pre-pregnancy BMI,
drinking before pregnancy,
passive smoking during
pregnancy, gravidity, parity, andfetus sex.
Effect modification: fetal sex
Other Bisphenols: BPA, BPB,
BPF, TBBPABKMR models used to estimate
effects of BPS as part of a
mixture of chemicals.

3-level BPS variable (tertiles)n = 446

Percentiles:

Second trimester thyroid hormones:
T2 vs T1, FT3 %Δ = 3.26 (95% CI: -1.43, 8.17)
T3 vs T1, FT3 %Δ = 6.07 (95% CI: 1.20, 11.17)

Mean ages (±SD): 27.21 ±5.44 years

25th: 0.096

No associations with TT3, TT4, FT4, TSH

50th: 0.097

Male fetus:
T2 vs T1, FT3 %Δ = 3.46 (95% CI: -1.00, 8.12)
T3 vs T1, FT3 %Δ = 6.53 (95% CI: 2.07, 11.19)

Inclusion criteria:

Bisphenol S

Geometric mean: 0.091

75th: 0.104

26

No clear patterns were found
for the other BPs. For firsttrimester hormone
measurements, BPA
associated with higher FT3,
BPB lower T4 and FT4 andhigher TSH, BPF associatedwith higher TSH, and TBBPA
associated with lower T3.
Second trimester hormone
measurements, BPB
associated with higher FT4,
BPF associated with higher
T4 and FT4 and lower TSH,

OEHHA
October 2023

Reference; study
location, period,
design, sample< 13 weeks gestational age,
singleton pregnancy,
Zhuang population (motheror father), no assisted
fertilization, no thyroid
disease during pregnancy

Exposure matrix
and concentration(ng/mL)

Outcomes

Results

95th: 0.198

No associations in pregnancies with female fetus

Timing of samples:
Samples taken in firsttrimesterIn restricted cubic spline and BKMR models, there
was an inverted-U shaped dose-response curve
between BPS and second trimester FT4, non-linear
p-value of 0.046.

Covariates, otherbisphenols analyzed

Comments
and TBBPA associated withhigher T4 and FT4.
Mixture analyses with FT3
showed non-linear effects of
BPs.
Ranges of BPS values
included in each tertile were
not reported.
Correlations between BPS
and other bisphenols: BPA (r= –0.09), BPF (r = 0.15), BPB(r = –0.01), TBBPA (r = 0.18)

Bisphenol S

27

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Outcomes

Results

Covariates, otherbisphenols analyzed

Comments

Sex Steroid Hormones and Related Proteins During Pregnancy
Aker et al. 2019

Urinary spot samples

Puerto Rico

Adjusted with SG

2012-2017

LOD: 0.1

Prospective cohort study% < LOD: 3.4 (16 to 20
weeks)), 8.6 (24 to 28
weeks)

Puerto Rico Testsite for
Exploring Contamination
Threats (PROTECT) Cohort
n = 602
Mean ages (±SD): 26.51 ±5.66 years
Inclusion criteria: singleton
fetus, 14 ± 2 weeks
gestation, 18 to 40 years, nooral contraceptive use within
3 months prior to getting
pregnant, no in vitro
fertilization, no known
medical health conditions(diabetes, hypertension,
etc.)

Values below the LOD
were calculated asLOD÷√2
Geometric mean ±geometric standard
deviation: 0.54 ± 3.15 (16to 20 weeks), 0.54 ± 5.2(24 to 28 weeks)
Percentiles (16 to 20
weeks, 24 to 28 weeks):

Reproductive andpregnancy hormones
via serum at visits 1
and 3: estriol,
progesterone,
testosterone, sexhormone-binding
globulin (SHBG),
corticotropin-releasing
hormone (CRH), TT3,
TT4, FT4, TSH,
TT3/TT4
Analyzed as %Δ

Per IQR increase in BPS%Δ CRH = –11.35, (95% CI: –18.71, – 3.33) (p <0.01)
The association with CRH in linear mixed models
was driven by findings at 16-20 weeks:%Δ CRH = –16.15, (95% CI: –26.6, –4.2) (p <0.01)
The association with CRH was weaker and nonsignificant at 24-28 weeks:%Δ CRH = –7.61, (95% CI: –16.3, 2.01) (p = 0.12)
Other outcomes:
Non-significant increase for testosterone, TT3, TT4,
TT3/TT4
Non-significant decrease for SHBG, progesterone,
estriol, progesterone/estriol, TSH, FT4

Models adjusted for: SG, study
visit, BMI at the first study visit,maternal age, the number ofhours of second-hand smoke
exposure per day, and socioeconomic status.
Other Bisphenols: BPA, BPF

At 24-28 weeks, BPA wasassociated with lower
testosterone, higher FT4 andT3: %Δ = 4.33, 95% CI: 0.11,
8.55.
At 16-20 weeks, BPF wasassociated with higher FT4.
2,4-dichlorophenol, 2,5dichlorophenol,
benzophenone-3, triclosanwere also analyzed, andweakly correlated with BPS(range of correlation
coefficients: –0.04 to 0.04).
Correlations between BPS
and other bisphenols: BPF (r= 0.11), BPA (r = 0.17).

25th: 0.23, 0.23
50th: 0.50, 0.47
75th: 1.07, 1.06
95th: 4.01, 4.23
Timing of samples
Visit 1: 16 to 20 weeks
Visit 2: 20 to 24 weeks
Visit 3: 24 to 28 weeks

Bisphenol S

28

OEHHA
October 2023

Reference; study
location, period,
design, sample
Wang et al. 2022
Shanghai, China
2012
Prospective birth cohort
Shanghai-Minhang Birth
Cohort Study (S-MBCS)n = 528 mother:child pairsn = 195 mother:girl childpairs
Mean ages (±SD): 28.5 ±3.4 years
Inclusion criteria: pregnant
women 12-16 gestational
weeks and receiving care at
the Minhang Maternal andChild Health Hospital in
Shanghai

Exposure matrix
and concentration(ng/mL)
Maternal urine spot
samples
Adjusted with creatinine
LOD: 0.004% < LOD: 75.8%
Values below the LOD
were calculated as LOD÷2(imputed in BKMR models)
Categorized into < LOD,
LOD to median, and ≥medianGeometric mean (95% CI)
creatinine corrected,
expressed as µg/g: 0.02
(0.01, 0.02)

Outcomes

Results

Total urinary
kisspeptin (ng/L),
including kisspeptin
54, 14, 13, and 10,
assessed in mothers
during the third
trimesterNo significant association between third trimester
urinary BPS and kisspeptin levels in pregnant
women.
Male fetus:
LOD-Median vs < LOD [β = –17.93 (95% CI:–78.23, 42.37)]≥ Median vs < LOD [β = 8.74 (95% CI: –61.42,
78.90)]
Female fetus:
LOD-Median vs < LOD [β = –14.92 (95% CI:–87.79, 57.96)]≥ Median vs < LOD [β = –56.89 (95% CI: –134.85,
21.07)]

Covariates, otherbisphenols analyzed
Models adjusted for: maternal
creatinine concentrations,household income, maternal
education, maternal age at
menarche, maternal age,maternal pre-pregnancy BMI,
parity, gestational age atmaternal urine collection, andfetus sex.
Effect modification: fetal sex
Other Bisphenols: BPA, BPF,
BPAF, tetrachlorobisphenol A(TCBPA)
BKMR models used to estimate
effects of BPS as part of a
mixture of chemicals.

Comments
2nd tertile maternal BPA andabove median maternal BPF
associated with decreasedmaternal kisspeptin levels inthird trimester for women witha male fetus.
Correlations between BPS
and other bisphenols: BPA (r= 0.095), BPF (r = 0.143),
BPAF (r = 0.036), TCBPA (r =-0.063).
Study also reviewed in thenext section, “Sex Steroid
Hormones and Related
Proteins in Young Girls”.

Percentiles (creatinine
corrected, expressed asµg/g):
25th: < LOD
50th: 0.01
75th: 0.03
Timing of samples:
third trimester

Bisphenol S

29

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Outcomes

Results

Covariates, otherbisphenols analyzed

Comments

Sex Steroid Hormones and Related Proteins in Young Girls
Hu et al. 2022

Urine spot samples

United States

Adjusted with creatinine

Cross-sectional study

Values below the LOD
were calculated asLOD÷√2National Health and Nutrition
Examination Survey(NHANES) 2013-2016
n = 1,179 participants, males
and females, with data on
urinary bisphenols, serum
sex hormones andcovariates

LOD: 0.1% < LOD: 11.7

No significant associations between BPS and sex
steroid hormone levels for either prepubertal or
pubertal females.
Prepubertal: non-significant increase E2, TT,
SHBG, FAI and non-significant decrease TT/E2
Pubertal: non-significant increase SHBG, TT/E2
and non-significant decrease E2, TT, FAI

Total for population (males
and females):Geometric mean: 0.52

6–19-year-olds

Percentiles:

Mean age, females (±SD):12.31 ± 3.93 years

50th: 0.37

6–11-year-olds wereclassified as children(females n = 273)

Serum sex steroid
hormone levels: TT(ng/dL), E2 (pg/mL),
SHBG (nmol/L),
TT/E2, Free androgenindex (FAI=TT/SHBG)

Other Bisphenols: BPA
BKMR models used to estimate
effects of BPS as part of a
mixture of chemicals.

For all individuals: BKMR
methods showed U-shaped
exposure for E2, linear for TT,
inverted U-shape for SHBG,
U-shape for FAI, and linear for
TT/E2 when reviewed for both
prepubertal and pubertal
individuals.
No associations found with
BPA for prepubertal andpubertal females.
Authors had an error in thepaper interchanging the BPA
and BPS descriptive statistics.

25th: 0.19

Correlations between BPS
and other bisphenols: BPA (r= 0.33).

75th: 0.74
95th: 2.91

Authors also examinedassociations between
parabens and sex steroid
hormone levels and thus
limited their sample toparticipants with bothbisphenols and parabensmeasured, resulting in aslightly different analytic
sample than Wang 2021
(another NHANES study).

12–19-year-olds Wereclassified as adolescents(females n = 301)
Puberty was defined as:
Total testosterone (TT) ≥50
ng/dL in boys, estradiol (E2)≥20 pg/mL in girls
Inclusion criteria: systematic
sample of US population,ages 6 - 19

Bisphenol S

Models adjusted for: age, BMI,
race/ethnicity, poverty index
ratio, serum cotinine level,
survey period, session of blood
sampling, NHANES cycle.

30

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Wang et al. 2021

Urine spot samples

United States

Adjusted with creatinine

2013-2016

Values below the LOD
were calculated asLOD÷√2

Cross-sectional study
NHANES
2 consecutive survey cycles(2013-2014 and 2015-2016)
n = 1,317 participants, males(n = 662) and females (n =655), with data on urinary
bisphenols, serum sex
hormones and covariates
6–19-year-olds mean age:
13 years

LOD: 0.1% < LOD: 11.6
Median (IQR):
Total population: 0.3 (0.6)
Females:

Outcomes
Serum sex steroid
hormone levels: TT(ng/dL), E2 (pg/mL),
SHBG (nmol/L),
TT/E2, FAI=TT/SHBG

Results
Female adolescents (ages 6 – 19):
Q2 vs Q1, higher TT/E2 ratio [ No values given;
results presented in graphs]
No associations with TT, E2
A non-linear, inverted U-shaped, dose-response
was observed for BPS and SHBG
(p for non-linearity = 0.013)
Suggestion of non-linear association with FAI (p for
non-linearity = 0.076)
No associations in female children (ages 6-11)

Covariates, otherbisphenols analyzed
Models adjusted for: age,race/ethnicity, ratio of family
income to poverty, BMI
categories, urinary creatinine,serum cotinine, session time ofvenipuncture, survey cycle, BPA,
BPF.
Other Bisphenols: BPA, BPF

Comments
Correlations between BPS
and other bisphenols: BPA (r= 0.33), BPF (r = 0.15),
BPA was negatively
associated with FAI and E2,
while being positively
associated with SHBG andTT/E2. BPF associated withlower TT in female
adolescents and non-linear
associations with FAI andSHBG

Children 0.28 (0.5)
Adolescent 0.4 (0.7)

6–11-year-olds wereclassified as children(females n = 274)
12–19-year-olds wereclassified as adolescents(females n = 381)
Inclusion criteria: systematic
sample of US population,ages 6-19

Bisphenol S

31

OEHHA
October 2023

Reference; study
location, period,
design, sample
Wang et al. 2022
Shanghai, China
2012
Prospective birth cohort
Shanghai-Minhang Birth
Cohort Study (S-MBCS)n = 528 mother:child pairsn = 195 mother:girl childpairs
Maternal mean ages (±SD):28.5 ± 3.4 years
Girls mean ages: 6 years
Inclusion criteria: pregnant
women 12-16 gestational
weeks and receiving care at
the Minhang Maternal andChild Health Hospital in
Shanghai

Bisphenol S

Exposure matrix
and concentration(ng/mL)
Maternal urine spot
samples
Adjusted with creatinine
LOD: 0.004% < LOD: 75.8%
Values below the LOD
were calculated as LOD÷2

Outcomes

Results

Total urinary
kisspeptin, including
kisspeptin 54, 14, 13,
and 10, assessed inmothers during thethird trimester and ingirls at age 6.

No significant association between maternal BPS
levels and kisspeptin levels in girls at 6 years ofage
LOD-Median vs < LOD β = –0.35 (95% CI -45.14,
44.44)≥ Median vs < LOD β = 23.22 (95% CI –23.23,
69.68)

Categorized into < LOD,
LOD to median, and≥ median

Covariates, otherbisphenols analyzed
Models adjusted for: maternal
creatinine concentrations,household income, maternal
education, maternal age at
menarche, maternal age,maternal pre-pregnancy BMI,
parity, children’s BMI z-score.
Other Bisphenols: BPA, BPF,
BPAF, TCBPABKMR models used to estimate
effects of BPS as part of a
mixture of chemicals.

Geometric mean (95% CI)
creatinine corrected,
expressed as µg/g: 0.02
(0.01, 0.02)

Comments
2nd tertile maternal BPA andabove LOD maternal BPF
associated with increased
kisspeptin in girls at age 6.
Correlations between BPS
and other bisphenols: BPA (r= 0.095), BPF (r = 0.143),
BPAF (r = 0.036), TCBPA (r =-0.063).
Study also reviewed in theprevious section, “Sex Steroid
Hormones and Related
Proteins During Pregnancy”.

Percentiles (creatinine
corrected), expressed asµg/g):
25th: < LOD
50th: 0.01
75th: 0.03
Timing of samples: third
trimester

32

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Outcomes

Results

Covariates, otherbisphenols analyzed

Comments

Polycystic Ovary Syndrome (PCOS)
Jurewicz et al. 2020
Poland2017
Case-control study, medical
practice-basedn = 357 (199 cases, 158
controls)
Mean case ages (±SD): 26.6
± 5.5 years

Serum measurements
Values below the LOD
were calculated asLOD÷√2
LOD: 0.022% < LOD: 5
Geometric mean: 0.14 forcases, 0.08 for controls
Tertiles:Mean control ages (±SD):32.1 ± 6.9 years

Lowest < 0.09

Inclusion criteria: ages 18-40
years

Highest >0.21

Cases: diagnosis of PCOS

Middle 0.09-0.21

PCOS diagnosed
prior to study

For continuous, per unit increase in ln BPS,
stratified by tertile of BPS
In T1 (< 0.09 ng/mL), OR: 1.12 (95% CI:1.03,
3.71) (p = 0.029).
In T2 (0.09-0.21 ng/mL), OR: 1.29 (95% CI: 0.79,
6.89) (p = 0.163)
In T3 (> 0.21 ng/mL), OR: 1.33 (95% CI: 0.58, 4.88)(p = 0.548)
BPS serum levels were higher in cases thancontrols (geometric mean 0.14 ng/mL vs. 0.08
ng/mL, p = 0.023); cases and controls differed inother characteristics, e.g., age, marital status,
education, smoking status.

Timing of samples: at
beginning of study

Other Bisphenols: BPA, BPF

BPA associated withdecreased HOMA-IR(Homeostasis Model ofAssessment-Insulin
Resistance) and testosterone.
The comparison group used
for the effect estimate was notclearly reported, assumed tobe per 1-unit increase in BPS.
No mention of matching
controls to cases, including
significant differences
between the groups based onage, marital status, education,and smoking status.
Correlations between BPS
and other bisphenols notreported.

Controls: without PCOS

Bisphenol S

Models adjusted for: age,education, BMI, income,smoking.

33

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Zhan et al. 2023

Urine spot samples

China

Adjusted with creatinine

2014-2016

Values below the LOD
were calculated asLOD÷√2Case-control study, hospitalbased
Cases = 321, controls = 412

LOD: 0.003

Mean age 29.0 years (14%35+)

% < LOD: 6.8
Percentiles:

Inclusion criteria cases:
women diagnosed withinfertility from PCOS;
controls as stated by author:“The controls consisted ofhealthy women with noendocrine disorders and had
AID”

25th: 0.02
50th: 0.12
75th: 0.49

Outcomes
PCOS identified priorto BPS sample
collection
Hormones: luteinizing
hormone (LH), follicle
stimulating hormone(FSH), E2, TT,prolactin (PRL)

Results
Odds of PCOS
For continuous, per unit increase in ln BPS
OR: 1.18 (95% CI: 1.10, 1.25)
4-level BPS variable (quartiles):Q2 vs Q1, OR: 1.49 (95% CI: 1.12, 1.91)
Q3 vs Q1, OR: 1.70 (95% CI: 1.28, 2.22)
Q4 vs Q1, OR: 2.12 (95% CI: 1.58, 2.79)
P trend: <0.001
Associations were stronger among overweight andobese women after stratification by BMI.
Within mixture analyses, BPS had the highest
weights in quantile-based g-computation models.
Hormones (in no-PCOS group)
For continuous BPS, per unit increase in ln BPS:
TT [β = 0.07 (95% CI: 0.02, 0.12)], (p < 0.05)

Covariates, otherbisphenols analyzed
Models adjusted for: age, BMI,
level of education, annual
household income, employment
status, study site, smoking
status, alcohol consumption.
Effect modification: BMI
Other Bisphenols: BPA,
Bisphenol P (BPP), BPB,
Bisphenol Z (BPZ), bisphenol AP(BPAP), and BPAF
Used quantile-based gcomputation to assess chemical
mixtures and extreme gradientboosting to assess interactions
between bisphenol analogs.

Comments
BPA, BPP, BPB, BPZ, andBPAF were associated withincreased odds of PCOS.
BMI was evaluated as aneffect modifier and could besignificant for otherbisphenols, but not for BPS.
Correlations between BPS
and other bisphenols:
positively correlated with BPA,
BPZ, BPAF, BPB, negatively
correlated with BPB (exact
correlation coefficients notreported).

Secondary analyses of BPS andhormones in no-PCOS group

No associations with LH, FSH, E2, PRL

Bisphenol S

34

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Outcomes

Results

Covariates, otherbisphenols analyzed

Comments

Other Female Reproductive Outcomes
Ao et al. 2022

Urine spot samples

Shanghai and Zhejiang
Providence, China

Adjusted with creatinine

2014-2016% < LOD: 17.5

Case-control study, hospitalbasedn = 1,180 cases, 571
controls
Mean ages (±SD) cases:
30.2 ± 4.1 years, controls:
30.3 ± 3.7 years
Inclusion criteria
Cases: ≥ 2 unexplained
recurrent miscarriage(URM), lack of genetic andinfectious disorders, normal
hormones, lack of known
fertility problems

LOD: 0.003
Values below the LOD
were calculated asLOD÷√2
Percentiles (adjusted withcreatinine expressed asµg/g):
25th: 0.01
50th: 0.11
75th: 0.48
Timing of samples:
measured after miscarriage(before 20 weeks
gestation)

Unexplained recurrent
miscarriage (URM)
identified prior to study
start

BPS quartiles
Q2 vs Q1, OR: 1.27 (95% CI: 1.22, 2.22)
Q3 vs Q1, OR: 1.37 (95% CI: 1.02,1.85)
Q4 vs Q1, OR: 0.92 (95% CI: 0.69,1.23)
P trend: 0.99
Effect modification by women < 30 and ≥ 30 yearsof age with BPS modeled as tertiles:
For women ≥ 30:
T2 vs T1, OR: 1.62 (95% CI: 1.13, 2.33)
T3 vs T1, OR: 1.19 (95% CI: 0.83, 1.71)
For women < 30:
T2 vs T1, OR: 1.01 (95% CI: 0.68, 1.50)
T3 vs T1, OR: 0.65 (95% CI: 0.44, 0.97)
Restricted cubic splines showed an upside-down U
shaped dose-response, p-nonlinear: <0.001
BPS was least weighted chemical in mixture.

Models adjusted for: age, BMI,
study site, education, study site.
Effect modification: age (above
or below age 30), study site
Other Bisphenols: BPA, BPAF,
BPAP, BPB, BPP
BKMR models and quantilebased g-computation used toestimate effects of BPS as partof a mixture of chemicals.

BPA, BPAF, BPAP, BPB, BPP
were associated withincreased odds of URM, effectmodified by those over 30
years for most BPs. Mixture
analyses showed that
BPAF/BPAP/BPA were thehighest contributors to theeffects of BPs on URM.
Ranges of BPS values
included in each tertile were
not reported.
Correlations between BPS
and other bisphenols: BPA (r= 0.23), BPAF (r = 0.46),
BPAP (r = 0.44), BPB (r =0.36), BPP (r = 0.45).

Controls: multiparous
women, no history of
miscarriage, preconception
visit

Bisphenol S

35

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Blaauwendraad et al. 2022

Urine spot samples

Rotterdam, Netherlands

Adjusted with creatinine

2004-2005

LOD: 0.15

Prospective cohort study% < LOD (per trimester
sampling time): 31.5, 70.3,
80.0

Generation R Studyn = 673 boys and n = 524
girls
Mean (±SD) maternal age inyears for: 30.7 (±4.8) boys
and 30.9 (±4.6) girls
Inclusion criteria: mothers
were ≤18 weeks gestation,
singleton

Values below the LOD
were calculated asLOD÷√2
Percentiles (per trimester
sampling time):

Outcomes
Reproductive
development of infants
and children: infant
reproductive tract
abnormalities, ovarianand testicular volume
at 10 years using
magnetic resonanceimaging and pubertal
development andonset of menstruation
at 13 years

Results
Per IQR increase in BPS
age at first menstruation (years) for girls[β = 0.17 (95% CI: 0.02, 0.31)] (p = 0.025)
Non-significant decrease in ovarian volume at age
10;
Non-significant differences in pubic hair and breast
development

Covariates, otherbisphenols analyzed
Models adjusted for child’smaternal age, ethnicity, prepregnancy BMI, education level,
parity, intake, smoking andalcohol use, breastfeeding,child’s gestational age adjusted
birthweight, age and age
adjusted BMI at time of
measurement.
Used quantile-based gcomputation to assess chemical
mixtures.
Other Bisphenols: BPA and BPF

25th: <LOD, < LOD, < LOD

Comments
BPF was not included in the
study with low detection levelsin the second trimester.
Exposure concentrations
converted from nmol/L tong/mL.
Correlations between BPS
and other bisphenols: BPA
range 0.03 – 0.06 bytrimester, BPF range 0.04 –0.28 by trimester.
Only analyses for female
offspring presented here.

50th: 0.2, <LOD, <LOD,
75th: 0.6, 0.08, <LOD,
Timing of samples: Threetime points as median(IQR): 12.9 (12.1-14.1),
20.4 (19.9-20.9), and 30.2
(29.9-30.8) weeks
gestation

Bisphenol S

36

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Peinado et al. 2020

Urine spot samples

Granada, Spain

Adjusted with creatinine

2018-2019

Values below the LOD
were calculated as LOD÷2

Case-control study, hospitalbased% < LOD: 88.6 for cases,
83.9 for controls

n = 124, n = 35 cases andn = 89 controls

Geometric mean: below
LOD

Mean ages (±SD): cases:
38.3 ± 9.3 years, controls:
35.8 ± 10.4 years

25th: <LOD

Bisphenol S

Endometriosis
diagnosis following
abdominal surgery attime of enrollment

Results

Covariates, otherbisphenols analyzed

No significant association between BPS andendometriosis

Models adjusted for: creatinine,age, BMI, parity and residence.

Above vs below LOD, OR: 2.0 (95% CI: 0.4, 10.2)

Other Bisphenols: BPA, BPF

LOD: 0.2

Endometriosis y Exposición
Ambiental (EndEA) study

Inclusion criteria:premenopausal women ages
20-54 years, undergoing
abdominal surgery, with BMI
below 35 kg/m2
Cases: diagnosed withendometriosis
Controls: without
endometriosis

Outcomes

Comments
BPA associated with increasein odds of endometriosis [OR
1.5 (95% CI: 1.0, 2.3)] per log
unit increase, and with thesecond tertile of exposure [OR
3.7 (95% CI: 1.3-10.3)].
Thiobarbituric acid reactive
substances (TBARS), a
marker of lipid peroxidation,
were also analyzed in urine.
Correlation between BPS andother bisphenols not reported.

75th: < LOD

LOD units given as mg/mL intext (assumed to be a typo for
ng/mL based on expected
range of values).

Timing of samples: First
morning urine samples atstart of study, prior tosurgery37

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Philips et al. 2018

Urine spot samples

Rotterdam, Netherlands

Adjusted with creatinine

2002-2006

Values below the LOD
were calculated asLOD÷√2

Prospective cohort study
Generation R Studyn = 877
Mean ages (±SD): 31.2 ±4.4
Inclusion criteria: singleton
pregnancy and live birth, noprior use of infertility
treatment

Bisphenol S

LOD: 0.05% < LOD: 29.5
Percentiles:

Outcomes

Results

Questionnaireanswers for time topregnancy(fecundability)
measured in months
and use of artificial
fertility treatments

No significant association of BPS with fecundability
For continuous, per unit increase in ln BPS:First trimester BPS fecundability ratio (FR) = 0.98
(95% CI: 0.94, 1.02)
By folic acid supplement use:
Inadequate folic acid FR = 0.94 (95% CI: 0.87,
1.01) (p <0.1)
Adequate folic acid FR = 1.02 (95% CI: 0.96, 1.09)

25th: 0.17
50th: 0.35

Covariates, otherbisphenols analyzed
Models adjusted for: maternalage, education, parity, folic acidsupplement use, and urinary
creatinine concentration.
Other Bisphenols: BPA, BPF,
BPZ, BPB, BPAP, BPP, BPAF
Effect modification: folic acid
Sensitivity analyses examined
BPS and BPA in same model(no change in estimates)

Comments
Correlation between BPS andother bisphenols not reported,
authors stated “correlations
between first trimester BPA
and BPS are weak”.
Fecundability ratio interpreted
as relative probability of
becoming pregnant in a single
menstrual cycle per log unit
increase in bisphenol (FR < 1
indicates longer time topregnancy).
Stratification by folic acid was
motivated by prior studies
showing folic acid buffers
adverse effects of BPA.

75th: 1.03
Timing of samples:First trimester (median 12.9
weeks, IQR 12.1-14.4
weeks)38

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Philips et al. 2019

Urine spot samples

Rotterdam, Netherlands

Adjusted with creatinine

2004-2005

LOD: 0.05

Prospective birth cohort% < LOD: 31.5

Generation R Study

Values below the LOD
were calculated asLOD÷√2n = 1,233
Mean age (±SD): 30.5 ± 4.8
years
Inclusion criteria: <18 weeks
gestation, singleton
pregnancy, no pre-existing
hypertension

Bisphenol S

Percentiles:
25th: 0.17
50th: 0.35
75th: 1.03
Timing of samples: Early
pregnancy (mediangestational age 13.1
weeks, IQR 12.1-14.5
weeks)

Outcomes
Blood measurements:
Placental angiogenic
markers measured inearly and midpregnancy: placental
growth factor, tyrosine
kinase ratios,
hemodynamic function

Results
No significant findings for BPS with placental
angiogenic markers, umbilical cord measures,
blood pressure or gestational hypertensive
disorders

Covariates, otherbisphenols analyzed
Models adjusted for: maternal
age at enrollment, educationlevel, ethnicity, parity, folic acidsupplementation, gestationalage, pre-pregnancy BMI (kg/m2),
smoking, and alcohol
consumption.
Other Bisphenols: BPA

Umbilical blood:umbilical artery
pulsatility index (PI),uterine artery
resistance index (RI),
notching and placental
weight,Comments
BPA associated withdecreasing slope of theumbilical artery PI Z-score,
increasing slope of the uterine
artery RI Z-score, but nothingelse. This was over time andwas not seen in the individualmodels.
Correlation between BPS and
BPA not reported.

Physical measures:
blood pressure
measured at each
visit, gestational
hypertensive disorders
via medical records

39

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Philips et al. 2020

Urine spot samples

Rotterdam, Netherlands

Adjusted with creatinine

2002-2006

Values below the LOD
were calculated asLOD÷√2

Prospective cohort study
Generation R Studyn = 1,213
Mean ages (±SD): 30.6 ±4.8 years
Inclusion criteria: singleton
pregnancy and live birth

LOD: 0.05% < LOD: 31.9 early and70.9 mid-pregnancy
Median (IQR) earlypregnancy: 0.35
(0.17,1.09)

Outcomes
Gestational weightgain measured during
study (grams)

Results
For continuous, per unit increase in ln BPS
measured in early pregnancy:
gestational weight gain [β: –261 grams (95% CI: –466, –56)] over total pregnancy
gestational weight gain [β: –142 (95% CI: –282, –2)] until late pregnancy (~ 30 weeks), not significant
after correction for multiple comparisons as statedby authors.
No associations with binary outcomes of insufficientweight gain and excessive weight gain

Median (IQR) midpregnancy: 0.24 (0.12,
0.49)

Covariates, otherbisphenols analyzed
Models adjusted for: maternalage, daily dietary caloric intake,parity, ethnicity, education,maternal smoking, maternal
alcohol, mid pregnancy totalbisphenols, and folic acidsupplementation.
Other Bisphenols: BPA, BPF

Comments
BPA showed a 132 gram
(95% CI: –231, –34) decreasein gestational weight gain
during mid to late pregnancy.
Correlations between BPS
and other bisphenols: Early
pregnancy BPS positively
associated with midpregnancy BPA (r = 0.192);
cross-sectional BPS-BPA
correlations not reported.
BPS-BPF correlation
coefficient not reported.
Mid pregnancy BPS not
analyzed due to low detection.

Timing of samples:
Early (median 13.1 weeks[IQR 12.1-14.5]) andmid-pregnancy (median
20.4 weeks [IQR 19.920.9])

Bisphenol S

40

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Wesselink et al. 2021

Urine spot samples

Detroit, Michigan

Adjustment: creatinine

2010-2012For values below the LOD,
authors used instrument
values.

Case-cohort study
Study of the Environment,
Lifestyle, and Fibroids
prospective cohort studyn = 754
Mean age: 28.6 ± 3.5 years
Inclusion criteria: Black
women, no previous uterine
leiomyomata (UL, fibroids),
no history of cancer, andhad an intact uterus

Bisphenol S

LOD: 0.1% < LOD: 4.6 (Baseline),
5.7 (20 months), 4.1 (40months)Percentiles (baseline, 20months, and 40 months):
50th: 0.5, 0.6, 0.8
90th: 2.6, 2.5, 3.5
Timing of samples:
Baseline measurement
followed by 20 and 40
monthsOutcomes
UL incidence,developed over thecourse of 60 months.
Fibroid growth,measured via
transvaginalultrasound on a subset
of fibroids, withmeasurements on up
to 6 fibroids per
participant.
Fibroid growthmeasured from 20-40
months (n = 67
fibroids), and from 4060 months (n = 145
fibroids).

Results

Covariates, otherbisphenols analyzed

Stratification by age:

Models adjusted for: age,education, parity, cigarettesmoking, alcohol intake, BMI,
age at menarche, depot
medroxyprogesterone acetate
use, time since last birth, andcreatinine.

Below age 30, HR of UL: 0.90 (95% CI: 0.82, 0.99)
30+ years: HR of UL: 0.95 (95% CI: 0.87, 1.04)

Effect modification: maternalage, parity, BMI

No evidence of effect modification by parity or BMI

Other Bisphenols: BPA, BPF

Per 50% increase in BPS:
Hazard ratios (HRs) of UL: 0.93 (95% CI: 0.87,
0.99)%Δ in UL volume over 18 months: 4.1% (95% CI:
0.3, 8.0)

BPS showed a nonlinear U-shaped dose-responsewith restricted cubic splines

Comments
Multiple imputation
approaches were used in the
study: (i) for missing urinary
phenol concentrations (10.1 –
18.1% of study participants
based on timepoint ofcollection), (ii) for missing
outcome status for those who
did not complete any follow-up(3.1% of study participants),
and (iii) for missing
confounder data (<1% of
study participants). Complete
case estimates presented insupplement are similar, butless precise.
Correlations between BPS
and other bisphenols: BPS
positively correlated with BPA
and BPS (exact coefficientsnot reported).

41

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)Yue et al. 2023a

Urine spot samples

Beijing, China

Adjusted with creatinine

2017

LOD: 0.002

Cross-sectional Study% < LOD: 2.3

n = 86

Values below LOD were
given the LOD

Mean (±SD) age: 40 (±10)
years

Outcomes
Thyroid hormones:
TT4, TT3, FT4, FT3,
and TSH

Results
No significant results with BPS and thyroid
hormones
Non-significant increase for TT3, FT3, TSH,
TT3/TT4 ratio, FT3/FT4 ratio
Non-significant decrease for TT4, FT4

Covariates, otherbisphenols analyzed
Models adjusted for: gender,
age, BMI, smoking status,
education levels, and urinary
iodine, and co-exposure to otherendocrine disrupting chemicals
with p -value < 0.15 in simple
linear regression models.

Inclusion criteria: adults
having health physicalexamination at Beijing
Hospital, China withoutmajor diseases/disorders of
the thyroid or other severe
diseases

Percentiles:

BKMR models used to estimate
effects of BPS as part of a
mixture of chemicals.

10th: 0.01

Other Bisphenols: BPA and BPF

Zhan et al. 2022

Urine spot samples

United States

Adjusted with creatinine

2013-2016

Values below the LOD
were calculated asLOD÷√2

Cross-sectional study
NHANESn = 857Median ages (IQR): 31.0
(23.0 – 39.0) years
Inclusion criteria: systematic
sample of US population,over 6 years of age, those
who did not havehysterectomy oroophorectomy

Bisphenol S

Mean ± SD: 0.23 ± 0.78

25th: 0.03
50th: 0.07

Comments
BPF associated withdecreases in TT3, TT4, fT3,
and FT4. BPF associated withincreases in TT3/TT4 ratio
and BPA associated withincreases in FT3/fT4 ratio.
Correlations between BPS
and other bisphenols: BPA (r= 0.29) and BPF (r = 0.18).
Only results for females arepresented here (sex-stratified
analyses were displayed
graphically in Figure 2).

75th: 0.15
90th: 0.41

LOD: 0.1% < LOD: 9.6
Percentiles:

History of infertility
either not conceiving
after one year of trying
or seeing a
reproductive specialist
based onquestionnaire

For continuous, per unit increase in ln BPS:
No significant associations of BPS with infertility
OR: 1.08 (95% CI: 0. 92, 1.28) (p = 0.35)
Exposure-response curve for BPS with risk ofinfertility by BKMR analysis showed a slight inverse
U-shape.
No evidence of interactions between various
pollutants

25th: 0.20

Models adjusted for: age,race/ethnicity, education level,
BMI, marital status, serum
cotinine, drinking status, povertyincome ratio, physical activity,
age at menarche, pregnancy
history.

BPA associated with anincreased odd of infertility.
Correlations between BPS
and other bisphenols: BPA (r= 0.11).

BKMR models used to estimate
effects of BPS as part of a
mixture of chemicals.
Other Bisphenols: BPA

50th: 0.60
75th: 1.30

42

OEHHA
October 2023

Reference; study
location, period,
design, sample

Exposure matrix
and concentration(ng/mL)

Zhang et al. 2023

Urine spot samples

Shenyang, China

Adjusted with creatinine

2020-2021

LOQ: 0.1

Cross-sectional study% < LOQ: 40

Shenyang Reproductive andEnvironmental (SYRAE)
study

Values below the LOQ
were calculated as LOQ÷2

n = 111
Median (IQR) age in years
32.0 (4.1)
Inclusion criteria: over 18
years old, seeking assisted
reproductive technology atstudy center, available
ovarian assessment data,
did not have PCOS

Bisphenol S

Geometric mean(creatinine adjusted,
expressed as µg/g): 0.37
Percentiles (creatinine
adjusted, expressed asµg/g):
25th: 0.19
50th: 0.32
75th: 0.55

Outcomes

Results

Covariates, otherbisphenols analyzed

Diminished ovarian
reserve (DOR) andindicators of ovarian
reserve: anti-müllerian
hormone (AMH,
ng/mL), FSH milliinternational units(mIU/mL), and E2(pg/mL)BPS modeled as categorical variable of higher(above the median) or lower (below the median):

DOR models adjusted for: age,
BMI, and household income.

odds of DOR

AMH, FSH, E2 models adjusted
for: age, BMI, smoking, passive
smoking, and infertility
diagnosis.

OR: 6.85 (95% CI: 1.24, 37.82) (p = 0.027)
Non-significant lower AMH, higher FSH, higher E2
For continuous, per unit increase in ln BPS:
AMH for all women:[β = –0.287 (95% CI: –0.505, –0.070)] (p = 0.01)
Non-significant higher FSH and lower E2
AMH excluding women with ovulation disorders:

Other Bisphenols: BPA and BPF

Comments
BPA associated withincreased odds of DOR anddecreased E2 levels. BPA
showed an inverse U-shapedose response with AMH
levels.
Imputation for continuous
values at 40% of BPS
samples.
Correlations between BPS
and other bisphenols: BPA (r= 0.585) and BPF (r = 0.237).

[β = –0.296 (95% CI: –0.518, –0.075)] (p <0.01)
BPS exposure showed subtle inverse U-shapedose response with AMH levels (non-linear term
not significant, p = 0.37)

Timing of samples:
Provided prior to study

43

OEHHA
October 2023

4.2 Studies in animals
OEHHA identified 43 studies that examined the effects of BPS on the female
reproductive system in mammalian and non-mammalian animal models. These included
18 studies in mice, 13 in rats, three in ewes, one in gerbils, one in hamsters, five inzebrafish, one in chickens, and one in C. elegans. A summary of each study ispresented in Table 4.2.1, and the evidence from these studies is discussed below.
4.2.1 Overview
The BPS related reproductive toxicity outcomes reported in the reviewed studies
included effects on weight and histology of the ovary, uterus, placenta, and – oneaccessory reproductive gland – the female prostate. Other reproductive toxicity
outcomes included effects on the endocrine system, puberty onset, estrous cycle,
mammary gland development, reproductive performance, and sex ratio. All resultspresented in this section were reported as statistically significant by pairwise
comparison with controls (p ≤ 0.05) unless otherwise noted (e.g., p ≤ 0.05 by trend test).
Differences in experimental design should be considered when comparing findings
across studies. The key features of each study noted in this section include the species
and strains of animals used, exposure regimen (dose, route, duration), timing of
exposure (e.g., gestational, perinatal, pubertal, adult), and timing of outcome
examination. In brief, most studies were conducted in laboratory mice and rats, of which
multiple strains were used. Most mammals were exposed to BPS orally by gavage, diet,or drinking water, although five studies in rats, three in mice, and one in ewes usedsubcutaneous (sc) injections, and one study in rats and two in mice used intraperitoneal(ip) injections. The duration of exposure in mammalian studies ranged from a few hours(in studies assessing the effects of exposure on hormone levels) to 90 days (in arepeated-dose oral toxicity study). Chickens were exposed by gavage, zebrafish were
exposed in tank water, and C. elegans were exposed directly via the nematode growth
medium. Daily exposure ranged from 0.001 µg/kg to 1,000 mg/kg in mammals andchickens, 0.5 µg/L to 200 µg/mL in zebrafish, and 31.2 to 125 µg/mL [125 to 500 μM] inC. elegans.
4.2.2 Organ weight and histology
Effects on the ovary
Alterations in ovarian weight may reflect changes in the physiology of the organ. For
example, faster or slower follicular development, inhibition of corpus luteum (CL)
Bisphenol S

44

OEHHA
October 2023

formation after ovulation, and ovarian cyst development are a few factors that can affectovarian weight. The following paragraphs focus specifically on the effects of BPS on
oocyte and follicle development.
A change in ovarian weight after exposure to BPS was assessed in several studies andresults were mixed. Decreased ovarian weight on postnatal day (PND) 75 was reported
in rats (strain not specified) subcutaneously injected with 5 and 50 mg/kg-day BPS on
PND 1 to 10 (Ahsan et al. 2018), in adult Sprague-Dawley (SD) rats intraperitoneallyinjected with 50 mg/kg-day BPS for 28 days (Ijaz et al. 2020), and in pubertal ICR mice
exposed to 0.001 to 100 μg/kg-day BPS in drinking water for 28 days (Nevoral et al.
2018). Two studies reported increased ovarian weight, one in Wistar rats dosed bygavage at 1,000 mg/kg-day BPS for 90 days (BASF 2014a) and in adult female Hy-Line
W-36 laying chickens dosed by gavage at 50 µg/kg-day for 90 days (Eldefrawy et al.
2021). Other studies did not detect significant changes in ovarian weight with daily BPS
doses ranging from 0.001 mg/kg-day to 1,000 mg/kg-day in SD and Wistar rats, gerbils,
and ewes (BASF 2020; Demacopulo and Kreimann 2019; Kaimal et al. 2021; Silva et al.
2019; Téteau et al. 2022), Anonymous Studies 12, 14, and 16 as described in (ECHA2019).
There are several studies that assessed the effects of BPS on oocyte and follicle
development at very early stages in fetal life. During gestation, fetal germ cells undergo
incomplete mitosis (lacking cytokinesis) and form syncytia, multinucleated structures
known as germ cell nests or cysts. Germ cell nests ultimately break down andgranulosa cells surround individual oocytes to form primordial follicles. At this point,
oocytes have started miotic division but are arrested in meiosis prophase I (diplotene
stage). Altered follicular or oocyte development can have downstream consequenceson ovulation and the formation of CL.
Prenatal exposure of ICR mice via treatment of pregnant dams with BPS at 2, 10, 50,
100, or 200 μg/kg-day from gestational day (GD) 12.5 to 15.5, targeting the period ofmeiosis, resulted in accelerated meiotic progression in fetal ovaries at GD 15.5 at all
doses, compared to controls. Accelerated meiotic progression was defined by a smaller
percentage of oocytes at earlier stages of meiosis prophase I (zygotene) and a larger
percentage of oocytes at later stages (pachytene and diplotene), and increased rate ofaberrant spindles and misaligned chromosomes at 2 and 10 µg/kg-day (Zhang et al.
2020). Because the effects on meiotic progression were observed at all BPS doses, theresearchers continued studying the effects of prenatal exposure to BPS on ovariandevelopment with only the two lower doses, 2 and 10 μg/kg-day. Both doses
accelerated germ cell nest (cyst) breakdown in F1 mice at PND 3, as evidenced by adecreased proportion of oocytes in nests and a relative increased proportion of oocytes
in primordial follicles, with no effect on the total number of oocytes, compared to
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controls. To test whether this accelerated nest breakdown affected subsequent folliculardevelopment, the researchers examined outcomes at PND 21, where they observed ahigher proportion of oocytes in primordial follicles and a lower proportion in secondary
and antral follicles in the prenatally exposed mice (Zhang et al. 2020). Acceleration ofcyst breakdown was similarly observed in the F2 generation 3 at PND 3 (Zhang et al.
2020).
In a study with a different strain of mice (CD-1) and longer duration of treatment (GD 11
to birth), BPS at 0.5 and 20 μg/kg-day inhibited germ cell nest breakdown, resulting in
more germ cells retained in nests at PND 4. The authors also reported decreases in theratio of primary follicles to total oocytes at doses of 0.5, 20, or 50 μg/kg-day, nonsignificant decreases for the ratio of secondary follicles to total oocytes, and no effects
on the ratio of primordial follicles to total oocytes (Shi et al. 2019).
Two studies investigated the effects of gestational BPS exposure on the number of CL.
One study reported a reduced number of CL in SD rats exposed orally to 5 µg/kg-day
BPS from GD 6 to 21 (Kaimal et al. 2021). The second study reported no changes in thenumber of CL in a different strain of rat (Wistar) exposed orally with up to 300 mg/kgday BPS from GD 6 to GD 19 (BASF 2014b).
In rodents, germ cell breakdown and formation of primordial follicles continues until
shortly after birth. Treatment of newborn CD-1 mice with 10 µg/kg-day by ip injection forthree days decreased the percentage of oocytes in cysts and increased the numbers ofoocytes in primordial follicles (Liu et al. 2021), a finding that is consistent with theaccelerated nest breakdown identified by Zhang et al. (2020). Similar to Zhang et al.
(2020), after following the animals to PND 21 to measure effects on downstream
folliculogenesis, the authors observed decreased numbers of secondary and antral
follicles in treated mice (Liu et al. 2021). The finding that BPS exposure decreased
numbers of antral follicles was observed, again, in a separate study that treated female
rats with 0.5, 5 or 50 mg/kg-day BPS by sc injections for 10 days postnatally (PND 1 to10). This treatment regimen also resulted in consequent decreases in the number of CL
at 5 or 50 mg/kg-day BPS and increases in the number of atretic cystic follicles at 50
mg/kg-day BPS (Ahsan et al. 2018). A final perinatal exposure study examined theinteraction between perinatal exposure to 200 µg/kg-day BPS in CD-1 mice from GD 8
to PND 19 followed by 1 µg/kg-day ethinyl estradiol (EE) from PND 19 to PND 21. BPS
treatment alone resulted in a higher number of secondary follicles with no effect onother follicle types. BPS + EE, however, showed that BPS inhibited the EE-induced

BPS was administered at doses of 2 or 10 μg/kg-day to F0 pregnant dams on GD 12.5 to 15.5 (Zhang etal. 2020).
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increases in total follicles. This finding suggests an endocrine mechanism of action for
BPS (Hill et al. 2017).
Two studies examined the effects of pubertal BPS exposure on ovarian histopathology.
At this developmental stage, the processes of resuming meiosis of oocytes, and follicle
recruitment and growth are tightly regulated by hormones. BPS treatment for 28 days inpubertal ICR mice decreased relative ovary weight at all tested doses (0.001, 0.1, 10,
100 μg/kg-day) and decreased total ovarian volume at 10 and 100 μg/kg-day. Thesechanges in ovary morphology paralleled reductions of CL volume and dose-dependent
decreases in primary, preantral, and antral follicles. The authors also observed
increased mean volume of antral follicles at 0.001 and 0.1 μg/kg-day, decreased atretic
follicle volume at 0.1 and 10 μg/kg-day, decreased medulla volume at all doses, anddecreased numbers of granulosa cell layers at 10 and 100 μg/kg-day (Nevoral et al.
2018). Abnormalities in oogenesis were also evident. The study reported increased
incidence of spindle malformation at 1 and 100 μg/kg-day, increased numbers ofabnormal oocytes at 0.1 and 10 μg/kg-day, and increased methylation of H3K27 (aspecific lysine residue in histone H3) at 10 and 100 μg/kg-day without abnormal
chromosome alignment (Nevoral et al. 2018). The second pubertal exposure study used
Wistar rats treated with 60 mg/kg-day dehydroepiandrosterone (DHEA) to induce
polycystic ovary-like syndrome (PCOS). In this study, co-treatment with sc injections of
BPS at 1 µg/kg-day for 20 days decreased the average number of cysts per ovarycompared to treatment with DHEA alone, providing more evidence of an endocrine
mechanism of action for BPS. BPS treatment alone induced a non-significant increasein follicular cysts (absent in controls) (Demacopulo and Kreimann 2019).
The last set of mammalian studies examined effects of BPS exposure on the ovary inadult mammals. Similar to the studies described above, there was evidence to suggestthat BPS exposure disrupts folliculogenesis and oocyte maturation. For example, adult(8-9 weeks old) CD-1 mice treated with BPS at 300 µg/kg-day for 28 days by oral
gavage had atrophy in immature follicles, an increased number of atretic follicles,
damaged oocyte structure, and more wrinkled and depressed zona pellucida compared
to controls (Yue et al. 2023b). In a separate study, seven days of oral BPS treatment in
adult ICR mice increased the proportion of abnormal mature oocytes at all doses (0.001
to 100 μg/kg-day) and increased abnormal spindles at 0.01, 10, and 100 μg/kg-day.
This study also reported non-significant increases in abnormal chromatin in in mature
metaphase II (MII) oocytes, increased apoptosis at 0.01, 10, and 100 µg/kg-day in, but
no effects on oocyte quantity or quality (Prokešová et al. 2020). A third study reported
that adult SD rats treated with BPS by ip injections at 50 µg/kg-day to 50 mg/kg-day for
28 days had changes in ovarian structure, reporting a reduced number of CL at ≥ 500µg/kg-day and reduced gonadosomatic index (GSI) and increased CL diameter at 50
mg/kg-day [50,000 µg/kg-day]. Also, antral and atretic follicles were decreased in rats
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treated with 5 and 50 mg/kg-day [5,000 and 50,000 µg/kg-day] and antral follicle
diameter was increased at all doses. Granulosa cell height was decreased at 5 mg/kgday but increased at 50 mg/kg-day with no effect on theca cell height (Ijaz et al. 2020).
In one study in adult female golden hamsters treated with BPS at a daily oral dose of
150 mg/kg-day for 28 days, a decrease in the number of growing follicles and CL (lackof ovulation) with an increase in the number of atretic follicles was reported (Pal et al.
2023). Lastly, one non-rodent study treated mature ewes with BPS at 4 or 50 µg/kg-day
and reported no effect on the total number of follicles available for aspiration(Desmarchais et al. 2022).
In wild-type zebrafish, five studies examined changes in ovarian maturation and ovarian
structures, including changes in oocyte morphology. In most studies, zebrafish (ABstrain) were exposed to BPS concentrations between 0.1 μg/L and 200 μg/L (with one
study using BPS concentration up to 200,000 µg/L) for a wide range of durations (7 to240 days). Two studies reported that BPS treatment reduced GSI in adult zebrafish
exposed for 21 days at 0.5, 5, and 50 μg/L (Ji et al. 2013) and in zebrafish embryos
exposed for 75 days at 100 μg/L (Naderi et al. 2014). Delays in oocyte and follicle
maturation were evident in other studies. For example, BPS treatment at 1, 10 and 100µg/L for 14 days increased the number of perinucleolar oocytes, decreased the numberof cortical alveolar oocytes, and decreased the percentage of late vitellogenic andspawning oocytes at all doses (Wang et al. 2020). Treatment at different days post
fertilization (dpf) from days 30 to 240 increased the number of cortical follicles at 1 μg/L,
increased the number of vitellogenic-stage oocytes at 100 μg/L, and increased egg
production at 1 and 100 μg/L. This study also reported fewer oocytes at the primary
growth stage and increased numbers of oocytes at the full-grown stage at 1 and 100
μg/L (Qin et al. 2021). A final study treated zebrafish at substantially higher
concentrations (8, 40, and 200 µg/mL) for 21 days and observed increased GSI andhepatic somatic index (HSI) at 40 µg/mL and oocyte degeneration at 40 and 200 µg/mL
compared to controls, as evidenced by regressed oocytes, perinucleolar oocytes, andyolk vesicle oocytes (Park et al. 2022).
Two final studies used chicken and C. elegans models. Chickens are a usefulexperimental model in which to study folliculogenesis considering the relative shortperiod of follicle development and maturation and daily ovulation in these animals, while
C. elegans, a hermaphroditic species, has been a useful animal model to study meiosis.
In adult Hy-Line W-36 chickens orally exposed to BPS at 50 µg/kg-day for three months
there was an increase in weight of the left ovaries (the active one in chickens), a
decrease in the number of preovulatory follicles, an increase in lymphocyte infiltration(presence of lymphocytes around the growing follicle and ovarian medulla), and anincrease in ovarian inflammation (Eldefrawy et al. 2021). In C. elegans exposed to BPS
at 125, 250 and 500 micromolar (µM) (internal dose = not detected, 0.21, and 0.39 µg/g,
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respectively) there were dose-dependent effects in germline cells such as irregular
distribution of nuclei along the distal to the proximal axis, nuclear loss, and cell death by
apoptosis (Chen et al. 2016b).
Effects on the uterus
Changes in uterine weight following exposure to BPS have been assessed in several
studies. SD rats treated with BPS (from 50 µg/kg-day to 50 mg/kg-day) by ip injection
for 28 days had reduced relative uterine weights at 5 and 50 mg/kg-day, compared tocontrols (Ijaz et al. 2020). The study by Ahsan et al. (2018) also reported decreases inrelative weight of paired uteri in rats treated from PND 1 to 10 with 50 mg/kg-day by sc
injection and assessed at PND 75. In two other rat studies, BPS treatment resulted inincreased uterine weight. One study treated SD rats with 30, 100, or 300 mg/kg-day
BPS by oral gavage for about 14 weeks from 6 weeks premating to weaning andobserved effects at 300 mg/kg-day Anonymous Study 14 as described in (ECHA 2019),
while the other treated SD rats with sc injections for three consecutive days andreported increased uterine weight at 20 and 500 mg/kg-day (Yamasaki et al. 2004).
Other studies reported no effects of BPS treatment on uterine weight. These included a
study that treated Wistar rats from GD 6 to GD 19 and evaluated dam uterine weight on
GD 20 (BASF 2014b), a study that treated SD rats with BPS for a total of 40-46 days
from premating to postpartum day 3 Anonymous Study 12 as described in (ECHA2019), and a study that treated SD rats with BPS by daily oral gavage at doses of 0,100, 300 or 600 mg/kg-day for 28 days (BASF 2020). A final study reported no changes
in uterine weight among adult ICR mice treated with BPS by oral gavage at 300 µg/kgday for 28 days (Yue et al. 2023b).
Other reported effects of BPS exposure on the uterus include histological and structural
alterations. The study by Yue et al. (2023) in adult ICR mice reported that BPS
treatment resulted in multiple changes in the shape and structure of the uterus in theabsence of an effect on uterine weight. These histopathological changes included anarrowed uterine cavity, increased number of uterine glands, reduced endometrial area,
and atrophied lamina propria, myometrium, and plasma layer (Yue et al. 2023b). Adult
female C57BL/6J mice treated with BPS by gavage at a dose of 30 mg/kg-day for 21
days developed “non-atypical” endometrial hyperplasia (increased number of normal
endometrial cells), increased number of glands in the endometrium, and increased
thickness of the myometrium, compared to controls. Migration of endometrial cells into
the myometrium (adenomyosis) was also observed in 2 out of 5 treated mice, comparedto 0 out of 5 controls (Benjamin et al. 2023).
In immature Wistar rats treated with 60 mg/kg-day DHEA for 20 days to induce PCOS,
cotreatment with BPS at 1 µg/kg-day resulted in the following alterations in the uterus:
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an increased intraepithelial lumen, increased cell proliferation in the endometrial
perimeter, and increased apicobasal polarity of the epithelium when compared to the
DHEA alone group. BPS treatment alone (no DHEA) increased cell vacuolization in theendometrium compared to controls (Demacopulo and Kreimann 2019). These findings
are similar to those presented by (Yamasaki et al. 2004) in rats and (Hill et al. 2017) inmice which showed that BPS treatment altered the effects of EE treatment. For
example, BPS at 20 mg/kg-day further increased the uterine weight of an EE treatedgroup of SD rats and at 500 mg/kg-day diminished the EE effect on increased uterine
weight (Yamasaki et al. 2004). In female CD-1 mice exposed to BPS from GD 8 until
PND 19 (via oral administration to dams during gestation at 200 µg/kg-day and direct
oral administration after birth at 200 µg/kg-day) there were no observed effects on
uterine tissue organization on PND 22, however, increased uterine epithelial height wasapparent at PND 22 in mice that received additional treatment with EE at 1 µg/kg-day
from PND 19 to PND 21 (Hill et al. 2017). Together, these findings suggest that BPS
alters uterine responsiveness to hormonal changes normally experienced duringpuberty.
The last histopathological uterine outcome reported was increased squamous
metaplasia of the uterus at a BPS dose of 1,000 mg/kg-day in 42-day-old female Wistar
rats treated for 90 days (BASF 2014a).
Effects on the female prostate
One study presented data on the effects of BPS on an accessory gland, the female
prostate. In this study adult female gerbils were orally treated with BPS at 40 µg/kg-day
for 28 days, which affected the three-dimensional structure of female prostatic tissue.
Compared to controls, BPS treated gerbils had increased relative frequency ofepithelium and muscular stroma and decreased ductal lumen. Prostatic histology
showed decreased percentage of normal tissue, increased hyperplasia, and increased
intraepithelial neoplasia without affecting prostate weight. There were no alterations inbody or gonadal weight (Silva et al. 2019).
Effects on placenta
Two studies, in different species, examined effects of BPS on placenta histopathology.
One study treated mice with BPS through oral ingestion of wafers at 200 μg/kg-day for 2
weeks prior to mating and through GD 12.5 and observed in placental cross-sections(through three placental layers: labyrinth, spongiotrophoblast, and giant cell) a
decreased ratio of spongiotrophoblast to giant cell area, which the authors propose may
be due to changes in gene expression (Mao et al. 2020). The second study treatedpregnant sheep with a sc injection of BPS at 0.5 mg/kg-day from GD 30 to 100. BPS
treatment resulted in a non-significant increase in placental weight at GD 120, and no
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effects on placental stereology, or number of placentomes per placenta (Gingrich et al.
2018).
4.2.3 Endocrine effects
Effects of BPS on the endocrine system involved variations in hormone levels, including
synthesis and/or metabolism, and alterations in hormone receptor activity and/or
expression. BPS effects on hormone levels were observed in various body fluids andtissues and included alterations in gonadotropins: luteinizing hormone (LH) and folliclestimulating hormone (FSH); steroid hormones: progesterone (P), testosterone (T),dihydrotestosterone (DHT), estradiol (E2) and estrone (E1); thyroid hormones; andprolactin.
In the female reproductive cycle, there are two well characterized ovarian phases:
follicular and luteal. During the follicular phase, the ovarian follicles are maturing, andthe surrounding granulosa cells are producing increasing amount of E2. In the middle ofthe cycle, levels of gonadotropins secreted by the pituitary gland surge in response topositive feedback from E2 and trigger ovulation of the fully mature antral follicle. The
remaining follicular cells (granulosa and theca) now form the CL that increases the
production of P in the luteal phase. Disruption of this hormone balance may affect, or
may be a consequence of, alterations in follicle development (e.g., FSH), ovulation(e.g., LH, E2), implantation (e.g., P) or other processes involved in female reproduction.
A small number of studies have reported effects of BPS on some steroidogenic enzyme
mRNA and protein levels, and other studies have reported effects on gene and/or
protein expression of the progesterone receptor (PR), both estrogen receptor alpha andbeta (ERα, ERβ) and related growth factors, the androgen receptor (AR), and theprolactin receptor (PrlR) and related factors.
Gonadotropins
Two studies in rats and one in mice reported the effects of BPS on gonadotropin levels.
In rats (unspecified strain), a decrease in plasma levels of LH and FSH was reported infemales treated with 50 mg/kg-day BPS by sc injection from PND 1 to 10 and assessedon the first day of estrus starting on PND 75 (Ahsan et al. 2018) and in adult female SD
rats treated with BPS at doses between 5 and 50 mg/kg-day by ip injection for 28 days
(Ijaz et al. 2020). In adult female NMRI mice, a dose-dependent decrease in serum LH
and FSH concentrations was observed following sc injection with BPS for 21
consecutive days at doses ranging from 1 to 100 µg/kg-day (Nourian et al. 2020).

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Steroid hormones
Progesterone
Five studies reported that exposure to BPS decreased levels of P in serum, plasma, or
follicular fluid in rats, mice, and ewes. Two studies reported increased levels of P inmice and zebrafish while one study in ewes reported no changes in P levels in plasma
after BPS treatment.
Decreases in plasma P levels at 50 mg/kg-day were found in neonatal female rats
(unknown strain) exposed to BPS (0.5, 5, or 50 mg/kg-day) by sc injection in the first 10
days of life (Ahsan et al. 2018). One study in CD-1 mice with BPS exposure during
gestation (GD 10.5 to GD 17.5) reported decreased serum P levels at 3 months of age
in the 5 mg/kg twice a day dose group, with no effects at PND 28 or at 14 months of age(Tucker et al. 2018). Decreases in plasma P levels were also observed in adult female
SD rats treated with BPS doses of 5 and 50 mg/kg-day via ip injection for 28 days (Ijazet al. 2020). In one study in adult female ewes, an interaction between BPS and diet
was observed on P levels. Two groups of animals – restricted feed and well fed – were
exposed to BPS to assess the effects on steroid hormones. Most of the BPS effects
were observed in the restricted feed group. In this study, BPS exposure at 50 µg/kg-day
for at least three months decreased P and 5α-dihydroprogesterone (P metabolite) levels
in serum, and 17α-hydroxyprogesterone levels in preovulatory follicular fluid in the
restricted feed group (Téteau et al. 2022). In pregnant ewes exposed to daily sc
injections of BPS at 0.5 mg/kg-day from GD 30 to 100, a non-significant decrease inserum P levels was observed between GD 75 -105 (Gingrich et al. 2018).
Increases in P levels following BPS exposure have also been reported in some studies.
In the mouse gestational exposure study discussed above by Tucker et al. (2018), an
increase in serum P levels was observed on PND 20 in the 0.05 mg/kg twice a day dosegroup; this dose is a notably smaller than the dose that resulted in decreased P at threemonths of age in the same study (5 mg/kg twice a day). In a zebrafish study, there wereincreased whole-body P levels observed after BPS exposure to 40 and 200 μg/mL for
21 days (Park et al. 2022). In the last study examining P, treating adult ewes orally with
BPS at 4 or 50 µg/kg-day had no effect on plasma or follicular fluid P levels(Desmarchais et al. 2022).
Androgens: T, DHEA, DHT
BPS effects on androgen levels were investigated in three studies in mice, two studies
in rats and one study in ewes. Increased serum T levels were reported after BPS
exposure during pregnancy or early in life in mice and rats, whereas decreased T levels
were observed following exposure later in life (PND 35 or at eight months of age).
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Increased serum T levels were observed in mice on PND 28 after gestational exposure
to BPS in the 0.05 mg/kg twice a day dose group (Tucker et al. 2018), at 9 months ofage after BPS treatment with 50 μg/kg-day [0.05 mg/kg-d] during gestation (Shi et al.
2019), and in newborn mice treated with BPS at 10 mg/kg (every three days) from PND
0 to PND 60 (Shi et al. 2017). Similarly, an increase in plasma T levels was observed inneonatal female rats exposed to BPS at 50 mg/kg-day by sc injection in the first 10 daysof life and evaluated on the first day of estrus starting on PND 75 (Ahsan et al. 2018).
Increased plasma T levels were also reported in adult female SD rats treated with BPS
doses of 5 and 50 mg/kg-day by ip injection for 28 days (Ijaz et al. 2020). In the BPS
gestational exposure study by Tucker et al. (2018) in mice, there were increased serum
DHEA (T precursor) levels on PND 20, increased serum T levels on PND 28, in the 0.05
mg/kg twice a day dose group, and no effects on serum DHEA on PND 28 or later.
However, there were decreased serum T levels on PND 35 (no effects on PND 56) in
the 0.5 and 5 mg/kg twice a day dose groups, and decreased serum T and DHEA levels
at eight months of age in the 0.5 mg/kg twice a day dose group, with no effects at 14
months of age (Tucker et al. 2018). Finally in the study by Téteau et al. (2022) in adultewes, exposure to 50 µg/kg-day BPS for at least three months resulted in decreased
levels of 11-dehydrocorticosterone and DHT and increased levels of 5βdihydrotestosterone (T metabolite) in follicular fluid in the restricted feed group.
Estrogens: E2 and E1
Three studies in mice, one in rats and one study in ewes reported decreased E2 levels
in different media. One study reported a decrease in serum E2 on lactation day (LD) 21
after BPS treatment at 2 or 200 µg BPS/kg-day during gestation and lactation in CD-1
mice (LaPlante et al. 2017). Mice treated during puberty for 4 weeks had decreased
plasma E2 levels and, as reported in Section 4.2.2, decreased number of granulosa cell
layers – the cells producing E2 – at BPS doses of 10 and 100 µg/kg-day and increased
volume of antral follicles at 0.001 and 0.1 µg/kg-day (Nevoral et al. 2018). After a single
sc injection to adult CF-1 mice, a decrease in urinary E2 levels was observed 10 hours
after BPS treatment at doses of 25.2 or 79.8 mg/kg, and a dose-dependent decrease inserum 3H-E2 levels was observed 90 minutes after treatment at doses of 78.5 or 252.7
mg/kg (Pollock et al. 2019). In adult female SD rats treated with BPS by ip injection for
28 days there was a decrease in plasma E2 levels at doses of 5 and 50 mg/kg-day and,as reported in Section 4.2.2, a decrease in the number of antral and atretic follicles was
also observed at these doses (Ijaz et al. 2020). In the study by Téteau et al. (2022), in
adult ewes treated with BPS at 50 µg/kg-day the plasma had decreased E2 andincreased E1 levels in the restricted feed group, the oviduct fluid had decreased E2 andE1 in the well fed and restricted feed group, respectively, and the follicular fluid had
increased E2 levels in the well fed group.
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Increased E2 levels after treatment with BPS were reported in two studies in mice andthree studies in zebrafish. Mice treated with BPS by sc injection (every three days) with50 μg/kg or 10 mg/kg starting at birth for 60 days had increased serum E2 levels at PND
60 at both doses (Shi et al. 2017). In the gestational exposure study in mice by Tucker
et al. 2018, serum E2 levels were increased on PND 20 in the 0.5 and 5 mg/kg twice a
day dose groups. In zebrafish exposed to BPS at 10 or 100 μg/L for 75 days, increases
in plasma E2 levels and vitellogenin (VTG, marker for estrogenic activity) production
were observed (Naderi et al. 2014). In another study, an increase in plasma E2 levels
and the E2/T ratio was reported in female zebrafish exposed to BPS at 50 μg/L, with noeffect on plasma T levels (Ji et al. 2013). The final zebrafish study reported that after 21days of BPS exposure there was an increase in whole-body E2 levels at allconcentrations tested (8, 40, and 200 µg/mL) (Park et al. 2022).
No effects of BPS on serum or plasma E2 levels were reported in several studies.
These included studies in mice that were exposed from GD 11 to birth at doses of up to
50 μg/kg-day and evaluated at three, six, or nine months of age (Shi et al. 2019), inmice that were exposed from GD 10.5 to GD 17.5 at doses of up to 5 mg/kg twice aday) and evaluated on PND 28, 35, or 56 (Tucker et al. 2018), and in adult mice orally
treated with BPS at 300 µg/kg-day for 28 consecutive days (Yue et al. 2023b). Other
studies observed no effects on plasma E2 levels in rats (unknown strain) treated withdaily BPS sc injections of up to 50 mg/kg-day from PND 1 to PND 10 (Ahsan et al.
2018) and no effects on plasma or follicular fluid E2 levels in ewes treated orally with
BPS at 4 or 50 µg/kg-day (Desmarchais et al. 2022). Finally, there was no effect onplacental E2, or E1 levels in mice treated orally with BPS at 200 μg/kg-day for two
weeks before mating (Mao et al. 2020).
Other hormones
Several studies assessed the effect of BPS exposure on levels of other hormones. In
adult female CD-1 (ICR) mice, no effects on serum levels of free triiodothyronine or freethyroxine were observed after treatment with 300 µg/kg-day BPS for 28 days (Yue et al.
2023b).
In zebrafish, one study reported decreased plasma T3 and T4 levels after exposure to
100 μg/L BPS for 75 days (Naderi et al. 2014), while another study found aconcentration-dependent increase in whole body T3 and T4 levels after BPS treatmentof adult zebrafish for 21 days at concentrations of 8, 40, and 200 µg/mL (Park et al.
2022).
In mice, BPS treatment at 200 μg/kg-day for 2 weeks prior to mating and through GD
12.5 resulted in an increase in placental dopamine concentrations and in thepercentage of dopamine-positive trophoblast giant cells, a decrease in placental
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serotonin (5-HT) concentrations and in the proportion of serotonin-positive giant cells,
and a substantial increase in the ratio of 5-hydroxyindoleacetic acid (primary 5-HT
metabolite) to 5-HT (Mao et al. 2020).
In ewes orally treated with BPS at 4 or 50 µg/kg-day, there were no reported effects on
plasma anti-müllerian hormone (AMH) levels (Desmarchais et al. 2022).
Steroidogenic enzymes
Steroidogenesis, i.e., the synthesis of steroid hormones from cholesterol, occurs
through metabolic processes involving cytochromes P450 and hydroxysteroid
dehydrogenases (HSD). Steroidogenesis can be modulated by many factors, includingchanges in gene expression, protein production and enzyme activity. Three studies
were identified that assessed the effects of BPS on steroidogenic enzyme gene and/or
protein expression.
In mice exposed to BPS during gestation from GD 11 to birth there was an increase in
ovarian mRNA for Cyp11a1 (P450scc) – the enzyme that catalyzes the initial step insteroidogenesis – at 3 months of age in the 0.5 μg/kg-day dose group (Shi et al. 2019),
while no effect on steroidogenic enzyme mRNA expression was observed in mice
exposed from birth to PND 60 (once every three days) at doses of up to 10 mg/kg (Shiet al. 2017).
In Wistar rats exposed to BPS during gestation and postnatally from birth through PND
21, there was a decrease in brain mRNA levels of 5α-reductase-3 – one of three
isozymes involved in the conversion of T into the non-aromatizable androgen, DHT–
with no changes observed in brain 5α-reductase protein levels for this isozyme (Castro
et al. 2015).
Hormone receptors
Effects of BPS exposure on hormone receptors were analyzed in several studies.
Results included effects on gene and/or protein expression of the P receptor (PR),estrogen receptors (ERs) and related growth factors, the androgen receptor (AR), and
the PrlR and related factors.
Progesterone receptor
Increases in PR protein levels were observed in mammary glands from pubertal (PND
32-35) female CD-1 mice exposed to BPS during gestation and lactation via maternal
oral administration (2 or 200 µg BPS/kg-day) from GD 9 to LD 20, but not from newly
weaned (PND 24) or adult (9-week-old) mice (Kolla et al. 2018). Another study reported
a decrease in mammary gland PR mRNA levels at 8 months of age in mice exposed to
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5 mg BPS/kg twice a day during gestation (via maternal oral administration from GD
10.5 – GD 17.5), with no changes observed at the two lower doses (Tucker et al. 2018).
No effects on mammary gland PR mRNA levels were found on PND 31 in CD-1 mice
exposed to BPS during gestation and lactation (GD 8 to PND 2) via maternal oral
administration (2, 200, or 2,000 µg/kg-day) (Kolla and Vandenberg 2019).
Estrogen receptor
In the gestational and lactational study in female CD-1 mice by Kolla et al. (2018), there
was a NS increase in estrogen receptor alpha (ERα) protein in the mammary gland at
puberty (PND 32-35), no effect at PND 24, and a decrease (200 µg/kg-day dose grouponly) at nine weeks of age. The gestational and lactational study in female CD-1 mice
by Kolla and Vandenberg (2019) reported an increase in Esr1 (Erα gene) mRNA levelsin the mammary gland on PND 31 in the 200 µg/kg-day dose group, with no effect atother doses. This study also reported that the effect of BPS on mammary gland Esr1
mRNA levels was not modified by a peripubertal estrogen (ethinyl estradiol, EE)
challenge (Kolla and Vandenberg 2019).
In the mammary glands of adult female CD-1 mice orally treated with BPS at 2 or 200
μg/kg-day during pregnancy and lactation (GD 8 to LD 21) and assessed on LD 21, an
increase in the percentage of Erα protein positive cells was observed at both doses.
Esr1 gene expression in the mammary gland was not significantly different from controlson LD 21 at 2 μg/kg-day; however, mRNA levels were lower in the 200 μg/kg-day dosegroup compared to the 2 μg BPS/kg-day dose group (LaPlante et al. 2017). In addition,
BPS exposure had no effect on the number of ERα positive cells in the arcuate nucleusof the hypothalamus (LaPlante et al. 2017).
A study in female CD-1 mice exposed to BPS during gestation and lactation via
maternal oral administration (200 µg BPS/kg-day from GD 8 to PND 19) reported nonsignificant increases in Esr1 (Erα) and Esr2 (Erβ) mRNA in the ovary and a nonsignificant decrease in Erα mRNA in the uterus on PND 22 (Hill et al. 2017).
Ovarian and uterine gene expression of epidermal growth factor-1 receptor (EGF-R)
and insulin growth factor-1 (IGF-1) can be regulated by estrogens and were also
assessed on PND 22. Ovarian EGF-R and IGF-1 mRNA levels and uterine IGF-1
mRNA levels were increased in the BPS exposed mice, compared to controls (Hill et al.
2017). The authors also investigated the effect of a peripubertal EE challenge on
ovarian and uterine gene expression in BPS exposed mice: ovarian Esr2 (ERβ) and
EGF-R mRNA levels were decreased and Esr1 (ERα) mRNA was non-significantly
decreased in EE-challenged mice exposed to BPS, compared to EE-challenged
controls. Uterine IGF-1 mRNA levels were increased in EE-challenged mice exposed to
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BPS compared to EE-challenged controls, while no differences were observed inuterine Esr1 (ERα) and EGF-R mRNA levels (Hill et al. 2017).
In adult female gerbils orally treated with BPS at 40 µg/kg-day per day for 28 days, anincrease in the percentage of ERα positive cells was observed in the stroma of thefemale prostate, compared to controls (Silva et al. 2019).
In adult female zebrafish exposed to BPS for 21 days (8, 40, 200 µg/mL) a nonsignificant increase in hepatic ERα mRNA levels was reported at 8 µg/mL, with nonsignificant decreases reported by the authors at the two higher concentrations. [Notethat the decrease at 200 µg/mL appears to be significant, based on visual inspection of
Figure 2B in the publication.] BPS exposure had no effect on hepatic ERβ mRNA levels,
while mRNA levels of the estrogen-responsive gene VTG were decreased at 200µg/mL, with non-significant decreases in VTG mRNA observed at the two lower
concentrations (Park et al. 2022).
Androgen receptor
In the adult female gerbil study by Silva et al. (2019) discussed above, an increase in
the percentage of AR positive cells was observed in the epithelium and stroma of thefemale prostate.
Prolactin receptor and related factors
In the study by LaPlante et al. (2017) of adult female CD-1 mice treated with BPS during
pregnancy and lactation and assessed on LD 21, mammary gland PrlR mRNA levels
were increased at 2 μg/kg-day and non-significantly decreased at 200 μg/kg-day, and anon-significant decrease was observed in the number of mammary epithelial cells
staining positive for signal transducer and activator of transcription 5 (stat5) protein, amarker of prolactin signaling and cell differentiation. BPS exposure had no effect on
PrlR mRNA levels in the pituitary, or on the number of stat5 positive cells in the arcuate
nucleus of the hypothalamus (LaPlante et al. 2017).
4.2.4 Puberty onset
Vaginal opening happens in concert with hormonal fluctuations in the hypothalamicpituitary-gonadal (HPG) axis and occurs as early as PND 35 (five weeks) in mice and
PND 32 in rats, and as late as PND 42 to PND 56 (six to eight weeks) in both species.
This wide variation in pubertal timing underscores the need for concurrent controls inexperimental designs. Accelerations or delays in pubertal timing, or deviations in HPG
hormonal fluctuations when comparing treated versus untreated rodents may indicate

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reproductive toxicity. Seven studies in rodents examined effects of BPS on puberty
onset measured by vaginal opening.
Two studies in rats and one in mice observed delays in puberty onset after early life
BPS exposure. One study treated newborn female rats with BPS by sc injections (0.5,5, or 50 mg/kg-day) on the first 10 postnatal days and observed delays in vaginal
opening in the 50 mg/kg-day dose group. These delays were accompanied by changes
in plasma hormone levels, including decreases in gonadotropins and progesterone(Ahsan et al. 2018). Delays in vaginal opening (PND 33.2), relative to the control group,
were also observed in female SD rats exposed during gestation and lactation via
maternal gavage administration of 60 mg/kg-day BPS (from 10 weeks premating
through mating, gestation, and lactation to PND 21), but not in rats exposed to lower orhigher doses (i.e., 20 and 180 mg/kg-day) (BASF 2019a). Puberty onset in this study
ranged from PND 28 to PND 40 (across all control and exposed animals) (BASF
2019a). A study in CD-1 mice reported a 2.5-day delay in vaginal opening following
exposure during gestation (GD 11 to birth) via maternal oral administration of 20 μg/kgday BPS, while puberty onset was 1-2 days earlier in the 0.5 μg/kg-day dose group, andno effects observed in the 50 μg/kg-day dose group (Shi et al. 2019).
Four studies reported no significant effects of BPS on puberty onset. These included agestational exposure study in CD-1 mice (maternal gavage administration of 0.05, 0.5,
or 5 mg/kg twice a day from GD 10.5 to GD 17.5) (Tucker et al. 2018), a gestational andearly postnatal exposure study in CD-1 mice (maternal oral administration of 2, 200, or
2,000 μg/kg-day from GD 8 to PND 2) (Kolla and Vandenberg 2019), a study thattreated CD-1 mice, starting as newborns, with sc injections of 50 μg/kg-day or 10
mg/kg-day BPS once every three days from PND 0 to 60 (Shi et al. 2017), and alactational exposure study in ICR mice (maternal administration of 0.375 or 37.5 ng/mL
BPS in drinking water from PND 0 to PND 15) (Nevoral et al. 2021).
4.2.5 Effects on estrous cycle
The rodent estrous cycle is a well-defined four-to-five-day cycle that includes fourphases (proestrus, estrus, metestrus, and diestrus). The preovulatory gonadotropin
surge occurs on the proestrus day followed by ovulation which occurs during estrus, theday that the female is receptive to mating. Altering the length of the estrous cycle,
especially more days in diestrus, may impair overall female reproductive success. One
study in mice and three in rats observed changes in estrous cyclicity after BPS
exposure.
In mice, gestational BPS exposure (maternal oral administration of 0.5, 20, or 50 μg/kgday from GD 11 to birth) resulted in irregular estrous cyclicity (several days in estrus
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and diestrus) at all doses tested (Shi et al. 2019). In adult SD rats, there were increases
in estrous cycle length observed after gestational and lactational exposure to BPS(treatment of dams with 180 mg/kg-day BPS by gavage for about 90 days frompremating to PND 21) (BASF 2019a), increases in the number of days in diestrus after
gestational and early postnatal lactational exposure to BPS (treatment of dams by
gavage with 300 mg/kg-day for 40-46 days from premating to PND 3) Anonymous Study
12 as described in (ECHA 2019), and increases in estrous cycle length after gestational
and lactational exposure to BPS (treatment of dams by gavage with 300 mg/kg-day forabout 14 weeks from 6 weeks premating to weaning) Anonymous Study 14 as
described in (ECHA 2019). No effect on the day of first estrus or on estrous cycle length
was observed in mice (Tucker et al. 2018).
4.2.6 Mammary gland development
Gestation is a sensitive period for exposure because mammary gland development
begins during this time. Development continues in puberty when production of ovarian
E2 and P stimulate the growth of terminal end buds (TEB). In adulthood, TEBs regress
and alveolar buds are formed, and in pregnancy, prolactin secreted from the pituitary
gland stimulates development of lobuloalveolar units in preparation for lactation. In thereviewed studies, effects of BPS exposure included alterations in the development andretention of TEBs, development of alveolar buds, and mammary gland proliferation. In
general, the reported effects of early life BPS exposure varied by timing of assessmentof the mammary gland (e.g., pre-pubertal, pubertal, adult). Four studies investigatedchanges in mammary gland development in mice exposed to BPS during gestation(Tucker et al. 2018) or during gestation and lactation (Kolla et al. 2018; Kolla andVandenberg 2019; LaPlante et al. 2017).
There were data to suggest that gestational and lactational exposure to BPS results inaccelerated development of TEBs in pre-pubertal mice, while no effect of exposure wasapparent in pubertal mice. One study observed dose-dependent increases in TEB
counts on PND 20 in (pre-pubertal) CD-1 mice exposed during gestation (maternal
administration of 0.05, 0.5, or 5 mg/kg twice a day from GD 10.5 to GD 17.5) (Tucker et
al. 2018). A second study observed the following changes on PND 24 in (pre-pubertal)
CD-1 mice exposed during gestation and lactation (maternal oral administration of 2 or
200 µg/kg-day from GD 9 to LD 20): larger TEB area in the low dose group; increased
average TEB size in both dose groups; and non-significant increases in TEB count inboth dose groups (Kolla et al. 2018). In this same study, no effects of early life BPS
exposure were apparent on TEBs when mice were assessed during puberty (PND 3235) (Kolla et al. 2018). A third study also reported no effects of early life BPS exposure(maternal oral administration of 2, 200, or 2,000 μg/kg-day from GD 8 to PND 2) on
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number or total area of TEBs in mice assessed on PND 31 (Kolla and Vandenberg2019).
Two of these studies examined TEB counts in adult mice, both showing increased
retention of TEBs in adult mammary glands of mice with early life BPS exposure.
Increased TEBs were observed at 3 months of age in mice exposed during gestation in
the 0.5 mg/kg twice a day dose group (Tucker et al. 2018), and increased TEB-likestructures were observed at 9 weeks of age in mice exposed during gestation and via
lactation in both the 2 and 200 µg/kg-day dose groups (Kolla et al. 2018). The latterstudy also reported increased numbers of mammary gland alveolar buds in the highdose group and increased incidence of intraductal hyperplasia in both dose groups at 9
weeks (Kolla et al. 2018) while the former study observed increased incidence of mixed
cell inflammation at 3 months of age (Tucker et al. 2018). At 14 months, there wasincreased incidence of all types of mammary gland inflammation, non-neoplastic
lesions, and lobuloalveolar hyperplasia in the 0.5 mg/kg twice a day dose group (Tucker
et al. 2018).
Additional effects on mammary gland development were reported in each of thesestudies. In the Kolla et al. (2018) study, Ki67 expression indicated a decrease in
mammary gland cell proliferation at 2 µg/kg-day BPS on PND 24, a non-significant
increase in proliferation at puberty in both dose groups and a significant increase inproliferation in adults at 2 µg/kg-day (Kolla et al. 2018). In the Kolla and Vandenberg(2019) study, a decrease in ductal area was observed in the 2 μg/kg-day dose group on
PND 31 (Kolla and Vandenberg 2019). And in the Tucker et al. (2018) study, increases
in the mammary gland developmental score were reported on PND 20 in the 5 mg/kg
twice a day dose group, on PND 35 in all dose groups, and on PND 56 in the 0.5 mg/kg
twice a day dose group, without any effects observed on mammary epithelial area
(Tucker et al. 2018).
A fourth study, in CD-1 mice, evaluated not only the anatomical effects on the mammary
gland of dams treated with BPS during pregnancy and lactation (2 or 200 μg/kg-day
from GD 9 to LD 20), but also effects on nursing behavior. The authors concluded from
histological examination of whole-mount mammary glands collected on either LD 2 or
LD 21 that there was the suggestion of an overall treatment-related trend of increased
functional mammary gland tissue at LD 2 and decreased functional mammary glandtissue on LD 21. A significantly reduced volume of lobules, a non-significant decrease inlobule size, and a significantly increased volume of adipose tissue were also reported in
the high dose group on LD 21. Regarding nursing behaviors, on LD 14, dams treatedwith 2 μg/kg-day BPS spent increased time in the “high-crouch” lactating position
compared to controls, while dams treated with 200 μg/kg-day BPS spent increased time

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nursing and a decreased percentage of pups initiated nursing (control: 91%; 200 μg/kgday: 50%) (LaPlante et al. 2017).
4.2.7 Reproductive performance
Reproductive performance or efficiency is closely related to fertility. Among anatomicaland physiological features in female reproduction, the ovary plays an important role
mostly because of the fixed number of oocytes at birth. After gamete production the
reproductive process continues through mating, conception, implantation, and deliveryof live offspring, where all these processes are closely controlled by the nervous andendocrine systems.
Reproductive performance was assessed in female CD-1 mice gestationally exposed to
BPS (maternal administration of BPS at 0.5, 20, or 50 μg/kg-day from GD 11 to birth) at
3, 6 and 9 months of age. No effects of BPS exposure were observed at 3 months,
while at 6 months “time to vaginal plug” (i.e., successful mating) was increased at all
doses, with further increases in “time to vaginal plug” observed for all doses at 9 months(Shi et al. 2019). A decrease in the percentage of successful mating was reported in the
low and mid dose groups at 6 months and in all dose groups at 9 months, with a greater
decrease at 9 (60%) compared to 6 months (25%). In addition, a decrease in thenumber of pups born per litter was observed at 9 months in the mid dose group (Shi etal. 2019).
Reproductive performance was assessed in female rats exposed as newborns to BPS
by sc injections (0.5, 5, 50 mg/kg-day) on the first 10 post-natal days. A decrease in thepercentage of females that conceived and a decrease in the number of pups born perlitter was observed in the high dose group (Ahsan et al. 2018). A non-significant
decreased fertility index (58.3 % vs 91.7 %) was reported in adult SD rats treated with
BPS at 300 mg/kg-day for a total of 40-46 days from premating to PND 3 Anonymous
Study 12 as described in (ECHA 2019).
Three studies in mice reported no effects of BPS on reproductive performance. The firststudy treated adult female mice orally with BPS daily at 200 μg/kg-day (from before
mating to GD 12.5) (Mao et al. 2020), and the second treated female mice with sc
injections of BPS early in life from birth to PND 60 (every three days at 50 μg/kg-day or
10 mg/kg-day) (Shi et al. 2017). These studies assessed time to conception (Shi et al.
2017), pregnancy success (Mao et al. 2020), gestational length (Shi et al. 2017),
maternal gestational weight (Mao et al. 2020; Shi et al. 2017), uterine and pup weights(Shi et al. 2017), implantation sites (Mao et al. 2020), and litter size (Mao et al. 2020;
Shi et al. 2017). No effect on litter size or body weight was observed in the F2

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generation of ICR mice in a study where BPS was administered orally to F0 dams on
GD 12.5-15.5 at doses of 2, 10, 50, 100, or 200 μg/kg-day (Zhang et al. 2020).
One study in SD rats reported no effects of BPS on reproductive performanceparameters. In rats treated orally during pregnancy on GD 6 to GD 21 with BPS at 5μg/kg-day, no statistically significant effects were reported on dam body weight gain,
proportion of dams with normal pregnancies or abortions, litter size, or offspring bodyweights and morphometric measurements. However, there were two stillborn pups (oneeach in two out of 13 litters) in the BPS group versus none in the control (Kaimal et al.
2021).
Female zebrafish exposed to BPS at 10 and 100 μg/L, starting as embryos, for 75 days
produced fewer numbers of eggs during a 7-day spawning period and the eggs had alower hatching rate with an increased time to hatching (Naderi et al. 2014).
BPS altered shoaling behavior, which is important during mating, of adult female
zebrafish exposed to BPS at 100 µg/L for 14 days, such that females spent less time inthe zone close to other fish and increased time far from other fish (Wang et al. 2020).
In C. elegans exposed to BPS at 125, 250 and 500 µM there was a concentrationdependent increase in embryonic lethality and a decrease in brood size at 125 and 500µM (Chen et al. 2016b).
Effects on fertilization
A decrease in fertilization rate was reported in ICR mice treated with BPS in drinking
water during puberty for four weeks at a dose of 10 µg/kg-day, with non-significant
decreases observed at lower doses (0.001 and 0.1 μg/kg-day), consistent with a doserelated effect. However, a significant increase in fertilization rate was reported at 100
μg/kg-day (Nevoral et al. 2018). F1 oocytes from female mice that were treated with
BPS at 2 or 10 μg/kg-day for 4 days from GD 12.5 to 15.5 were used for in vitro
fertilization with sperm from untreated animals. Reproductive outcomes observed in thisstudy included a dose-dependent decrease in the proportion of two- and four- cells
embryos, and lower proportion of blastocysts (Zhang et al. 2020).
BPS also decreased the in vitro fertilization rate in mouse (strain not specified) oocytes
harvested from females treated by ip injection for 21 days during adulthood with dosesof 10, 50 and 100 µg/kg-day BPS (Nourian et al. 2017). Dose-dependent decreases in
the percentages of two-cell embryos and blastocysts were observed. In addition, type I
embryo arrests were increased at 50 and 100 µg/kg-day (Nourian et al. 2017). In adultewes on either a restricted or well-fed diet, BPS treatment at doses of 4 or 50 µg/kg-day

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for three months had no effect on early embryo development (e.g., 2-cells, 4-cells, or
blastocyst), following in vitro fertilization (Desmarchais et al. 2022).
Effects on implantation
A decreased number of implantation sites was reported in two studies in adult female
SD rats treated with BPS at 300 mg/kg-day by gavage, either from premating to PND 3
in Anonymous Study 12 as described in ECHA (2019) or for a total of approximately 14
weeks, from 6 weeks premating to weaning in Anonymous Study 14 as described in
ECHA (2019). In the latter study, an increased rate of post implantation loss was also
observed in the 300 mg/kg-day dose group in the Anonymous Study 14 as described in
ECHA (2019). A dose-related decreased trend in the number of implantation sites, withno effect on fertility index, was reported in adult female SD rats treated by gavage with
BPS at doses of 20, 60 or 180 mg/kg-day for about 90 days from premating throughpregnancy (and weaning on PND 21) (BASF 2019a). No effects on mean number of
implantation sites or percentage of post implantation loss was reported in adult female
Wistar rats treated with BPS by gavage at doses of 30, 100, or 300 mg/kg-day during
pregnancy (GD 6 to GD 19) and evaluated on GD 20 (BASF 2014b), or in mice treatedorally with BPS at 200 μg/kg-day for 2 weeks prior to mating and through GD 12.5 (Maoet al. 2020).
4.2.8 Sex ratio
No effect on sex ratio was seen in the F2 generation of CD-1 mice in a study where
BPS was administered orally to F0 dams on GD 11 to birth at doses of 0.5, 20, or 50
μg/kg-day (Shi et al. 2019) or in the F2 generation of ICR mice in a study where BPS
was administered orally to F0 dams on GD 12.5-15.5 at doses of 2, 10, 50, 100, or 200
μg/kg-day (Zhang et al. 2020). In zebrafish exposed to BPS from 2 to 75 dpf atconcentrations of 0.1, 1, 10, or 100 μg/L, sex ratio was skewed towards females at 10
and 100 μg/L BPS (Naderi et al. 2014).
Summary Table
Data discussed above on female reproductive toxicity from whole animal studies are
summarized in Table 4.2.1 below. The results presented are statistically significant(p<0.05), unless otherwise stated (i.e., non-significant [NS] change). The studies areorganized alphabetically by first author.

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Table 4.2.1 BPS: Evidence on the female reproductive toxicity in animal studies
Study Design

Outcomes Assessed

Major Findings

Ahsan et al. 2018
Female rats (no strain provided), fifteen rats per group.
Treatment: BPS (purity not reported) in castor oil, daily
subcutaneous (sc) injections (50 µL total volume) at doses of0, 0.5, 5. or 50 mg/kg-day for 10 days (postnatal day (PND) 1
to 10).
Eight females/group were sacrificed on morning of the first
estrus (starting on PND 75). Organs and blood samples
collected for further analysis.
Methods not very well described

Day of vaginal opening
Start of estrous cyclicity (assessed from PND
60 to 70)
Recorded weights of ovary and uterus on PND
75
Gonadosomatic index (GSI) = (100 × [gonad
weight (g)/body weight (g)])
Starting on PND 75, blood was collected forplasma hormone levels of luteinizing hormone(LH), follicular stimulating hormone (FSH),
testosterone (T), estradiol (E2), andprogesterone (P) on the first estrus day.
Histologic analysis of ovaries.
Five adult females/group were assessed for
fertility. Each female was mated with one fertile
adult male: Litter size, litter weight and sex ratio
were registered.
General toxicity:
Body weight

Puberty:
Delayed vaginal opening at 50 mg/kg-day
Decreased mean GSI at 50 mg/kg-day
Decreased paired ovarian weight at 5 and 50 mg/kg-day
Decreased absolute and relative weight of paired uteri at 50 mg/kg-day
Hormonal analysis (at first estrus from PND 75):
Decreased plasma levels of LH, FSH and P at 50 mg/kg-day
Increased T levels at 50 mg/kg-day. No effect on E2
Histology:
Decreased number of corpora lutea (CL) and antral follicles at 5 and 50 mg/kg-day
Increased number of atretic follicles at 50 mg/kg-day, non-significant (NS) difference in thenumber of preovulatory follicles
Estrus cycle:
Altered estrus cyclicity at 50 mg/kg-day.
Fertility:
Percentage of females that conceived: 60% BPS and vs 100% in controls)
Reduced number of pups born/female (5.33 ± 0.67) at 50 mg/kg-day (8.80 ± 0.58 in controls)
General toxicity
Increased body weight on PND 30, 45, and 60 at 50 mg/kg-day

BASF 2014a
Female Wistar rats, 42 days old at the start of treatment, 10
rats per group.
Treatment: BPS (99.4% purity) in water containing 1%
carboxymethylcellulose (CMC) at a final volume of 10 mL/kg,
daily oral gavage at doses of 0, 100, 300 or 1,000 mg/kg-day
for a total of 90 days.
Animals were sacrificed after 16 hours of fasting.

Reproductive organ weights
Histopathology in ovary and uterus
General toxicity:
Body weight, organ weight, red blood cell(RBC) count

Increased ovarian weight at 1,000 mg/kg-day
Squamous metaplasia of uterus at 1,000 mg/kg-day.
General toxicity:
No effect on body weight, increased adrenal and liver weight (absolute and relative),decreased RBC count, lower hematocrit at 1,000 mg/kg-day.

BASF 2014b
Pregnant Wistar rats, 42 days old at the start of treatment, 25
rats per group.
Treatment: BPS (99.3% purity) in water containing 1% CMC ata final volume of 10 mL/kg, daily oral gavage at doses of 0, 30,
100, or 300 mg/kg-day from gestational day (GD) 6 to GD 19.
Animals were sacrificed on GD 20.

Number of corpora lutea
Uterus weight
Reproductive parameters: implantation sites,
post implantation loss
General toxicity:
body weightNo effects on number of corpora lutea, gravid uterus weight, mean number of implantationsites or in % of post implantation loss
General toxicity:
Decreased net body weight gain at 300 mg/kg-day.

BASF 2019a)

F0: Estrous cycle duration, fertility index
F1: Vaginal opening from PND 27

F0: Increased estrous cycle duration at 180 mg/kg-day
F0: No effect on fertility index; decreased trend in number of implantation sites.

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Study Design

Outcomes Assessed

Major Findings

Adult female Sprague-Dawley (SD) (Crl:CD) rats, 24 rats pergroup (F0 rats).
Treatment: BPS (99.9% purity) in 0.5% CMC suspension, daily
oral gavage (10 mL/kg-day) at doses of 0, 20, 60 and 180
mg/kg-day for about 90 days (from 10 weeks premating
through mating, gestation, and lactation until weaning, PND
21).
Mating of F0 females with treated males (with the same doseas females) after 10 weeks of treatment
F0 females were sacrificed after weaning their offspring (PND
21).

General toxicity:
Body weight

F1: Delay in vaginal opening (PND 33.2) at 60 mg/kg-day (range PND 28 – PND 40 for all
doses)
General toxicity:
F0: Increased body weight at 60 and 180 mg/kg-day on days 14 and 28 of the premating
period. No effect on the final body weight.

BASF 2020
SD female rats, five rats per group.
Treatment: BPS (purity not reported) in 0.5 % methylcellulose
in water, daily oral gavage (10 mL/kg) at dose of 0, 100, 300
or 600 mg/kg-day for 28 days.
Animals were sacrificed at the end of exposure period.

Ovary and uterus weight
General toxicity:
Body weightNo effect on ovarian or uterus weight
General toxicity:
Reduced body weight at 600 mg/kg-day

Benjamin et al. 2023
7-month-old C57BL/6J female mice, five animals per group.
Treatment: BPS (purity not reported) diluted in distilled water(vehicle). BPS at doses of 0 (control) or 30 mg/kg-day, daily byoral gavage for 21 days
Estrous cycle monitored by daily vaginal smears and animals
were sacrificed at the estrus after the 21-day treatment.

Mouse uteri were collected and fixed in 10%
neutral buffered formalin for histopathologicalanalysis (number of endometrial glands anduterine wall thickness)

Uterus histology:
Developed non-atypical (increased number of normal cells) endometrial hyperplasia
Increased thickness of the myometrium
Increased number of glands in endometrium
Adenomyosis (endometrial cells found in myometrium) in 40% of treated mice.

Castro et al. 2015
Female Wistar rats, ten rats per group.
Treatment: BPS (98% purity) in 0.1% dimethyl sulfoxide(DMSO)/99.9% olive oil, daily sc injections at doses of 0 or 10µg/kg-day from GD 12 to parturition.
Female pups were treated similarly via sc injection from PND 1
to PND 21. Pups were sacrificed 30 min after last injection andbrains saved for neuro-steroidogenesis analysis.

Effects on 5α-reductase (5α-R) isozymes (1, 2,
and 3), dopamine, and serotonin (5-HT) related
genes in the prefrontal cortex
RT-PCR, western blot, and quantitative PCR
array

Genes involved in steroidogenesis:
No effect on 5α-R1 or 5α-R2 mRNA levels (no data on protein expression for these two
isozymes)
Decreased 5α-R3 mRNA levels, no effect on protein expression.
Genes associated with the dopamine and serotonin systems:
5-HT: upregulation of 5 genes, including Cyp2d4 (involved in 5-HT synthesis) anddownregulation of 1 gene involved in metabolism and 2 genes involved in signal transduction
pathways.
Dopamine: NS downregulation of two dopamine receptor genes: Drd1 and Drd3.

Chen et al. 2016bC. elegans hermaphrodite worms cultured at 20°C on
nematode growth medium (NGM) plates without cholesterol, atleast five worms per group.

Viable egg number and rate of embryoniclethality, number of adults (brood size),
Germline effects

Female-like reproductive toxicity in hermaphrodites:
No effect on the number of fertilized eggs
Increased rate (5-fold) of embryonic lethality (dose-dependent)
Decreased brood size (-25%) at 125 and 500 µM.
Germline effects:
Irregular distribution of germline nuclei

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Study Design

Outcomes Assessed

Treatment: BPS (purity not reported) in 0.1% ethanol at 0,125, 250, and 500 micromolar (µM) (internal concentrations
for BPS were ND, 0.21 µg/g and 0.39 µg/g, respectively)
Exposure from the L1 larval stage to adulthood: sodium
hypochlorite-treated eggs plus BPS or control, followed byincubation at 20°C for 4 days; analysis at 20 and 24 hours
post-L4 (first day of adulthood).

Major Findings
Dose-dependent germline nuclear loss
Increased apoptosis and checkpoint activation at all doses
Increased (about 3x) germline nuclear gap phenotype
Alteration of SYP-1 disassembly (chromosome misalignment)

Demacopulo and Kreimann 2019
Immature Wistar rats (19–22 days old), five to six rats pergroup
Exp 1. Treatment: BPS (purity not reported) in 20% ethanol /80% sesame oil, daily sc injections (0.1 ml total volume) at 0
or 1 µg/kg-day for 20 days
Exp 2. Polycystic ovary-like syndrome induction by
dehydroepiandrosterone (DHEA).
Treatment: daily sc injections of BPS at 1 µg/kg-day plus 60
mg/kg-day of DHEA for 20 days
After treatment, estrous cycle day was determined for eachanimal and then euthanized for organ/tissue collection.

Estrous cycle stage
Histologic analysis of ovaries and uterus
Cell apoptosis, cell proliferation by measuring
proliferation marker: proliferating cell nuclear
antigen (PCNA), and EZRIN (marker ofapicobasal polarity in endometrial cells)
expression by immunohistochemistry

Exp 1.
Ovary
No difference in ovarian weightOvaries had normal follicles and corpora lutea
NS increase in ovarian cysts (no cysts in controls)
UterusNS effect on uterus morphology: decreased diameter and endometrial luminal height(increased variability in the data), increased cell vacuolization.
Exp 2. BPS-DHEA interaction:
Decreased average number of cysts in the BPS+DHEA group (compared to DHEA alonegroup)
Increased intraepithelial lumens in the BPS+DHEA group (compared to DHEA alone group)
Increased PCNA positive cells in endometrial perimeter in the BPS+DHEA group whencompared to DHEA alone.
Increased EZRIN expression in areas of the epithelium and increased apicobasal polarity ofthe epithelium in the BPS+DHEA group compared to DHEA alone.

Desmarchais et al. 2022
2.5 year old ewes (Ile-de-France)
Study BPS and diet interaction. Two diet groups, restricted(RF) and well fed (WF), thirty ewes per group, further divided
into ten animals per dose group.
Treatment: BPS (purity not reported), daily in the diet (vehicle
not reported) at doses of 0, 4 or 50 µg/kg-day for at least threemonths.
Animals were estrous synchronized and stimulated forsuperovulation with FSH. Then, mature follicles were aspirated
under anesthesia on all 60 ewes at 7-day intervals during thebreeding season (September to December).
The recovered cumulus–oocyte complexes (COCs) were used
for in vitro maturation, in vitro fertilization (IVF), and embryo
culture procedures. Oocytes were fertilized with sperm from
untreated ram donors.

Plasma hormones measured: P and E2 (also inpreovulatory follicular fluid), anti-müllerian
hormone (AMH), and thyroid hormones, free
triiodothyronine (FT3), free thyroxine (FT4) andtotal thyroxine (TT4)
Embryo production data
Metabolic effects: plasma glucose, and nonesterified fatty acidHormone levels: No effect of BPS on P or E2 levels in plasma or follicular fluid. No effect on
plasma AMH, or any thyroid hormones.
Ovary: No difference in the total number of follicles available for aspiration
IVF: No significant effect on any of the parameters in terms of number of embryos anddevelopmental rates, however, the following NS effects were observed:
NS decreases (approximately 40%) at 4 µg/kg-day in the numbers of cleaved embryos, >4cell embryos, blastocysts, and early blastocysts in the RF diet group.
NS increases (approximately 200%) at 4 µg/kg-day in these parameters in the WF diet group.
Significant diet and BPS dose interaction.
There was a statistically significant interaction between dose and diet for the number and rateof cleaved embryos, the number of >4-cell embryos, the number of blastocysts and thenumber of early blastocysts.

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ECHA 2019 - Anonymous Study 12, 2000, (report following
OECD TG 421 protocol)
Adult female SD (F0) rats, 12 rats per group.
Treatment: BPS (purity not stated) in 0.5 % aqueous sodium
CMC solution with 0.1 % Tween 80, daily oral gavage at doses
of 0, 10, 60 or 300 mg/kg-day for a total of 40-46 days
(premating, gestation, and to lactation day (LD) 3).

Reproductive performance: Copulation,
Implantation sites / index
Reproductive tract weightOvarian histology: number of corpora lutea
General toxicity:
Body weightNo effects on reproductive performanceNo effect on ovary or uterus weightNo effect on number of corpora lutea
Increased duration of estrous cycle (5.57 days vs 4.08 days); more days in diestrus (5 females
vs 0 in control) at 300 mg/kg-day.
Females in diestrus had a NS decreased fertility index (58.3 % vs 91.7 %) at 300 mg/kg-day
Decreased implantation index at 300 mg/kg-day.
General toxicity:
Decreased final body weight at 60 mg/kg-day.

ECHA 2019 - Anonymous Study 14, 2017
Adult female SD rats, 10 rats per group.
Treatment: BPS (purity not reported) in 0.5% CMC daily oral
gavage (vehicle and route was according to other study in thereport – Anonymous Study 16, 1999) at doses of 0, 30, 100 or
300 mg/kg-day for about 14 weeks (6 weeks premating, 2
weeks mating, gestation, and lactation)

Organ weights
Estrus cyclicity
Reproductive performance
General toxicity:
Mortality, body weight(All effects at 300 mg/kg-day)
No changes in ovarian weight
Increased uterus weight
Longer estrous cycles
Decreased number of implantation sites
Increased rate of post implantation loss
General toxicity:
No premature death, decreased body weight by 6% at 300 mg/kg-day

ECHA 2019 - Anonymous Study 16, 1999
Adult female SD rats, 6 rats per group.
Treatment: BPS (purity not reported) in 0.5% CMC, daily oral
gavage at doses of 0, 40, 200 or 1,000 mg/kg-day for a total of
28 days.
Animals sacrificed at the end of dosing period

Ovary weight
General toxicity:
Body weightNo effect on ovary weight. No data reported on other reproductive organs
General toxicity:
Decreased body weight gain

Eldefrawy et al. 2021
Adult female Hy-Line W-36 laying chickens, six animals pergroup.
Treatment: BPS (purity not reported) in corn oil, daily by
gavage (from a 300 μg/mL solution) at doses of 0, or 50µg/kg-day for 3 months.

Organ weight of ovary, oviduct, and liver after
necropsy
Histopathology: Numbers of preovulatory
follicles and diameters of the five largest
preovulatory follicles (F1, F2, F3, F4, and F5)
were recorded.
Oophoritis was scored using a 1–5 scaleaccording to lymphocyte or plasma cell
infiltration.
RNA isolation and RNA-sequencing (RNA-seq)
in theca and granulosa layers from the largest
mature preovulatory follicle (F1)
Bioinformatics analysis of RNA seq data andpathway analysis

Increased left ovary weight (the right ovary in chickens regresses and is not functional).
Decreased numbers of preovulatory follicles.
Lymphocytes present around the growing follicle, with ovarian inflammation (oophoritis). No
oophoritis observed in controls.
DEGs in largest mature preovulatory follicle:
Examples of downregulated genes: STAR (Steroidogenic acute regulatory protein), CHRNA4(Cholinergic receptor nicotinic alpha 4), ZAR1L (Zygote arrest 1 like), and LOC416931(uncharacterized protein). Among the examined hens, only one gene was upregulated: AVD(avidin gene, can affect ovarian function)

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RNA seq analysis was used to identify
Differentially expressed genes (DEGs).
Gingrich et al. 2018
Pregnant sheep (cross between Polypay and Dorset breeds),
six to seven animals per group
Treatment: BPS (purity not reported) in corn oil, daily sc
injection (injection volume not reported), at doses of 0, or 0.5
mg/kg-day from GD 30 to 100
Biweekly serum samples from GD 30–120 for hormonal,
protein, and biochemical analyses
Animals were sacrificed, and placenta samples were collected
on GD 120.

Placental endocrine function
Pregnancy-associated glycoprotein 1 andpregnancy-specific protein B
Serum P
Placental morphology

Decreased serum maternal pregnancy-associated glycoprotein1 on GD 75 to GD 90
Decreased serum pregnancy-specific protein B on GD 60, 75, and 90
NS decrease in serum P at GD 75-105 (P=0.09)
Placenta
NS increase in placental weight (P=0.07)
No effect on placental tissue distribution, or number of placentomes per placenta
50% lower e-cadherin protein expression in GD 120 placentomes
20% decrease in trophoblast derived binucleate cells
Fusogenic genes: Decreased expression of enJSRV and HYAL2
Increased transcription factor glial cell missing factor 1.

Hill et al. 2017
Adult female CD-1 mice, five dams per group
Treatment: BPS (99% purity) in tocopherol-stripped corn oil,orally in animals trained to drink the BPS containing solution (1µL/g body weight) at a dose of 0 or 200 µg/kg-day from GD 8
until PND 19.
On PND 19 (prepubertal), two female offspring from each litter
were weaned from their mothers. One female was fed
tocopherol-stripped corn oil (control) and the second was fed 1
μg/kg-day ethinyl estradiol (EE) in stripped corn oil from PND
19 to PND 21. Animals were sacrificed 24h after.
Uterine responsiveness to EE was confirmed visually via
change in uterine size on PND 22.

On PND 22, female reproductive tissues
including the ovary, oviduct and uterus werecollected.
All organs were examined for grossmalformations including ovarian cysts.
Expression of several estrogenic activity
related genes was measured in the uterus andovary:
Esr1 (estrogen receptor α (ERα)),Esr2 (ERβ),
Igf-1 (insulin growth factor 1) andEgf-r (epidermal growth factor 1)
BPS and EE interaction

No effect on tissue organization of oviduct or uterus including endometrial cell height.
Uterus: NS decrease in Ki67 (marker for cell proliferation), or in cells undergoing apoptosis in
the uterine stroma or epithelium
Ovary:
Increase in number of secondary follicles
No effects on Ki67 or on apoptosis in granulosa or theca cells.
Gene expression (mRNA levels):
Uterus: increased Igf-1;
NS decrease of Esr1; no effect on Egf-r
Ovary: increased Igf-1 and Egf-r
NS increase of Esr1 and Esr2
BPS and EE challenge:
BPS + EE increased uterine epithelial height compared to unchallenged controls
BPS inhibited the EE-induced increases in the number of total follicles.
BPS + EE increased theca cells apoptosis compared to EE vehicle control
BPS + EE increased uterine IGF-1 mRNA levels, compared to EE-challenged controls,
BPS inhibited the effect of EE on expression of IGF-1 (NS decrease) and EGF-R (decrease),and on the Esr1 (NS decrease) and Esr2 (decrease) in the ovary.

Ijaz et al. 2020
Adult female SD rats, ten animals per group.
Treatment: BPS (99% purity) in saline, daily intraperitoneal (ip)
injection (1 mL/kg final volume) at doses of 0, 50, 500 µg/kgday and 5, 50 mg/kg-day for 28 days
On day 29, animals were sacrificed at the estrus day of thecycle

Ovaries and uteri weights; GSI
Plasma concentration for T, E2, P, luteinizing
hormone (LH) and follicle stimulating hormone(FSH)
Biochemical and histological analysis of ovary
and uterus
Determination of oxidative stress in ovary:
catalase (CAT), peroxidase, superoxide
dismutase (SOD) activity, reactive oxygen
species (ROS) assay, and thiobarbituric acid

Decreased GSI at the high dose (50 mg/kg-day)Hormones:
Increased plasma T at 5 and 50 mg/kg-day
Decreased plasma E2, P, LH, FSH at 5 and 50 mg/kg-day
Ovaries:
Decreased paired ovarian weight at 50 mg/kg-day
Decreased number of CL at doses ≥ 500 µg/kg-day; increased CL diameter at 50 mg/kg-day
Decreased number of antral and atretic follicles at 5 and 50 mg/kg-day
Increased antral follicle diameter at all doses
Decreased granulosa cell height at 5 mg/kg-day, and increased at 50 mg/kg-day
No effect on theca cell height
Uterus: decreased relative uterus weight at 5 and 50 mg/kg-day

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reactive substance (TBARS)-lipid peroxidationassessment

Oxidative stress:
No significant change in peroxidase activity
Decrease in:
CAT (U/mg protein) at 50 mg/kg-day
SOD (U/mg protein) at 50 µg/kg-day and at 50 mg/kg-day
Increase in:
ROS (U/min tissue) at 500 µg/kg-day and at 5 and 50 mg/kg-day
TBARS (nmol/mg tissue) at 50 mg/kg-day

Ji et al. 2013
Adult zebrafish (AB strain), four male and six female fish (andtwo replicates) per group.
Treatment: BPS (purity not reported) in 0.1% methanol in 6 L
exposure medium (filtered water) to a final concentration of
0.5, 5, and 50 μg/L for 21 days.
After the exposure, fish were euthanized, and organs werecollected for further analysis.

GSI, hepatic somatic index(HSI) = (100 × [liver weight (g)/body weight(g)])
Brain-somatic index(BSI) = (100 × [brain weight (g)/body weight(g)])
Plasma hormones: E2 and T
Transcription of 21 genes related to functional
processes of the hypothalamic-pituitarygonadal axis (brain and ovary)

Decreased GSI at all doses, NS decreased BSI and NS increased HSI.
Plasma hormone levels: Increased E2 at 50 μg/L, no effect on T and increase in E2/T ratio at
50 μg/L.
Down-regulation of gene expression at 50 μg/L:
Brain: gnrh3 (gonadotropin releasing hormone) and fshβ.
Ovary: hmgr a and hmgr b (Hydroxy methyl glutaryl CoA reductase). No effect on aromatasegene expression or on other genes analyzed.

Kaimal et al. 2021
Adult female Sprague-Dawley rats, at least nine animals pergroup
Treatment: BPS (98% purity) in phosphate buffered saline(PBS), daily oral by micro pipetting the solution into the mouth(about 15 μL total volume) at doses of 0, or 5 μg/kg-day for 15
days (GD 6 to 21)
F1 female rats were sacrificed at 16 to 24 weeks of age on the
day of diestrus.

Number of pregnant dams, abortions per dam,
stillbirths, and live births (gestational index
In F1:
Organ weight for pituitary, ovary, and uterus
Morphometric analysis of ovaries: Follicle
characterization

No effects on proportion of dams with normal pregnancies or abortions, litter size, or offspring
body weight
Two stillborn pups (one each in two out of 13 litters) and none in the control group
F1: No effect on pituitary or ovary plus uterus weight.
F1: Decreased number of CL in offspring ovaries; no effects on number of follicles at any
stage (primordial, primary, secondary, antral, Graafian, or atretic).

Kolla et al. 2018
Pregnant CD-1 mice, at least eight animals per group
Treatment: BPS (purity not reported) in 70% ethanol appliedand allowed to dry on a small wafer, fed daily to dams at
doses of 0, 2 or 200 µg/kg-day from GD 9 to LD 20
One female from each litter was selected for necropsy at eachage: PND 24 (prepubertal), PND 32–35 (pubertal), and 9
weeks of age (adult)

Pregnancy loss
Mammary gland analysis:
Whole mount morphometric and histologicalanalyses, and immunohistochemical analyses
for Ki67 (marker of proliferation), and protein
expression of ERα and progesterone receptor(PR)

Dams delivered naturally
No effect on pregnancy loss
Mammary gland morphology
At PND 24: Larger TEB area in the low dose group, increased average TEB size in both dosegroupsAt puberty: No significant effect at any doseAt adulthood: Increased number of alveolar buds at 200 µg/kg-day; increased terminal ends at
2 µg/kg-day; increased incidence of TEB-like structures at both doses.
Ki67 expression
At PND 24: Decreased at 2 µg/kg-day (NS decrease at 200 µg/kg-day)
At puberty: NS increase at both doses.

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At adulthood: Increased at 2 µg/kg-day, no effect at 200 µg/kg-day.
Increased incidence of intraductal hyperplasia at both doses (related to the K67 increase)
Mammary gland steroid receptor protein expression
At PND 24: No effects on ERα or PR
At puberty: Increased expression of PR at both doses; NS increase in ERα at both doses.
At adulthood: Decreased ERα at 200 µg/kg-day
No effects on PR expression

Kolla and Vandenberg 2019
Pregnant CD-1 mice, at least seven animals per group
Treatment: BPS (purity not reported) in tocopherol stripped
corn oil, daily oral administration to dams with a pipette to themouth (1 μg oil/g body weight) at doses of 0, 2, 200, or 2,000μg/kg-day from GD 8 to PND 2
EE challenge: Two female pups per litter from each of theexperimental groups described above were treated with EE atdoses of 0, or 1 μg//kg-day, for 10 days (from PND 21 to PND
30)
Animals were sacrificed on PND 31

Time to vaginal opening; anogenital index(AGI); uterus weight
Mammary gland histology.
Gene expression for ER and PR

NS effect on time to vaginal opening, AGI, or uterine weight.
BPS effects on mammary gland:
Decreased ductal area at 2 μg/kg-day
No effect on the number or total area of TEBs
Increased gene expression of Esr1 (ERα) at 200 μg/kg-day
No effect on Pgr (PR) gene expression.
EE – BPS interaction:
No effect of EE challenge on vaginal opening, AGI, uterine weight, mammary gland ductal
area (BPS effects remained), number or total area of TEBs.
No effect of EE challenge on gene expression of Esr1 (BPS effects remained).

LaPlante et al. 2017
Timed pregnant CD-1 mice, at least 5 animals per group.
Treatment: BPS (> 99% purity) in 70% ethanol applied andallowed to dry on a small wafer, fed daily at doses of 0, 2 or
200 μg/kg-day.
Exp 1: Exposure from GD 8 until weaning on LD 21. Dams
were euthanized on LD 21.
Exp 2: BPS from GD 9 through LD 1. Dams euthanized on LD
2.

Histoarchitecture in dams whole-mountmammary gland on LD 2 and 21.
On LD 21:
Mammary gland receptor protein expression byimmunohistochemistry: Prolactin receptor(PrlR), signal transducer and activator oftranscription 5 (Stat5, a PrlR signaling marker),ERα, and Ki67
Mammary gland gene expression: Prlr and
Esr1
Serum E2
Brain immunohistochemistry: ER, Stat5 and
PrlR in the arcuate nucleus of the
hypothalamus
Nursing behaviors on LD 2, 7, and 14
General toxicity: Dam and pup body weights

Whole-mount mammary gland
Trend to increased functional mammary tissue at LD 2, and decreased functional mammary
gland tissue on LD 21
LD 2: NS increase in lobule size (23% at 2 μg/kg-day; 30% at 200 μg/kg-day).
LD 21: Decreased volume fraction of lobules, and increase in the volume fraction of adipose
tissue at 200 μg/kg-day
NS decrease in lobule size (8% and 24% at 2 and 200 μg/kg-day, respectively)
Prolactin signaling:
NS decrease in Stat5 positive epithelial cells
Increased Prlr gene expression at 2 μg/kg-day and NS decrease at 200 μg/kg-day.
Gene and protein expression on LD 21:
Esr1: NS increase in mRNA at 2 μg/kg-day (compared to control) and decrease at 200 μg/kgday (compared to the 2 μg/kg-day dose group).
No effect on number of ERα positive cells in the arcuate nucleus of the hypothalamus
Ki67: No effect on protein expression
Endocrine effects on LD 21:
Decreased serum concentrations of E2 at both doses
Increased ERα protein expression at both doses
No effects on Prlr, or Stat5 positive cells in the brain.
Nursing behavior:
No effect on the time spent nursing at LD 2 and LD 7
Increased time nursing on LD 14 at 200 μg/kg-day
Increased time in the high-crouch position on LD 14 at 2 μg/kg-day

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Major Findings
Decreased percentage of pup-initiated nursing on PND 14 (91% in controls; 50% in 200 μg/kgday dose group)
General toxicity: Decreased pup wt on PND 14 at 2 μg/kg-day; NS decrease at 200 μg/kg-day.

Liu et al. 2021
Newborn CD-1 mice, number of animals not reported but thestatistical analysis section states that each experiment was
repeated at least three times.
Treatment: BPS (purity not reported) in saline solution, daily ip
injection (volume not specified) at doses of 0, 2 or 10 μg/kgday for 3 days
Animals were sacrificed after treatment (not clear how longafter treatment) and ovaries saved for follicle analysis.
Another set of animals (with the same treatments) weresacrificed at PND 21.

Ovaries were collected and oocytes in germ
cell nests (or cysts) and primordial follicles
were counted using mouse vasa homolog(MVH)—a germ cell marker.
Immunofluorescence andimmunohistochemistry for proliferation markers
Folliculogenesis at PND 21 and measurement
gene and protein expression of GDF9 and
BMP15 (oocyte specific markers)

Dose-dependent increase in the percentage of MVH-positive oocytes in primordial follicles
Decreased percentage of oocytes in germ cell cysts.
No effect on total number of oocytes
Increased number of cells positive for Ki67 (proliferation marker) at both doses.
Dose dependent increase in proliferation of granulosa cell precursors.
Folliculogenesis at PND 21
Increased number of primary follicles and decreased number of secondary and antral follicles
at 10 μg/kg-day.
Decreased gene and protein expression of GDF9 at 10 μg/kg-day and BMP15 at both doses.

Mao et al. 2020
Female C57BL6J mice, at least six animals per group.
Exposure: BPS (> 98% purity) in 70% ethanol (8 to 24 μL/kg)
applied and allowed to dry on a small wafer, fed daily at doses
of 0, or 200 μg/kg-day for two weeks.
After treatment, females were paired with untreated males.
Euthanized on GD 12.5; each uterine horn was incised; theuterine position of each conceptus was delineated, andplacenta samples were obtained and saved half in liquid
nitrogen and the other half was fixed for histological analyses.

Pregnancy outcomes
Placental histology: measurement of thelabyrinth, spongiotrophoblast, and trophoblast
giant cell (GC) areas
DEGs analyses in placental tissues on GD 12.5
Metabolomics: Effects on fatty acids
Neurotransmitters: Effects on 5-HT anddopamine tissue distribution byimmunohistochemistry
Placental hormone levels: E2, estrone (E1),
Corticosterone, T, and P
Placenta morphometric analyses

No effects on pregnancy success, maternal gestational weight, implantation sites, and number
or sex ratio of fetuses
Effects in GD 12.5 placenta:
Decreased ratio of spongiotrophoblast zone to GC area.
DEGs: 11 genes were differentially expressed with upregulation of Actn2 (actinin α2) with a
2.17-fold change. The other 10 genes were downregulated, such as Calm4 (calmodulin 4) with
-22.52-fold change and Guca2a (guanylate cyclase activator 2a, guanylin) with -26.72-foldchange.
Metabolomics:
Effect on fatty acids: Decreased stearic acid, palmitic acid, docosahexaenoic acid,octadecenoic acid, and hexadecanoic (palmitic) acid.
Neurotransmitters: Increase in dopamine concentration
Increased percentage of dopamine-positive GCs.
Decrease in serotonin concentration and the percentage of serotonin-positive GCs
Ten-fold increase in the ratio of 5-hydroxyindoleacetic acid (primary 5-HT metabolite) to 5-HT.
Decreased percentage of serotonin-positive GCs.
Placental steroids: NS effect on any steroid hormone analyzed

Naderi et al. 2014
Zebrafish (wild-type) embryos at blastula stage 2 days post
fertilization (dpf), 700 embryos in total.
Treatment: BPS (> 98% purity) in 0.01% acetone was added
to 5 L tanks to a final concentration of 0, 0.1, 1, 10 and 100
μg/L (50% of exposure solution was renewed daily) for 75
days
Males and females from the same treatments were randomly
assigned to spawning tanks; 2 males per 2 female colonies)

Fish development, reproduction, plasma
vitellogenin (VTG), GSI, and HSI
Plasma hormones: E2, T, T4 and T3
Numbers of eggs spawned
Hatching rate and time to hatching (hours)
General toxicity: Survival rate, body weight andlengthSkew in the sex ratio towards females in adults: percentage of females at 10 and 100 μg/L
BPS was 58.8% and 66.7%, respectively (46.3% in the control)
Decreased GSI at 100 μg/L (in females), (males lower at 10 μg/L)
Increased HSI at 10 and 100 μg/L
Increased VTG induction at 10 and 100 μg/L
Endocrine effects:
Increased plasma E2 at 10 and 100 μg/L
Decreased plasma T3 and T4 at 100 μg/L
Reproductive effects:

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Blood samples from 4 to 6 fish of the same sex and same
treatment were pooled to generate a total of three replicate
pooled blood samples for each treatment.

Major Findings
Decreased number of eggs during the 7-day spawning period at 10 and 100 μg/L.
Lower hatching rate and increased time to hatching of embryos at 10 and 100 μg/L.
General toxicity: Decreased survival rate at 100 μg/L, no effect on body length or body weight.

Nevoral et al. 2018
Female ICR mice at puberty (five weeks-old), at least 4
animals per group.
Treatment: BPS (purity not reported) in drinking water (solvent
vehicle not reported) at doses of 0, 0.001, 0.1, 10, or 100µg/kg-day for 4 weeks
Seven to eight animals were sacrificed at the end of treatment,
ovaries were fixed for histological evaluation (left ovary) andproteomic analysis (right ovary).
Other group of animals was treated with gonadotropin to
retrieve the mature oocytes or mated with untreated males forembryo analysis.

Ovarian histology on seven left ovaries:
primary, preantral, antral and atretic follicles
and their volumes
Ovaries from BPS treated at (0.1 µg/kg--day)were used for proteomic analysis by liquid
chromatography-mass spectrometry.
Number and percentage of matured oocytes
were counted and used for
immunocytochemistry for α-tubulin, pericentrin,
H3K27me2, and methyl cytosine (5meC).
Embryo quality assessed after in vivo
fertilization.
Serum hormone levels

Decreased relative ovary weight at all doses and total ovarian volume at 10 and 100 µg/kgday.
Decreased medulla volumes at all doses
Decreased corpora lutea volume (no data presented)
Increased volume of antral follicles at 0.001 and 0.1 µg/kg-day
Decreased volume of atretic follicles at 0.1 and 10 µg/kg-day
Dose-dependent decreased numbers of primary, preantral, and antral follicles
Decreased quantity of granulosa cell layers and decreased serum E2 at 10 and 100 µg/kg-day
Increased incidence of spindle malformation at 1 and 100 µg/kg-day associated with α-tubulin
and pericentrin alterations
Increased numbers of abnormal oocytes at 0.1 and 10 µg/kg-day
NS increase in number of metaphase II (MII) oocytes retrieved (flushed).
Increased methylation of H3K27em2 at 10 and 100 µg/kg-day; no effects on 5meC
Decreased fertilization rate in apparent dose-dependent manner at the three lower doses but
only significant at 10 µg/kg-day
Increased fertilization rate at 100 µg/kg-day

Nevoral et al. 2021
Pregnant ICR mice, at least 4 animals per group.
In vivo (lactational exposure)
Treatment: BPS (purity not reported) in 0.1% ethanol added todrinking water at a concentration of 0, 0.375 ng/mL or 37.5
ng/mL to a calculated dose of 0, 0.2 or 20 µg/kg-day duringlactation for 15 days (PND 0 to PND 15).
Assessments on F1 female ovaries isolated on PND 15 and
PND 60In vitro maturation studies
Immature oocytes at the germinal vesicle (GV) stage (from F1
females treated perinatally) that were in proestrus or on estrus
day (ovulation). GV-oocytes were used for in vitro maturation
to obtain matured MII oocytes.
Another group of F1 females were gonadotropin stimulated toretrieve mature oocytes that were used for in vitro fertilization
or immunocytochemistry.

AGD and day of vaginal opening (puberty)
Histology on PND 15 and 60 ovaries
Stereology and follicle classification count(primordial, primary, preantral, antral, andatretic follicles)
In vitro GV oocyte maturation
Immunocytochemistry: α-tubulin for spindle
visualization, and demethylated histone H3 onlysine K27 (H3K27me2) heterochromatinmarker were measured in matured oocytes (invitro and in vivo)
Parthenogenetic activation: in vivo-matured
oocytes without the cumulus cells. Cultured for
24 hours (cleaved embryos) and 96 hours(blastocysts)

No effect on:
AGD, or day of vaginal opening
Primordial follicle reserve in young ovaries (not measured on PND 60)
Primary, preantral, and antral follicles pool on PND 60
Oocyte maturation:
No effects on number of GV oocytes per ovary, germinal vesicle breakdown, maturation stage,
number of flushed in vivo matured oocytes, and the number of atretic or fragmented oocytes.
Immunocytochemistry: Increased occurrence of spindle malformation and frequency ofchromosome misalignment in in vitro-matured oocytes at 0.2 µg/kg-day
Decreased H3K27me2 in mature ovulated oocytes at 0.2 µg/kg-day
In vivo mature oocytes:
No effect on number of flushed oocytes
Increased proportion of oocytes with spindle damage (α-tubulin) and chromosome
misalignment at 0.2 µg/kg-day
No effect on histone heterochromatin marker methylation status (H3K27me2)
Parthenogenetic activation: Increased rate of activated eggs and increased cleavage rate;
decreased blastocyst rate at 0.2 µg/kg-day

Nourian et al. 2017
Adult, same cycling stage, female mice (strain not specified),
at least five animals per group.

Ovaries were used for biochemical analyses,
and measurement of lipid peroxidation, using
malondialdehyde (MDA) as an indicator.

Decreased fertilization and blastocyst development rates at 10, 50 and 100 µg/kg-day.
Dose-dependent decreases in the percentages of two-cell embryos and blastocysts
Increased type I embryo arrests at 50 and 100 µg/kg-day

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Treatment: BPS (99% purity) in ethanol (ethanol % andvehicle not specified), daily ip injections (volume not specified)at doses of 0, 1, 5,10, 50 or 100 µg/kg-day for 21 consecutivedays.
24 hours after the last treatment, five mice in each groupwhere euthanized and the ovaries were collected.
Oocytes collected for IVF assays were obtained from fivesuper-ovulated female mice randomly selected from eachgroup. Mouse caudal epididymis sperm were capacitated andused for metaphase II arrested oocyte insemination.

Zygotes were evaluated 24 hours, 48 hours,
and five days after insemination.

Lipid peroxidation: dose dependent increase in MDA

Nourian et al. 2020
Adult, regular estrus cycle, female NMRI mice, at least 5
animals per group.
Treatment: BPS (99% purity) in PBS solution (0.1 to 0.5%ethanol), daily sc injections (volume not specified) at doses of0, 1, 5, 10, 50 and 100 µg/kg-day for 21 consecutive days
Blood samples from five mice per group were collected at one
day after last treatment. Animals were sacrificed and ovaries
collected for analysis.
IVF: BPS treated females were super-ovulated bygonadotropin treatment (5 mice per group) and oocytes wereinseminated with capacitated sperm from one untreatedmouse. Blastocysts (10-20 per group) were collected four days
later

Serum hormone levels of LH and FSH
Ovary
RNA quantity and integrity
Oxidative stress: total antioxidant capacity,
SOD, glutathione peroxidase, CAT
IVF embryos
Gene expression: estrogen signaling pathway(Erα and Erβ) and apoptosis induction pathway(P53, Bax, Nrf2 and E2F1)

Dose-dependent decreased serum LH and FSH levels
Ovary: Induced oxidative stress at all doses (except for glutathione peroxidase that issignificant at doses ≥ 5 µg/kg-day)Blastocysts: Increased gene expression of P53, Bax, E2f1, Erα, and Erβ at doses > 5 µg/kgday
Decreased gene expression of P53 and F2f1 at 50 and 100 µg/kg-day
IVF parameters: Positive correlation with antioxidant enzymes but not MDA, the marker for
lipid peroxidation
Negative correlation with all genes except ERα
Positive correlation for LH, FSH, ERα
Hatch percentage: associated with Nrf2, E2f1, MDA, and CAT

Pal et al. 2023
Adult female golden hamsters, 12 animals per group
Treatment: BPS (purity not reported) dissolved in corn oil,daily oral exposure at doses of 0 (vehicle), or 150 mg/kg-day
for 28 days
Twenty-four hours after treatment, animals were sacrificed.
Serum samples were used for further analysis.
Ovaries of one side: weighed and fixed for histological andimmunohistochemical studies.
The other ovaries were used for western blot, qPCR andbiochemical analyses.

Serum hormone levels: E2, P, LH, FSH, T3,
T4, melatonin
Serum inflammatory factor levels: TNFα, CRP,
and nitrite–nitrate
Histology in one ovary
Immunohistochemistry for PCNA
Western blot in ovary:ERα, aromatase, and TRα; Silent information
regulator 1 (SIRT-1), FOXO-1, Nrf2, PI3K and
pAkt, PCNA, connexin-43
qPCR in ovary:
Melatonin receptor (MT-1), SIRT-1 [involved inredox balance and ovarian functions], NFkB
[involved in redox and inflammatory function in
ovary], and inducible nitric oxide synthase

Serum hormone levels:
Decrease in LH, FSH, E2, P4, T3, T4, and melatonin
Increase in TNFα, CRP, and nitrite–nitrate.
Ovarian histology:
Decreased number of growing follicles and CL; increased number of atretic follicles.
Immunohistochemistry:Low immunoreactivity signal for PCNA
Western blot analysis in ovary:
Decreased expression of ERα, aromatase, and TRα
Decreased expression of SIRT-1, FOXO-1, Nrf2, PI3K and pAkt
Increased expression of NFkB, COX-2, caspase-3
Decreased PCNA and connexin-43
qPCR in ovary:
Decreased mRNA levels for MT-1 and SIRT-1

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Ovarian oxidative load
General toxicity: Body and organs weightOvarian oxidative load:
Decrease in SOD and CAT
Increase in lipid peroxidation.
General toxicity: Increased body weight and decrease in relative ovarian weight.

Park et al. 2022
Zebrafish (AB strain), 20 embryos per replicate and fivefemale fish per group depending on experiment.
Exp 1: Embryo-larval toxicity tests on normal fertilized
embryos at 4–6 hpf (20 embryos per replicate) in 60 x 15 mm
cell culture dishes.
Treatment: BPS (purity not reported) in 0.1 % DMSO, (15 mL
final volume) at concentrations of 0, 1.6, 3.1, 6.3, 12.5, 25, 50,
100, 250, 500, 1,000 μg/mL for seven days.
Exp 2: Short-term reproductive toxicity test. Female fish (ABstrain, 3–4 months old), five female fish per group.
Treatment: BPS (purity not reported) in 0.1 % DMSO added towater (350 mL final volume) at concentrations of 0, 8, 40, 200µg/mL (fresh test solution at 3-day intervals) for 21 days. Afterthe treatment fish were recovered for further analysis.

Fish were sampled and prepared to determine
gonadal and liver development: GSI, and HSI,
hepatic VTG and ER mRNA levels,
Steroid and thyroid hormones (T3 and T4), andresidual BPS concentration
General toxicity: Embryo-larval toxicity tests,
toxicity was determined by recording lethality
and teratogenic effects.

Increased HSI and GSI at 40 µg/mL
Decreased GSI value at 200 µg/mL relative to the 40 µg/mL group.
Oocyte maturation:
Normal mature oocytes in control and at 8 µg/mL
Regressed oocytes, numerous perinucleolar oocytes and yolk vesicle oocytes at 40 and 200µg/mL
Gene transcription:
Decreased liver VTG mRNA at 200 µg/mL and a NS decrease at the two lower concentrations
Hepatic ERα mRNA: NS increase at 8 µg/mL and a NS decrease at the two highest doses.
No effects on ERβ mRNA.
Hormone effects in whole body (Exp 2):
Increased P levels at 40 and 200 μg/mL, and increased E2 levels at all doses
Concentration-dependent increased thyroid hormone levels at all doses
Concentration-dependent increase in mean whole-body BPS concentrations
General toxicity: EC50 for embryo toxicity test was 332.6 μg/mL and the LOEL was 250 μg/mL(supplemental information).

Pollock et al. 2019
Female CF-1 mice 3–4 months old, at least seven animals pergroup
Treatment: BPS (≥ 98% purity) in peanut oil (0.1 mL final
volume), single sc injection on diestrus day at doses of 0, 1, 3,
or 9 mg/animal (final doses specific for each experiment were
provided by author and they are below)
Exp. 1: BPS at doses of 0, 36.7, 114.6, or 330 mg/kg.
Exp. 2: BPS at doses of 0, 27.7, 78.5, or 252.7 mg/kg.
30 min after BPS injection, animals were fed 0.2 g peanutbutter containing 50 μg/kg 14C-BPA (Exp. 1) or 14.5 ng of 5
μCi 3H-E2 (Exp. 2).
One hour after oral administration (in food) of radiolabeled
compound (Exp. 1 and 2), blood was collected by cardiac
puncture from anesthetized animals, and tissues werecollected for further analysis of the radiolabel.
Exp. 3: BPS at doses of 0, 25.2, or 79.8 mg/kg (only the twolowest BPS doses (1 and 3 mg/animal) tested in thisexperiment))

Exp 1 and 2: Distribution of 3H-E2 and 14C-BPA
in various organs (heart, lung, muscle, adipose,
uterus, ovaries), and serum
Exp 3: Urinary E2 levels

Exp. 1: Increased radiolabeled BPA levels in uterus at 114.6, or 330 mg/kg and in serum at
330 mg/kg. No increased radiolabel detected in the other tissues and organs analyzed.
Exp.2: Dose-dependent decrease in serum 3H-E2 significant at 78.5 and 252.7 mg/kg and NS
increase in 3H- E2 levels in the tissue and organs analyzed above control
Exp 3: Decreased urinary E2 at 10 h after treatment at 25.2 or 79.8 mg/kg.

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Prokešová et al. 2020
Adult (8 weeks old), female ICR mice, 15 animals per group
Treatment: BPS (purity not reported) in 50 μL 50% glycerol
containing 0.1 % DMSO, daily by oral gavage at doses of 0,0.001, 0.1, 10, or 100 µg/kg-day for seven days.
After exposure animals were sacrificed and immature oocytes
in the GV stage were collected from ovaries for further
analysis or cultured for 16 hours to obtain matured MII oocytes

Oocyte development
Chromosome misalignment (DNA) and spindle
malformations (tubulin) in in vitro mature MII
oocytes.
DNA integrity (apoptosis) by terminal
deoxynucleotidyl transferase dUTP nick endlabelling (TUNEL) assay on mature oocytes
Immunocytochemistry: heterochromatinmarkers, 5'-methyl cytosine (5meC) andmethylation of histone H3 on lysine K27(H3K27me2) in both GV and mature MII
oocytes.
RNA isolation, effects of BPS on gene
expression in transcriptionally silenced GV
oocytes.

No effect on oocyte quantity or quality
Effects on MII oocytes:
Increased proportion of abnormal oocytes at all doses based on chromatin and spindle
malformations
Increased abnormal spindles at 0.01, 10, and 100 µg/kg-day.
NS increase in abnormal chromatin.
Increased apoptosis at 0.01, 0.1, and 100 µg/kg-day
Methylation in immature GV oocytes:
Increased H3K27 dimethylation at 10 ng/g-day relative to control (other variations amonggroups)No effect on 5meC relative to the control group. Increased 5meC levels at 10 and 100 µg/kgday compared to the 0.1 µg/kg-day group.
Methylation in mature MII oocytes:
Increased 5meC in metaphase chromosomes at 0.1 µg/kg -day
NS effect on H3K27me2 levels at any dose.
Transcription effects in GV oocytes at 0.1 µg/kg -day: 102 genes affected: 89 genes wereupregulated (cellular stress, and markers of preimplantation and embryonic development) and13 genes were downregulated

Qin et al. 2021
Healthy adult wild-type AB zebrafish, 200 fertilized (2 hpf)
eggs per group.
Treatment: BPS (99.9% purity) in 0.002% DMSO, added to 1.5
L test solution at a final concentration of 0, 1 or 100 μg/L for 20
days (two-thirds of the exposure solution was renewed everyday).
At 20 dpf, larvae were transferred to 4.5 L volumes of test
solutions and then moved to 40 L, in a 50 L glass tank, from30 to 240 dpf
After 235 days of treatment, the proportion of males andfemales was approximately equal, with about 80 females pergroup. Four female fish per group were allowed to mate withuntreated males on the morning (8:30 - 10:30) of the 240th
day, and eggs were collected within 2 hpf.
After exposure, body sized of female fish was measured, andplasma collected by pooling blood samples per group.
Ovaries and liver were collected for further analysis

GSI, HSI
Histopathological analysis on left ovaries todetermine percentage of oocytes at different
developmental stages.
Lipid profiles in tissue homogenates:
triacylglycerol (TAG), total cholesterol (TC),high-density lipoprotein (HDL), low-density
lipoprotein (LDL), and VTG content from liver,left ovary and plasma.
Gene expression from right ovary
General toxicity: Body length and weight,
survival rate 24 hpf.

Increased HSI and GSI at 100 μg/L
Lipid accumulation in interstitial fibrosis tissue (tissue that surrounds the oocyte, necessary forsupplying nutrients to oocytes).
Follicle maturation:
Fewer primary growth stage and more full-grown stage oocytes at both concentrations
Increased number of cortical follicles at 1 μg/L
Increased vitellogenic (VS)-stage oocytes 100 μg/L
Increased egg production at both concentrations
VTG and lipid profiles:
VTG: Increased at 1 μg/L in plasma and at both doses in ovary; no effect in liver
TAG: Increased at both doses in liver and plasma and at low dose in ovary
TC: Increased at 1 μg/L in plasma and at both doses in ovary, no effect in liver
LDL: Decreased in liver, increased in plasma and ovary at 1 μg/L
HDL: Increased HDL at 1 μg/L in plasma, no effect in liver or ovary
Ovarian gene transcription:
Upregulation of pparα and pparγ at 100 μg/L
Upregulation of metabolic enzyme-related genes:
acc, fasn, acsl1, at 100 μg/L
Increased fatty acid β-oxidation gene expression (hsl, cpt2, and hsd17β4) at bothconcentrations; and acadm, acadl at 100 μg/L
General toxicity: Increased body weight and BMI at both concentrations, no effect on survival
rate: 87% in the controls, 75% at 1 μg/L and 82% at 100 μg/L

Urine was collected at 2, 4, 6-, 8-, 10-, and 12-hours postinjection for hormone measurement.

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Shi et al. 2017
CD-1 mouse pups, 5-6 litters per group.
Treatment: BPS (98% purity) in corn oil, (sc) injection (20 to 50μl) every three days at doses of 0, 50 μg/kg or 10 mg/kg, alldoses for 60 days (PND 0 to 60).
Litter size was adjusted to 10 pups (5 per sex) and all pups inthe same litter received the same treatment.
Half the females were sacrificed on diestrus day (after PND60). The remaining females were mated with untreated males

Vaginal opening monitored from PND 15
Monitor estrous cyclicity from PND 60
Body and uterine weights on half the F1
femalesSerum hormone levels: E2 and testosterone (T)on PND 60
One ovary for RNA isolation for steroidogenic
enzymes: Star, Cyp11a1, Cyp17a1, Hsd3b1,
Hsd17b1, and Cyp19a1
The other ovary for histological analysis
Mated females (n = 10–12/group): gestational
length, pup numbers, weights, and sex ratio.

No effect on vaginal opening, time to plug, or body and uterine weight.
Increased serum E2 at both doses
Increased serum T at 10 mg/kg
No effect on steroidogenic enzyme mRNA expression.
Three animals in the BPS 50 μg/kg dose group took over 10 days (up to 16 days) to become
pregnant.
No effects on gestational days, litter size, sex ratio or pup weights

Shi et al. 2019CD-1 pregnant mice, number of animals not specified but
analysis was made on 4–5 litters/group.
Treatment: BPS (purity not reported) in corn oil, daily oral
administration by placing a pipette tip in the mouth at doses of0, 0.5, 20, or 50 μg/kg-day from GD 11 to birth
One female pup from each litter was euthanized on PND 4
and ovaries fixed for histological analysis.
All other F1 females were mated at 3, 6, and 9 months of age
with untreated CD-1 males
Dams were sacrificed on PND 14 and ovaries saved for RNA
and histological analysis

Histological analysis of ovaries on one female
pup per litter at PND 4: germ cells in nests andfolliculogenesis
Puberty: vaginal opening assessed from PND
17
Estrous cyclicity for 30 days after vaginal
opening
Serum E2 and T in F1 females at 3, 6, and 9months of age
Ovarian steroidogenic enzymes: Star,
Cyp11a1, Cyp17a1, Hsd3b1, Hsd17b1, andcyp19a1 at 3, 6 and 9 months of age
In F1 mated females:
Time to vaginal plug
Pregnancy rate, pup numbers, weights, andsex ratio.

Ovarian histology: Retention of germ cells in nests at 0.5 and 20 μg/kg-day
Decreased ratio of primary follicles to total oocytes at 0.5, 20, and 50 ug/kg; no effects on
secondary follicles
NS decreases in the ratio of secondary follicles to total oocytes
No effects on the ratio of primordial follicles to total oocytes.
Vaginal opening (F1) and estrus cycle:
1–2 days earlier at 0.5 μg/kg-day
2.5 days delayed at 20 μg/kg-day
Irregular estrous cyclicity: several days in estrus and diestrus at all doses.
F1 female serum hormones:
Increased T at 50 μg/kg-day at 9 months
No effect on T at other doses or on E2 levels at any dose or age
Steroidogenic enzymes:
Increased Cyp11a1 mRNA at 0.5 μg/kg-day (at 3 months of age); no effect on other enzymes
analyzed.
Mating F1 at 6 and 9 months of age: Increased time to vaginal plug for all dose groups.
Decreased mating rate:
At 6 months: 25% in the low and mid dose groups
At 9 months: 34% at the low dose, and 60% at the two higher doses (100% in control)
Decreased number of F2 pups/litter at 20 μg/kg-day (NS decrease at 6 months)
No effects on F2 pup body weight or sex ratio.

Silva et al. 2019
Adult female gerbils (Meriones unguiculatus; 90 days old), 10
animals per group
N = 5 – 8 / analysis

Serum levels of E2 and T
Stereological (three-dimensional
measurements) analysis
Histology of prostatic complexes
Histopathological analysis of prostate:frequency of prostatic disorders

No effects on serum E2 or T levels
Three-dimensional analysis of prostate:
Increased epithelium and muscular stroma; decreased ductal lumen.
Histology of alveolar prostatic morphology:
Decreased % normal cells; increased hyperplasia; increased intraepithelial neoplasia.
No effects on PAS-positive prostatic secretion area.

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Treatment: BPS (purity not reported) in mineral oil, daily oral
exposure (100 µl total volume), at doses of 0 or 40 µg/kg-day
for 28 days
Twenty-four hours after treatment animals were sacrificed.
Blood samples and prostatic complexes (vaginal segment,
corresponding urethral segment and prostatic tissue) andovaries were saved for further analysis.

Histological sections stained by periodic acidSchiff test (PAS) reaction (secretionassessment).
Immunohistochemistry analysis: AR, ERα and
PCNA protein expression
General toxicity: Body and organs weight

Immunohistochemistry:
Increased frequency of AR-positive epithelial and stromal cells
Increased ERα immunolabeling in stromal cells
Increased PCNA-positive cell frequency in the epithelium and stroma
General toxicity: No alterations in body, prostate, or gonadal weight

Téteau et al. 2022
2.5 years old ewes (Ile-de-France), divided in two diet groups:
Restricted feed (RF) and well fed (WF, 165 % of foodrequirement). Twenty animals per BPS group
Treatment: BPS (purity not reported) daily administration in thediet at doses of 0 or 50 µg/kg-day for at least three months.
Ewes were synchronized for estrus and treated withgonadotropins (pregnant mare serum globulin, PMSG) to
induce follicular growth. Two days after PMSG administration(preovulatory time), ewes were sacrificed. Blood, andreproductive tract samples were collected and saved forfurther analysis.

BPS and BPS glucuronide measured in plasma
and in follicular fluid (FF).
Steroids in plasma, FF, and oviduct fluid (OF).
General toxicity: Body weight, body condition
score (BCS)

NS increase in ovary weight (p=0.051)
No effect on follicle number (any size)
BPS effects on steroid levels in plasma:
Decreased P and 5α-dihydroprogesterone (P metabolite) in RF group
Decreased E2 and increased E1 in RF group
BPS effects on steroid levels in preovulatory FF:
Decreased dihydrotestosterone (DHT) and 11-dehydrocorticosterone levels in RF group.
Increased E2 levels in WF (no effect in the RF group)
Decreased 17α-hydroxyprogesterone and increased 5β-dihydrotestosterone in RF group.
There was an interaction between BPS and diet for: E2 (p =0.020), hydroxy pregnenolone (p =0.028) and DHT (p = 0.001), and E2 (when measured separate p= 0.002)
BPS effects on steroid levels in OF:
Decreased E2 in the WF group and E1 in the RF group
General toxicity:
WF ewes had higher body weight and BCS compared to RF (but they were not overweight).
There was a BPS effect on BCS (p = 0.026).

Tucker et al. 2018
Pregnant CD-1 mice (GD 8.5), at least 11 animals per group
Treatment: BPS (≥ 97:5% purity) in sesame oil, twice daily(every 7h) by oral gavage (10 μL/g body weight) at doses of 0,0.05, 0.5, or 5 mg/kg twice a day (equivalent to 0.10, 1.0, and10 mg/kg-day, respectively) from GD 10.5 to GD 17.5
(Parturition GD 18.5 –- 19)
Analysis on F1 females at PND 20 to 14 months (1 or 2 perlitter)Observations were made on F1 females at
PND 20, 35, 56, and at 3, 8, and 14 months ofage.
Female pubertal indices: vaginal opening and
estrus cyclicity; mammary development/scorechanges in mammary gland whole mounts. In
addition, changes in gene expression andmammary gland histopathology were analyzed
at 3, 8 and 14 months.
Serum hormone levels: E2, P, T, and DHEA
mRNA for steroid receptor gene expression at
8 and 14 months
General toxicity: Body weight

No differences in age at vaginal opening; first estrus (PND 25 – 27) or estrus cyclicity (3 – 4
days)
Mammary gland
PND 20: Dose-dependent increases in TEB length and TEB counts
Increased branching density at 0.05 and 5 mg/kg twice a day
Increased developmental score at 5 mg/kg twice a day
No effects on mammary epithelial area
PND 35: Increased developmental score at all doses
PND 56: Increased developmental score at 0.5 mg/kg twice a day.
3 months of age:
Increased TEB counts at 0.5 mg/kg twice a day
Increased incidence of diagnoses of mixed cell inflammation at 0.5 mg/kg twice a day (25%inflammation and 50% increased TEB)
14 months of age:
Increased incidence of perivascular lymphoplasmacytic inflammation at 0.5 mg/kg twice a day
Increases in all types of inflammation diagnoses at 0.5 mg/kg twice a day
Increased non-neoplastic lesions at 0.5 mg/kg twice a day
Increased incidence of lobuloalveolar hyperplasia at 0.5 mg/kg twice a day
Serum hormone analysis:

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Major Findings
Increased E2 at the mid and high doses on PND 20 (no effects at other doses/times)
Increased P and DHEA at 0.05 mg/kg twice a day on PND 20 (NS increase at mid and highdose on PND 56); decreased P at 5 mg/kg twice a day at 3 months of age
No effect on P, DHEA or E2 levels on PND 28
Increased T at 0.05 mg/kg twice a day on PND 28
Decreased T at mid and high dose on PND 35 (no effects on PND 56)
Decreased E2 at 0.05 mg/kg twice a day. Decreased T and DHEA at 0.5 mg/kg twice a day at
8 months of age
No effects at 14 months of age on any hormone analyzed
Mammary gland nuclear receptor gene expression:
Decreased Pgr expression at 5 mg/kg twice a day at 8 months. No other gene expression
changes at 8 or 14 months
General toxicity: Decreased body weight on PND 56 at 5 mg/kg twice a day (no effects onbody weight at other times or doses)

Wang et al. 2020
Six-month-old adult female wild-type (AB strain) zebrafish, 40
females/group
Treatment: BPS (≥ 98% purity) in 0.002% DMSO added to 4 L
of water (refreshed every 24 hours) at doses of 0, 1, 10, and100 µg/L for 14 days

Test 1: adult female behavioral analysis andobservation of body pigment and gonad
structure
Test 2: 4 adult females and 4 adult males werepaired and spawned for 0.5 d.
Oocyte classification: perinucleolar oocytes,
cortical alveolar oocytes, late vitellogenic
oocytes, and spawning oocytes.
General toxicity: Body weight and length

Debilitated female shoaling at 100 µg/L: Decreased proportion of time spent in the zone close
to fish (1.23-fold reduction), increased time spent in the zone far from fish (3.01-fold)
Ovarian histology (effects at all doses):
Increased perinucleolar oocytes
Decreased cortical alveolar oocytes
Decreased late vitellogenic and spawning oocytes
General toxicity: No effects on body weight or body length. Females presented depigmentation
at all concentrations.
.

Yamasaki et al. 2004
Twenty-day-old SD female rats, six rats per group
Treatment: BPS (99.4% purity) in olive oil (vehicle control),daily sc injection at doses of 0, 20, 100, and 500 mg/kg-day on
three consecutive days
Positive control group: EE in olive oil was also sc injected tosome rats at a dose of 0 or 0.6 μg/kg-day on three consecutive
days right after BPS administration.
Another group was injected with tamoxifen at 1 mg/kg-day
plus EE on three consecutive days

Uterus weight (uterotrophic assay)
Estrogen receptor assay

Effects on uterine relative weights:
Increased at 20 and 500 mg/kg-day BPS
Increased with EE positive control
Tamoxifen reduced the EE effect on uterine weight
BPS + EE effects:
BPS at 20 mg/kg-day increased uterine weight further than the EE observed effect
BPS at 500 mg/kg-day decreased the EE effect on uterine weight
BPS - Estrogen binding affinity assay = 0.0055 % of E2

Yue et al. 2023b
Mature female (8 to 9 weeks old) CD-1 (ICR) mice, 9 mice per
group

Serum hormone levels: E2, P, FT3, FT4.
Histological examination:
Ovary follicle analysis
Uterus, measurement of endometrial area andglandsNo effects on serum hormones analyzed (E2, P, FT3, and FT4)
Effects on follicles: (author description) Damaged oocyte structure, wrinkled and depressedzona pellucida, atrophy in immature follicles, increased number of atretic follicles
Effects in uterus: Reduced endometrial area, atrophied lamina propria, myometrium andplasma layer, and narrowed uterine cavity.
Increase in number of uterine glands

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Treatment: BPS (purity not reported) in corn oil (volume notspecified) daily by oral gavage at doses of 0 or 300 µg/kg-day
for 28 consecutive days.
After exposure, blood samples and uterus and ovary werecollected for further analysis.

RNA sequencing: Identification of DEG
Gene Ontology enrichment analysis.
Associations among BPS, hub genes, anddiseases using the Comparative
Toxicogenomic Database online website(http://ctdbase.org/)

No effects on uterus weight.
Gene Ontology analysis in uterus and ovary:
Affected metabolic hormone related pathways in uterus and ovary (steroids and sterol
metabolic process), immune and inflammatory related pathways in uterus and ovary.
Gene - Disease associations: Uterine genes included were associated with mammary glandneoplasms and breast cancer. Ovarian genes were associated with polycystic ovary
syndrome, hypercholesterolemia, ovarian neoplasm, ovarian and breast carcinoma,
endometriosis, estrogen resistance, and others.

Zhang et al. 2020
ICR 8-week-old pregnant female mice, at least 3 animals pergroup.
Treatment: BPS (purity not reported) in 1% DMSO diluted innormal saline, daily oral administration (volume not specified)at doses of 0, 2, or 10 μg/kg-day for 4 days (GD 12.5 to 15.5)
NOTE: In a dose finding study, six animals per group weretreated with BPS at doses of 0, 2, 10, 50, 100, or 200 μg/kgday for 3 days (GD 12.5 to 15.5) following the same treatment
protocol. All doses had effect on meiosis prophase I
progression, therefore all subsequent studies used BPS dosesof 2 or 10 μg/kg-day.
Complete oocytes were isolated from female mice aged 5
weeks and used for oocyte culture.
MII oocytes from COCs from 5-week old F1 oviducts used for
IVF and blastocyst culture

meiosis prophase I progression in ovaries from
GD 15.5 fetuses.
Gene expression and protein levels formeiosis-related genes in PND 15.5 ovaries:
deleted in azoospermia-like (Dazl), stimulated
by retinoic acid 8 (Stra8), synaptonemal
complex protein 3 (Scp3), dosage suppressorof mck1 homolog (Dmc1) and its related protein
levelsGene expression and protein levels for genes
related to oocyte quality in PND 21 ovaries:
bone morphogenetic protein 15 (Bmp15) andgrowth differentiation factor 9 (Gdf9) and its
related protein levels.
Germ cell cyst breakdown assessed in F1
ovaries at PND 3, and folliculogenesis
assessed in F1 ovaries at PND 21
Oocyte maturation, fertilization (IVF), and early
embryo development of F1 females assessedat five weeks
Epigenetic alterations in F1 oocytes
Ovarian effects in F2 generation

Acceleration in meiotic progression (at all doses):
Decreased percentage of oocytes at zygotene stage,increased percentage at pachytene and diplotene stage
Meiosis related gene and protein expression (PND 15.5):
Increased expression of Stra8, Scp3, and Dmc1 at both doses and Dazl at 10 μg/kg-day
Increased protein levels of DAZL and STRA8 at both doses and SCP3 at 10 μg/kg-day
Oocyte quality related genes and protein expression:
Dose related decreases in gene expression and protein levels of BMP15 and GDF9Germ cell cyst breakdown (PND 3):
Increased MVH positive oocytes in primordial follicles at both doses
Follicular development (F1, PND 21):
Increased oocytes in primordial follicles and decrease in secondary and antral follicles on PND
21 at both doses
No effect on total number of oocytes
Decreased rates of germinal vesicle breakdown and release of first polar body at both doses
Increased rates of aberrant spindles and misaligned chromosomes at both doses
Effects on IVF:
Decreased rates of cleavage and blastocysts after in vitro fertilization at both doses
Decreased rates of 4-cell embryos (dose-dependent manner) at both doses
Decreased rate of blastocysts at both doses
Epigenetic effects on F1 oocytes at both doses:
Decreased levels of trimethyl-histone H3 (Lys4) (H3K4me3)
Increased levels of H3K9me3
Effects on F2 generation:
No effects on litter size, body weight or sex ratio
Acceleration of germ cell cyst breakdown on PND 3 ( no change in total number of follicles)
No effects at PND 21

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4.3 Mechanistic considerations and other data relevant to female reproductive
toxicity
Findings from studies on the possible mechanisms of action of bisphenol S (BPS)
relevant to considerations of female reproductive toxicity are discussed in this section.
These findings, which come from in vivo and in vitro studies, are then discussed in thecontext of the key characteristics of female reproductive toxicants. Table 4.3.1, at theend of this section, summarizes key elements of the mechanistic studies, except for
those in vivo studies already presented in Table 4.2.1.
Several possible mechanisms of action of BPS relevant to its effects on the female
reproductive system have been investigated, including mechanisms related to its
estrogenic activity. Like bisphenol A (BPA), BPS can mimic and interfere with the actionof estrogen. BPS modulates estrogen receptor (ER)-mediated effects through several
different ER subtypes, such as ERα, ERβ, and the plasma membrane G-proteincoupled estrogen receptor (GPER), as well as their downstream signaling pathways.
These effects have been observed in various studies conducted on human cells in vitro,
as well as in animal studies conducted in vivo and in vitro. The evidence supportingthese findings has been extensively discussed in multiple reviews (Chen et al. 2016a;
NTP 2017; Pelch et al. 2019; Rochester and Bolden 2015).
4.3.1 Effects on ovarian development and maturation of oocytes
The effects of BPS on ovarian physiology and follicle development, including oocyte
maturation, were presented in section 4.2. Studies reporting mechanistic data relevantto these physiologic effects are briefly summarized below:
In vitro studies
Oocyte maturation••

Newborn mouse ovaries cultured for 3 days in vitro in the presence of BPS at 10
to 100 μM underwent abnormal germ cell cyst breakdown, starting at 10 μM. Thiseffect was blocked by co-incubation with 1 μM tamoxifen, a selective estrogen
receptor modulator (SERM) with the potential for both ER agonism andantagonism, suggesting an estrogenic mode of action for BPS (Liu et al. 2021).
Pig ovary cumulus–oocyte complexes (COCs) incubated with BPS in vitro
resulted in alterations in oocyte maturation. A concentration dependent decreasein oocytes reaching metaphase I (MI) and a failure to resume meiosis occurred
after 24 hours at 300 nanomolar (nM) and 30 micromolar (µM) BPS,

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concentrations that had no effect on oocyte or cumulus cell viability after 24 or 48
hours. There was also a concentration dependent decrease in progression tometaphase II (MII) at 48 and 72 hours. All BPS-treated oocytes matured to atleast the MI stage by 48 hours but by 72 hours the maturation process wasdelayed, disrupted and/or blocked at all concentrations (Žalmanová et al. 2017).
Spindle morphology and chromosome alignment•••

Cow oocytes treated with BPS in vitro from 1 femtomolar (fM) to 50 µM resulted
in alterations in spindle morphology and chromosome alignment in MII oocytes at
BPS concentrations greater than 1 nM without altering the proportion of oocytes
entering MII (Campen et al. 2018a).
Pig ovary COCs incubated with BPS at 3 nM, 300 nM or 30 µM for 24 or 48
hours resulted in α-tubulin assembly alterations, including a concentration
dependent decrease in the number of tubulin filaments. This translates into
swollen chromosomes and irregular organization, spindles in circular formation,elongated metaphase plates, and reduced spindle size at all doses in MI and MII
oocytes (Žalmanová et al. 2017).
Pig ovary COCs incubated with BPS at 300 picomolar (pM), 30 nM or 3 µM for 48
hours resulted in a concentration dependent decrease in the proportion ofoocytes reaching MII, with increases in degenerated oocytes, retention in thegerminal vesicle (GV) stage, arrest in the MI stage, spindle disorganization, andchromosome misalignment at all concentrations (Žalmanová et al. 2023).

Follicular cells (granulosa and theca cells)•••

Adult ewe ovarian granulosa cells incubated with BPS at 200 µM for 48 hours
resulted in a decrease in granulosa cell proliferation (Téteau et al. 2020).
In human and sheep primary cultures of ovarian theca cells, BPS treatment for
24 hours at up to 1000 ng/mL in sheep cells, and at 200 ng/ml in human cells
increased theca cell gap junction intercellular communication (GJIC) atconcentrations above 100 ng/mL (Gingrich et al. 2021).
Signaling pathways such as those involving protein kinase C (PKC), protein
kinase A (PKA), mitogen-activated protein kinase (MAPK), phosphatidyl choline
specific phospholipase C (PC-PLC), and phosphatidylinositol-specificphospholipase C (PI-PLC) may be required for proper cellular proliferation.
Incubating human and sheep theca cells with specific inhibitors for thesepathways to assess effects on BPS stimulated GJIC resulted in full inhibition(back to control values) by a MAPK inhibitor (human and sheep theca cells);
partial inhibition with inhibitors of PI-PLC, PC-PLC, and ERK-MAPK; and no

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•

effect with PKA or PKC inhibitors (sheep theca cells only). These findings
indicate that BPS can modulate GJIC through the MAPK pathway and partiallythrough PC-PLC and PI-PLC and that these effects are independent of PKA and
PKC activation (Gingrich et al. 2021).
Cow ovary COCs, denuded oocytes, and cumulus cells (comprised of granulosa
cells) incubated with BPS at 0, or 0.05 mg/mL for 24 hours resulted in changes insome COC cells in the expression of connexins (Cx – gap junctional proteins):
Cx37 and Cx43. There was an increase in Cx37 mRNA expression in cumulus
cells with a NS increase in Cx37 protein expression, and no effects on Cx43 orCx37 mRNA or protein expression in COCs as a whole (Sabry et al. 2021).

Mitochondria•

Pig ovary COCs incubated with BPS at 30 nM and 3 µM for 48 hours resulted inlower relative intensity in fluorescence (about a 30% decrease) for mitochondria
fusion protein markers, MFN1 and MFN2, and a relative increase in fluorescenceat 30 nM (Žalmanová et al. 2023).

In vivo studies
Relevant mechanistic data from in vivo studies of BPS on ovarian development andmaturation of oocytes presented in Section 4.2 and Table 4.2.1 include the following:• Chromosome misalignment and/or incidence of spindle malformation in mice
(Nevoral et al. 2018; Zhang et al. 2020), and C. elegans (Chen et al. 2016b)• Altered gene expression in mice (Liu et al. 2021; Prokešová et al. 2020; Yue etal. 2023b; Zhang et al. 2020), chickens (Eldefrawy et al. 2021), and zebrafish(Qin et al. 2021)• Apoptosis in mice (Nourian et al. 2020) and C. elegans (Chen et al. 2016b)• Epigenetic effects in mice (Nevoral et al. 2018; Prokešová et al. 2020; Yue et al.
2023b; Zhang et al. 2020)• Effects on the signaling pathway involving Notch2 and Jagged1, which isimportant in cyst breakdown and primordial follicle assembly in mice (Liu et al.
2021)• Effects on lipid profiles in mice (Yue et al. 2023b), and zebrafish (Qin et al. 2021)• Indicators of oxidative stress in mice (Nevoral et al. 2018; Nourian et al. 2017,2020) and rats (Ijaz et al. 2020).

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4.3.2 Effects in placenta
The effects of BPS on the placenta were presented in section 4.2. Studies reporting
mechanistic data relevant to these physiologic effects are briefly summarized below:
In vitro studies••

BPS treatment of a human cell line (HTR-8/SVneo) derived from extravillous
trophoblast cells from a first trimester pregnancy at concentrations ranging from
10-9 M to 10-7 M resulted in increased cell proliferation (at all concentrations).
This increased cell proliferation was mediated through the ER and extracellularsignal-regulated kinase 1/2 (ERK1/2) pathways. Inhibition of ERK1/2
phosphorylation by BPS induced secretion of the inflammatory cytokinesinterleukins (IL) 6 and 8 (Profita et al. 2021).
In the human trophoblastic 3A placental cell line CRL-1584 BPS altered ABCB1
promoter activity in a haplotype-dependent manner. Changes in ABCB1
promoter activity affects P-glycoprotein levels (P-gp, placental transporter); in thehuman placenta, efflux of P-gp affects trophoblasts and fluctuations in P-gp
levels affect fetal development (Speidel et al. 2018).

In vivo studies
Relevant mechanistic data from in vivo studies of BPS on the placenta presented in
Section 4.2 and Table 4.2.1 include the following:•

Findings in mouse placenta (Mao et al. 2020)• Gene expression: 11 genes were differentially expressed with one
upregulated, Actn2 (actinin α2), and 10 downregulated including Calm4(calmodulin 4) and Guca2a (guanylate cyclase activator 2a, guanylin)• Effect on fatty acids: Decreased stearic acid, palmitic acid, docosahexaenoic
acid, octadecenoic acid, and hexadecanoic (palmitic acid)• Effects on neurotransmitters: Increased dopamine levels, increased
percentage of dopamine-positive giant cells; decreased serotonin levels anddecreased percentage of serotonin-positive giant cells•

Findings in sheep placenta (Gingrich et al. 2018)• 50% lower e-cadherin protein expression in GD 120 placentomes• 20% decrease in trophoblast derived binucleate cells• Decreased expression of fusogenic genes: JSRV and HYAL2Increasedtranscription factor glial cell missing factor 1

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4.3.3 Effects on the endocrine system
Endocrine effects of BPS observed in vivo in animal studies were presented in section
4.2. Endocrine effects observed in vitro, as well as other relevant mechanistic data arebriefly summarized below:
In vitro studies
Progesterone
•
•

•
•

•

Decreases in progesterone (P) levels were reported in primary cultures of humangranulosa cells, at BPS doses of 10 and 50 µM (Amar et al. 2020).
A concentration dependent decrease in P secretion was reported in adult ewe
granulosa cells incubated with BPS exposure at concentrations ranging from 10
to 200 µM (Téteau et al. 2020).
Decreases in P levels were reported in ewe granulosa cells treated with BPS at
10 or 50 µM for up to 48 hours (Téteau et al. 2023).
Increased P and 17-OH-progesterone levels were reported in H295R cells (ahuman adrenal derived cell line) treated with BPS from 0.8 to 50 μM (Rosenmai
et al. 2014).
In cow theca cells isolated from antral follicles, in vitro treatment with BPS from 1
fM to 100 µM did not affect P secretion (Campen et al. 2018b).

Androgens••

In cow theca cells isolated from antral follicles, in vitro treatment with BPS from 1
fM to 100 µM did not affect androstenedione secretion (Campen et al. 2018b).
In the human adrenal cell derived cell line H295R there was a decrease intestosterone (T), androstenedione and dehydroepiandrosterone (DHEA) after
BPS exposure (Rosenmai et al. 2014).

Estradiol•••

Decreases in estradiol (E2) levels were reported in primary cultures of humangranulosa cells exposed to 50 µM BPS for 48 hours (Amar et al. 2020).
Decreases in E2 levels were reported in ewe granulosa cells treated with BPS at
10 and 50 µM for 48 hours (Téteau et al. 2023).
A concentration dependent increase in E2 levels was reported in ewe granulosa
cells treated with BPS from 1 nM to 200 µM for 48 hours (Téteau et al. 2020).

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••

Increases in E2 secretion were reported in cow granulosa cells isolated fromantral follicles after BPS exposure at 100 µM (Campen et al. 2018b; Žalmanová
et al. 2017).
No effects on E2 or estrone (E1) levels were reported in H295R cells (a humanadrenal derived cell line) after treatment with BPS from 0.8 μM to 50 μM(Rosenmai et al. 2014).

Estrogen receptor
•

•
•

•

•
•

In pig ovary COCs treated with BPS at 3 nM, 300 nM or 30 µM there weredecreases in Erα gene expression in oocytes at all doses, decreases in ErβmRNA in cumulus cells at 30 µM, and increases in ER (α and β) protein levels at
300 nM (Žalmanová et al. 2017).
Exposure of Ishikawa cells (a human endometrial epithelial cell line) to 100 nM
BPS increased gene expression of ESR2, but not ESR1 (Benjamin et al. 2023).
In ovaries from newborn mice that were cultured for 3 days in the presence of
BPS at 10 μM increases in ER (α and β) mRNA and protein levels were observed(Liu et al. 2021).
Granulosa cells from adult ewes treated with BPS from 1 nM to 200 µM for 48
hours increased Esr2 expression at 50 μM and a two-fold increase in Esr1 andEsr2 expression at 100 µM (Téteau et al. 2020).
In H295R cells treated with BPS from 0.8 μM to 50 μM, there was an increase in
ER activity (Rosenmai et al. 2014).
No effect on the in vitro response of ewe granulosa cells to BPS was reported in
the presence of the GPER antagonist G-15, suggesting the decreases in P andE2 levels observed in BPS treated cells (discussed above) were not mediated by
GPER (Téteau et al. 2023).

Other hormone receptors•••

In granulosa cells from adult ewes treated with BPS from 1 nM to 200 µM for 48
hours no effects were observed on androgen receptor or progesterone receptor
gene expression (Téteau et al. 2020).
In H295R cells treated with BPS from 0.8 μM to 50 μM, there was an apparent
concentration-related decrease (NS) in androgen receptor (AR) activity(Rosenmai et al. 2014).
In cow COCs and in vitro fertilized cow embryos incubated with BPS at 0.05
mg/mL there was an increase in anti-müllerian hormone receptor II (AMHRII)mRNA levels and a non-significant increase in protein levels reported in COCs,
and an increase in AMHRII protein levels in day-8 blastocysts (Saleh et al. 2021).

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Steroidogenic enzymes••

In pig ovary COCs, treatment with BPS at 3 nM, 300 nM or 30 µM resulted indecreased aromatase gene expression in oocytes (at 3 nM and 300 nM) andcumulus cells (at 30 µM) and decreased aromatase protein levels in oocytes (at 3nM) (Žalmanová et al. 2017).
In primary cultures of human granulosa cells there was a non-significant
decrease in CYP11A1 (P450 side chain cleavage [P450scc]) protein levels(37.7%; p= 0.09) at 10 µM (Amar et al. 2020).

In vivo studies
Relevant mechanistic data from in vivo studies of BPS and its effects on the endocrine
system presented in Section 4.2 and Table 4.2.1 include the following:
Hormone levels
•

•
•

•
•
•
•

•
•

Decreased gonadotropin levels (follicle stimulating hormone and luteinizing
hormone) in rats (Ahsan et al. 2018; Ijaz et al. 2020) and mice (Nourian et al.
2020).
Decreased P plasma levels in rats (Ahsan et al. 2018; Ijaz et al. 2020) and mice(at 5 mg/kg twice a day) (Tucker et al. 2018)
Increased P levels in zebrafish (Park et al. 2022) and in mice at 0.05 mg/kg twice
a day a dose that it was notably smaller than the dose that resulted in decreased
P in the same study (Tucker et al. 2018).
Increased plasma T levels in rats (Ahsan et al. 2018; Ijaz et al. 2020) and mice
(Shi et al. 2017; Shi et al. 2019) (Tucker et al. 2018)
Decreased plasma T levels in mice (at 0.5 and 5 mg/kg twice a day on PND 35)(Tucker et al. 2018).
Decreased preovulatory follicular fluid dihydrotestosterone and 11dehydrocorticosterone levels in ewes (restricted feed diet) (Téteau et al. 2022).
Decreased plasma E2 levels in mice (LaPlante et al. 2017; Nevoral et al. 2018;
Pollock et al. 2019), rats (Ijaz et al. 2020), and ewes (restricted feed diet) (andincreased plasma E1) (Téteau et al. 2022).
Increased E2 levels in mice (Shi et al. 2017; Tucker et al. 2018) and zebrafish (Jiet al. 2013; Naderi et al. 2014; Park et al. 2022).
Increased preovulatory follicular fluid E2 levels in ewes (well fed diet) (Téteau etal. 2022)

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Hormone receptors
•
•
•
•

•

•

•

•

•

Increase in P receptor (PR) protein levels in mammary glands of pubertal mice
(Kolla et al. 2018)
Decrease in PR gene expression in mammary glands of adult mice (Tucker et al.
2018)
Increase in AR positive epithelial and stromal cells in the female prostate in
gerbils (Silva et al. 2019).
Increased ERα gene or protein expression in the mammary glands of mice (Kolla
and Vandenberg 2019; LaPlante et al. 2017), stromal cells of the female prostatein gerbils (Silva et al. 2019), and in vitro fertilized embryos from BPS exposed
mice (Nourian et al. 2020).
An ethinyl estradiol (EE) challenge altered ovarian ERα and Erβ gene expression
in BPS exposed mice: non-significant increases in ERα and Erβ mRNA levels
occurred with BPS alone, while decreases in Erβ mRNA and non-significant
decreases in ERα mRNA occurred with BPS + EE (Hill et al. 2017).
BPS exposure of mice was associated with a greater number of secondary
follicles, with no effects on other follicle types. Co-exposure with BPS + EE wasobserved to inhibit the increases in total follicles induced by EE alone (Hill et al.
2017).
BPS exposure was associated with a classical response in female rats indicativeof ER-mediated estrogenic activity, increasing uterine weight. Co-exposure with
EE and BPS at 20 mg/kg-day was observed to increase the uterotrophic effect of
EE alone, while co-exposure with EE and BPS at 500 mg/kg-day was observed
to diminish the uterotrophic effect of EE alone (Yamasaki et al. 2004).
In a rat model of induction of PCOS by DHEA, BPS + DHEA was observed todecrease the average number of cysts per ovary compared to treatment with
DHEA alone (Demacopulo and Kreimann 2019).
BPS exposure was observed to alter a classical ER-mediated response in fish,i.e., hepatic vitellogenin (VTG) expression. BPS was observed to decrease
hepatic VTG mRNA levels in zebrafish, indicating altered ER signaling (Park et
al. 2022).

Steroidogenic enzymes••

Increased ovarian Cyp11a1 (P450scc) mRNA levels in mice (Shi et al. 2019)
Decreased brain mRNA levels of one isozyme of 5α-reductase in rats (Castro etal. 2015).

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4.3.4 Key characteristics of female reproductive toxicants
The key characteristics (KCs) of female reproductive toxicants, as described in Luderer
et al. 2019, are listed in Table 4.3.1. One or more of these key characteristics are
frequently exhibited by exogenous agents that cause female reproductive toxicity(Luderer et al. 2019).
Table 4.3.1 Key characteristics of female reproductive toxicants (source: Luderer
et al. (2019)
 KC 1 Alters hormone receptor signaling; alters reproductive hormone
production, secretion, or metabolism
 KC 2 Chemical or metabolite is genotoxic
 KC 3 Induces epigenetic alterations
 KC 4 Causes mitochondrial dysfunction
 KC 5 Induces oxidative stress KC 6 Alters immune function
 KC 7 Alters cell signal transduction
 KC 8 Alters direct cell–cell interactions
 KC 9 Alters survival, proliferation, cell death, or metabolic pathways
 KC 10 Alters microtubules and associated structures

A related set of key characteristics, those of endocrine-disrupting chemicals, has been
described in La Merrill et al. (2020), and is presented in Table 4.3.2.
Table 4.3.2 Key characteristics of endocrine-disrupting chemicals (source: (LaMerrill et al. 2020).











EDC KC 1 Interacts with or activates hormone receptors
EDC KC 2 Antagonizes hormone receptors
EDC KC 3 Alters hormone receptor expression
EDC KC4 Alters signal transduction in hormone-responsive cells
EDC KC5 Induces epigenetic modifications in hormone-producing or hormoneresponsive cells
EDC KC6 Alters hormone synthesis
EDC KC7 Alters hormone transport across cell membranes
EDC KC8 Alters hormone distribution or circulating hormone levels
EDC KC9 Alters hormone metabolism or clearance
EDC KC10 Alters fate of hormone-producing or hormone-responsive cells

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Relevant findings from studies of BPS can be considered in the context of the key
characteristics of female reproductive toxicants and of endocrine-disrupting chemicals.
Here we focus on the key characteristics of female reproductive toxicants, andsummarize findings related to KC 1, KC 2, KC 8, and KC 10.
KC 1 Alters hormone receptor signaling; alters reproductive hormone production,
secretion, or metabolism.
•

A large number of studies, all of which are presented in section 4.3.3, above,report findings relevant to KC 1. Please see section 4.3.3.

KC 2 Chemical or metabolite is genotoxic.
•

In pig ovary COCs, BPS treatment resulted in spindle disorganization,
chromosome misalignment, and increased aneuploidy in oocytes (Žalmanová etal. 2023).

KC 8 Alters Direct Cell-Cell Interactions•••

In cow ovary cumulus cells (i.e., granulosa cells), incubation with BPS alteredexpression of connexin (Cx) 37, one of multiple connexins that regulate gap
junctional intracellular communication. Specifically, an increase in Cx37 mRNA
expression and a non-significant increase in Cx37 protein expression wasreported, with no effect of BPS on Cx43 mRNA or protein expression (Sabry et
al. 2021).
Mice ovaries cultured for 3 days in vitro with BPS from 10 to 100 μM resulted inabnormal germ cell cyst breakdown starting at 10 μM. Germ cell cyst (or nest)
breakdown is a normal physiological step in the development of the ovary, and it
is mediated by activation of the ER. Activated ER stimulates the c-Jun N-terminal
kinases (JNK) signaling pathway to modulate the expression of E-cadherin,
which is a cell adhesion protein and is involved in cell-to-cell interactions
between oocytes in the germ cell cysts. The BPS effect on the normal cyst
breakdown, was blocked by co incubation with SERM tamoxifen, suggesting an
estrogenic mode of action for BPS (Liu et al. 2021).
In human and sheep primary ovarian theca cell cultures, BPS treatment
increased of theca cell gap junction intercellular communication (GJIC) (Gingrichet al. 2021).

KC 10 Alters microtubules and associated structures.
•

BPS treatment of oocyte from cows in vitro from 1 fM to 50 µM resulted inalterations on spindle morphology and chromosome alignment in MII oocytes at

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•

•

•

BPS concentrations greater than 1nM without altering the proportion of oocytes
entering MII (Campen et al. 2018a).
Pig ovary COCs incubated with BPS at 3 nM, 300 nM or 30 µM for 24 hours or
48 hours, resulted in α-tubulin assembly alterations including a concentration
dependent decrease in the number of tubulin filaments. This translates into
swollen chromosomes and irregular organization, spindles in a circular formation,elongated metaphase plates, and reduced spindle size at all doses in metaphase
I and II (Žalmanová et al. 2017).
In COCs from pig ovaries, there was a concentration dependent decrease inproportion of cells reaching MII: retention in GV stage, degenerated oocytes,
arrested in MI stage, spindle disorganization and chromosome misalignment at
all concentrations (Žalmanová et al. 2023).
Chromosome misalignment and incidence of spindle malformation have beenreported in BPS exposed mice (Nevoral et al. 2018; Zhang et al. 2020), and C.
elegans (Chen et al. 2016b).

Summary Table
Data discussed above on female reproductive toxicity from studies with mechanistic
data not already included in Table 4.2.1 are summarized in Table 4.3.3 below. The
results presented are statistically significant (p<0.05), unless otherwise stated (i.e., nonsignificant [NS] change). The studies are organized alphabetically by first author.

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Table 4.3.3 Female reproductive toxicity: studies with mechanistic data not already included in Table 4.2.1
Study Design

Outcomes Assessed

Amar et al. 2020

Hormone levels: progesterone (P), estradiol (E2)

Human granulosa cells in vitro from 59 women

Steroidogenesis:
Western Blot Analysis of BPS 10 and 50 µM treatment groups for 3beta-hydroxysteroid dehydrogenase (HSD3B1), cytochrome P450
family 11 subfamily (CYP11A1), and cytochrome P450 family 19(CYP19A1) proteins

Treatment: BPS (purity not reported) in the culture
medium at 10 nanomolar (nM), 100 nM, 1 micromolar(µM), 10 µM or 50 µM
Cell cultures were assessed 48 hours after treatment
began for viability, steroidogenesis, cell proliferation andgene and protein expression

Gene expression
Mitogen-activated protein kinase (MAPK 3/1) signaling pathway
assays (at 5, 10, 30 or 60 min) in the presence or absence of 10 µM
BPS
BPS glucuronide was quantified in follicular fluid
General toxicity: Viability, cell proliferation

Benjamin et al. 2023
Ishikawa (ECACC 99040201) cells (human endometrial
epithelial cells). 250,000 cells/well on a 6-well plate
Treatment: BPS (purity not reported) dissolved indimethyl sulfoxide (DMSO, vehicle control) then diluted in
distilled water to a concentration of 0, 1 nM, or 100 nM
for 72 hours

Major Findings
Hormone levels: Decreased P at 10 µM (by 16%) and at 50 µM BPS (by 64%)
Decreased E2 at 50 µM BPS (by 46%)
Steroidogenesis NS decrease in CYP11A1 expression levels (37.7%; p=0.0947)
at 10 µM BPS
No effect on protein levels of HSD3B1 (at 10 µM) or CYP19A1 (at 50 µM)
Gene Expression
Increased ESRRG (estrogen-related receptor γ) expression (1.9-fold) at 50 µM.
BPS treatment had no effect on the other analyzed genes (STAR, CYP17A1,
androgen receptor (AR), progesterone receptor (PR), and ERs)
Signaling Pathways
No effect on MAPK3/1 phosphorylation (at 10 µM)
General toxicity: No effects on cell viability or cell proliferation

Gene expression:
Estrogen receptors (ESR1, ESR2)
Proliferation genes (CCND1, CCNB1, GREB1A, CMYC)
Migratory gene (VIM)
Wound healing assay: estimation of time for cells to reachconfluence, assessed 2 hours after scraping the cell monolayer inthe presence of mitomycin C (DNA synthesis inhibitor)

Increase in cell number: 3 to 8-fold increase in cell proliferation
Wound healing assay: Increased cell migration capacity at 1 nM
Increased gene expression of ESR2 at 100 nM
No effect on proliferation genes
Increased expression of migratory gene VIM
General toxicity: Increased cell viability

General toxicity: Cell viability
Campen et al. 2018a
In vitro: Cumulus–oocyte complexes (COCs) from cow
ovaries (Holstein-Friesian and Jersey heifers).
Treatment: BPS (purity not reported) in culture medium
at concentration of 0 or from 1 femtomolar (fM) to 100
nM (in 10-fold increments), or 50µM

Number of oocytes that reached MII
Analysis of spindle and chromosome configuration. Microfilaments,
microtubules, and DNA were detected by immunocytochemistry
analyses of each oocyte

A total of 432 oocytes were analyzed for meiotic stage,
including 110 from the vehicle- control group. Of the 432
oocytes analyzed, 350 were at metaphase II (MII) (91 MII

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No effect on the proportion of oocytes that had matured to MII by 24 hours.
In the control group, 85.7% of MII oocytes had a bipolar spindle and 93.4% hadaligned chromosomes.
BPS had overall effects on spindle morphology and chromosome alignment:
Reduced proportion of oocytes with a bipolar spindle at 10 fM, 1 picomolar (pM),
100 pM, 1 nM,10 nM, 100 nM or 50 µM.
(No effect on spindle morphology at 1 fM, 100 fM or 10 pM).
Increased flattened spindles with broad poles representing 17% of abnormalities
at 100 nM.

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Study Design

Outcomes Assessed

oocytes were in the control group). MII oocytes wereanalyzed for spindle and chromosome configuration.
Campen et al. 2018b
Antral follicles (3–7 mm in diameter) from cow ovaries
with an active corpus luteum. Pooled cells from two to
three animals per replicate.
Granulosa and theca cells were obtained from antral
follicles.

Decreased proportion of oocytes with properly aligned chromosomes at 10 fM,
100 pM, 10 nM, 100 nM and 50 µM
P, measured in conditioned medium collected from either granulosa
or theca cell cultures.
E2 from granulosa cells
Androstenedione from theca cells

Sheep primary ovarian theca cells
Isolation of theca cells was performed on eightmultiparous Polypay × Dorsett breed sheep at
gestational day 120 of pregnancy. The theca internal cell
layer from antral follicles was isolated by microdissection
and plated.
N=8 primary cell cultures with 3 replicates per
experiment
Treatment: BPS (99.7% purity) in 0.1% DMSO (vehicle
control) at concentrations of 0, 1, 10, 100, 200, 500, or
1,000 ng/mL, incubated for 24 hours
Human ovarian theca cells
Human ovaries collected from healthy non pregnant
women (n=3, aged 31-46 years)

Granulosa cells:
Increased basal E2 secretion at 100 µM
No effect of BPS under FSH -stimulated conditions
No effect on P secretion under any condition (including FSH)
Theca cells:
No effect on secretion of P or androstenedione in basal condition or in thepresence of LH

General toxicity: Cell viability

Treatment: BPS (purity not reported) in culture medium
containing 0.05% DMSO, at concentrations of 0, andfrom 1 fM to 100 µM (in 10-fold increments), in thepresence or absence of 0.33 ng/mL follicle stimulating
hormone (FSH) (granulosa cells) or 100 pg/mL
luteinizing hormone (LH) (theca cells)
Gingrich et al. 2021

Major Findings

General toxicity: No effect on granulosa or theca cell viability

Theca cell GJIC quantification:
Rhodamine-dextran dye used to measure GJIC at different stagesof theca cell development (pre-luteinized, luteinizing, and luteinized)
Signaling pathway analysis using specific signal transduction
pathway inhibitors (D609, ET-18-OCH3, GF109203X, H89,
SB202190, and U0126) for:phosphatidyl choline specific phospholipase C (PC-PLC),
phosphatidyl inositol specific phospholipase C (PI-PLC),
protein kinase C (PKC),
protein kinase A (PKA),
extracellular receptor kinase-mitogen activated protein kinase (ERKMAPK)
General Toxicity: Cell viability via MTT assay

Theca cell GJIC quantification in both cell types (sheep and human)increased GJIC >100 ng/mL (pre-luteinized)
Increased GJIC >100 ng/mL (luteinizing)
Non monotonic response (luteinized)
Signaling pathway analysis:
None of the pathway inhibitors tested individually reduced GJIC below that of thevehicle control.
Effects of protein kinase inhibitors on BPS-stimulated GJIC:
Sheep theca cells:
No effect of PKA or PKC inhibitors
Full inhibition (= to control): MAPK inhibitor
Partial inhibition with PI-PLC, PC-PLC, and ERK-MAPK inhibitors
Human theca cells:
Full inhibition (= to control): MAPK inhibitor
General toxicity: No effect on cell viability or morphology in sheep or human cells.

Treatment: BPS (99.7% purity) in 0.1% DMSO (vehicle
control) at concentrations 0, or 200 ng/mL, incubated for
24 hours, in the presence or absence of MAPK inhibitor (1µM)

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Outcomes Assessed

Major Findings

PKA activator (CW008) used to evaluate molecularpathways involved in BPS-induced upregulation of gap
junctional intercellular communication (GJIC)
Nonspecific negative control: Phorbol 12-myristate 13acetate (TPA) (inhibits GJIC) by internalizing gap
junction channels through PCK/ER1/2) dependent
mechanisms
Liu et al. 2021

Follicular cell proliferation in in vitro treated ovaries at 10 μM

In vitro:
PND 0 CD-1 mice ovaries in culture medium, number ofanimals (or ovaries) was not reported but in the statisticalanalysis section it is stated that each experiment was
repeated at least three times.
Treatment: BPS (purity not reported) in culture medium
at concentrations of 0, 10, 50, or 100 μM for 3 days.

Immunofluorescence and immunohistochemistryCo-treatment:
A selective estrogen receptor (ER) modulator, tamoxifen,
in culture medium at a final concentration of 0 or 1 μM.
JNK inhibitor SP600125 in culture medium at a final
concentration of 0 or 5 μM for 3 days.

Gene and protein expression:
ER (α and β); c-Jun N-terminal kinases (JNKs), Notch2, Jagged1,
and Cx43 (genes involved in germ cell nest (cyst) breakdown andthe formation of primary follicles)
Western blot analysis for JNK (phosphorylation) and E-cadherin (akey protein in the JNK signaling pathway)

BPS treated ovaries (10 μM BPS) were incubated with athymidine analog (proliferation marker) in the culture
medium at 50 nM for 2 hours before assessment.
Profita et al. 2021
HTR-8/SVneo human trophoblast cell line (derived fromhuman first-trimester placenta), used to measureadhesion, migration, and invasion
Treatment: BPS (purity not reported) in DMSO atconcentrations of 0 or 0.1 µM
Co-exposure studies:
Tamoxifen at 0.01 µM
U0126 (MEK1,2 inhibitor) at 0.01 µM

Bisphenol S

Effects in follicles:
Dose-dependent increase in percentage of MVH-positive cells (oocytes) inprimary follicles
BPS at all doses increased germ cell cyst breakdown, this effect was blocked by 1
μM tamoxifen and by 5 μM SP600125
Decreased percentage of oocytes in germ cell cysts
No effect on total number of oocytes
Gene and protein expression at 10 μM BPS:
Increased Er-α and Er-β mRNA and protein levels
Increased protein and gene expression of Notch2, Jagged1, and Cx43 (granulosa
cell proliferation)
Decreased E-cadherin protein detection, effect reverted by 5 μM JNK inhibitor
SP600125
Activation of JNK phosphorylation (counteracted by 1 μM tamoxifen)
Components of JNK pathway: increased expression of MDM2 and decreased
expression of E-cadherin; adding SP6000125 had the opposite effect

Cell proliferation: E-SCREEN assay used to evaluate proliferative
effect of BPS on HTR-8/SV cells
Cell Migration

E-SCREEN: Increased cell proliferation at 10-11-10-7 M
Increase in ERK1/2 phosphorylation at 10-9 M and 10-7 M
Increase in IL-6 and IL-8 secretion at 10-11 M that was counteracted with U0126
inhibitor

Phosphorylation
Inflammatory cytokine analyses
Viability: MTT assay
Western blot analysis to measure MAPK activity as phosphorylated /total ERK1/2 (% of phosphorylation with respect to control)
Luminex assay (analysis of inflammatory cytokines and cytokineinduced vascular endothelial growth factor (VEGF)

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Outcomes Assessed

Pre-exposure to E2:
Incubated with E2 (10-12M-10-8 M) and then exposed to
BPS for 3 days

Secretions of IL-6 (interleukin 6), IL-8 (interleukin 8), IL-1β(interleukin 1β), TNF-α (tumor necrosis factor-α), chemokine CCL2
(Chemokine Ligand 2), and VEGF

Rosenmai et al. 2014

Cell viability

No effect on cell viability

H295R cell line (human adrenal cell line)

H295R steroidogenesis assay

Treatment: BPS (purity not reported) in DMSO atconcentrations of 0, or from 0.8 to 50 μM in 2-folddilutionsTime-resolved fluorimmunoassay: E2, testosterone (T), and P

Steroidogenesis:
Increased P and 17-OH-P levels
Decreased levels of T, androstenedione, and DHEA
Decreased cortisol and corticosterone levels
No effect on E2 or E1 levels

QSAR modeling

HPLC/MS: P, 17β-OH P, dehydroepiandrosterone (DHEA),
androstenedione T, dihydrotestosterone, corticosterone, cortisol,
E2, and estrone (E1)

Major Findings

Aryl hydrocarbon receptor reporter gene assay

Receptor activity assays:
Increased ER activity
NS trend on decreased AR activity; predicted negative in the QSAR modeling for
AR activity.

Sabry et al. 2021

Confocal microscopy examination of COCs.

No change in COC morphology

In vitro: cumulus–oocyte complexes (COCs), denuded
oocytes, and cumulus cells from bovine ovaries.
n= 40/group.

IVF

Accelerated embryo development, with hatched blastocysts at 8 days

Gene and protein expression of connexins (Cx), i.e., gap junctional
proteins: Cx37 and Cx43 in COCs, oocytes, and cumulus cells by
mRNA and western blot analyses.

Increased Cx37 mRNA levels, with NS increase in protein levels in cumulus cells

On COCS, oocytes and cumulus cells:
Gene and protein detection for anti-müllerian hormone (AMH) and
its receptor (AMHRII) on each frozen sample.

COCs, oocytes and cumulus cells

ER activity: luciferase reporter gene assay.
AR reporter gene assay

Exp. 1 Oocyte culture. Treatment: BPS (purity notreported) in media with 0.1% ethanol at concentrations of
0 or 0.05 mg/mL for 24 hours.
After 24 hours maturation, oocytes were used for
imaging or for in vitro fertilization (IVF) incubation withuntreated bovine sperm.
After 18 hours, likely zygotes were separated fromcumulus cells and incubated until at the 8-cell blastocyst
stage.

No effect on Cx43 or Cx37 mRNA levels in COCs
No effect on Cx43 mRNA levels in oocytes or cumulus cells
No effect on Cx37 mRNA levels in oocytes
No changes in Cx37 protein levels in COCs or localization in cumulus cells(confocal microscopy)

Exp. 2 Cumulus cell culture. Treatment: BPS in media.
After 24 hours cells were used for mRNA and protein
detection.
Saleh et al. 2021
In vitro: COCs from bovine ovaries, 60 COCs per group
Treatment: BPS (purity not stated) in 2.5 µl of 0.1%
ethanol added to the in vitro maturation medium

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94

Increased AMHRII mRNA levels in COCs; no effects in oocytes or cumulus cells.
NS increase in AMHRII protein expression in COCs; no effects in oocytes orcumulus cells

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Study Design
supplemented with gonadotropins and E2 (2.5 mL final
volume) at a BPS final concentration of 0 or 0.05 mg/mL
for 24 hours.
After incubation, 30 COCs were removed and washedwith PBS and snap frozen.
30 denuded oocytes and the cumulus cells were isolated
and snap frozen for further analysis.

Outcomes Assessed
In IVF embryos:
Rates of development determined for:cleavage rate at 40–45 hpf
2–4 cells between 45 and 50 hpf
8–16 cells between 75 and 80 hpf
blastocyst rates at 8 days post-fertilization
Apoptosis rate

Major Findings
No effect on AMH mRNA or protein level in COCs; no effects on protein levels in
oocytes or cumulus cells
AMH:
No effect in mRNA levels on cumulus cells, oocytes or COCs
IVF embryos
No effects on development rates in IVF embryos.
Apoptosis:
Increased average percentage of DNA fragmentation (a marker for apoptosis) inarrested 2–4 cells; 8-16 cells and day-8 blastocysts

IVF produced embryos: After maturation, COCs fromeach group were incubated with bovine sperm for 18
hours.
Presumptive zygotes were incubated until day-8
blastocysts.

Gene and protein expression of AMH and AMHRII

Speidel et al. 2018

Cell viability

No effect on cell viability after acute exposure

Human trophoblastic 3A placental cell line (CRL-1584)

Acute and chronic effects of BPS on ABCB1 promoter activity using
4 haplotypes: 1, 4, 29 and 30

Acute exposure
Decrease in ABCB1 promoter activity of haplotypes 4, 29 and 30(This cell type represents the human placenta and is wellcharacterized, and capable of synthesizing humanchorionic gonadotropin in culture.)

Determination of sex ratio
via real-time polymerase chain reaction (qPCR): expression of two
transcripts that are only expressed in males (DDX3Y and USP9Y),
blastocysts that did not express any of these transcripts wereclassified as female

Increased protein levels of AMH and AMHRII in day-8 blastocysts.
No effect on sex ratio

Chronic exposure
Increase in ABCB1 promoter activity of haplotypes 1, 29 and 30

Treatment: BPS (99% purity) dissolved in 50% ethanol in
culture medium, final ethanol concentration of 0.01%, atconcentrations as described below:
Acute BPS exposure (0 or 0.5 nM) for 90 min withsamples at 0, 15, 30, 45, 60, and 90 mins.
Chronic BPS exposure (0 or 0.3 nM) for 12 days.
After 12 days of BPS exposure, cells were transferred to
6-well plates with exposure for 24 hours after
transfection. Cells were harvested 36-48 hours after
transfection with ABCB1 promoter haplotype luciferase
reporter generation in CRL-1584 cells (luciferase
constructs represents different haplotypes)
Téteau et al. 2020

Cell proliferation

In vitro: Granulosa cells isolated from adult ewes; 1 × 105

Hormone secretion: P and E2 levels were measured in culture
medium.

viable cells/well
Treatment: BPS (purity not reported) in ethanol added to

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95

Decreased cell proliferation at 200 µM.
Hormone secretion:
Dose related decreased P secretion from 10 µM to 200 µM
Increased E2 secretion at concentrations ≥ 10 μM.

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Outcomes Assessed

culture medium (150 µL/well) at concentrations of 0, 1
nM, 10 nM, 100 nM, 1 µM, 10 µM, 50 µM, 100 µM or
200 µM for 48 hours

Immunoblotting on protein lysates in six independent cultures for
identification of steroidogenic enzymes (CYP19A1, CYP11A1 and
HSD3B1)

Cell proliferation: BPS treatment (as above) was
supplemented with 10 µM bromodeoxyuridine/5-bromo2'-deoxyuridine (BrdU) for 48 hours to determine effects
on cell proliferation

Gene expression in six independent cultures by qPCR for AR, PR,
ER1, ER2, CYP19A1, CYP11A1, HSD3B1 and STAR

Gene expression of hormone receptors:
No effect on AR or PR mRNA levels
Increased ESR2 at 50μM and a significant two-fold increase in ESR1 and ESR2
at 100 µM

Téteau et al. 2023

Cell viability at 1 and 48 hours

No effect on cell viability

In vitro: Granulosa cells isolated from adult ewes,
100,000 viable cells/well in 150 µL medium/well

Hormone levels: P and E2 were measured in the supernatant after
48 hours of incubation with BPS (with or without 10 µM G-15)

Treatment: BPS (purity not reported) in ethanol (1/1111)
added to culture medium (150 µL/well) at concentrationsof 0, 10 or 50 µM for 1, 12, 24, or 48 hours (timedepending on the experiment).

qPCR and gene expression on differentially expressed genes (DEG)
on granulosa cells treated for 12, 24 or 48 hours with BPS and/or 10µM G-15

Decreased P levels at both BPS concentrations compared to control andcompared to G-15 + BPS
Decreased E2 levels at both BPS doses compared to control and compared to G15 + BPS.

Granulosa cells were cultured with the plasma
membrane ER (GPER) agonist, G-1, at 1 or 10 μM(positive control) or co-cultured with the GPER
antagonist, G-15, at 10 μM in ethanol (same ethanol
concentration as used for BPS treatment)
At the end of culture time, supernatant and cells wereseparated for further analysis

Gene expression after 1 hour incubation with 50 μM BPSŽalmanová et al. 2017

Oocyte maturation stages: progression to MI and MII

In vitro: COCs from pig ovary

α-tubulin assembly

Treatment: BPS in 01% DMSO added to the culture
medium at concentrations of 0, 3 nM, 300 nM or 30 µM
for 24, 48 or 72 hours.

Gene expression (mRNA) of ERα, ERβ, and aromatase in oocytes
and cumulus cells treated for 48 hours.

After treatment, oocytes were separated from cumulus
cells and fixed and the oocyte stage determined:
germinal vesicle (GV), late diakinesis, metaphase I (MI),anaphase I, telophase I , and MII

Content of hyaluronic acid (abundant extracellular compound
produced by cumulus cells, and an indicator of chemical damage tocumulus cells or of cumulus cell expansion)
Protein detection of ERα, ERβ, and aromatase in oocytes byimmunofluorescence
Cell viability

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96

Major Findings
Steroidogenic enzymes: No effects on mRNA or protein levels for the three
enzymes analyzed.

Gene expression: 12 DEGs were identified at 50 μM BPS.
Four genes were downregulated, and eight genes were upregulated.
These DEG are involved mostly in cell communication.

Oocyte maturation:
Decreased MI stage achievement after 24 hours (dose dependent)
Meiosis was not resumed after 24 hours at 300 nM and 30 µM
Decreased MII stage achievement after 48 and 72 hours (concentration
dependent)
At 48 hours: all BPS-treated oocytes mature to at least MI
At 72 hours: delayed, disrupted and blocked oocyte maturation at allconcentrations tested
α-tubulin assembly:
Swollen chromosomes and irregular organization, Decreased numbers of tubulin
filaments, spindles in a circular formation, elongated metaphase
plates, and reduced spindle size at all doses in MI and MII oocytes
Gene expression:
Decreased Erα mRNA levels in oocytes at all concentrations; no effect on

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Outcomes Assessed

Major Findings
cumulus cells
Decreased Erβ mRNA levels at 30 μM in cumulus cells; no changes in oocytes
Decreased aromatase mRNA levels at 3 and 300 nM in oocytes and at 30 μM in
cumulus cells
Increased hyaluronic acid content at 300 nM at 24 and 48 hours
Immunofluorescence in oocytes:
Increased ERα at 3 and 300 nM (24 and 48 hours)
Increased ERβ at 300 nM and 30 μM (24 hours)
Decreased ERβ at 300 nM (48 hours)
Decreased aromatase at 3 nM (24 hours)
No effects on viability of oocytes or cumulus cells after 24 and 48 hours

Žalmanová et al. 2023

Cell viability

In vitro: COCs from pig ovaries. Oocytes were cultured inmodified M199 medium in the presence of gonadotropins
to stimulate oocyte maturation.

Meiotic progression of porcine oocytes

Treatment: BPS (purity not reported), dissolved in 0.1%
DMSO, (control) at concentrations of 0, 300 pM, 30nM,
or 3 µM for 48 hours.
MII oocytes with extrusion of first polar body wereselected for IVF. 15-20 MII oocytes were incubated inmodified Tris-buffered medium with sperm from
untreated males.

Spindle organization and chromosome alignment
Chromosome spreads of MII pig oocytes (aneuploidy)
TUNEL assay (DNA integrity, apoptosis)
Mitochondria status (by immunofluorescence):
number and distribution
Mitochondrial fusion proteins (mitofusins, MFN1 and MNF2)
Glycolytic activity of cumulus cells

Concentration dependent decrease in proportion of cells reaching MII: retention in
GV stage, degenerated oocytes, arrested in MI stage.
No effect on the number or quality of blastocysts that completed MII maturation.
Spindle organization:
Spindle disorganization and chromosome misalignment at all concentrations.
Number of chromosomes:
Increased number of aneuploidy oocytes at all concentrations.
DNA integrity:
No effects on DNA damage or apoptosis
No effects on the quality of blastocysts or in the number of double strand breaks(DSBs) in blastocysts
Mitochondria number and distribution:
Lower relative intensity (about 30% decrease) of the marker for mitochondria at
30 nM and 3 µM
Relative increase in fluorescence signal for MFN1 and MFN2 at 30 nM
No effects on glucose metabolism
Increased cell viability at both BPS doses (in cultures without serum)

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5. REFERENCES
Abrantes-Soares F, Lorigo M, Cairrao E. 2022. Effects of BPA substitutes on the prenatal andcardiovascular systems. Crit Rev Toxicol 52:469-498.
Ahsan N, Ullah H, Ullah W, Jahan S. 2018. Comparative effects of Bisphenol S and Bisphenol A
on the development of female reproductive system in rats; a neonatal exposure study.
Chemosphere 197:336-343.
Aker AM, Johns L, McElrath TF, Cantonwine DE, Mukherjee B, Meeker JD. 2018. Associations
between maternal phenol and paraben urinary biomarkers and maternal hormones during
pregnancy: A repeated measures study. Environ Int 113:341-349.
Aker AM, Ferguson KK, Rosario ZY, Mukherjee B, Alshawabkeh AN, Calafat AM, et al. 2019. A
repeated measures study of phenol, paraben and triclocarban urinary biomarkers andcirculating maternal hormones during gestation in the Puerto Rico PROTECT cohort. Environ
Health 18:28.
Amar S, Binet A, Téteau O, Desmarchais A, Papillier P, Lacroix MZ, et al. 2020. Bisphenol S
impaired human granulosa cell steroidogenesis in vitro. Int J Mol Sci 21.
Ao J, Huo X, Zhang J, Mao Y, Li G, Ye J, et al. 2022. Environmental exposure to bisphenol
analogues and unexplained recurrent miscarriage: A case-control study. Environ Res
204:112293.
BASF. 2014a. DHDPS repeated-dose 90-day oral toxicity study in wistar rats administration bygavage.
BASF. 2014b. DHDPS prenatal developmental toxicity study in wistar rats oral administration(gavage) part I of III.
BASF. 2019a. DHDPS modified extended one-generation reproduction toxicity study in Sprague
Dawley rats oral administration (gavage).
BASF. 2019b. 14C-4,4’-sulphonyldiphenol study on kinetics in vistar rats. (part I of II).
BASF. 2020. DHDPS test study in male and female Sprague Dawley rats oral administration(gavage).
Benjamin K, Marquez CM, Morta M, Reyes EM, Aragones L, Velarde M. 2023. Bisphenol S
increases cell number and stimulates migration of endometrial epithelial cells. J ASEAN Fed
Endocr Soc 38:13-22.
Biomonitoring California. 2020. Bisphenol S (BPS). Available:
https://biomonitoring.ca.gov/chemicals/bisphenol-s-bps [accessed 09/07/2023.

Bisphenol S

98

OEHHA
October 2023

Blaauwendraad SM, Jaddoe VW, Santos S, Kannan K, Dohle GR, Trasande L, et al. 2022.
Associations of maternal urinary bisphenol and phthalate concentrations with offspring
reproductive development. Environ Pollut 309:119745.
Campen KA, Kucharczyk KM, Bogin B, Ehrlich JM, Combelles CMH. 2018a. Spindle
abnormalities and chromosome misalignment in bovine oocytes after exposure to low doses ofbisphenol A or bisphenol S. Hum Reprod 33:895-904.
Campen KA, Lavallee M, Combelles CMH. 2018b. The impact of bisphenol S on bovine
granulosa and theca cells. Reprod Domest Anim 53:450-457.
Cao XL, Zhou S, Popovic S, Dabeka R. 2022a. Bisphenol S in individual and composite meatand meat products and implication for its sources. Food Addit Contam Part A Chem Anal
Control Expo Risk Assess 39:572-579.
Cao XL, Zhou S, Popovic S, Dabeka R. 2022b. Temporal trend of bisphenol S occurrence inmeat samples from 2008-2020 Canadian total diet study. Food Addit Contam Part A Chem Anal
Control Expo Risk Assess 39:1634-1639.
Castro B, Sánchez P, Torres JM, Ortega E. 2015. Bisphenol A, bisphenol f and bisphenol S
affect differently 5α-reductase expression and dopamine-serotonin systems in the prefrontal
cortex of juvenile female rats. Environ Res 142:281-287.
ChemAnalyst. 2021. Bisphenol S market analysis: Plant capacity, production, operating
efficiency, technology, demand and supply, end-user industries, distribution channel, regional
demand, 2015-2030. Decode the future of Bisphenol S. Last updated November 2021.
Available: htps://www.chemanalyst.com/industry-report/bisphenol-s-290.
Chen D, Kannan K, Tan H, Zheng Z, Feng YL, Wu Y, et al. 2016a. Bisphenol analogues otherthan BPA: Environmental occurrence, human exposure, and toxicity-a review. Environ Sci
Technol 50:5438-5453.
Chen Y, Shu L, Qiu Z, Lee DY, Settle SJ, Que Hee S, et al. 2016b. Exposure to the BPAsubstitute Bisphenol S causes unique alterations of germline function. PLoS Genet
12:e1006223.
Demacopulo B, Kreimann EL. 2019. Bisphenol S increases ezrin expression and the detrimental
effects induced by dehydroepiandrosterone in rat endometrium. Mol Cell Endocrinol 483:64-73.
Derakhshan A, Shu H, Peeters RP, Kortenkamp A, Lindh CH, Demeneix B, et al. 2019.
Association of urinary bisphenols and triclosan with thyroid function during early pregnancy.
Environ Int 133:105123.
Derakhshan A, Philips EM, Ghassabian A, Santos S, Asimakopoulos AG, Kannan K, et al.
2021. Association of urinary bisphenols during pregnancy with maternal, cord blood andchildhood thyroid function. Environ Int 146:106160.

Bisphenol S

99

OEHHA
October 2023

Desmarchais A, Téteau O, Kasal-Hoc N, Cognié J, Lasserre O, Papillier P, et al. 2022. Chronic
low BPS exposure through diet impairs in vitro embryo production parameters according tometabolic status in the ewe. Ecotoxicol Environ Saf 229:113096.
Dodson RE, Boronow KE, Susmann H, Udesky JO, Rodgers KM, Weller D, et al. 2020.
Consumer behavior and exposure to parabens, bisphenols, triclosan, dichlorophenols, andbenzophenone-3: Results from a crowdsourced biomonitoring study. Int J Hyg Environ Health230:113624.
ECHA. 2019. (European Chemicals Agency) CLH report proposal for harmonised classificationand labelling based on regulation (EC) No 1272/2008 (CLP regulation), annex VI, part 2
international chemical identification: 4,4’-sulphonyldiphenol; bisphenol S. Available:
https://echa.europa.eu/documents/10162/21282a0f-29b9-712a-8004-b6f659248268.
ECHA. 2022. 4,4'-sulphonyldiphenol as a substance of very high concern because of its toxic
for reproduction (article 57c), endocrine disrupting properties (article 57(f) - environment),
endocrine disrupting properties (article 57(f) - human health) properties. Available:
https://echa.europa.eu/documents/10162/cffe2e0d-2c25-bfb1-f5ff-eca0b3b30f32.
ECHA. 2023. Substance infocard for Bisphenol S, june 9, 2023 update. Available:
https://echa.europa.eu/substance-information/-/substanceinfo/100.001.137.
Ecology Center. 2019. Receipt paper study 2018 findings. Available:
https://www.ecocenter.org/our-work/healthy-stuff-lab/reports/more-you-bargained-receipt-paperstudy-2018/receipt-paper-study [accessed 9/14/2023.
Eldefrawy F, Xu HS, Pusch E, Karkoura A, Alsafy M, Elgendy S, et al. 2021. Modulation offolliculogenesis in adult laying chickens by bisphenol A and bisphenol S: Perspectives on
ovarian morphology and gene expression. Reprod Toxicol 103:181-190.
Fan X, Katuri GP, Caza AA, Rasmussen PE, Kubwabo C. 2021. Simultaneous measurement of
16 bisphenol A analogues in house dust and evaluation of two sampling techniques. Emerging
Contaminants 7:1-9.
Fays F, Palazzi P, Hardy EM, Schaeffer C, Phillipat C, Zeimet E, et al. 2020. Is there an optimal
sampling time and number of samples for assessing exposure to fast elimination endocrine
disruptors with urinary biomarkers? Sci Total Environ 747:141185.
Gingrich J, Pu Y, Roberts J, Karthikraj R, Kannan K, Ehrhardt R, et al. 2018. Gestational
bisphenol S impairs placental endocrine function and the fusogenic trophoblast signaling
pathway. Arch Toxicol 92:1861-1876.
Gingrich J, Pu Y, Upham BL, Hulse M, Pearl S, Martin D, et al. 2021. Bisphenol S enhances
gap junction intercellular communication in ovarian theca cells. Chemosphere 263:128304.
Grandin FC, Lacroix MZ, Gayrard V, Gauderat G, Mila H, Toutain PL, et al. 2018. Bisphenol S
instead of Bisphenol A: Toxicokinetic investigations in the ovine materno-feto-placental unit.
Environ Int 120:584-592.

Bisphenol S

100

OEHHA
October 2023

Hill CE, Sapouckey SA, Suvorov A, Vandenberg LN. 2017. Developmental exposures tobisphenol S, a BPA replacement, alter estrogen-responsiveness of the female reproductive
tract: A pilot study. Cogent Med 4.
Hu P, Pan C, Su W, Vinturache A, Hu Y, Dong X, et al. 2022. Associations between exposure toa mixture of phenols, parabens, and phthalates and sex steroid hormones in children 6-19 years
from NHANES, 2013-2016. Sci Total Environ 822:153548.
Huang H, Liang J, Tang P, Yu C, Fan H, Liao Q, et al. 2022. Associations of bisphenol exposure
with thyroid hormones in pregnant women: A prospective birth cohort study in China. Environ
Sci Pollut Res Int 29:87170-87183.
Ijaz S, Ullah A, Shaheen G, Jahan S. 2020. Exposure of BPA and its alternatives like bpb, BPF,
and BPS impair subsequent reproductive potentials in adult female Sprague Dawley rats.
Toxicol Mech Methods 30:60-72.
Iribarne-Durán LM, Peinado FM, Freire C, Castillero-Rosales I, Artacho-Cordón F, Olea N.
2022. Concentrations of bisphenols, parabens, and benzophenones in human breast milk: A
systematic review and meta-analysis. Sci Total Environ 806:150437.
Ji K, Hong S, Kho Y, Choi K. 2013. Effects of bisphenol s exposure on endocrine functions andreproduction of zebrafish. Environ Sci Technol 47:8793-8800.
Jurewicz J, Majewska J, Berg A, Owczarek K, Zajdel R, Kaleta D, et al. 2020. Serum bisphenol
A analogues in women diagnosed with the polycystic ovary syndrome - is there an association?
Environ Pollut:115962.
Kaimal A, Al Mansi MH, Dagher JB, Pope C, Varghese MG, Rudi TB, et al. 2021. Prenatal
exposure to bisphenols affects pregnancy outcomes and offspring development in rats.
Chemosphere 276:130118.
Karrer C, Roiss T, von Goetz N, Gramec Skledar D, Peterlin Mašič L, Hungerbühler K. 2018.
Physiologically based pharmacokinetic (pbpk) modeling of the bisphenols BPA, BPS, BPF, andbpaf with new experimental metabolic parameters: Comparing the pharmacokinetic behavior of
BPA with its substitutes. Environ Health Perspect 126:077002.
Khmiri I, Côté J, Mantha M, Khemiri R, Lacroix M, Gely C, et al. 2020. Toxicokinetics ofbisphenol-S and its glucuronide in plasma and urine following oral and dermal exposure involunteers for the interpretation of biomonitoring data. Environ Int 138:105644.
Kim K, Shin HM, Busgang SA, Barr DB, Panuwet P, Schmidt RJ, et al. 2021. Temporal trends ofphenol, paraben, and triclocarban exposure in California pregnant women during 2007-2014.
Environ Sci Technol 55:11155-11165.
Kolla S, Morcos M, Martin B, Vandenberg LN. 2018. Low dose bisphenol S or ethinyl estradiol
exposures during the perinatal period alter female mouse mammary gland development.
Reprod Toxicol 78:50-59.

Bisphenol S

101

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October 2023

Kolla SDD, Vandenberg LN. 2019. Data describing effects of perinatal exposure to bisphenol S
on a peripubertal estrogen challenge in intact female CD-1 mice. Data Brief 25:103862.
Kumar A, Gupta K, Tomer V, Kaur A, Kumar V. 2018. Bisphenols as human health hazard: A
systematic review on potent sources, route of exposure, harmful effects and safe alternatives.
Toxicology International 25:78-92.
La Merrill MA, Vandenberg LN, Smith MT, Goodson W, Browne P, Patisaul HB, et al. 2020.
Consensus on the key characteristics of endocrine-disrupting chemicals as a basis for hazard
identification. Nat Rev Endocrinol 16:45-57.
LaPlante CD, Catanese MC, Bansal R, Vandenberg LN. 2017. Bisphenol S alters the lactating
mammary gland and nursing behaviors in mice exposed during pregnancy and lactation.
Endocrinology 158:3448-3461.
Lehmler HJ, Liu B, Gadogbe M, Bao W. 2018. Exposure to Bisphenol A, Bisphenol F, andBisphenol S in U.S. Adults and children: The National Health and Nutrition Examination Survey
2013-2014. ACS Omega 3:6523-6532.
Liao C, Kannan K. 2013. Concentrations and profiles of bisphenol A and other bisphenol
analogues in foodstuffs from the United States and their implications for human exposure. J
Agric Food Chem 61:4655-4662.
Liu J, Li J, Wu Y, Zhao Y, Luo F, Li S, et al. 2017. Bisphenol A metabolites and Bisphenol S inpaired maternal and cord serum. Environ Sci Technol 51:2456-2463.
Liu J, Martin JW. 2019. Comparison of Bisphenol A and Bisphenol S percutaneous absorption
and biotransformation. Environ Health Perspect 127:67008.
Liu WX, Donatella F, Tan SJ, Ge W, Wang JJ, Sun XF, et al. 2021. Detrimental effect ofBisphenol S in mouse germ cell cyst breakdown and primordial follicle assembly. Chemosphere264:128445.
Luderer U, Eskenazi B, Hauser R, Korach KS, McHale CM, Moran F, et al. 2019. Proposed key
characteristics of female reproductive toxicants as an approach for organizing and evaluating
mechanistic data in hazard assessment. Environ Health Perspect 127:75001.
Luo D, Pan Y, Zeng L, Du B, Li J, Mei S. 2021. Occurrence of multiple Bisphenol S derivatives
in breast milk from chinese lactating women and implications for exposure in breast-fed infants.
Environmental Science & Technology Letters 8:176-182.
Mao J, Jain A, Denslow ND, Nouri MZ, Chen S, Wang T, et al. 2020. Bisphenol A and bisphenol
S disruptions of the mouse placenta and potential effects on the placenta-brain axis. Proc Natl
Acad Sci U S A 117:4642-4652.
Mao W, Mao L, Zhao N, Zhang Y, Zhao M, Jin H. 2022. Disposition of Bisphenol S metabolites
in Sprague-Dawley rats. Sci Total Environ 811:152288.

Bisphenol S

102

OEHHA
October 2023

Morgan MK, Clifton MS. 2021. Exposure to triclosan and bisphenol analogues b, f, P, S and z inrepeated duplicate-diet solid food samples of adults. Toxics 9.
Naderi M, Wong MY, Gholami F. 2014. Developmental exposure of zebrafish (Danio rerio) tobisphenol-S impairs subsequent reproduction potential and hormonal balance in adults. Aquat
Toxicol 148:195-203.
Nevoral J, Kolinko Y, Moravec J, Žalmanová T, Hošková K, Prokešová Š, et al. 2018. Long-term
exposure to very low doses of bisphenol S affects female reproduction. Reproduction 156:4757.
Nevoral J, Havránková J, Kolinko Y, Prokešová Š, Fenclová T, Monsef L, et al. 2021. Exposure
to alternative bisphenols BPS and BPF through breast milk: Noxious heritage effect during
nursing associated with idiopathic infertility. Toxicol Appl Pharmacol:115409.
Niu Y, Wang B, Yang R, Wu Y, Zhao Y, Li C, et al. 2021. Bisphenol analogues and theirchlorinated derivatives in breast milk in China: Occurrence and exposure assessment. J Agric
Food Chem 69:1391-1397.
Nourian A, Soleimanzadeh A, Shalizar Jalali A, Najafi G. 2017. Effects of bisphenol-S low
concentrations on oxidative stress status and in vitro fertilization potential in mature female
mice. Vet Res Forum 8:341-345.
Nourian A, Soleimanzadeh A, Shalizar Jalali A, Najafi G. 2020. Bisphenol-a analogue(bisphenol-S) exposure alters female reproductive tract and apoptosis/oxidative gene
expression in blastocyst-derived cells. Iran J Basic Med Sci 23:576-585.
NTP. 2015. Handbook for preparing report on carcinogens monographs. (Morrisville, NC: US
National Toxicology Program).
NTP. 2017. NTP research report on biological activity of bisphenol A (BPA) structural analogues
and functional alternatives.
Oh J, Choi JW, Ahn YA, Kim S. 2018. Pharmacokinetics of bisphenol S in humans after single
oral administration. Environ Int 112:127-133.
Pal S, Sahu A, Verma R, Haldar C. 2023. BPS-induced ovarian dysfunction: Protective actionsof melatonin via modulation of sirt-1/nrf2/nfĸb and ir/pi3k/pakt/glut-4 expressions in adult golden
hamster. J Pineal Res 75:e12869.
Pan Y, Deng M, Li J, Du B, Lan S, Liang X, et al. 2020. Occurrence and maternal transfer ofmultiple bisphenols, including an emerging derivative with unexpectedly high concentrations, inthe human maternal-fetal-placental unit. Environ Sci Technol 54:3476-3486.
Park CB, Kim GE, On J, Pyo H, Park JW, Cho SH. 2022. Sex-specific effects of bisphenol S
with tissue-specific responsiveness in adult zebrafish: The antiandrogenic and antiestrogenic
effects. Ecotoxicol Environ Saf 229:113102.

Bisphenol S

103

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Peinado FM, Lendínez I, Sotelo R, Iribarne-Durán LM, Fernández-Parra J, Vela-Soria F, et al.
2020. Association of urinary levels of bisphenols a, f, and S with endometriosis risk: Preliminary
results of the endea study. Int J Environ Res Public Health 17.
Pelch K, Wignall JA, Goldstone AE, Ross PK, Blain RB, Shapiro AJ, et al. 2019. A scoping
review of the health and toxicological activity of bisphenol A (BPA) structural analogues andfunctional alternatives. Toxicology 424:152235.
Pelch K, Wignall J, Goldstone A, Ross P, Blain R, Shapiro A, et al. 2017. NTP research reporton biological activity of bisphenol A (BPA) structural analogues and functional alternatives.
Philips EM, Kahn LG, Jaddoe VWV, Shao Y, Asimakopoulos AG, Kannan K, et al. 2018. First
trimester urinary bisphenol and phthalate concentrations and time to pregnancy: A populationbased cohort analysis. J Clin Endocrinol Metab 103:3540-3547.
Philips EM, Trasande L, Kahn LG, Gaillard R, Steegers EAP, Jaddoe VWV. 2019. Early
pregnancy bisphenol and phthalate metabolite levels, maternal hemodynamics and gestational
hypertensive disorders. Human Reproduction 34:365-373.
Philips EM, Santos S, Steegers EAP, Asimakopoulos AG, Kannan K, Trasande L, et al. 2020.
Maternal bisphenol and phthalate urine concentrations and weight gain during pregnancy.
Environ Int 135:105342.
Pollock T, Greville LJ, Weaver RE, Radenovic M, deCatanzaro D. 2019. Bisphenol S modulates
concentrations of bisphenol A and oestradiol in female and male mice. Xenobiotica 49:540-548.
Profita M, Fabbri E, Spisni E, Valbonesi P. 2021. Comparing effects and action mechanisms of
BPA and BPS on HTR-8/SVneo placental cells†. Biol Reprod 105:1355-1364.
Prokešová Š, Ghaibour K, Liška F, Klein P, Fenclová T, Štiavnická M, et al. 2020. Acute lowdose bisphenol S exposure affects mouse oocyte quality. Reprod Toxicol 93:19-27.
Qin JY, Ru S, Wang W, Hao L, Wei S, Zhang J, et al. 2021. Unraveling the mechanism of longterm bisphenol S exposure disrupted ovarian lipids metabolism, oocytes maturation, andoffspring development of zebrafish. Chemosphere 277:130304.
Reale E, Vernez D, Hopf NB. 2021. Skin absorption of Bisphenol A and its alternatives inthermal paper. Ann Work Expo Health 65:206-218.
Rochester JR, Bolden AL. 2015. Bisphenol S and f: A systematic review and comparison of thehormonal activity of Bisphenol A substitutes. Environ Health Perspect 123:643-650.
Rosenmai AK, Dybdahl M, Pedersen M, van Vugt-Lussenburg BMA, Wedebye EB, Taxvig C, etal. 2014. Are structural analogues to bisphenol a safe alternatives? Toxicological Sciences
139:35-47.
Sabry R, Apps C, Reiter-Saunders JA, Saleh AC, Balachandran S, St John EJ, et al. 2021. BPA
and BPS affect connexin 37 in bovine cumulus cells. Genes (Basel) 12.

Bisphenol S

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Saleh AC, Sabry R, Mastromonaco GF, Favetta LA. 2021. BPA and BPS affect the expressionof anti-mullerian hormone (amh) and its receptor during bovine oocyte maturation and early
embryo development. Reprod Biol Endocrinol 19:119.
Segovia-Mendoza M, Palacios-Arreola MI, Monroy-Escamilla LM, Soto-Piña AE, Nava-Castro
KE, Becerril-Alarcón Y, et al. 2022. Association of serum levels of plasticizers compounds,
phthalates and bisphenols, in patients and survivors of breast cancer: A real connection? Int J
Environ Res Public Health 19.
Shi M, Sekulovski N, MacLean JA, 2nd, Hayashi K. 2017. Effects of bisphenol A analogues on
reproductive functions in mice. Reprod Toxicol 73:280-291.
Shi M, Sekulovski N, MacLean JA, Whorton A, Hayashi K. 2019. Prenatal exposure to
Bisphenol A analogues on female reproductive functions in mice. Toxicol Sci 168:561-571.
Shin HM, Moschet C, Young TM, Bennett DH. 2020. Measured concentrations of consumer
product chemicals in California house dust: Implications for sources, exposure, and toxicity
potential. Indoor Air 30:60-75.
Silva JPA, Ramos JG, Campos MS, da Silva Lima D, de Azevedo Brito PV, Mendes EP, et al.
2019. Bisphenol-S promotes endocrine-disrupting effects similar to those promoted bybisphenol-a in the prostate of adult gerbils. Reprod Toxicol 85:83-92.
Šimková M, Vítků J, Kolátorová L, Vrbíková J, Vosátková M, Včelák J, et al. 2020. Endocrine
disruptors, obesity, and cytokines - how relevant are they to pcos? Physiol Res 69:279-293.
Sonker AK, Bhateria M, Karsauliya K, Singh SP. 2021. Investigating the glucuronidation andsulfation pathways contribution and disposition kinetics of Bisphenol S and its metabolites usinglc-MS/MS-based nonenzymatic hydrolysis method. Chemosphere 273:129624.
Speidel JT, Xu M, Abdel-Rahman SZ. 2018. Bisphenol A (BPA) and bisphenol S (BPS) alter thepromoter activity of the abcb1 gene encoding P-glycoprotein in the human placenta in a
haplotype-dependent manner. Toxicol Appl Pharmacol 359:47-54.
Tang P, Liang J, Liao Q, Huang H, Guo X, Lin M, et al. 2021. Associations of bisphenol
exposure with the risk of gestational diabetes mellitus: A nested case-control study in guangxi,
China. Environ Sci Pollut Res Int.
Téteau O, Jaubert M, Desmarchais A, Papillier P, Binet A, Maillard V, et al. 2020. Bisphenol A
and S impaired ovine granulosa cell steroidogenesis. Reproduction 159:571-583.
Téteau O, Liere P, Pianos A, Desmarchais A, Lasserre O, Papillier P, et al. 2022. Bisphenol S
alters the steroidome in the preovulatory follicle, oviduct fluid and plasma in ewes with
contrasted metabolic status. Front Endocrinol (Lausanne) 13:892213.
Téteau O, Vitorino Carvalho A, Papillier P, Mandon-Pépin B, Jouneau L, Jarrier-Gaillard P, et al.
2023. Bisphenol A and bisphenol S both disrupt ovine granulosa cell steroidogenesis but
through different molecular pathways. J Ovarian Res 16:30.

Bisphenol S

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Tucker DK, Hayes Bouknight S, Brar SS, Kissling GE, Fenton SE. 2018. Evaluation of prenatal
exposure to bisphenol analogues on development and long-term health of the mammary glandin female mice. Environ Health Perspect 126:087003.
US EPA. 2020. Chemical data reporting database. Access: Https://www.Epa.Gov/chemicaldata-reporting/access-cdr-data.
US FDA. 2014. Bisphenol A (BPA): Use in food contact application. Available:
https://www.fda.gov/food/food-additives-petitions/bisphenol-bpa-use-food-contactapplication#regulations) [accessed May 4 2018].
Verner MA, Salame H, Housand C, Birnbaum LS, Bouchard MF, Chevrier J, et al. 2020. How
many urine samples are needed to accurately assess exposure to non-persistent chemicals?
The biomarker reliability assessment tool (brat) for scientists, research sponsors, and risk
managers. Int J Environ Res Public Health 17.
Waidyanatha S, Black SR, Snyder RW, Yueh YL, Sutherland V, Patel PR, et al. 2018.
Disposition and metabolism of the bisphenol analogue, bisphenol S, in harlan Sprague Dawley
rats and B6C3F1/N mice and in vitro in hepatocytes from rats, mice, and humans. Toxicol Appl
Pharmacol 351:32-45.
Waidyanatha S, Black SR, Silinski M, Sutherland V, Fletcher BL, Fernando RA, et al. 2020.
Comparative toxicokinetics of bisphenol S in rats and mice following gavage administration.
Toxicol Appl Pharmacol 406:115207.
Wang W, Ru S, Wang L, Wei S, Zhang J, Qin J, et al. 2020. Bisphenol S exposure alters
behavioral parameters in adult zebrafish and offspring. Sci Total Environ 741:140448.
Wang Y, Aimuzi R, Nian M, Zhang Y, Luo K, Zhang J. 2021. Bisphenol A substitutes and sex
hormones in children and adolescents. Chemosphere 278:130396.
Wang Z, Miao M, Xu J, Chen Y, Liang H, Yang L, et al. 2022. Gestational exposure to bisphenol
analogues and kisspeptin levels in pregnant women and their children: A pregnancy-birth cohort
study. Sci Total Environ 848:157720.
Wesselink AK, Weuve J, Fruh V, Bethea TN, Claus Henn B, Harmon QE, et al. 2021. Urinary
concentrations of phenols, parabens, and triclocarban in relation to uterine leiomyomata
incidence and growth. Fertil Steril 116:1590-1600.
Xu Z, Tian L, Liu L, Goodyer CG, Hales BF, Bayen S. 2023. Food thermal labels are a source of
dietary exposure to Bisphenol S and other color developers. Environ Sci Technol 57:4984-4991.
Yamasaki K, Noda S, Imatanaka N, Yakabe Y. 2004. Comparative study of the uterotrophic
potency of 14 chemicals in a uterotrophic assay and their receptor-binding affinity. Toxicol Lett
146:111-120.
Yue B, Ning S, Miao H, Fang C, Li J, Zhang L, et al. 2023a. Human exposure to a mixture of
endocrine disruptors and serum levels of thyroid hormones: A cross-sectional study. J Environ
Sci (China) 125:641-649.

Bisphenol S

106

OEHHA
October 2023

Yue H, Tian Y, Wu X, Yang X, Xu P, Zhu H, et al. 2023b. Exploration of the damage andmechanisms of BPS exposure on the uterus and ovary of adult female mice. Sci Total
Environ:161660.
Žalmanová T, Hošková K, Nevoral J, Adámková K, Kott T, Šulc M, et al. 2017. Bisphenol S
negatively affects the meotic maturation of pig oocytes. Sci Rep 7:485.
Žalmanová T, Hošková K, Prokešová Š, Nevoral J, Ješeta M, Benc M, et al. 2023. The
bisphenol S contamination level observed in human follicular fluid affects the development of
porcine oocytes. Frontiers in Cell and Developmental Biology 11.
Zhan W, Yang H, Zhang J, Chen Q. 2022. Association between co-exposure to phenols andphthalates mixture and infertility risk in women. Environ Res:114244.
Zhan W, Tang W, Shen X, Xu H, Zhang J. 2023. Exposure to bisphenol A and its analogs andpolycystic ovarian syndrome in women of childbearing age: A multicenter case-control study.
Chemosphere 313:137463.
Zhang MY, Tian Y, Yan ZH, Li WD, Zang CJ, Li L, et al. 2020. Maternal Bisphenol S exposure
affects the reproductive capacity of F1 and F2 offspring in mice. Environ Pollut 267:115382.
Zhang N, Zhao Y, Zhai L, Bai Y, Jia L. 2023. Urinary bisphenol A and S are associated withdiminished ovarian reserve in women from an infertility clinic in northern China. Ecotoxicol
Environ Saf 256:114867.
Zhang W, Xia W, Liu W, Li X, Hu J, Zhang B, et al. 2019. Exposure to bisphenol a substitutes
and gestational diabetes mellitus: A prospective cohort study in China. Front Endocrinol(Lausanne) 10:262.
Zhu Y, Hedderson MM, Calafat AM, Alexeeff SE, Feng J, Quesenberry CP, et al. 2022. Urinary
phenols in early to midpregnancy and risk of gestational diabetes mellitus: A longitudinal study
in a multiracial cohort. Diabetes 71:2539-2551.

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APPENDIX A. LITERATURE SEARCH APPROACH ON THE FEMALEREPRODUCTIVE TOXICITY OF BPS
Searches of the published scientific literature on the developmental and reproductive
toxicity (DART) of bisphenol S (BPS) were conducted in January 2022 and updates tothe original search were conducted in February and July 2023. The goal was to identify
peer- open source and proprietary journal articles, print and digital books, reports, andgray literature that potentially reported toxicological and epidemiological information on
the DART of this chemical.
The searches were conducted using the following three approaches:•••

Primary searches in major biomedical databases, conducted by OEHHA librarian
Nancy Firchow, MLS
Searches in other data sources, including authoritative reviews and reports, anddatabases or web resources, conducted by OEHHA scientists
Additional focused searches, conducted by OEHHA scientists

In addition to information identified from these searches, OEHHA also considered thefollowing:•

One submission received during the data call-in period (March 4 – April 18, 2022)(https://oehha.ca.gov/proposition-65/comments/comment-submissions-requestrelevant-information-reproductive-toxicity)

Primary Search Process
1. Data Sources
The biomedical literature databases used as data sources that were searched to findinformation on BPS are listed below.
PubMed (National Library of Medicine) (https://www.ncbi.nlm.nih.gov/pmc/)
Embase (https://www.embase.com/)
Scopus (https://www.scopus.com/)
SciFinder-n (https://scifinder-n.cas.org/)

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Google Scholar (https://scholar.google.com)
2. Search term identification
•

•

•

•

The US EPA's CompTox Chemicals Dashboard(https://comptox.epa.gov/dashboard) was used to identify synonyms for BPS. The
PubMed MeSH database (https://www.ncbi.nlm.nih.gov/mesh/) was used to findadditional synonyms, subject headings and other index terms related to thechemical.
The PubMed DART filter(https://www.nlm.nih.gov/bsd/pubmed_subsets/dart_strategy.html) was used fordevelopmental and reproductive toxicity-related terminology.
National Toxicology Program’s Standard Search Strings for Literature Database
Searches: Appendix to the Draft Handbook for Preparing Report on Carcinogens
Monographs (NTP 2015)(https://ntp.niehs.nih.gov/ntp/roc/handbook/rochandbookappendix_508.pdf) wasused to identify search strategies for Experimental Animals and ADME concepts.
Additional strategies for DART and Key Characteristics of Female Reproductive
Toxicity and Key Characteristics of Male Reproductive Toxicity were developed by
OEHHA.

3. Primary search execution
Searches were executed in PubMed, Embase, Scopus, SciFindern, and Google Scholar
in January 2022.
Three separate searches were done in PubMed, Embase and Scopus. Searches inthese databases were divided into evidence streams as:•••

Human DART Studies
Animal DART Studies
ADME Studies

The basic structure used for each search and search dates are shown in Tables A.1
through A.3.

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Table A.1 Human DART studies search structure (PubMed, Embase, Scopus)
SearchDates
1/2022
1/20237/2023

SearchSearch Concepts
step#1
BPS terms#2

DART terms (PubMed DART Filter)#3

Additional DART terms (OEHHA strategy)#4

Key Characteristics of Male Reproductive Toxicity(OEHHA Strategy)#5

Key Characteristics of Female Reproductive Toxicity(OEHHA Strategy)#6#2 OR #3 OR #4 OR #5#7#1 AND #6

Table A.2 Animal DART studies search structure (PubMed, Embase, Scopus)
SearchDates
1/2022

Searchstep#1

1/2023#2

DART terms (PubMed DART Filter)#3

Additional DART terms (OEHHA strategy)#4

Key Characteristics of Male Reproductive Toxicity
terms (OEHHA Strategy)#5

Key Characteristics of Female Reproductive
Toxicity terms (OEHHA Strategy)#6#2 OR #3 OR #4 OR #5#7#1 AND #6#8

Experimental Animals terms (RoC strategy)#9#7 AND #8

7/2023

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Search Concepts
BPS terms

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Table A.3 ADME studies search structure (PubMed, Embase, Scopus)
SearchDate
8/2022

Searchstep#1

Search Concepts
BPS terms#2

ADME terms (RoC Strategy)#3#1 AND #2

The searches were run first in PubMed. The search terms and syntax were then tailoredaccording to the search features unique to the other databases. For example, Embase
uses different subject headings than PubMed, so the Emtree subject heading list was
searched to identify equivalent terms to replace the MeSH terms used in the PubMed
searches.
Two separate searches were run in SciFindern. Searches in this database were divided
into Human and Animal evidence streams. The basic structure used in each search isshown in Tables A.4 and A.5.
Table A.4 Human DART studies search structure (SciFindern)
SearchDates
1/2022

Searchstep#1

Search Concepts
BPS terms#2

Limit to Journal Article#3

Limit to human concept#4

Limit to Database "CA Plus"#5

Search within results: DART terms

CAplus (chemical abstract plus) is a database of chemical information that can beaccessed via SciFinder-N

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Table A.5 Animal studies search structure (SciFindern)
SearchDates
1/2022

Searchstep#1

Search Concepts
BPS terms#2

Limit to Journal Article#3

Limit to animals concept#4

Limit to Database "CA Plus"#5

Search within results: DART terms

CAplus, chemical abstract plus.
One search was run in Google Scholar. The basic structure used in the search is shownin Table A.6.
Table A.6 DART studies search structure (Google Scholar)
SearchDates
1/2022
1/2023

Searchstep#1

Search Concepts(BPS terms) AND (DART terms)

Results from all databases were uploaded to EndNote, maintaining separate libraries foreach of the three concepts searched (human DART studies, animal DART studies, andADME studies). Duplicates were removed. The results of the primary searches for BPS
are shown in Table A.7.
Table A.7 BPS DART studies search results (totals from all search dates)
SearchHuman
DART
AnimalDART
ADME

Bisphenol S

PubMedResults

EmbaseResults

ScopusResults

SciFindern
Results

GoogleScholarResults

643

583

596

131

9

816

422

499

397

102

16

584

236

489

505

none

none

759

112

Unique Results
After
Deduplication

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4. Other data sources
Additional databases and websites of governmental and other authoritative entities(e.g., US EPA, European Chemicals Agency) were searched for data and reports that
may have been missed in the primary literature search. Other relevant studies wereidentified from citations in individual articles, and through alert services (e.g.,
ScienceDirect, Google Scholar, etc.).
Literature Screening Process
Use of health assessment workspace collaborative
The EndNote libraries containing the literature search results (citations) for BPS were
uploaded to HAWC (Health Assessment Workspace Collaborative,(https://hawcproject.org). HAWC is a tool used for multi-level screening of literaturesearch results. Using HAWC, the references were screened and tagged.
In Level 1 screening, each citation was first screened by at least one OEHHA scientist,
based solely on titles and abstracts, using specific inclusion and exclusion criteria toeliminate studies or articles that did not contain information on BPS and studies of its
DART or other key related topics (e.g., pharmacokinetics, mechanisms of action). Thisinitial screen (Level 1) was intended to identify all studies deemed to have a reasonable
possibility of containing information relevant to DART that could be useful for the review
process, and to further identify (i.e., tag in HAWC) studies relevant to specific aspects of
DART (e.g., male reproductive toxicity, female reproductive toxicity, developmental
toxicity).
Screening the literature relevant to female reproductive toxicity
For purposes of identifying the available evidence on the female reproductive toxicity of
BPS, citations identified as having a reasonable possibility of containing information
relevant to female reproductive toxicity underwent Level 2 screening. In the Level 2
screening of this subset of citations, the full text was obtained. These full papers werescreened independently by one OEHHA scientist, using similar inclusion/exclusion
criteria as was used in the Level 1 screening. However, Level 2 reviewers could makemore accurate judgements about the relevance of the articles because they werereviewing the full text in addition to the title and abstract. Following Level 2 screening,
the tagging of articles according to key topics was updated in HAWC. Level 1 and 2
screenings were repeated as search results were updated, and with additional relevant

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studies identified from citations in individual articles and alert services (e.g.,
ScienceDirect, Google Scholar).
One hundred and twenty-four references were cited in this document.

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Detailed PubMed Literature Search Strategies – Primary Searches
Table A.8 PubMed search strategy for human DART studies
SET
STRATEGY#(80-09-1[rn] OR "Bisphenol S"[tiab] OR "bis(4-hydroxyphenyl)sulfone"[nm] OR"bis(4-hydroxyphenyl)sulfone"[tiab] OR "4,4'-Sulfonyldiphenol"[tiab] OR "Phenol,
1 4,4'-sulfonylbis-"[tiab] OR "BPS-monoglucuronide"[tiab] OR "BPS-1G"[tiab] OR"bisphenol S dicyanate ester"[tiab] OR (bps[tiab] NOT ("bps"[tiab] NOT"bisphenol"[tiab])))2

CONCEPT GROUP

Chemical Terms(abnormalities, drug-induced [mh] AND (fetus [mh] or pregnancy [mh])) OR((abnormalities, multiple/chemically induced [mh] OR abnormalities,
multiple/epidemiology [mh] OR abnormalities, multiple/etiology[mh] ORabnormalities, multiple/genetics [mh] OR abnormalities, multiple/pathology [mh])AND (pregnancy [mh] OR fetus [mh]))OR (abnormalities, radiation-induced [mh]AND (fetus [mh] or pregnancy [mh])) OR abortion, habitual/chemically induced[mh] OR abortion, habitual/etiology [mh] OR abortion, spontaneous/chemicallyinduced [mh] OR abortion, spontaneous/etiology [mh] OR (alcoholic
intoxication[mh] AND (fetus [mh] or pregnancy [mh])) OR (alcohol drinking[mh:noexp] AND (fetus [mh] or pregnancy [mh])) OR Birth Defects Res B Dev
Reprod Toxicol [TA] OR birth weight/drug effects [mh] OR birth weight/radiation
effects [mh] OR breast feeding/drug effects [mh] OR (carcinogens, environmental[mh] AND (fetus [mh] or pregnancy [mh])) OR (carcinogens [mh] AND (fetus [mh]OR pregnancy [mh])) OR (cardiovascular abnormalities/ci [mh] AND fetus [mh]) OR(cardiovascular abnormalities/et [mh] AND fetus [mh]) OR (cocaine[mh] AND(fetus [mh] or pregnancy [mh])) OR (congenital abnormalities [mh] AND (fetus[mh] or pregnancy [mh])) OR (dna damage [mh] AND (pregnancy [mh] OR fetus[mh])) OR embryo/de [mh] OR embryo/re [mh] OR embryo loss/ci [mh] ORembryonic and fetal development/drug effects [mh] OR embryonic and fetal
development/radiation effects [mh] OR embryonic structures/drug effects [mh]OR embryonic structures/pathology [mh] OR embryonic structures/radiation
effects [mh] OR (environmental exposure[mh] AND (pregnancy [mh] OR fetus[mh])) OR fertility/drug effects [mh] OR fertility/radiation effects [mh] OR fetal
alcohol syndrome[mh:noexp] OR fetal death/chemically induced [mh] OR fetal
death/etiology [mh] OR fetal death/genetics [mh] OR fetal death/pathology [mh]OR fetal diseases/chemically induced [mh] OR fetal diseases/etiology [mh] OR
fetal diseases/genetics [mh] OR fetal growth retardation/et [mh] OR fetal growth
retardation/ci [mh] OR fetal resorption/chemically induced [mh] OR fetal
resorption/etiology [mh] OR fetal resorption/genetics [mh] OR
fetus/abnormalities [mh] OR fetus/drug effects [mh] OR fetus/radiation effects
Bisphenol S

115

PubMed DART
strategy

OEHHA
October 2023

SET
STRATEGY#[mh] OR (fetus*[tw] AND expos*[tw]) OR (genetic diseases, inborn/CI [mh] AND(fetus [mh] OR pregnancy [mh])) OR germ cells/drug effects [mh] OR germ
cells/radiation effects [mh] OR (hazardous substances [mh] AND (fetus [mh] or
pregnancy [mh])) OR heavy metal poisoning[mh] OR lactation/drug effects [mh]OR lactation/radiation effects [mh] OR (lead [mh:noexp] AND (fetus [mh] or
pregnancy [mh])) OR (lead poisoning[mh] AND (fetus [mh] or pregnancy [mh]))OR
maternal exposure [mh] OR maternal-fetal exchange/genetics [mh] OR maternalfetal exchange/drug effects [mh] OR maternal-fetal exchange/radiation
effects[mh] OR (mutagens [mh] AND (pregnancy [mh] OR fetus [mh])) OR neonatal
abstinence syndrome[mh] OR "neonatal abstinence syndrome"[ti] OR neonatal
sepsis [mh] OR ovary/drug effects [mh] OR ovary/radiation effects [mh] ORpaternal exposure [mh] OR placenta diseases/chemically induced [mh] OR
placenta diseases/etiology [mh] OR placenta/abnormalities [mh] OR
placenta/drug effects [mh] OR placenta/radiation effects [mh] OR pregnancy
Complications, Infectious/epidemiology [mh] OR pregnancy Complications/ci [mh]OR pregnancy outcome/ge [mh] OR (prenatal*[tw] AND expos*[tw]) OR prenatal
exposure delayed effects [mh] OR (protein deficiency[mh:noexp] AND (fetus [mh]or pregnancy [mh])) OR reproduction/drug effects [mh:noexp] OR
reproduction/radiation effects [mh] OR rubella/congenital[mh:noexp] OR rubella
syndrome, congenital/etiology[mh:noexp] OR (teratogens [mh] AND (pregnancy[mh] OR fetus [mh]))OR Teratology [Journal] OR teratology [mh] OR testis/drug
effects [mh] OR testis/radiation effects [mh]3"abortion, spontaneous"[mh] OR "abortion*"[tiab] OR "Acrosome"[mh] OR"Acrosome"[tiab] OR "Adrenarche"[tiab] OR "androgen antagonists"[mh] OR"androgen*"[tiab] OR "androgens"[mh] OR "Androstenedione"[tiab] OR"anogenital distance"[tiab] OR "ano genital distance"[tiab] OR "anovulat*"[tiab]
OR "Aspermia"[tiab] OR "atretic follicle*"[tiab] OR "Azoospermia"[tiab] OR "birthdefect*"[tiab] OR "birth weight"[mh] OR "birth weight"[tiab] OR "breast
feed*"[tiab] OR "breast feeding"[mh] OR "breastfeed*"[tiab] OR "chorionicvilli"[tiab] OR "conception*"[tiab] OR "congenital abnormalities"[mh] OR"Congenital"[tiab] OR "corpus luteum"[tiab] OR "cumulus cell*"[tiab] OR"cytotrophoblast*"[tiab] OR "decidua"[tiab] OR "deciduum"[tiab] OR "dnadamage"[mh] OR "ductus deferens"[tiab] OR "efferent duct*"[tiab] OR"ejaculat*"[tiab] OR "Embryo"[tiab] OR "Embryoes"[tiab] OR "embryonic and fetal
development"[mh] OR "embryonic structures"[mh] OR "Embryonic"[tiab] OR"embryotoxic*"[tiab] OR "endometri*"[tiab] OR "Epididymis"[mh] OR"Epididymis"[tiab] OR "erecti*"[tiab] OR "Estradiol"[tiab] OR "estrogen

Bisphenol S

116

CONCEPT GROUP

Additional DART
terms (OEHHA
Strategy)

OEHHA
October 2023

SET
STRATEGY#
antagonists"[mh] OR "estrogen receptor modulators"[mh] OR "estrogen*"[tiab]
OR "estrogens"[mh] OR "Estrus"[tiab] OR "fallopian tube*"[tiab] OR "fallopian
tubes"[mh] OR "fecund*"[tiab] OR "Fertility"[mh] OR "Fertility"[tiab] OR"Fertilization"[tiab] OR "Fetal"[tiab] OR "Fetus"[mh] OR "Fetus"[tiab] OR"foetal"[tiab] OR "foetus"[tiab] OR "follicle stimulating hormone"[tiab] OR"FSH"[tiab] OR "genetic diseases, inborn"[mh] OR "genital diseases, female"[mh]OR "genital diseases, male"[mh] OR "genital*"[tiab] OR "genitalia"[mh] OR "germcell*"[tiab] OR "germ cells"[mh] OR "gestat*"[tiab] OR "gonad*"[tiab] OR "gonadal
disorders"[mh] OR "gonadal hormones"[mh] OR "gonadotropins"[mh] OR"gonads"[mh] OR "graafian follicle*"[tiab] OR "granulosa cell*"[tiab] OR "humandevelopment"[mh] OR "Implantation"[tiab] OR "in utero"[tiab] OR "infant*"[tiab]
OR "infant, newborn"[mh] OR "infertil*"[tiab] OR "Inhibin"[tiab] OR"Intrauterine"[tiab] OR "Lactation"[tiab] OR "lactation disorders"[mh] OR "leydig
cell*"[tiab] OR "leydig cells"[mh] OR "LH"[tiab] OR "luteal cell*"[tiab] OR"luteinizing hormone"[tiab] OR "maternal exposure"[mh] OR "Maternal"[tiab] OR"Menses"[tiab] OR "menstrua*"[tiab] OR "miscarriage*"[tiab] OR "neonat*"[tiab]
OR "Oligospermia"[tiab] OR "oocyte*"[tiab] OR "Oogonia"[tiab] OR "Ova"[tiab] OR"ovarian follicle*"[tiab] OR "Ovarian"[tiab] OR "Ovaries"[tiab] OR "Ovary"[mh] OR"Ovary"[tiab] OR "oviduct*"[tiab] OR "oviducts"[mh] OR "ovulat*"[tiab] OR"Ovum"[mh] OR "Ovum"[tiab] OR "paternal exposure"[mh] OR "Paternal"[tiab] OR"peripubert*"[tiab] OR "pituitary hormones"[mh] OR "placenta*"[tiab] OR"placenta"[mh] OR "placental hormones"[mh] OR "preconception*"[tiab] OR "preconception*"[tiab] OR "pregnan*"[tiab] OR "pregnancy complications"[mh] OR"pregnancy"[mh] OR "prenatal exposure delayed effects"[mh] OR "prenatal"[tiab]
OR "Pre-natal"[tiab] OR "Preterm"[tiab] OR "Pre-term"[tiab] OR "primaryfollicle*"[tiab] OR "Progesterone"[tiab] OR "progestin*"[tiab] OR "progestins"[mh]OR "Prostate"[mh] OR "Prostate"[tiab] OR "reproduct*"[tiab] OR "reproductive
physiological phenomena"[mh] OR "secondary follicle*"[tiab] OR "Semen"[mh] OR"Semen"[tiab] OR "seminal vesicle*"[tiab] OR "seminal vesicles"[mh] OR"Seminal"[tiab] OR "seminiferous epithelium"[tiab] OR "seminiferous
tubule*"[tiab] OR "seminiferous tubules"[mh] OR "Seminiferous"[tiab] OR "sertoli
cells"[mh] OR "sertoli cell*"[tiab] OR "sexual development"[mh] OR "Sperm"[tiab]
OR "spermatid*"[tiab] OR "spermatocyte*"[tiab] OR "spermatogenesis"[tiab] OR"Spermatogonia"[tiab] OR "Spermatozoa"[mh] OR "Spermatozoa"[tiab] OR"Sterile"[tiab] OR "Sterility"[tiab] OR "stillbirth*"[tiab] OR "Stillborn"[tiab] OR"syncytiotrophoblast*"[tiab] OR "teratogen*"[tiab] OR "teratogens"[mh] OR"tertiary follicle*"[tiab] OR "Testes"[tiab] OR "testic*"[tiab] OR "Testis"[mh] OR"Testis"[tiab] OR "Testosterone"[tiab] OR "theca cell*"[tiab] OR "thyroid
hormones"[mh] OR "trophoblast*"[tiab] OR "urogenital abnormalities"[mh] OR

Bisphenol S

117

CONCEPT GROUP

OEHHA
October 2023

SET
STRATEGY#"urogenital*"[tiab] OR "Uterine"[tiab] OR "Uterus"[mh] OR "Uterus"[tiab] OR"vagina*"[tiab] OR "vas deferens"[mh] OR "vas deferens"[tiab] OR "zygote*"[tiab]

4"spermatozoa"[mh] OR "acrosome reaction"[mh] OR "Sperm Capacitation"[mh]OR "sperm transport"[mh] OR "sperm-ovum interactions"[mh] OR"acrosome"[tiab] OR "spermatozoa"[tiab] OR "sperm"[tiab] OR"spermatogonia"[tiab] OR "spermatophore*"[tiab] OR "spermatocyte*"[tiab] OR"spermatid*"[tiab] OR "spermatogenesis"[tiab] OR "capacitation"[tiab] OR(("Germ cells"[mh] OR "germ cell*"[tiab]) AND ("male"[mh] OR "male"[tiab])) OR"Leydig cells"[mh] OR "leydig cell*"[tiab] OR "sertoli cells"[mh] OR "sertoli
cell*"[tiab] OR "cytoskeleton"[tiab] OR "gonadal somatic cells"[tiab] OR "folliclestimulating hormone"[mh] OR "testosterone congeners"[mh] OR "luteinizing
hormone"[mh] OR "androgens"[mh] OR "follicle stimulating hormone*"[tiab] OR"FSH"[tiab] OR "luteinizing hormone"[tiab] OR "LH"[tiab] OR "Inhibin"[tiab] OR"testosterone"[tiab] OR "prolactin"[tiab] OR "androgen*"[tiab] OR "gonadotropin
releasing hormone"[tiab] OR "GnRH"[tiab] OR "steroidogenic"[tiab] OR"reproductive hormone*"[tiab] OR "reproductive steroid hormone*"[tiab] OR "sexsteroid*"[tiab] OR "hypothalamic pituitary gonadal axis"[tiab] OR "hpg axis"[tiab]
OR "hypothalamic pituitary adrenal axis"[tiab] OR "hypothalamic pituitary thyroid
axis"[tiab] OR "CYP3A4"[tiab] OR "CYP17A1"[tiab] OR "aromatase"[tiab] OR"receptors, gonadotropin"[mh] OR "receptors, pituitary hormone"[mh] OR"receptors, estrogen"[mh] OR "receptors, oxytocin"[mh] OR "receptors,
androgen"[mh] OR "hormone receptor*"[tiab] OR "lh receptor*"[tiab] OR"gonadotropin receptor*"[tiab] OR "estrogen receptor*"[tiab] OR "oestrogen
receptor*"[tiab] OR "fsh receptor*"[tiab] OR "androgen receptor*"[tiab] OR"testosterone receptor*"[tiab] OR "prolactin receptor*"[tiab] OR "DNA
Adducts"[mh] OR "comet assay"[mh] OR "Germ-line mutation"[mh] OR

Bisphenol S

118

CONCEPT GROUP

Key
Characteristics ofMale
Reproductive
Toxicity

OEHHA
October 2023

SET
STRATEGY#"Mutagenesis"[mh] OR "Mutagenicity tests"[mh] OR "sister chromatid
exchange"[mh] OR "Mutation"[mh] OR "DNA Repair"[mh] OR "genomic
instability"[mh] OR "Aneuploidy"[mh] OR "ames assay"[tiab] OR "ames test"[tiab]
OR "bacterial reverse mutation assay"[tiab] OR "clastogen*"[tiab] OR "genetic
toxicology"[tiab] OR "hyperploid"[tiab] OR "micronucleus test"[tiab] OR"tetraploid"[tiab] OR "chromosome aberrations"[tiab] OR "mutation*"[tiab] OR"chromosome translocation*"[tiab] OR "dna protein crosslink*"[tiab] OR "dnadamag*"[tiab] OR "dna inhibit*"[tiab] OR "Micronuclei"[tiab] OR"Micronucleus"[tiab] OR "Mutagens"[tiab] OR "strand break*"[tiab] OR"unscheduled dna synthes*"[tiab] OR "chromosomal aberration*"[tiab] OR"chromosome aberration"[tiab] OR "chromosomal abnormalit*"[tiab] OR"chromosome abnormalit*"[tiab] OR "chromosome damage*"[tiab] OR"genotoxic*"[tiab] OR "adduct formation"[tiab] OR "dna adduct*"[tiab] OR "dnabreak*"[tiab] OR "dsdna break*"[tiab] OR ("DNA"[tiab]AND "Crosslink"[tiab]) OR"microsatellite-instability"[tiab] OR "chromosomal-instability"[tiab] OR"binucleation"[tiab] OR "binucleated"[tiab] OR (("comet assay"[tiab] OR"Mutagenic"[tiab] OR "Mutagenicity"[tiab] OR "mutations"[tiab] OR"chromosomal-aberration-test"[tiab] OR "sister chromatid exchange"[tiab] OR"SOS-response"[tiab] OR "polyploid*"[tiab] OR "genomic instability"[tiab] OR "dnarepair*"[tiab] OR "aneuploid*"[tiab]) NOT "Medline"[Filter]) OR "epigenesis,
genetic"[mh] OR "epigenomics"[mh] OR "DNA methylation"[mh] OR "genesilencing"[mh] OR "histone deacetylases"[mh] OR "RNA Interference"[mh] OR"microRNAs"[mh] OR "rna, small interfering"[mh] OR "cpg islands/genetics"[mh]OR "cpg island methylator"[tiab] OR "cpg island methylation"[tiab] OR"epigenotype"[tiab] OR "epimutation*"[tiab] OR "methylation associatedsilencing"[tiab] OR "histone tail modifications"[tiab] OR "histone tailmodification"[tiab] OR "chromatin organization"[tiab] OR "chromatin
packag*"[tiab] OR "histone modification"[tiab] OR "histone retention"[tiab] OR"epigenetic*"[tiab] OR "epigenomic*"[tiab] OR "RNA Interference"[tiab] OR"noncoding rna"[tiab] OR "ncRNA"[tiab] OR "gene activation"[tiab] OR"proteasome"[tiab] OR "DNA methylation"[tiab] OR "Methylation"[Title] OR"CIMP"[tiab] OR "Free Radicals"[mh] OR "Reactive Oxygen Species"[mh] OR"Oxidative stress"[mh] OR "Electron Transport"[mh] OR "free radical*"[tiab] OR"oxygen radical*"[tiab] OR "Oxidative stress"[tiab] OR "redox balance"[tiab] OR"oxidative damage*"[tiab] OR "Reactive Oxygen Species"[tiab] OR "reactive
nitrogen species"[tiab] OR "superoxide radical*"[tiab] OR "hydroxyl radical"[tiab]
OR "glutathione deplet*"[tiab] OR "oxidative protein damage"[tiab] OR "Creactive protein"[mh] OR "eosinophils"[mh] OR ("fibrinogen"[tiab]AND"Inflammation"[tiab]) OR "chronicinflammation"[tiab] OR "chronicallyinflamed"[tiab] OR "acute inflammat*"[tiab] OR "infiltrating leukocyt*"[tiab] OR
Bisphenol S

119

CONCEPT GROUP

OEHHA
October 2023

SET
STRATEGY#"inflammatory-leukocyte"[tiab] OR "inflammatory-leukocytes"[tiab] OR "leukocyte
infiltrat*"[tiab] OR "pro-inflammatory"[tiab] OR "proinflammatory"[tiab] OR"macrophage-recruitment"[tiab] OR "macrophage inflammatory proteins"[tiab]
OR "macrophage colony stimulating factor*"[tiab] OR "urethritis"[tiab] OR"prostatitis"[tiab] OR "seminal vesiculitis"[tiab] OR "epididymitis"[tiab] OR"orchitis"[tiab]

5("hypothalamic pituitary ovarian axis"[tiab] OR "hpo axis"[tiab] OR "gonadal
hormones"[mh] OR "pituitary hormones"[mh] OR "gonadotropin releasing
hormone"[mh] OR "follicle stimulating hormone"[mh] OR "testosterone"[mh] OR"receptors, gonadotropin"[mh] OR "receptors, pituitary hormone"[mh] OR"receptors, estrogen"[mh] OR "receptors, oxytocin"[mh] OR "estrogens"[mh] OR"estradiol"[mh] OR "estriol"[mh] OR "estrone"[mh] OR "luteinizing hormone"[mh]OR "androgens"[mh] OR "hydroxyprogesterones"[mh] OR"Dehydroepiandrosterone"[mh] OR "Androstenedione"[mh] OR"Androstenediol"[mh] OR "Dihydrotestosterone"[mh] OR "androgen
receptor*"[tiab] OR "estradiol receptor*"[tiab] OR "estrogen receptor*"[tiab] OR"follicle stimulating hormone"[tiab] OR "fsh receptor*"[tiab] OR "gonadotropin
releasing hormone"[tiab] OR "hormone receptor*"[tiab] OR "lh receptor*"[tiab]
OR "luteinizing hormone"[tiab] OR "oestrogen receptor*"[tiab] OR "ovarian
hormone*"[tiab] OR "ovarian steroid*"[tiab] OR "oxytocin receptor*"[tiab] OR

Bisphenol S

120

CONCEPT GROUP

Key
Characteristics ofFemaleReproductive
Toxicity

OEHHA
October 2023

SET
STRATEGY#"plasma membrane receptor*"[tiab] OR "prolactin receptor*"[tiab] OR"reproductive hormone*"[tiab] OR "sex hormone*"[tiab] OR "testosterone
receptor*"[tiab] OR "activin"[tiab] OR "estradiol"[tiab] OR "estriol"[tiab] OR"estrogen"[tiab] OR "estrone"[tiab] OR "FSH"[tiab] OR "gnrh"[tiab] OR"gonadotropin*"[tiab] OR "gonadotropin receptor*"[tiab] OR "hcg"[tiab] OR"inhibin"[tiab] OR "LH"[tiab] OR "LHRH"[tiab] OR "oestriol"[tiab] OR"oestradiol"[tiab] OR "oestrogen"[tiab] OR "oestrone"[tiab] OR "Oxytocin"[tiab]
OR "progesterone"[tiab] OR "prolactin"[tiab] OR "Steroidogenic"[tiab] OR"testosterone"[tiab] OR "Pregnenolone"[tiab] OR "17alpha hydroxy 6 methylene
progesterone"[Supplementary Concept] OR "Dehydroepiandrosterone"[tiab] OR"DHEA"[tiab] OR "DHEAS"[tiab] OR "Androstenedione"[tiab] OR"Androstenediol"[tiab] OR "Dihydrotestosterone"[tiab] OR "steroidogenic acuteregulatory protein"[Supplementary Concept] OR "steroidogenic acute regulatory
protein"[tiab] OR "star protein"[tiab] OR "cholesterol side chain cleavage
enzyme"[mh] OR "cholesterol side chain cleavage enzyme"[tiab] OR "cholesterol
desmolase"[tiab] OR "cytochrome p 450 scc"[tiab] OR "P450scc"[tiab] OR"CYP11A"[tiab] OR "CYP11A1"[tiab] OR "17alpha hydroxylase"[tiab] OR "17,20
lyase"[tiab] OR "P450c17"[tiab] OR "CYP17"[tiab] OR "aromatase"[mh] OR"aromatase"[tiab] OR "cytochrome p450 family 19"[mh] OR "cytochrome p450
family 19"[tiab] OR "P450arom"[tiab] OR "CYP19"[tiab] OR "3 or 17 betahydroxysteroid dehydrogenase"[Supplementary Concept] OR "3betahydroxysteroid dehydrogenase"[tiab] OR "3beta hsd"[tiab] OR "17betahydroxysteroid dehydrogenase"[tiab] OR "17beta hsd*"[tiab] OR "5alphareductase"[tiab] OR ("DNA Adducts"[mh] OR "Comet Assay"[mh] OR "Germ-line
mutation"[mh] OR "Mutagenesis"[mh] OR "Mutagenicity tests"[mh] OR "Sisterchromatid exchange"[mh] OR "Mutation"[mh] OR "Ames-Assay"[tiab] OR "Amestest"[tiab] OR "Bacterial-Reverse-Mutation-Assay"[tiab] OR "clastogen*"[tiab] OR"dna repair*"[tiab] OR "Genetic-toxicology"[tiab] OR "hyperploid"[tiab] OR"micronucleus-test"[tiab] OR "tetraploid"[tiab] OR "Chromosomeaberrations"[tiab] OR "DNA-damage"[tiab] OR "mutation*"[tiab] OR"chromosome-translocations"[tiab] OR "dna protein crosslink*"[tiab] OR "dnadamag*"[tiab] OR "dna inhibit*"[tiab] OR "Micronuclei"[tiab] OR"Micronucleus"[tiab] OR "Mutagens"[tiab] OR "strand break*"[tiab] OR"unscheduled dna synthes*"[tiab] OR "chromosomal aberration*"[tiab] OR"chromosome aberration*"[tiab] OR "chromosomal abnormalit*"[tiab] OR"chromosome abnormalit*"[tiab] OR "genotoxic*"[tiab] OR "adductformation"[tiab] OR "dna adduct*"[tiab] OR "dna break*"[tiab] OR "dsdna
break*"[tiab]) OR ("epigenesis, genetic"[mh] OR "epigenomics"[mh] OR "DNAmethylation"[mh] OR "gene silencing"[mh] OR "histone deacetylases"[mh] OR"RNA Interference"[mh] OR "microRNAs"[mh] OR "rna, small interfering"[mh] OR
Bisphenol S

121

CONCEPT GROUP

OEHHA
October 2023

SET
STRATEGY#"cpg islands/genetics"[mh] OR "cpg island methylator"[tiab] OR "cpg island
methylation"[tiab] OR "epigenotype"[tiab] OR "epimutation*"[tiab] OR"methylation associated silencing"[tiab] OR "histone tail modifications"[tiab] OR"histone tail modification"[tiab] OR "chromatin organization"[tiab] OR "chromatin
packag*"[tiab] OR "histone modification"[tiab] OR "histone retention"[tiab] OR"epigenetic*"[tiab] OR "epigenomic*"[tiab] OR "RNA Interference"[tiab] OR"noncoding rna"[tiab] OR "ncRNA"[tiab] OR "gene activation"[tiab] OR"proteasome"[tiab] OR "DNA methylation"[tiab] OR "Methylation"[Title] OR"CIMP"[tiab]) OR ("mitochondria"[mh] OR "oxidative phosphorylation"[mh] OR"mitochrondria*"[tiab] OR "oxidative phosphorylation"[tiab] OR "oxidative
damage"[tiab] OR "fatty acid beta oxidation"[tiab] OR "calcium buffering"[tiab] OR"ca2 buffering"[tiab] OR "mitochondrial dna mutation*"[tiab] OR "mtdna
mutation*"[tiab] OR "mtDNA"[tiab]) OR ("Free Radicals"[mh] OR "Reactive Oxygen
Species"[mh] OR "Oxidative stress"[mh] OR "Electron Transport"[mh] OR "freeradical*"[tiab] OR "oxygen radical*"[tiab] OR "Oxidative stress"[tiab] OR "redox
balance"[tiab] OR "oxidative damage*"[tiab] OR "Reactive Oxygen Species"[tiab]
OR "reactive nitrogen species"[tiab] OR "superoxide radical*"[tiab] OR "hydroxyl
radical"[tiab] OR "glutathione deplet*"[tiab] OR "oxidative protein damage"[tiab])
OR ("cytotoxicity, immunologic"[mh] OR "Immunologic Factors"[mh] OR"Immunomodulation"[mh] OR "B-Cell Activation Factor Receptor"[mh] OR"Antigenic Modulation"[mh] OR "B-Cell Activating Factor"[mh] OR "Immunologic
Factors"[Pharmacological Action] OR "b-cell-activation"[tiab] OR "immune
surveillance"[tiab] OR "immune suppress*"[tiab] OR "immunostimulant"[tiab] OR"immune-activation"[tiab] OR "immunodeficien*"[tiab] OR "somatichypermutation"[tiab] OR "immune-activation"[tiab] OR "immune-systemactivation"[tiab] OR "Chronic-antigenic-stimulation"[tiab] OR"immunosuppress*"[tiab] OR "immune dysregulation"[tiab]) OR ("signaltransduction"[mh] OR "signal transduction"[tiab] OR "signal pathway*"[tiab] OR"signaling pathway*"[tiab] OR "ion channel"[tiab] OR "signaling system*"[tiab] OR"cell signal*"[tiab] OR "cellular signal*"[tiab] OR "intracellular signal*"[tiab] OR"signal cascade*"[tiab] OR "signaling cascade*"[tiab] OR "second
messenger*"[tiab] OR "calcium signal*"[tiab]) OR ("cell communication"[mh] OR"gap junctions"[mh] OR "connexins"[mh] OR "connexins"[Supplementary Concept]
OR "cell communication*"[tiab] OR "cellular communication*"[tiab] OR"intracellular communication*"[tiab] OR "cell interaction*"[tiab] OR "gapjunction*"[tiab] OR "connexin*"[tiab]) OR ("Apoptosis"[mh] OR "cytotoxicity,
immunologic"[mh] OR "Caspases"[mh] OR "autophagy"[mh] OR "necrosis"[mh] OR"Autolysis"[mh] OR "Angiogenesis Modulating Agents"[mh] OR "Angiogenesis
Inducing Agents"[Pharmacological Action] OR "Angiogenesis Inducing Agents"[mh]OR "neovascularization, pathologic"[mh] OR "Cell Proliferation"[mh] OR
Bisphenol S

122

CONCEPT GROUP

OEHHA
October 2023

SET
STRATEGY#"homeostasis"[mh] OR "Cyclin-Dependent Kinases"[mh] OR "Cyclin-Dependent
Kinase Inhibitor Proteins"[mh] OR "Mitogens"[mh] OR"Mitogens"[Pharmacological Action] OR "cell hypoxia"[mh] OR "angiogenic"[tiab]
OR "Apoptosis"[tiab] OR "autophagy"[tiab] OR "Caspases"[tiab] OR "cell cycle
control*"[tiab] OR "cell cycle arrest"[tiab] OR "cell hypoxia"[tiab] OR "Cell
Proliferation"[tiab] OR "cellular-energetics"[tiab] OR "cellular-hypoxia"[tiab] OR"cellular proliferation"[tiab] OR "cellular replication*"[tiab] OR "Cytogenesis"[tiab]
OR "Cytogenic"[tiab] OR "Cytotoxin"[tiab] OR "hepatocellular-proliferation"[tiab]
OR "hyperplasia"[tiab] OR "hypoxic cell*"[tiab] OR "mitogenesis"[tiab] OR "mitotic
checkpoint*"[tiab] OR "Neoplasia"[tiab] OR "p53 delet*"[tiab] OR "p53inactivat*"[tiab] OR "p53 inhibit*"[tiab] OR "prb delet*"[tiab] OR "prbinactivat*"[tiab] OR "prb inhibit*"[tiab] OR "programmed cell death"[tiab] OR("Rb"[All Fields] AND "p16ink4a inactiv*"[tiab]) OR "retinoblastoma-protein"[tiab]
OR "senescence"[tiab] OR "senescent"[tiab] OR "survivin"[tiab]) OR"microtubules"[mh] OR "spindle apparatus"[mh] OR "microtubule organizing
center"[mh] OR "microtubule*"[tiab] OR "spindle formation"[tiab] OR "spindle
apparatus"[tiab] OR "meiotic spindle*"[tiab] OR "mitotic spindle*"[tiab])
6#2 OR #3 OR #4 OR #5

7#1 AND #6

8#7 NOT (animals[mh] NOT humans[mh])CONCEPT GROUP

Combine DART
concept groups
CombineChemical + DART
Remove animalstudies
FINAL

Table A.9 PubMed search strategy for animal DART studies
SET
STRATEGY#

CONCEPT
GROUP

1(80-09-1[rn] OR "Bisphenol S"[tiab] OR "bis(4-hydroxyphenyl)sulfone"[nm]OR "bis(4-hydroxyphenyl)sulfone"[tiab] OR "4,4'-Sulfonyldiphenol"[tiab] OR"Phenol, 4,4'-sulfonylbis-"[tiab] OR "BPS-monoglucuronide"[tiab] OR "BPS1G"[tiab] OR "bisphenol S dicyanate ester"[tiab] OR (bps[tiab] NOT("bps"[tiab] NOT "bisphenol"[tiab])))Chemical Terms

2(abnormalities, drug-induced [mh] AND (fetus [mh] or pregnancy [mh])) OR((abnormalities, multiple/chemically induced [mh] OR abnormalities,
multiple/epidemiology [mh] OR abnormalities, multiple/etiology[mh] ORabnormalities, multiple/genetics [mh] OR abnormalities, multiple/pathology

PubMed DART
strategy

Bisphenol S

123

OEHHA
October 2023

SET
STRATEGY#

CONCEPT
GROUP[mh]) AND (pregnancy [mh] OR fetus [mh]))OR (abnormalities, radiationinduced [mh] AND (fetus [mh] or pregnancy [mh])) OR abortion,
habitual/chemically induced [mh] OR abortion, habitual/etiology [mh] ORabortion, spontaneous/chemically induced [mh] OR abortion,
spontaneous/etiology [mh] OR (alcoholic intoxication[mh] AND (fetus [mh]or pregnancy [mh])) OR (alcohol drinking [mh:noexp] AND (fetus [mh] or
pregnancy [mh])) OR Birth Defects Res B Dev Reprod Toxicol [TA] OR birthweight/drug effects [mh] OR birth weight/radiation effects [mh] OR breast
feeding/drug effects [mh] OR (carcinogens, environmental [mh] AND (fetus[mh] or pregnancy [mh])) OR (carcinogens [mh] AND (fetus [mh] ORpregnancy [mh])) OR (cardiovascular abnormalities/ci [mh] AND fetus [mh])OR (cardiovascular abnormalities/et [mh] AND fetus [mh]) OR (cocaine[mh]AND (fetus [mh] or pregnancy [mh])) OR (congenital abnormalities [mh]AND (fetus [mh] or pregnancy [mh])) OR (dna damage [mh] AND(pregnancy [mh] OR fetus [mh])) OR embryo/de [mh] OR embryo/re [mh]OR embryo loss/ci [mh] OR embryonic and fetal development/drug effects[mh] OR embryonic and fetal development/radiation effects [mh] ORembryonic structures/drug effects [mh] OR embryonic structures/pathology[mh] OR embryonic structures/radiation effects [mh] OR (environmental
exposure[mh] AND (pregnancy [mh] OR fetus [mh])) OR fertility/drug effects[mh] OR fertility/radiation effects [mh] OR fetal alcohol syndrome[mh:noexp]
OR fetal death/chemically induced [mh] OR fetal death/etiology [mh] OR
fetal death/genetics [mh] OR fetal death/pathology [mh] OR fetal
diseases/chemically induced [mh] OR fetal diseases/etiology [mh] OR fetal
diseases/genetics [mh] OR fetal growth retardation/et [mh] OR fetal growth
retardation/ci [mh] OR fetal resorption/chemically induced [mh] OR fetal
resorption/etiology [mh] OR fetal resorption/genetics [mh] OR
fetus/abnormalities [mh] OR fetus/drug effects [mh] OR fetus/radiation
effects [mh] OR (fetus*[tw] AND expos*[tw]) OR (genetic diseases,
inborn/CI [mh] AND (fetus [mh] OR pregnancy [mh])) OR germ cells/drug
effects [mh] OR germ cells/radiation effects [mh] OR (hazardous
substances [mh] AND (fetus [mh] or pregnancy [mh])) OR heavy metal
poisoning[mh] OR lactation/drug effects [mh] OR lactation/radiation effects[mh] OR (lead [mh:noexp] AND (fetus [mh] or pregnancy [mh])) OR (leadpoisoning[mh] AND (fetus [mh] or pregnancy [mh]))OR maternal exposure[mh] OR maternal-fetal exchange/genetics [mh] OR maternal-fetal
Bisphenol S

124

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SET
STRATEGY#

CONCEPT
GROUP

exchange/drug effects [mh] OR maternal-fetal exchange/radiation
effects[mh] OR (mutagens [mh] AND (pregnancy [mh] OR fetus [mh])) OR
neonatal abstinence syndrome[mh] OR "neonatal abstinence syndrome"[ti]OR neonatal sepsis [mh] OR ovary/drug effects [mh] OR ovary/radiation
effects [mh] OR paternal exposure [mh] OR placenta diseases/chemicallyinduced [mh] OR placenta diseases/etiology [mh] OR
placenta/abnormalities [mh] OR placenta/drug effects [mh] OR
placenta/radiation effects [mh] OR pregnancy Complications,
Infectious/epidemiology [mh] OR pregnancy Complications/ci [mh] ORpregnancy outcome/ge [mh] OR (prenatal*[tw] AND expos*[tw]) OR
prenatal exposure delayed effects [mh] OR (protein deficiency[mh:noexp]
AND (fetus [mh] or pregnancy [mh])) OR reproduction/drug effects[mh:noexp] OR reproduction/radiation effects [mh] OR
rubella/congenital[mh:noexp] OR rubella syndrome,congenital/etiology[mh:noexp] OR (teratogens [mh] AND (pregnancy [mh]OR fetus [mh]))OR Teratology [Journal] OR teratology [mh] OR testis/drug
effects [mh] OR testis/radiation effects [mh]3"abortion, spontaneous"[mh] OR "abortion*"[tiab] OR "Acrosome"[mh] OR"Acrosome"[tiab] OR "Adrenarche"[tiab] OR "androgen antagonists"[mh] OR"androgen*"[tiab] OR "androgens"[mh] OR "Androstenedione"[tiab] OR"anogenital distance"[tiab] OR "ano genital distance"[tiab] OR"anovulat*"[tiab] OR "Aspermia"[tiab] OR "atretic follicle*"[tiab] OR"Azoospermia"[tiab] OR "birth defect*"[tiab] OR "birth weight"[mh] OR "birthweight"[tiab] OR "breast feed*"[tiab] OR "breast feeding"[mh] OR"breastfeed*"[tiab] OR "chorionic villi"[tiab] OR "conception*"[tiab] OR"congenital abnormalities"[mh] OR "Congenital"[tiab] OR "corpus
luteum"[tiab] OR "cumulus cell*"[tiab] OR "cytotrophoblast*"[tiab] OR"decidua"[tiab] OR "deciduum"[tiab] OR "dna damage"[mh] OR "ductus
deferens"[tiab] OR "efferent duct*"[tiab] OR "ejaculat*"[tiab] OR"Embryo"[tiab] OR "Embryoes"[tiab] OR "embryonic and fetal
development"[mh] OR "embryonic structures"[mh] OR "Embryonic"[tiab] OR

Bisphenol S

125

Additional
DART terms(OEHHA
Strategy)

OEHHA
October 2023

SET
STRATEGY#

CONCEPT
GROUP"embryotoxic*"[tiab] OR "endometri*"[tiab] OR "Epididymis"[mh] OR"Epididymis"[tiab] OR "erecti*"[tiab] OR "Estradiol"[tiab] OR "estrogenantagonists"[mh] OR "estrogen receptor modulators"[mh] OR"estrogen*"[tiab] OR "estrogens"[mh] OR "Estrus"[tiab] OR "fallopian
tube*"[tiab] OR "fallopian tubes"[mh] OR "fecund*"[tiab] OR "Fertility"[mh]OR "Fertility"[tiab] OR "Fertilization"[tiab] OR "Fetal"[tiab] OR "Fetus"[mh]OR "Fetus"[tiab] OR "foetal"[tiab] OR "foetus"[tiab] OR "follicle stimulating
hormone"[tiab] OR "FSH"[tiab] OR "genetic diseases, inborn"[mh] OR"genital diseases, female"[mh] OR "genital diseases, male"[mh] OR"genital*"[tiab] OR "genitalia"[mh] OR "germ cell*"[tiab] OR "germ cells"[mh]OR "gestat*"[tiab] OR "gonad*"[tiab] OR "gonadal disorders"[mh] OR"gonadal hormones"[mh] OR "gonadotropins"[mh] OR "gonads"[mh] OR"graafian follicle*"[tiab] OR "granulosa cell*"[tiab] OR "humandevelopment"[mh] OR "Implantation"[tiab] OR "in utero"[tiab] OR"infant*"[tiab] OR "infant, newborn"[mh] OR "infertil*"[tiab] OR "Inhibin"[tiab]
OR "Intrauterine"[tiab] OR "Lactation"[tiab] OR "lactation disorders"[mh] OR"leydig cell*"[tiab] OR "leydig cells"[mh] OR "LH"[tiab] OR "luteal cell*"[tiab]
OR "luteinizing hormone"[tiab] OR "maternal exposure"[mh] OR"Maternal"[tiab] OR "Menses"[tiab] OR "menstrua*"[tiab] OR"miscarriage*"[tiab] OR "neonat*"[tiab] OR "Oligospermia"[tiab] OR"oocyte*"[tiab] OR "Oogonia"[tiab] OR "Ova"[tiab] OR "ovarian follicle*"[tiab]
OR "Ovarian"[tiab] OR "Ovaries"[tiab] OR "Ovary"[mh] OR "Ovary"[tiab] OR"oviduct*"[tiab] OR "oviducts"[mh] OR "ovulat*"[tiab] OR "Ovum"[mh] OR"Ovum"[tiab] OR "paternal exposure"[mh] OR "Paternal"[tiab] OR"peripubert*"[tiab] OR "pituitary hormones"[mh] OR "placenta*"[tiab] OR"placenta"[mh] OR "placental hormones"[mh] OR "preconception*"[tiab] OR"pre conception*"[tiab] OR "pregnan*"[tiab] OR "pregnancy
complications"[mh] OR "pregnancy"[mh] OR "prenatal exposure delayed
effects"[mh] OR "prenatal"[tiab] OR "Pre-natal"[tiab] OR "Preterm"[tiab] OR"Pre-term"[tiab] OR "primary follicle*"[tiab] OR "Progesterone"[tiab] OR"progestin*"[tiab] OR "progestins"[mh] OR "Prostate"[mh] OR"Prostate"[tiab] OR "reproduct*"[tiab] OR "reproductive physiologicalphenomena"[mh] OR "secondary follicle*"[tiab] OR "Semen"[mh] OR"Semen"[tiab] OR "seminal vesicle*"[tiab] OR "seminal vesicles"[mh] OR"Seminal"[tiab] OR "seminiferous epithelium"[tiab] OR "seminiferous
tubule*"[tiab] OR "seminiferous tubules"[mh] OR "Seminiferous"[tiab] OR
Bisphenol S

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SET
STRATEGY#

CONCEPT
GROUP"sertoli cells"[mh] OR "sertoli cell*"[tiab] OR "sexual development"[mh] OR"Sperm"[tiab] OR "spermatid*"[tiab] OR "spermatocyte*"[tiab] OR"spermatogenesis"[tiab] OR "Spermatogonia"[tiab] OR "Spermatozoa"[mh]OR "Spermatozoa"[tiab] OR "Sterile"[tiab] OR "Sterility"[tiab] OR"stillbirth*"[tiab] OR "Stillborn"[tiab] OR "syncytiotrophoblast*"[tiab] OR"teratogen*"[tiab] OR "teratogens"[mh] OR "tertiary follicle*"[tiab] OR"Testes"[tiab] OR "testic*"[tiab] OR "Testis"[mh] OR "Testis"[tiab] OR"Testosterone"[tiab] OR "theca cell*"[tiab] OR "thyroid hormones"[mh] OR"trophoblast*"[tiab] OR "urogenital abnormalities"[mh] OR "urogenital*"[tiab]
OR "Uterine"[tiab] OR "Uterus"[mh] OR "Uterus"[tiab] OR "vagina*"[tiab] OR"vas deferens"[mh] OR "vas deferens"[tiab] OR "zygote*"[tiab]

4"spermatozoa"[mh] OR "acrosome reaction"[mh] OR "Sperm
Capacitation"[mh] OR "sperm transport"[mh] OR "sperm-ovum
interactions"[mh] OR "acrosome"[tiab] OR "spermatozoa"[tiab] OR"sperm"[tiab] OR "spermatogonia"[tiab] OR "spermatophore*"[tiab] OR"spermatocyte*"[tiab] OR "spermatid*"[tiab] OR "spermatogenesis"[tiab] OR"capacitation"[tiab] OR (("Germ cells"[mh] OR "germ cell*"[tiab]) AND("male"[mh] OR "male"[tiab])) OR "Leydig cells"[mh] OR "leydig cell*"[tiab]
OR "sertoli cells"[mh] OR "sertoli cell*"[tiab] OR "cytoskeleton"[tiab] OR"gonadal somatic cells"[tiab] OR "follicle-stimulating hormone"[mh] OR"testosterone congeners"[mh] OR "luteinizing hormone"[mh] OR"androgens"[mh] OR "follicle stimulating hormone*"[tiab] OR "FSH"[tiab] OR"luteinizing hormone"[tiab] OR "LH"[tiab] OR "Inhibin"[tiab] OR"testosterone"[tiab] OR "prolactin"[tiab] OR "androgen*"[tiab] OR"gonadotropin releasing hormone"[tiab] OR "GnRH"[tiab] OR"steroidogenic"[tiab] OR "reproductive hormone*"[tiab] OR "reproductive
steroid hormone*"[tiab] OR "sex steroid*"[tiab] OR "hypothalamic pituitary
gonadal axis"[tiab] OR "hpg axis"[tiab] OR "hypothalamic pituitary adrenalaxis"[tiab] OR "hypothalamic pituitary thyroid axis"[tiab] OR "CYP3A4"[tiab]
OR "CYP17A1"[tiab] OR "aromatase"[tiab] OR "receptors,
gonadotropin"[mh] OR "receptors, pituitary hormone"[mh] OR "receptors,
estrogen"[mh] OR "receptors, oxytocin"[mh] OR "receptors, androgen"[mh]OR "hormone receptor*"[tiab] OR "lh receptor*"[tiab] OR "gonadotropin
receptor*"[tiab] OR "estrogen receptor*"[tiab] OR "oestrogen receptor*"[tiab]
OR "fsh receptor*"[tiab] OR "androgen receptor*"[tiab] OR "testosterone
receptor*"[tiab] OR "prolactin receptor*"[tiab] OR "DNA Adducts"[mh] OR
Bisphenol S

127

Key
Characteristicsof MaleReproductive
Toxicity

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October 2023

SET
STRATEGY#

CONCEPT
GROUP"comet assay"[mh] OR "Germ-line mutation"[mh] OR "Mutagenesis"[mh]OR "Mutagenicity tests"[mh] OR "sister chromatid exchange"[mh] OR"Mutation"[mh] OR "DNA Repair"[mh] OR "genomic instability"[mh] OR"Aneuploidy"[mh] OR "ames assay"[tiab] OR "ames test"[tiab] OR "bacterial
reverse mutation assay"[tiab] OR "clastogen*"[tiab] OR "genetic
toxicology"[tiab] OR "hyperploid"[tiab] OR "micronucleus test"[tiab] OR"tetraploid"[tiab] OR "chromosome aberrations"[tiab] OR "mutation*"[tiab]
OR "chromosome translocation*"[tiab] OR "dna protein crosslink*"[tiab] OR"dna damag*"[tiab] OR "dna inhibit*"[tiab] OR "Micronuclei"[tiab] OR"Micronucleus"[tiab] OR "Mutagens"[tiab] OR "strand break*"[tiab] OR"unscheduled dna synthes*"[tiab] OR "chromosomal aberration*"[tiab] OR"chromosome aberration"[tiab] OR "chromosomal abnormalit*"[tiab] OR"chromosome abnormalit*"[tiab] OR "chromosome damage*"[tiab] OR"genotoxic*"[tiab] OR "adduct formation"[tiab] OR "dna adduct*"[tiab] OR"dna break*"[tiab] OR "dsdna break*"[tiab] OR ("DNA"[tiab]AND"Crosslink"[tiab]) OR "microsatellite-instability"[tiab] OR "chromosomalinstability"[tiab] OR "binucleation"[tiab] OR "binucleated"[tiab] OR (("cometassay"[tiab] OR "Mutagenic"[tiab] OR "Mutagenicity"[tiab] OR"mutations"[tiab] OR "chromosomal-aberration-test"[tiab] OR "sister
chromatid exchange"[tiab] OR "SOS-response"[tiab] OR "polyploid*"[tiab]
OR "genomic instability"[tiab] OR "dna repair*"[tiab] OR "aneuploid*"[tiab])
NOT "Medline"[Filter]) OR "epigenesis, genetic"[mh] OR "epigenomics"[mh]OR "DNA methylation"[mh] OR "gene silencing"[mh] OR "histone
deacetylases"[mh] OR "RNA Interference"[mh] OR "microRNAs"[mh] OR"rna, small interfering"[mh] OR "cpg islands/genetics"[mh] OR "cpg island
methylator"[tiab] OR "cpg island methylation"[tiab] OR "epigenotype"[tiab]
OR "epimutation*"[tiab] OR "methylation associated silencing"[tiab] OR"histone tail modifications"[tiab] OR "histone tail modification"[tiab] OR"chromatin organization"[tiab] OR "chromatin packag*"[tiab] OR "histone
modification"[tiab] OR "histone retention"[tiab] OR "epigenetic*"[tiab] OR"epigenomic*"[tiab] OR "RNA Interference"[tiab] OR "noncoding rna"[tiab]
OR "ncRNA"[tiab] OR "gene activation"[tiab] OR "proteasome"[tiab] OR"DNA methylation"[tiab] OR "Methylation"[Title] OR "CIMP"[tiab] OR "FreeRadicals"[mh] OR "Reactive Oxygen Species"[mh] OR "Oxidative
stress"[mh] OR "Electron Transport"[mh] OR "free radical*"[tiab] OR"oxygen radical*"[tiab] OR "Oxidative stress"[tiab] OR "redox balance"[tiab]
Bisphenol S

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SET
STRATEGY#

CONCEPT
GROUP

OR "oxidative damage*"[tiab] OR "Reactive Oxygen Species"[tiab] OR"reactive nitrogen species"[tiab] OR "superoxide radical*"[tiab] OR "hydroxyl
radical"[tiab] OR "glutathione deplet*"[tiab] OR "oxidative protein
damage"[tiab] OR "C-reactive protein"[mh] OR "eosinophils"[mh] OR("fibrinogen"[tiab]AND "Inflammation"[tiab]) OR "chronicinflammation"[tiab]
OR "chronically inflamed"[tiab] OR "acute inflammat*"[tiab] OR "infiltrating
leukocyt*"[tiab] OR "inflammatory-leukocyte"[tiab] OR "inflammatoryleukocytes"[tiab] OR "leukocyte infiltrat*"[tiab] OR "pro-inflammatory"[tiab]
OR "proinflammatory"[tiab] OR "macrophage-recruitment"[tiab] OR"macrophage inflammatory proteins"[tiab] OR "macrophage colony
stimulating factor*"[tiab] OR "urethritis"[tiab] OR "prostatitis"[tiab] OR"seminal vesiculitis"[tiab] OR "epididymitis"[tiab] OR "orchitis"[tiab]

5("hypothalamic pituitary ovarian axis"[tiab] OR "hpo axis"[tiab] OR "gonadal
hormones"[mh] OR "pituitary hormones"[mh] OR "gonadotropin releasing
hormone"[mh] OR "follicle stimulating hormone"[mh] OR "testosterone"[mh]OR "receptors, gonadotropin"[mh] OR "receptors, pituitary hormone"[mh]OR "receptors, estrogen"[mh] OR "receptors, oxytocin"[mh] OR"estrogens"[mh] OR "estradiol"[mh] OR "estriol"[mh] OR "estrone"[mh] OR"luteinizing hormone"[mh] OR "androgens"[mh] OR"hydroxyprogesterones"[mh] OR "Dehydroepiandrosterone"[mh] OR"Androstenedione"[mh] OR "Androstenediol"[mh] OR"Dihydrotestosterone"[mh] OR "androgen receptor*"[tiab] OR "estradiol
receptor*"[tiab] OR "estrogen receptor*"[tiab] OR "follicle stimulating
hormone"[tiab] OR "fsh receptor*"[tiab] OR "gonadotropin releasing
hormone"[tiab] OR "hormone receptor*"[tiab] OR "lh receptor*"[tiab] OR
Bisphenol S

129

Key
Characteristics
of Female
Reproductive
Toxicity

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October 2023

SET
STRATEGY#

CONCEPT
GROUP"luteinizing hormone"[tiab] OR "oestrogen receptor*"[tiab] OR "ovarian
hormone*"[tiab] OR "ovarian steroid*"[tiab] OR "oxytocin receptor*"[tiab] OR"plasma membrane receptor*"[tiab] OR "prolactin receptor*"[tiab] OR"reproductive hormone*"[tiab] OR "sex hormone*"[tiab] OR "testosterone
receptor*"[tiab] OR "activin"[tiab] OR "estradiol"[tiab] OR "estriol"[tiab] OR"estrogen"[tiab] OR "estrone"[tiab] OR "FSH"[tiab] OR "gnrh"[tiab] OR"gonadotropin*"[tiab] OR "gonadotropin receptor*"[tiab] OR "hcg"[tiab] OR"inhibin"[tiab] OR "LH"[tiab] OR "LHRH"[tiab] OR "oestriol"[tiab] OR"oestradiol"[tiab] OR "oestrogen"[tiab] OR "oestrone"[tiab] OR"Oxytocin"[tiab] OR "progesterone"[tiab] OR "prolactin"[tiab] OR"Steroidogenic"[tiab] OR "testosterone"[tiab] OR "Pregnenolone"[tiab] OR"17alpha hydroxy 6 methylene progesterone"[Supplementary Concept] OR"Dehydroepiandrosterone"[tiab] OR "DHEA"[tiab] OR "DHEAS"[tiab] OR"Androstenedione"[tiab] OR "Androstenediol"[tiab] OR"Dihydrotestosterone"[tiab] OR "steroidogenic acute regulatory
protein"[Supplementary Concept] OR "steroidogenic acute regulatory
protein"[tiab] OR "star protein"[tiab] OR "cholesterol side chain cleavage
enzyme"[mh] OR "cholesterol side chain cleavage enzyme"[tiab] OR"cholesterol desmolase"[tiab] OR "cytochrome p 450 scc"[tiab] OR"P450scc"[tiab] OR "CYP11A"[tiab] OR "CYP11A1"[tiab] OR "17alphahydroxylase"[tiab] OR "17,20 lyase"[tiab] OR "P450c17"[tiab] OR"CYP17"[tiab] OR "aromatase"[mh] OR "aromatase"[tiab] OR "cytochrome
p450 family 19"[mh] OR "cytochrome p450 family 19"[tiab] OR"P450arom"[tiab] OR "CYP19"[tiab] OR "3 or 17 beta hydroxysteroid
dehydrogenase"[Supplementary Concept] OR "3beta hydroxysteroid
dehydrogenase"[tiab] OR "3beta hsd"[tiab] OR "17beta hydroxysteroid
dehydrogenase"[tiab] OR "17beta hsd*"[tiab] OR "5alpha-reductase"[tiab]
OR ("DNA Adducts"[mh] OR "Comet Assay"[mh] OR "Germ-line
mutation"[mh] OR "Mutagenesis"[mh] OR "Mutagenicity tests"[mh] OR"Sister-chromatid exchange"[mh] OR "Mutation"[mh] OR "AmesAssay"[tiab] OR "Ames-test"[tiab] OR "Bacterial-Reverse-MutationAssay"[tiab] OR "clastogen*"[tiab] OR "dna repair*"[tiab] OR "Genetictoxicology"[tiab] OR "hyperploid"[tiab] OR "micronucleus-test"[tiab] OR"tetraploid"[tiab] OR "Chromosome-aberrations"[tiab] OR "DNAdamage"[tiab] OR "mutation*"[tiab] OR "chromosome-translocations"[tiab]
OR "dna protein crosslink*"[tiab] OR "dna damag*"[tiab] OR "dnaBisphenol S

130

OEHHA
October 2023

SET
STRATEGY#

CONCEPT
GROUP

inhibit*"[tiab] OR "Micronuclei"[tiab] OR "Micronucleus"[tiab] OR"Mutagens"[tiab] OR "strand break*"[tiab] OR "unscheduled dna
synthes*"[tiab] OR "chromosomal aberration*"[tiab] OR "chromosome
aberration*"[tiab] OR "chromosomal abnormalit*"[tiab] OR "chromosome
abnormalit*"[tiab] OR "genotoxic*"[tiab] OR "adduct-formation"[tiab] OR"dna adduct*"[tiab] OR "dna break*"[tiab] OR "dsdna break*"[tiab]) OR("epigenesis, genetic"[mh] OR "epigenomics"[mh] OR "DNAmethylation"[mh] OR "gene silencing"[mh] OR "histone deacetylases"[mh]OR "RNA Interference"[mh] OR "microRNAs"[mh] OR "rna, smallinterfering"[mh] OR "cpg islands/genetics"[mh] OR "cpg island
methylator"[tiab] OR "cpg island methylation"[tiab] OR "epigenotype"[tiab]
OR "epimutation*"[tiab] OR "methylation associated silencing"[tiab] OR"histone tail modifications"[tiab] OR "histone tail modification"[tiab] OR"chromatin organization"[tiab] OR "chromatin packag*"[tiab] OR "histone
modification"[tiab] OR "histone retention"[tiab] OR "epigenetic*"[tiab] OR"epigenomic*"[tiab] OR "RNA Interference"[tiab] OR "noncoding rna"[tiab]
OR "ncRNA"[tiab] OR "gene activation"[tiab] OR "proteasome"[tiab] OR"DNA methylation"[tiab] OR "Methylation"[Title] OR "CIMP"[tiab]) OR("mitochondria"[mh] OR "oxidative phosphorylation"[mh] OR"mitochrondria*"[tiab] OR "oxidative phosphorylation"[tiab] OR "oxidative
damage"[tiab] OR "fatty acid beta oxidation"[tiab] OR "calcium
buffering"[tiab] OR "ca2 buffering"[tiab] OR "mitochondrial dna
mutation*"[tiab] OR "mtdna mutation*"[tiab] OR "mtDNA"[tiab]) OR ("FreeRadicals"[mh] OR "Reactive Oxygen Species"[mh] OR "Oxidative
stress"[mh] OR "Electron Transport"[mh] OR "free radical*"[tiab] OR"oxygen radical*"[tiab] OR "Oxidative stress"[tiab] OR "redox balance"[tiab]
OR "oxidative damage*"[tiab] OR "Reactive Oxygen Species"[tiab] OR"reactive nitrogen species"[tiab] OR "superoxide radical*"[tiab] OR "hydroxyl
radical"[tiab] OR "glutathione deplet*"[tiab] OR "oxidative protein
damage"[tiab]) OR ("cytotoxicity, immunologic"[mh] OR "Immunologic
Factors"[mh] OR "Immunomodulation"[mh] OR "B-Cell Activation Factor
Receptor"[mh] OR "Antigenic Modulation"[mh] OR "B-Cell Activating
Factor"[mh] OR "Immunologic Factors"[Pharmacological Action] OR "b-cellactivation"[tiab] OR "immune surveillance"[tiab] OR "immune
suppress*"[tiab] OR "immunostimulant"[tiab] OR "immune-activation"[tiab]
OR "immunodeficien*"[tiab] OR "somatic-hypermutation"[tiab] OR "immuneBisphenol S

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October 2023

SET
STRATEGY#

CONCEPT
GROUP

activation"[tiab] OR "immune-system-activation"[tiab] OR "Chronicantigenic-stimulation"[tiab] OR "immunosuppress*"[tiab] OR "immune
dysregulation"[tiab]) OR ("signal transduction"[mh] OR "signaltransduction"[tiab] OR "signal pathway*"[tiab] OR "signaling pathway*"[tiab]
OR "ion channel"[tiab] OR "signaling system*"[tiab] OR "cell signal*"[tiab]
OR "cellular signal*"[tiab] OR "intracellular signal*"[tiab] OR "signal
cascade*"[tiab] OR "signaling cascade*"[tiab] OR "second
messenger*"[tiab] OR "calcium signal*"[tiab]) OR ("cell communication"[mh]OR "gap junctions"[mh] OR "connexins"[mh] OR"connexins"[Supplementary Concept] OR "cell communication*"[tiab] OR"cellular communication*"[tiab] OR "intracellular communication*"[tiab] OR"cell interaction*"[tiab] OR "gap junction*"[tiab] OR "connexin*"[tiab]) OR("Apoptosis"[mh] OR "cytotoxicity, immunologic"[mh] OR "Caspases"[mh]OR "autophagy"[mh] OR "necrosis"[mh] OR "Autolysis"[mh] OR"Angiogenesis Modulating Agents"[mh] OR "Angiogenesis Inducing
Agents"[Pharmacological Action] OR "Angiogenesis Inducing Agents"[mh]OR "neovascularization, pathologic"[mh] OR "Cell Proliferation"[mh] OR"homeostasis"[mh] OR "Cyclin-Dependent Kinases"[mh] OR "CyclinDependent Kinase Inhibitor Proteins"[mh] OR "Mitogens"[mh] OR"Mitogens"[Pharmacological Action] OR "cell hypoxia"[mh] OR"angiogenic"[tiab] OR "Apoptosis"[tiab] OR "autophagy"[tiab] OR"Caspases"[tiab] OR "cell cycle control*"[tiab] OR "cell cycle arrest"[tiab] OR"cell hypoxia"[tiab] OR "Cell Proliferation"[tiab] OR "cellular-energetics"[tiab]
OR "cellular-hypoxia"[tiab] OR "cellular proliferation"[tiab] OR "cellular
replication*"[tiab] OR "Cytogenesis"[tiab] OR "Cytogenic"[tiab] OR"Cytotoxin"[tiab] OR "hepatocellular-proliferation"[tiab] OR"hyperplasia"[tiab] OR "hypoxic cell*"[tiab] OR "mitogenesis"[tiab] OR"mitotic checkpoint*"[tiab] OR "Neoplasia"[tiab] OR "p53 delet*"[tiab] OR"p53 inactivat*"[tiab] OR "p53 inhibit*"[tiab] OR "prb delet*"[tiab] OR "prbinactivat*"[tiab] OR "prb inhibit*"[tiab] OR "programmed cell death"[tiab] OR("Rb"[All Fields] AND "p16ink4a inactiv*"[tiab]) OR "retinoblastomaprotein"[tiab] OR "senescence"[tiab] OR "senescent"[tiab] OR"survivin"[tiab]) OR "microtubules"[mh] OR "spindle apparatus"[mh] OR"microtubule organizing center"[mh] OR "microtubule*"[tiab] OR "spindle
formation"[tiab] OR "spindle apparatus"[tiab] OR "meiotic spindle*"[tiab] OR"mitotic spindle*"[tiab])
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SET
STRATEGY#

CONCEPT
GROUP

6#2 OR #3 OR #4 OR #5

Combine DART
concept groups

7#1 AND #6

CombineChemical +DART

8("Animals, Genetically Modified"[mh] OR "Animals, Inbred Strains"[mh] OR"Chimera"[mh] OR "Animals, Laboratory"[mh] OR animals[mh:noexp]) OR(animal-stud*[tiab] OR wood-mouse[tiab] OR murinae[tiab] ORmuridae[tiab] OR cricetinae[tiab] OR rodentia[tiab] OR rodent[tiab] ORrodents[tiab] OR ferrets[tiab] OR ferret[tiab] OR polecat*[tiab] OR mustelaputorius[tiab] OR cavia[tiab] OR callithrix[tiab] OR marmoset*[tiab] ORchinchilla*[tiab] OR jird[tiab] OR jirds[tiab] OR merione[tiab] ORmeriones[tiab] OR cats[tiab] OR cat[tiab] OR felis[tiab] OR canis[tiab] ORsheep[tiab] OR sheeps[tiab] OR goats[tiab] OR goat[tiab] OR capra[tiab]
OR saguinus[tiab] OR tamarin*[tiab] OR leontopithecus[tiab] OR ape[tiab]
OR apes[tiab] OR pan-paniscus[tiab] OR bonobo*[tiab] OR pantroglodytes[tiab] OR gibbon*[tiab] OR siamang*[tiab] OR nomascus[tiab]
OR symphalangus[tiab] OR chimpanzee*[tiab] OR orangutan*[tiab] ORhorse[tiab] OR horses[tiab] OR equus[tiab] OR cow[tiab] OR cows[tiab] ORchicken[tiab] OR chickens[tiab] OR wistar[tiab] OR balb[tiab] OR C57[tiab]
OR C57bl[tiab] OR quail[tiab] OR long-evans[tiab] OR guppy[tiab] OR
medaka[tiab] OR zebrafish[tiab] OR flying-fox[tiab] OR Fruit-bat[tiab] OR
non-human-primate*[tiab] OR capuchin*[tiab] OR rhesus[tiab] OR
macaque*[tiab] OR cattle[tiab] OR bovine[tiab] OR pigs[tiab] OR pig[tiab]
OR swine[tiab] OR swines[tiab] OR piglet*[tiab] OR Sprague-Dawley[tiab]
OR vervet*[tiab]) OR ((mice[tiab] OR mouse[tiab] OR murine[tiab] ORrats[tiab] OR rat[tiab] OR hamster[tiab] OR hamsters[tiab] OR guineapig*[tiab] OR gerbil*[tiab] OR rabbits[tiab] OR rabbit[tiab] OR dogs[tiab] ORdog[tiab] OR monkey[tiab] OR monkeys[tiab] OR pongo-pygmaeus[tiab] ORsow[tiab] OR sows[tiab] OR boar[tiab] OR boars[tiab]) NOT medline[sb])OR ((in vitro[tiab] OR in vitro techniques[mh] OR cell line*[tiab]) AND
animals[mh:noexp])

Experimental
Animals (RoC
modified)

9#7 AND #8

CombineChemical +Bisphenol S

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SET
STRATEGY#

CONCEPTGROUPDART +
Animals

Table A.10 PubMed search strategy for ADMESETSTRATEGY#

CONCEPT
GROUP

1(80-09-1[rn] OR "Bisphenol S"[tiab] OR "bis(4-hydroxyphenyl)sulfone"[nm]OR "bis(4-hydroxyphenyl)sulfone"[tiab] OR "4,4'-Sulfonyldiphenol"[tiab] OR"Phenol, 4,4'-sulfonylbis-"[tiab] OR "BPS-monoglucuronide"[tiab] OR "BPS1G"[tiab] OR "bisphenol S dicyanate ester"[tiab] OR (bps[tiab] NOT("bps"[tiab] NOT "bisphenol"[tiab])))Chemical Terms

2(("Volume of Distribution"[tiab] OR "Toxicokinetics"[mh] OR "tissue
distribut*"[tiab] OR "Renal Elimination"[mh] OR "protein bound"[tiab] OR"protein bind*"[tiab] OR "plasma protein"[tiab] OR "Pharmacokinetics"[mh]OR "Metabolism"[mh] OR "kinetic"[tiab] OR "Intestinal Elimination"[mh] OR"Hepatobiliary Elimination"[mh] OR "Hepatobiliary"[tiab] OR"enterohepatic"[tiab] OR "entero-hepatic"[tiab] OR "Distribution
volume"[tiab] OR "cellular clearance"[tiab] OR "cell clearance"[tiab] OR"Biotransformation"[tiab] OR "bioavailability"[tiab] OR "ADME"[tiab] OR"absorptive"[tiab] OR "PBPK"[tiab] OR "toxicodynamic*"[tiab] OR("Skin"[tiab] AND "absorption"[tiab]) OR ("Oral"[tiab] AND "absorption"[tiab])
OR ("Injection"[tiab] AND "absorption"[tiab]) OR ("Gavage"[tiab] AND"absorption"[tiab]) OR ("Dietary"[tiab] AND "absorption"[tiab]) OR("Dermal"[tiab] AND "absorption"[tiab])) OR (("urine"[tiab] OR"Urination"[tiab] OR "toxicokinetic*"[tiab] OR "Pharmacokinetic*"[tiab] OR"Metabolite*"[tiab] OR "metabolism"[tiab] OR "Metabolic*"[tiab] OR"feces"[tiab] OR "fecal"[tiab] OR "excretion"[tiab] OR "defecation"[tiab] OR"biliary"[tiab] OR "Bile"[tiab]) NOT Medline[sb]))

ADME Terms#1 AND #2

CombineChemical +ADME terms

3

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APPENDIX B
Table B.1 BPS: Summary of epidemiologic studies of female reproductive toxicity.
Study/
Design*Date ofCollection/
Sample size

LOD (ng/mL)/
Adjustment for<LOD% sampleswith BPS
detected

Matrix, MedianBPS(ng/mL) **25th, 75th Percentile(ng/mL)

Results***

Gestational Diabetes Mellitus
Tang et al. 2021
Case-control

2015-2021
n = 500

0.046
LOD÷√2

82.2

Serum, 0.097

0.05, 0.107

Higher odds of GDM for women withfemale fetuses or high BMI

Zhang et al. 2019
Cohort2013-2015
n = 1,841

0.2
LOD÷√2

90.1

Urine, 0.31

0.14, 0.81

No significant results with GDM

0.1
LOD÷√2

Cases:
83.8 (TM1)
88.2 (TM2)
Controls:
84.2 (TM1)
81.7 (TM2)Urine,
Cases:
0.6 (TM1)
0.5 (TM2)
Controls:
0.4 (TM1)
0.4 (TM2)

Cases:
0.3, 1.3 (TM1)
0.2, 1.2 (TM2)
Controls:
0.2, 0.9 (TM1)
0.2, 1.1 (TM2)

26.2 ‡

Urine, <LOD

<LOD, no mention

Exposure dichotomized above/below
LOD; FT4 lower at < 15 weeks, TSH
higher for term births at < 15 weeks

80.2

Urine, 0.08

No mention

No significant results with thyroid
hormones

66.6 (TM1)
29.8 (TM2)
18.9 (TM3)Urine,
0.34 (TM1)0.24 (TM2)
NA (TM3)< LOD, 8.83 (TM1)< LOD, 1.69 (TM2)
NA (TM3)

86.3

Serum, 0.097

0.096, 0.104

Zhu et al. 2022
Case-control

2015-2017
n = 333

Higher odds of GDM for total
population, with even higher odds for
women identifying as nonAsian/Pacific Islanders

Thyroid Hormones During Pregnancy
Aker et al. 2018
Case-control

2006-2008
n = 439

Derakhshan et al. 2019
Cohort2007-2010
n = 1,996

0.03
Reported
concentrations< LOD used

Derakhshan et al. 2021
Cohort2002-2006
n = 1,267

0.05
LOD÷√2

Huang et al. 2022
Cohort2015-2018
n = 4460.046
LOD÷√2

Bisphenol S

0.1 ‡LOD÷√2135

Higher TT4 in early pregnancy
Lower TT3 in TM1, and higher FT3 in
TM2

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Study/Design*Date ofCollection/
Sample size

LOD (ng/mL)/
Adjustment for<LOD% sampleswith BPS
detected

Matrix, MedianBPS(ng/mL) **96.6 (TM2a)91.4 (TM2b)

Urine0.50 (TM2a)0.47 (TM2b)0.23, 1.07 (TM2a)0.23, 1.06 (TM2b)

Lower CRH

24.2

Urine, 0.01 µg/g

< LOD, 0.03 µg/g

No significant results with kisspeptin

25th, 75th Percentile(ng/mL)

Results***Sex Steroid Hormones and Related Proteins During Pregnancy
Aker et al. 2019
Cohort2012-2017
n = 6020.1
LOD÷√2

Wang et al. 2022
Cohort2012
n = 528

0.004
LOD÷2

Sex Steroid Hormones and Related Proteins in Young Girls
Hu et al. 2022
Cross-sectional

2013-2016
n = 1,179

0.1
LOD÷√2

88.3

Urine, 0.37

0.19, 0.74

No significant results with sex
hormones

Wang et al. 2021
Cross-sectional

2013-2016
n = 1,317

0.1LOD÷√2

88.4

Urine, 0.3

IQR = 0.6

Higher TT/E2 ratio, inverted U-shapedose-response

Wang et al. 2022
Cohort2012
n = 195

0.004
LOD÷2

24.2

Urine, 0.01 µg/g

< LOD, 0.03 µg/g

No significant results with kisspeptin

0.003LOD÷√2

93.2

Urine, 0.12

0.02, 0.49

Higher levels of TT

Sex Steroid Hormones in Women
Zhan et al. 2023
Case-control

2014-2016
n = 733

Polycystic Ovary Syndrome
Jurewicz et al. 2020
Case-control

2017
n = 357

0.022
LOD÷√2

95

Urine,
GM 0.14 (cases)
GM 0.08 (controls)

Not mentioned

Higher odds of PCOS (inference notinterpretable)

Zhan et al. 2023
Case-control

2014-2016
n = 7330.003
LOD÷√2

93.2

Urine, 0.12

0.02, 0.49

Higher odds of PCOS

0.003
LOD÷√2

82.5

Urine, 0.11 µg/g

0.01, 0.48 µg/g

Higher odds of unexplained recurrent
miscarriage for women at or over 30
years of age

Other Female Reproductive Outcomes
Ao et al. 2022
Case-control

Bisphenol S

2014-2016
n = 1,751

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Study/Design*Date ofCollection/
Sample size

LOD (ng/mL)/
Adjustment for<LOD% sampleswith BPS
detected

Matrix, MedianBPS(ng/mL) **25th, 75th Percentile(ng/mL)

Results***

Blaauwendraad et al.
2022
Cohort2004 – 2005
n = 5240.15
LOD÷√268.5 (TM1)
29.7 (TM2)
20 (TM3)

Urine,
0.6 (TM1)< LOD (TM2)< LOD (TM3)< LOD, 2.4 (TM1)< LOD, 0.3 (TM2)< LOD, < LOD (TM3)

Increased age at first menstruation

Peinado et al. 2020
Case-control

2018-2019
n = 124

0.2
LOD÷2

14.8

Urine, < LOD< LOD, < LOD

No significant results withendometriosis

Philips et al. 2018
Cohort2002-2006
n = 8770.05
LOD÷√2

70.5

Urine, 0.35

0.17, 1.03 ‡‡No significant results withfecundability

Philips et al. 2019
Cohort2004-2005
n = 1,233

0.05
LOD÷√2

69.5

Urine, 0.35

0.17, 1.03 ‡‡No significant results with gestational
hypertension measures

Philips et al. 2020
Cohort2002-2006
n = 1,213

0.05
LOD÷√268.1 (TM1)
29.1 (TM2)Urine,
0.35 (TM1)0.24 (TM2)

0.17, 1.09 (TM1)0.12, 0.49 (TM2)

Lower gestational weight gain in earlypregnancy

Wesselink et al. 2021
Case-cohort

2010-2012
n = 754

0.1
Reported
concentrations <LOD used

95.4 (Visit 1)
94.3 (Visit 2)
95.9 (Visit 3)

Urine,
0.5 (Visit 1)
0.6 (Visit 2)
0.8 (Visit 3)

IQR not reported
90th 2.6 (Visit 1)90th 2.5 (Visit 2)
90th 3.5 (Visit 3)

Reduced fibroid risk

Yue et al. 2023a
Cross-sectional

2017
n = 860.002
LOD value used

97.7

Urine, 0.07

0.03, 0.15

No significant results with adult thyroid
hormones

Zhan et al. 2022
Cross-sectional

2013-2016
n = 8570.1
LOD÷√2

90.4

Urine, 0.60

0.20, 1.30

No significant results with infertility

Zhang et al. 2023
Cross-sectional Study

2020-2021
n = 1110.1
LOD÷√260 > LOQ

Urine, 0.32 µg/g

0.19, 0.55 µg/g

Decreased AMH and increased odds
of diminished ovarian reserve

Abbreviations: AMH = Anti-Müllerian hormone, BPS = bisphenol S, BMI = body mass index, CRH = corticotropin-releasing hormone, GDM = gestational diabetes mellitus, GM = geometric mean, FT3= free triiodothyronine , FT4 = free thyroxine, E2 = estradiol, IQR = interquartile range, LOD = limit of detection, LOQ = level of quantification, PCOS= polycystic ovary syndrome , NA = not applicable,
TSH = thyroid-stimulating hormone, TM1 = trimester 1, TM2 = trimester 2, TM3 = trimester 3, TT = total testosterone, TT4 = total thyroxine, TT3 = total triiodothyronine.
*Studies ordered by outcome, then by author.
** Mean is reported only when median wasn't available.
*** Brief overview of summary results focused on statistically significant findings. See Table 4.1 for complete reporting of methodology and results for each study.
‡ Mistakes were noted in the methods section where the BPS LOD is given as 0.1 ng/mL in text and 0.4 ng/mL in Table 2 of the study. The percentage of samples with BPS above the LOD was given
as <25% in text and given as 26.2% in Table 2.
‡‡ Median, 25th, and 75th percentile values reported to be the same in Phillips et al (2018) and Phillips et al (2019) despite differing analytic sample size

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