Study ID
RefID (DistillerSR)	230
Reference (authors, year, title, other info)	Waalkens‐Berendsen DH, Kuilman‐Wahls MEM & Bär A, 2004. Embryotoxicity and teratogenicity study with neohesperidin dihydrochalcone in rats. Regulatory Toxicology and Pharmacology, 40(1), 74–79. https://doi.org/10.1016/j.yrtph.2004.05.007
Source (published/unpublished)	Published
Type of study and guideline
Good laboratory practice (yes/no/no, but before establishment of GLP)	Yes
Guideline studies (if yes, specify)	OECD TG 414 (1981)
Type of study	Prenatal developmental toxicity
Animal model
Species and strain	Rat, Wistar strain
Disease models (e.g. diabetes, allergy, obesity)	Not applicable
Housing conditions
Housing condition	21 days, during gestation, the dams were housed individually in suspended stainless steel cages fitted with wire‐mesh fronts and floors. Throughout the study, the temperature of the animal room ranged from 19 to 25°C and the relative humidity from 30% to 70%. A 12‐h light/dark cycle was maintained.
Diet name and source (if reported)	RM3 Diet (SDS Special Diets Services, Witham, UK)
Treatment
Test material	Neohesperidine dihydrochalcone (Zoster S.A., Zeneta‐Murcia, Spain)
Provider	Sponsor
Compound purity	96.9%
Vehicle used	None (direct admixture with powder rodent diet)
Dose regimen (dose level or concentration per group, and frequency) and achieved doses if available	0%, 1.25%, 2.5%, and 5% neohesperidine dihydrochalcone in the diet. Equal to: 0, 800–900, 1,600–1,700, 3,100–3,400 mg/kg bw per day
Route of administration (diet, drinking water, gavage)	Diet
Period of exposure (pre‐mating, mating, gestation, lactation, adult)	Gestation
Duration of the exposure	21 Gestation Days (GDs)
Study design
Sex and age at the start of the treatment	Females (12–13 weeks)
Number animals/sex/group	28 positively mated females/group. 4 groups.
Measured endpoints	Body weight, weight gain, food consumption, ovaries weight, gravid and empty uterus weight, cecum weight, number of corpora lutea, fetuses weight and sex, placentas weight, number of early and late resorptions, number of dead fetuses and number of implantations.
Time of measurement/observation period	The general condition and behaviour of the animals were observed twice daily. Body weight was determined on days 0, 7, 14, and 21 of gestation. Food consumption was determined during three consecutive periods (days 0–7, 7–14, and 14–21 of gestation). On day 21 of gestation the females were decapitated and examined for macroscopic abnormalities.
Methods to measure the endpoints	In line with OECD TG 414
Statistical analysis
Statistical methods	ANOVA followed by Dunnett's test, Kruskal–Wallis followed by the Mann–Whitney U test and Fisher's test.
Results
Findings reported by the study author/s	The consumption of NHDC was well tolerated. No signs of ill health, abnormal behaviour or intolerance were noted in any treatment group. Mean maternal body weights and body weight changes during gestation were similar in all groups. Maternal food consumption, when expressed in g/kg bw per day, was slightly yet significantly increased in the mid and high‐dose group during the last week of pregnancy. Except for caecal enlargement, there were no changes observed at necropsy which could be related to the NHDC treatment. All dams had viable fetuses. There were no differences for the mean weight of the gravid and empty uterus, ovaries and placenta between the NHDC treatment groups and the controls. The fecundity and gestation index, the number of corpora lutea, implantation sites, live fetuses, early and late resorptions, pre‐ and post‐implantation losses, and sex‐ratio were not affected by the treatment. Mean fetal body weights of the viable fetuses were similar in all groups. Examination of the fetuses for external, visceral and skeletal changes did not reveal any fetotoxic, embryotoxic or teratogenic effects of NHDC. The observed caecal enlargement is a well‐known physiological, adaptive response to the ingestion of high doses of a low‐digestible substance and is generally accepted to lack toxicological relevance.
No observed adverse effect level, lowest observed adverse effect level, benchmark dose/benchmark dose lower bound	NOAEL: 5% neohesperidine dihydrochalcone. This level corresponded to an intake of about 3,300 mg/kg bw per day.
Further information
None