Safety evaluation of certain food additives - ipcs inchem

196 STEVIOL GLYCOSIDES (addendum)
on days 11, 14, 18 and 21 of lactation, all of the F1 juveniles were weighed on days
11, 14, 17 and 21 of age, and selected juveniles were weighed on days 24, 27, 31
and 35 of age. For the dams, food consumption was measured on days 11–13
(pretreatment), 14–17 and 18–20 of lactation. For the selected F1 juveniles, food
consumption was recorded on days 21, 24, 27, 31 and 35 of age. Clinical signs,
litter size, litter body weights, individual juvenile body weights and sex ratio were
recorded for each litter. The dams were killed on day 21 of lactation, the litters were
culled to four females and four males per litter on day 11 of age, unselected offspring
were killed on day 21 of age, and selected offspring were killed on day 35 of age
(10 per sex per group were selected showing good health and having body weights
within the median range). All F0 and all selected F1 juvenile animals underwent a
detailed necropsy with full macroscopic examination of internal and external tissues
in situ. Any abnormalities were recorded, and these tissues were retained. No
deaths or clinical signs of toxicity were observed in any of the groups. Body weight
changes and food consumption were unaffected by treatment. Enlarged parotid
salivary glands were found in a small number of animals in each of the treatment
groups, but the occurrence was not dose related, and the relationship between this
finding and treatment was considered to be uncertain. Based on reduced body
weight gains noted in offspring up to 35 days of age at 50 000 and 37 500 mg/kg
feed, the 25 000 mg/kg dietary concentration was considered suitable for use as
the high dose in the two-generation reproductive toxicity study (Stamp, 2007a; Curry
et al., 2008).
In the main study, three groups of 30 female and 30 male HsdRcc Han
Wistar rats (F0 generation) received a diet containing 0, 7500, 12 500 or 25 000 mg
rebaudioside A (98.5–98.7% purity)/kg for 10 weeks prior to mating and throughout
mating, gestation and lactation until termination. The mean doses in the premating
stage of treatment were equal to 0, 586, 975 and 2048 mg/kg bw per day in
males and 0, 669, 1115 and 2273 mg/kg bw per day in females. The mean doses
expressed as steviol in each of the groups were 0, 193, 321 and 674 mg/kg bw
per day in the males and 0, 220, 367 and 748 mg/kg bw per day in the females.
During the gestation period, F0 females received mean doses of 0, 614, 1034 and
2017 mg/kg bw per day (steviol content equivalent to 0, 202, 340 and 664 mg/kg
bw per day); during the lactation period, the F0 females received mean doses of 0,
1052, 1817 and 3832 mg/kg bw per day (steviol content equivalent to 0, 346, 598
and 1262 mg/kg bw per day). The F1 generation comprised 25 males and 25 females
from each group receiving treated diets on weaning and throughout mating, gestation
and lactation until termination at the same point as the F0 generation. The mean
doses received at the premating stage were equal to 0, 734, 1254 and 2567 mg/kg
bw per day for males (steviol content equivalent to 0, 242, 413 and 845 mg/kg bw
per day) and 0, 798, 1364 and 2768 mg/kg bw per day for females (steviol content
equivalent to 0, 263, 449 and 911 mg/kg bw per day). For F1 females, the mean
doses were 0, 600, 1004 and 2145 mg/kg bw per day during gestation (steviol
content equivalent to 0, 198, 331 and 706 mg/kg bw per day) and 0, 1170, 2023
and 4066 mg/kg bw per day during lactation (steviol content equivalent to 0, 385,
666 and 1339 mg/kg bw per day). The F2 generation was weaned on a treated diet,
which continued until termination at 30 days of age. At 25 days of age, the offspring
were weaned and separated from the dam; in the case of the F1 generation, 25