Safety evaluation of certain food additives - ipcs inchem

138 PHYTOSTEROLS, PHYTOSTANOLS AND THEIR ESTERS
Table 5. Pup weights in two-generation reproductive study in rats
administered diets containing phytostanol esters
PND and sex Pup weights (g): mean values [standard error] a
% of plant phytostanol
esters (phytostanols) in the
diet
Offspring of F0 generation
2.4.5 Special studies on estrogenic activity
0% (control) 1.75% (1.0%) 4.38% (2.5%) 8.76% (5%)
1 M + F 4.84 [0.144] 4.89 [0.165] 4.76 [0.153] 4.81 [0.117]
4 M + F (preculling) 7.50 [0.319] 7.75 [0.281] 7.36 [0.225] 7.40 [0.183]
4 M + F (postculling) 7.50 [0.314] 7.72 [0.283] 7.42 [0.239] 7.37 [0.1941
7 M + F 12.01 [0.413] 12.08 [0.442] 11.63 [0.303] 11.28 [0.235]
14 M + F 26.45 [0.586] 26.24 [0.773] 25.34 [0.522] 23.96 [0.497]*
21 M 43.90 [0.753] 42.78 [1.245] 42.38 [0.789] 39.87 [0.700]*
21 F 42.21 [0.760] 42.00 [1.144] 41.11 [0.819] 38.49 [0.646]*
21 M + F
Offspring of F1 generation
43.11 [0.720] 42.36 [1.168] 41.81 [0.797] 39.27 [0.636]*
1 M + F 4.98 [0.140] 5.13 [0.166] 4.81 [0.113] 4.99 [0.110]
4 M + F (preculling) 7.44 [0.293] 7.82 [0.353] 7.27 [0.147] 7.48 [0.192]
4 M + F (postculling) 7.44 [0.290] 7.80 [0.353] 7.30 [0.141] 7.47 [0.188]
7 M + F 11.63 [0.332] 12.15 [0.520] 11.20 [0.193] 11.01 [0.247]
14 M + F 25.83 [0.502] 26.42 [1.035] 24.35 [0.452] 23.49 [0.478]*
21 M 42.12 [0.765] 42.36 [1.400] 40.80 [0.803] 38.14 [0.903]*
21 F 41.68 [0.807] 41.13 [1.298] 39.46 [0.682] 36.76 [0.884] #
21 M + F 41.88 [0.772] 41.78 [1.340] 40.10 [0.722] 37.47 [0.883]**
F, female; M, male.
a # Significantly different from controls: * P < 0.05, ** P < 0.01; P < 0.001; analysis of variance
(ANOVA) and Dunnett test.
The potential estrogenic activity of four mixtures of phytostanols, derived
from vegetable oil, was tested in vitro. The relative percentage of -sitostanol in the
mixtures varied from 58% to 67%, and the percentage of campestanol varied from
29% to 32%. The phytosterol content was below 4%. Estrogenic activity was
measured as the ability to induce proliferation of human mammary adenocarcinoma
(MCF-7) cells. Proliferation was measured by staining the cells with the protein stain
sulforhodamine B and measuring the optical density (modified E-screen test).
MCF-7 cells were cultured for 6 days with 0, 1, 10 or 100 μmol phytostanol mixture/l.