Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Endpoint:

multi-generation reproductive toxicity

Remarks:

based on test type (migrated information)

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication/study report which meets basic scientific principles

Qualifier:

no guideline followed

Principles of method if other than guideline:

Rats of both sexes were fed with 10% sucrose (test group) or 10% maize starch (control group) throughout 3 successive generations. For each generation, two litters were reared until they were at least 3 weeks old. From the second litter of the F3 generation, 10 animals were used for autospy and histopathology at the age of 4 weeks.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

other: Cpb:WU (Wistar Random)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Central Institute for the Breeding of Laboratory Animals TNO, Zeist, The Netherlands
- Age at study initiation: newly weaned rats, not further specified
- Housing: 5 animals of the same sex per cage in steel-cages with wire-mesh fronts and floors (premating periods)
- Diet: Institute´s powdered cereal based open-formula diet supplemented with 2.5% casein, 0.1% D,L-methionine and 10% maize starch (control groups) or 10% sucrose (test groups), ad libitum
- Water: ad libitum
- Acclimation period: 9 - 16 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 1
- Humidity (%): approx. 50%
- Photoperiod (hrs dark / hrs light): 12 / 12

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Mixing appropriate amounts with: stock diet

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: 3 weeks
- After 1 week, the first male was replaced by another male so that three different males were available for each female.
- After successful mating each pregnant female was caged: individually

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

(P) Males and Females: 12 and 21 weeks before the 1st and 2nd mating, respectively and throughout the study
(F1): Selected rats were kept on the control or test diet of their parents for 12 and 21 weeks before the 1st and 2nd mating, respectively, and throughout the study.
(F2): Selected rats were kept on the control or test diet of their parents for 12 and 21 weeks before the 1st and 2nd mating, respectively, and throughout the study.

Frequency of treatment:

continous

Details on study schedule:

- F1 parental animals not mated until 15 weeks after selected from the F1 litters.
- Selection of parents from F1 generation when pups were at weaning age (not further specified).
- Age at mating of the mated animals in the study: 24 weeks

- F2 parental animals not mated until 15 weeks after selected from the F2 litters.
- Selection of parents from F2 generation when pups were at weaning age (not further specified).
- Age at mating of the mated animals in the study: 24 weeks

Remarks:

Doses / Concentrations:
10%
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
P: 7.5 g/kg bw/day (male), 8.0 g/kg bw/day (female);
Basis:
other: mean dose value calculated from the reported body weight and food intake values (see attached Table 1)

Remarks:

Doses / Concentrations:
F1: 8.4 g/kg bw/day (male), 9.0 g/kg bw/day (female)
Basis:
other: mean dose value calculated from the reported body weight and food intake values (see attached Table 1)

Remarks:

Doses / Concentrations:
F2: 8.5 g/kg bw/day (male), 8.9 g/kg bw/day (female)
Basis:
other: mean dose value calculated from the reported body weight and food intake values (see attached Table 1)

Remarks:

Doses / Concentrations:
F3: 12 g/kg bw
Basis:
other: mean dose value calculated from the reported body weight and food intake values (see attached Table 1)

No. of animals per sex per dose:

100 P males and females
20 F1 males and females
20 F2 males and females
10 F3 males and femals

Control animals:

other: yes, diet supplemented with 10% maize starch

Details on study design:

- Other:
Study design:
The P generation consisted of 100 males and females in the control and test group. After 12 weeks on the test and control diets, the first mating occurred. Pups were weaned at day 21 and 50 males and females were used for a 8 week feeding study. The remaining pups were discarded at 3 – 5 weeks of age. The mating procedure was repeated 9 weeks after the first mating by using the same combination of mating partners. From this F1 generation, 20 males and females were used to produce the F2 generation. Therefore, the selected rats were weaned at day 21 and kept on the diets of their parents for further 12 weeks before mating. The offspring of the first mating was discarded and 20 females and males of the second mating were used for the production of the F3 generation. As with the F1-generation, the selected F2 generation rats were fed with the various diets for 12 weeks after weaning and subsequently mated. After 21 weeks of exposure, the second mating procedure started. Pups from the first F3 offsping were discarded 3 weeks after birth. Weanlings from the second mating procedure were used for gross and microscopic evaluation after receiving the diets for 4 weeks (10/sex).

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes, daily

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights were recorded weekly during the premating and mating period. No weighing was performed during pregnancy and lactation periods.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: NO
- Food consumption for each animal determined as g food/rat/week: YES
For each generation, food consumption was recorded weekly during the premating period of 12 weeks. Food intake was not measured during the mating and lactation periods.

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 1 postpartum: yes
- If yes, maximum of 8 pups/litter.

PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring:
number and sex of pups, live births, postnatal mortality, presence of gross anomalies, weight gain, other: viability index at birth and day 4, litter size
F3: haemoglobin level

Postmortem examinations (offspring):

SACRIFICE
- The F1 and F2 offspring not selected as parental animals were sacrificed when they were 3 weeks old.
- The F3 offspring of the first mating was sacrificed at the age of 3 weeks whereas 10 weanlings of the second mating were sacrificed 4 weeks after weaning (males: 28 days after weaning, females: 29 days after weaning) for detailed gross and microscopic examinations.

GROSS NECROPSY
- The following organs were weighed: adrenals, brain, caecum, heart, kidneys, liver, ovaries, spleen, testes, thymus and thyroid.

HISTOPATHOLOGY / ORGAN WEIGTHS
Tissues of all the major systems from the F3 generation including kidneys, liver, lungs, heart, stomach, epididymides, prostate, urinary bladder, pituitary, thyroid were examined microscopically.

Statistics:

Mean values and standard errors were calculated from the examined parameters. Statistical analyses were performed using ANOVA and Dunnett´s test (changes in body weights, food intake, haemoglobin levels and relative organ weights), Student´s t test (mean litter size at birth and pup weight) or chi-square test (viability and lactation indices).

Reproductive indices:

The following parameter were determined in the P, F1 and F2 generation for the first and second mating:
1. fertility index (%): (no. of pregnant females / no. of mated females) x 100
2. gestation index (%): (no. of females with live fetuses / no. of pregnant females) x 100
3. lactation index: (no. of live pups at day 21 / no. of live pups at day 4)

Offspring viability indices:

viability index on day 1 (%): (no. of pups born alive / total no. of born pups) x 100
viability index on day 4 (%) : (no. of live pups at day 4 / total no. of live pups on day 1) x 100%

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

7 out of 100 females died due to delivery problems (non-adverse)

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

F1 females: decreased body weight on day 21; F2 males: increased body weight on day 21 (non-adverse)

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

F1 females: decreased body weight on day 21; F2 males: increased body weight on day 21 (non-adverse)

Organ weight findings including organ / body weight ratios:

not examined

Histopathological findings: non-neoplastic:

not examined

Other effects:

no effects observed

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

not examined

Reproductive performance:

effects observed, treatment-related

Description (incidence and severity):

The fertility index of parental dams was significantly reduced in the first and second mating to 82% and 92% of controls, respectively. As the effect was not reproducible in the F1 and F2 matings, the effect is not considered as treatment-related.

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
No abnormalities of condition or behaviour were observed. As the mortality of 7 out of 100 animals is due to delivery problems, the effect on mortality is considered as not-treatment related.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
A significant but slight decrease in body weight was observed in F1 females at day 21 (control group: 143 ± 1.9; test group: 138.5 ± 2.6) whereas a significant increase in body weight was determined in F2 males (control group: 170.0 ± 2.9; test group: 179.8 ± 3.6). As the effect was not consistent over the observation period and not reproducible in all generations, the effect is not considered to be treatment-related.

TEST SUBSTANCE INTAKE (PARENTAL ANIMALS)
No substance-related changes were observed in food intake.

GROSS PATHOLOGY (PARENTAL ANIMALS)
Gross pathology did not reveal any alterations in the parental animals.

Dose descriptor:

NOAEL

Effect level:

>= 7 455 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: mortality; body weight; food consumption and compound intake; gross pathology; organ weights; histopathology; mating index; fertility index; pregnancy index; pup weight; viability index; lactation index; corresponding to 10%

Dose descriptor:

NOAEL

Effect level:

>= 7 998 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: mortality; body weight; food consumption and compound intake; gross pathology; organ weights; histopathology; mating index; fertility index; pregnancy index; pup weight; viability index; lactation index; corresponding to 10%

Dose descriptor:

NOAEL

Effect level:

>= 11 816 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: mortality; body weight; food consumption and compound intake; gross pathology; organ weights; histopathology; mating index; fertility index; pregnancy index; pup weight; viability index; lactation index; corresponding to 10%

Remarks on result:

other: Generation: F3 (migrated information)

Dose descriptor:

NOAEL

Effect level:

>= 12 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: mortality; body weight; food consumption and compound intake; gross pathology; organ weights; histopathology; mating index; fertility index; pregnancy index; pup weight; viability index; lactation index; corresponding to 10%

Remarks on result:

other: Generation: F3 (migrated information)

Clinical signs:

no effects observed

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

Significant differences in pup viability were observed (non-adverse).

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Increases in body weights during lactation and after weaning in pups from F0 and F1 (non-adverse).

Sexual maturation:

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Male F3 pups showed a statistically significant increase in relative kidney weight (non-adverse).

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Different abnormalities were observed in one pup each of the F1, F2 and F3 generation (non-adverse).

Histopathological findings:

no effects observed

VIABILITY (OFFSPRING)
Viability index: Several statistically significant differences in pup survival indices were determined including both, increases and decreases in the test group. As the differences from the controls were only small and never appeared to be consistent in successive generations, the effects are considered as non-adverse.
Lactation index: After the 1st and 2nd mating of the P generation, the survival of pups during lactation was generally lower in all groups including controls than in breedings of the following generations. No treatment-related alterations were determined.

CLINICAL SIGNS (OFFSPRING)
No signs of toxicity were observed throughout the study.

BODY WEIGHT (OFFSPRING)
Effects during lactation: Pups from the first mating of the F0 generation showed a significant higher body weight on day 4. Further, pups from both matings of the F1 generation showed a significant higher body weight at day 14 (first mating) and at days 4, 14 and 21 (second mating). In contrast, no alterations were determined in pups from the F2 generation. As the effects are not consistent through the litters and the generations, no effect on body weight of sucrose seems to be apparent during lactation.

Effects after lactation and exposure via feed: Females of the F1 and males of F2 generation showed a significant higher body weight on day 21. As the effect is not consistent in the successive generations, the effect is considered as non-adverse.

SEXUAL MATURATION (OFFSPRING)
No effects on sexual maturation were observed in exposed animals.

ORGAN WEIGHTS (OFFSPRING)
A statistically significant increase in the relative kidney weight was observed in male F3 pups which was not accompanied by histological changes. Thus, the effect is not interpreted as adverse.

GROSS PATHOLOGY (OFFSPRING)
Malformations were observed in one F1 pup (anophthalmia of one eye), one F2 pup (thick and paralysed hind leg) and one F3 pup (missing foot, which was most probably amputated shortly after birth). All these abnormalities were considered fortuitous and not treatment-related.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 8 377 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: mortality; body weight; food consumption and compound intake; gross pathology; organ weights; histopathology; mating index; fertility index; pregnancy index; pup weight; viability index; lactation index; corresponding to 10%

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 8 997 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: mortality; body weight; food consumption and compound intake; gross pathology; organ weights; histopathology; mating index; fertility index; pregnancy index; pup weight; viability index; lactation index; corresponding to 10%

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

>= 8 475 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: mortality; body weight; food consumption and compound intake; gross pathology; organ weights; histopathology; mating index; fertility index; pregnancy index; pup weight; viability index; lactation index; corresponding to 10%

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

>= 8 892 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: mortality; body weight; food consumption and compound intake; gross pathology; organ weights; histopathology; mating index; fertility index; pregnancy index; pup weight; viability index; lactation index; corresponding to 10%

Reproductive effects observed:

not specified

Conclusions:

In the conducted study, exposure to 10% sucrose of 3 successive generations did not induce adverse effects on fertility, reproductive performance or development compared with control animals fed with 10% maize starch. Therefore, sucrose is not considered to induce toxic effects on reproduction.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 2) from a reference substance with similar structure and intrinsic properties. Read-across is justified based on common functional group(s), common breakdown products, similarities in PC/ECO/TOX properties(refer to endpoint discussion for further details).
The selected study is thus sufficient to fulfil the standard information requirements set out in Annex VIII-IX, 8.7, in accordance with Annex XI, 1.5, of Regulation (EC) No 1907/2006.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Reproductive Toxicity

Justification for read-across

There are no data available on reproductive toxicity of Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose. In accordance with Regulation (EC) No 1907/2006, Annex XI, 1.5, read-across from structurally related substances is conducted to fulfill the standard information requirements set out in Regulation (EC) No 1907/2006, Annex X, 8.7.

According to Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met.” In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across) “to avoid the need to test every substance for every endpoint”.

All substances contained in Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose represent mono- or disaccharides which all consist of glucose and/or fructose. Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose is the aqueous solution (syrup) of the reaction mass of isomaltulose (CAS 13718-94-0), trehalulose (CAS 51411-23-5), fructose (CAS 57-48-7), glucose (CAS 50-99-7), sucrose (CAS 57-50-1), isomaltose (CAS 499-40-1) and oligosaccharides.

All ingredients are substances naturally occurring in fruits, vegetables and other crops or honey.

Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006, whereby physicochemical, toxicological and ecotoxicological properties may be predicted from data for reference substance(s) by interpolation to other substances on the basis of structural similarity, sucrose (CAS 57-50-1) is selected as source substances for assessment of reproductive toxicity.

The read-across is based on the presence of common functional groups and common breakdown products via biological processes, which result in structurally similar chemicals. In general, disaccharides like isomaltulose, trehalulose and sucrose are enzymatically hydrolysed at the glycosidic bond between the monosaccharide units to equal parts in glucose and fructose (Cheetham, 1982; Goda and Hosoya, 1983; MacDonald and Daniel, 1983; Yamada et al., 1985; Ziesenitz, 1986; Goda et al., 1991; Würsch, 1991; Günther and Heymann, 1998), which subsequently enter well-characterized carbohydrate metabolic pathways (Lina et al ., 2002) as essential energy substrate or they are converted to storable glycogen (see Toxicokinetics). A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

Reproductive toxicity of sucrose was evaluated in a multigeneration study. Rats of both sexes were fed continuously with 10% sucrose throughout 3 successive generations corresponding to more than 7 500 mg sucrose/kg bw/day (Waalkens-Berendsen et al., 1990). Control animals received a diet supplemented with 10% maize starch. The first mating occurred after 12 weeks of exposure with 100 parental animals for each sex. The administration of sucrose or maize starch was further continued throughout the study. Pups of the first mating were weaned at day 21 and discarded at 3 – 5 weeks of age. The mating procedure of the parental generation was repeated 9 weeks after the first mating by using the same combination of mating partners. From the F1 generation, 20 males and females were selected for further matings. The selected rats were weaned at day 21 and kept on the diets of their parents for further 12 weeks before mating. The offspring of the first mating was discarded and 20 females and males of the second mating were used for the production of the F3 generation. As with the F1-generation, the selected F2 generation rats were fed with the various diets for 12 weeks after weaning and subsequently mated. After 21 weeks of exposure, the second mating procedure started. All litters were standardized to 8 pups. For the parental animals, cage side observations, body weight gain, food consumption and gross pathology data were reported. Litter observations included number and sex of pups, live births, postnatal mortality, presence of gross anomalies, weight gain, viability index at birth and day 4, litter size and the evaluation of the haemoglobin level in the F3 litters. Further, 10 weanlings of each sex from the second mating procedure were used for gross and microscopic evaluation after receiving sucrose for 4 weeks.

In the parental generation, no abnormalities of condition or behaviour were observed. Further, no treatment-related mortality was reported. A significant but slight decrease in body weight was observed in F1 females at day 21 whereas a significant increase in body weight was determined in F2 males. As the effect was not consistent over the observation period and not reproducible in all generations, the effect was not considered to be treatment-related. Further, no substance-related changes were observed in food intake and gross pathology. The fertility index of parental dams was significantly reduced in the first and second mating to 82% and 92% of maize starch exposed animals, respectively. However, this effect was not reproducible in the F1 and F2 matings, and was thus not considered as treatment-related.

For the offspring, several statistically significant differences in pup survival indices were determined including both increases and decreases in the test group exposed to sucrose compared to animals receiving maize starch. As the differences to the maize starch group were only small and never appeared to be consistent in successive generations, the effects were considered as non-adverse. No treatment-related alterations were determined on the lactation index or sexual maturation and and no signs of toxicity were observed in the offspring. Inconsistent effects on the body weight were reported through the litters and generations during and after lactation thereby pointing rather to incidental findings than treatment-related effects. A statistically significant increase in the relative kidney weight was observed in male F3 pups which was not accompanied by histological changes. No further changes in relative organ weights were described. Thus, sucrose was considered to not induce adverse effects on the organ weights. The incidence and degree of histopathological changes including anophthalmia of the eye and malformations on the limbs was similar in all test groups. In summary, exposure to 10% sucrose of 3 successive generations did not induce adverse effects on fertility, reproductive performance or development compared with animals fed with 10% maize starch in the conducted study. Therefore, NOAELs ≥ 7 500 mg sucrose/kg bw/day were defined for reproductive toxicity in the parental and following generations.  

The multigeneration study revealed no adverse effects on reproductive performance, and hence, sucrose is considered as non-hazardous to reproduction. Thus, Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose is considered as safe in regard to reproductive toxicity.

 

References not included in IUCLID:

Blainey M, Avila Cd, van der Zandt P. Review of REACH Annex IV--establishing the minimum risk of a substance based on its intrinsic properties. Regul Toxicol Pharmacol. 2010 Feb;56(1):111-20.

Cheetham, P.S.J. 1982. The human sucrase-isomaltase complex: Physiological, biochemical, nutritional and medical aspects. In: Lee, C.K.; Lindley, M.G. (Eds.). Developments in Food Carbohydrate - 3. Disaccharidases. Applied Science Publishers; London, Engl./Englewood, New Jersey, pp. 107-140.

Goda, T.; Hoyosa, N. 1983. Hydrolysis of palatinose by rat intestinal sucrase-isomaltase complex. Nihon Eiyo Shokuryo Gakkaishi 36:169-173. Cited In: Würsch, 1991.

Goda, T.; Takase, S.; Hosoya, N. 1991. Hydrolysis of palatinose condens ates by rat intestinal disaccharidases. Nihon Eiyo Shokuryo Gakkaishi 44(5):395-398.

Günther, S.; Heymann, H. 1998. Di- and oligosaccharide substrate specificities and subsite binding engergies of pig intestinal glycoamylase-maltase.Arch Biochem Biophys 354(1):111-116.

Lina, B.A.R.; Jonker, D.; Kozianowski, G. 2002.Isomaltulose (Palatinose®): A review of biological and toxicological studies. Food Chem Toxicol 40(10):1375-1381

MacDonald, I.; Daniel, J.W. 1983. The bioavailability of isomaltulose in man and rat. Nutr Rep Int 28(5):1083-1090.

Würsch, P. 1991. Metabolism and tolerance of sugarless sweeteners. In: Rugg-Gunn, A.J.(Ed.). Sugarless: The Way Forward. Else vier Applied Science; New York, pp. 32-51.

Yamada, K.; Shinohara, H.; Hosoya, N. 1985. Hydrolysis of 1-O-α-D-glucopyranosyl-D-fructofuranose (Trehalulose) by rat intestinal surcrase-isomaltase complex. Nutrition Reports International 32 (5): 1211 - 1220

Ziesenitz, S.C. 1986. Carbohydrasen aus jejunalmucosa des Menschen = [Carbohydrases from the human jejunal mucosa]. Z Ernährungswiss 25(4):253-258. Cited In: Würsch, 1991.

Short description of key information:

Key studies on toxicity to reproduction are available for the following read-across analogues:

Multigeneration Study: NOAEL, oral: (P, female) ≥ 7998 mg/kg bw/day; CAS 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (P, male) ≥ 7455 mg/kg bw/day; CAS 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F1, female) ≥ 8997 mg/kg bw/day; CAS 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F1, male) ≥ 8377 mg/kg bw/day; CAS 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F2, female) ≥ 8892 mg/kg bw/day; CAS 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F2, male) ≥ 8475 mg/kg bw/day; CAS 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F3, female) ≥ 12000 mg/kg bw/day; CAS 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F3, male) ≥ 11816 mg/kg bw/day; CAS 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Developmental Toxicity Study: NOAEL, oral (maternal toxicity) ≥ 6930 mg/kg bw/day; CAS 13718-94-0, isomaltulose (Lina et al., 1997)

Developmental Toxicity Study: NOAEL, oral (developmental toxicity) ≥ 6930 mg/kg bw/day; CAS 13718-94-0, isomaltulose (Lina et al., 1997)

No data are available for repeated dose toxicity after dermal exposure and inhalation, respectively.

Justification for selection of Effect on fertility via oral route:

Hazard assessment is conducted by means of a read-across from a structural surrogate. The selected study is an adequate and reliable study based on the identified similarities in structure and intrinsic properties between source and target substance and overall assessment of quality, duration and dose descriptor level (refer to endpoint discussion for further details).

Justification for selection of Effect on fertility via inhalation route:

No study required since exposure of humans via inhalation is unlikely taking into account the physicochemical properties of the substances and the lack of exposure to aerosols, particles or droplets of inhalable size under normal conditions of use.

Justification for selection of Effect on fertility via dermal route:

No study required since sufficient weight of evidence is available to exclude that Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose is toxic after acute dermal exposure considering the available data on read-across to the main components, which have been shown not to be toxic after acute or repeated oral exposure. Moreover, sufficient information is known for the ingredients glucose, fructose and sucrose to consider them as non-hazardous and to include them in Annex IV of Regulation (EC) No. 987/2008.  Therefore, in accordance with Annex XI, Section 1.2 of Regulation (EC) 1907/2006 further testing on vertebrate animals for that property shall be omitted and further testing not involving vertebrate animals may be omitted.  

Effects on developmental toxicity

Description of key information

Key studies on toxicity to reproduction are available for the following read-across analogues:

Multigeneration Study: NOAEL, oral: (P, female) ≥ 7998 mg/kg bw/day; CAS# 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (P, male) ≥ 7455 mg/kg bw/day; CAS# 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F1, female) ≥ 8997 mg/kg bw/day; CAS# 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F1, male) ≥ 8377 mg/kg bw/day; CAS# 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F2, female) ≥ 8892 mg/kg bw/day; CAS# 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F2, male) ≥ 8475 mg/kg bw/day; CAS# 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F3, female) ≥ 12000 mg/kg bw/day; CAS# 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Multigeneration Study: NOAEL, oral: (F3, male) ≥ 11816 mg/kg bw/day; CAS# 57-50-1, sucrose (Waalkens-Berendsen et al., 1990)

Developmental Toxicity Study: NOAEL, oral (maternal toxicity) ≥ 6930 mg/kg bw/day; CAS# 13718-94-0, isomaltulose (Lina et al., 1997)

Developmental Toxicity Study: NOAEL, oral (developmental toxicity) ≥ 6930 mg/kg bw/day; CAS# 13718-94-0, isomaltulose (Lina et al., 1997)

No data are available for repeated dose toxicity after dermal exposure and inhalation, respectively.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 414 (Prenatal Developmental Toxicity Study)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

other: Wistar (Hsd/Cpb:WU)

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan-CPB, Zeist, The Netherlands
- Age at study initiation (at mating): 13 weeks
- Housing: mated females were housed individually in suspended stainless-steel wire-screen cages
- Diet: institute's cereal-based powdered stock diet (during treatment period supplemented with 2.5% casein, 0.1% DL-methionine and 0-10% maize starch), ad libitum
- Water: ad libitum
- Acclimation period: 5 days before mating

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-24
- Humidity (%): 50-60
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on exposure:

DIET PREPARATION
- Mixing appropriate amounts with (Type of food): institute's cereal-based powdered stock diet supplemented with 2.5% casein, 0.1% DL-methionine and 10% maize starch; 0, 2.5, 5 or 10% test substance was incorporated at the expense of maize starch.
- Storage temperature of food: freezer

Fresh portions of diet were provided to the rats every 4-6 days.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

All diets were analysed for test substance content by HPLC. The adequacy of the mixing procedure was investigated by analysis of 5 samples of each test diet from the first batch. The stability of the test substance in this batch of diets was examined after storage for 7 days at room temperature.
The test substance was found to be stable in the diet under the experimental conditions applied.
The actual levels of the test substance were as follows:
low-dose diet: 87-96% of the intended level
mid-dose diet: 98-102% of the intended level
high-dose diet: 98-103% of the intended level
The analyses of the 5 samples of each of the diets from the first batch showed coefficients of variation of 4% or less, indicating satisfactory homogeneity.

Details on mating procedure:

- Impregnation procedure: cohoused
- M/F ratio per cage: 1:2
- Proof of pregnancy: sperm in vaginal smear referred to as day 0 of pregnancy

Duration of treatment / exposure:

Day 0 - 21 of gestation

Frequency of treatment:

continuously

Duration of test:

21 days of gestation

Remarks:

Doses / Concentrations:
2.5, 5 and 10%
Basis:
nominal in diet

Remarks:

Doses / Concentrations:
1760, 3520 and 6930 mg/kg bw/day
Basis:
other: overall means of periodical calculations from data on body weight, food intake and nominal levels of test substance

No. of animals per sex per dose:

24 females

Control animals:

yes, plain diet

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Body weight gain during days 0-6, 6-16 and 16-21 of gestation was determined.

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Water intake (g/rat/day) during days 0-6, 6-16 and 16-21 of gestation was determined.

POST-MORTEM EXAMINATIONS: Yes
- Sacrifice on gestation day 21
- Organs examined: major abdominal and thoracic maternal organs

Ovaries and uterine content:

The ovaries and uterine content was examined after termination: Yes
Examinations included:
- Gravid uterus weight: Yes
- Number of corpora lutea: Yes
- Number of implantations: Yes
- Number of early resorptions: Yes
- Number of late resorptions: Yes
- Other: Number of live and dead foetuses; sex of live foetuses; foetal and placental weights; foetal length;

Fetal examinations:

- External examinations: Yes: [all per litter]
- Soft tissue examinations: Yes: [half per litter]
- Skeletal examinations: Yes: [half per litter]
- Head examinations: No

Statistics:

Body weight gain, food and water intake were evaluated by ANOVA/Dunnett's test (P < 0.05). Maternal survival and pregnancy status, maternal and foetal external macroscopic observations, and skeletal and visceral examinations were evaluated by Fisher's exact probability test. The percentages of pre- and post-implantation loss were subjected to Kruskal-Wallis one-way ANOVA followed by the Mann-Whitney U test.

Indices:

Fertility index (%) = (No. of pregnant females/No. of mated females) x 100
Preimplantation loss (%) = [(No of corpora lutea-No. of implantation sites)/No. of corpora lutea] x 100
Postimplantation loss (%) = [(No of implantation sites-No. of live foetuses)/No. of implantation sites] x 100

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
No mortality occured and no clinical signs attributabe to the treatment were noted. Body weight gain, food and water consumption did not reveal treatment-related differences during pregnany. There were no severe macroscopic abnormalities at gross pathology of the maternal organs and tissues.
No significant differences in fertility and gestation indices, the numbers of corpura lutea and implantations, live and dead foetuses, foetal sex ratios and early and late resorptions were noted. There were no differences in ovaries, gravid and empty uterus weights in any of the dose groups compared to control animals.

Dose descriptor:

NOAEL

Effect level:

>= 6 930 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: No adverse effects were noted up to the highest dose tested.

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
Placental weight, foetal weight and foetal length were comparable between treatment and control groups.
Examination of foetal soft tissues did not reveal treatment related changes. There were no visceral malformations in any of the groups. The few visceral anomalies or visceral variations (Table 1) were spread about equally among the treatment and control groups or occurred occationally in one or a few foetuses. No significant differences between treated and control animals were found at examination of the placentas during foetal screening.
No foetal skeletal malformations occurred. Among skeletal variations, the incidence of foetuses with a supernumerary rudimentary 14th rib was higher in the controls than in any of the treatment groups (Table 2). The incidence of crooked toes varied among the treatment groups and was slightly increased in foetuses of the top-dose group. Similar variations have been noted regularly in control rats in previous studies and therefore the occurence of crooked toes in the present study is considered to reflect a processing artefact rather than a treatment-related effect. A statistically significant increased incidence of foetuses showing incomplete ossification of the skull bones was found in the low-dose group and was considered to be accidental, since it was not confirmed in the higher dose groups (table 2). Almost every foetus, including those in the control group, showed delayed ossification of the phalanges (Table 2). The delayed ossification was more prominent in the treatment groups. However, there was no dose-response relationship and no relevant effects on ossification of other bones were observed. Thus, the delayed ossification of the phalanges was not considered treatment-related.

Dose descriptor:

NOAEL

Effect level:

>= 6 930 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No adverse effects were noted up to the highest dose tested.

Abnormalities:

not specified

Developmental effects observed:

not specified

Table 1: Visceral alterations in foetuses

Parameters	Dose groups (dietary levels)
	0	2.5%	5%	10%
No of foetuses examined	126	120	119	130
Visceral anomalies
Eyes: folded retin	1	0	0	1
Brain: meningeal haemorrhage	3	1	1	2
Heart: pericard filled with haemorrhagic fluid	3	2	2	3
Spleen: heamorrhage	0	1	0	0
Stomach: heamorrhage	2	2	5	5
Thoracic activity: filled with haemorrhagic fluid	0	0	0	1
Abdominal cavity: containing haemorrhagic fluid	1	2	0	1
Extremeties: flexure of one limb	0	0	0	1
Pancreas: haemorrhage	5	10	7	11
Skin subcutaneous haemorrhage	0	0	1	0
Total incidence of visceral anomalies	13	16	15	21
Visceral variations
Oesophagus: oesophagectasia	7	8	9	3
Thymus: small haemorrhage(s), petechiae	3	1	0	0
Stomach: hypertrophy	13	5	5	12
Stomach: hypotrophy	0	1	0	0
Kidneys: increased renal pelvic cavitation	11	11	19	14
Urether: hydrourether	1	2	0	4
Urether: bent urether	0	1	0	0
Tail: crooked	3	5	4	2
Tail: curly	1	2	2	1
Tail: ringtail	0	1	0	0
Extremeties: crooked	0	2	0	0
Extremeties: bent	0	1	0	0
Salivary gland: haemorrhage	0	0	0	2
Total incidence of visceral variations	30	34	31	32

 

 

Table 2: Skeletal alterations in foetuses

 

Parameters	Dose groups (dietary levels)
	0	2.5%	5%	10%
No of foetuses examined	137	134	137	145
Skeletal variations
Sternum: dislocated sternebra(e)	37	31	30	51
Sternum: supernumerary sternebra(e)	0	1	1	1
Sternum: absent	0	0	1	0
Ribs: 14^thrib present	9	3	4	1*
Ribs: irregular shaped ribs	0	1	0	0
Ribs: wavy rib(s)	1	0	0	1
Ribs: 13^thrib rudimentary	0	0	1	0
Forelimbs: crooked fingers	1	2	1	2
Hindlimbs: crooked toes	1	7	4	10*
Total skeletal variations	44	43	38	61
Skeletal ossification
Skull bones: two or more bones incompletely ossified	36	58***	48	49
Skull bones: hyoid bone absent	0	1	0	2
Sternum: sternebra(e) reduced or incompletely ossified	59	70	63	63
Phalanges front: 10-20 digits incompletely ossified	4	1	3	5
Phalanges front: 10-20 digits not ossified	136	118***	132	133**
Phalanges front: more than 20 digits not ossified	1	15***	4	10*
Phalanges hind: 10-20 digits incompletely ossified	3	1	2	0
Phalanges hind: 10-20 digits not ossified	136	111	129*	127***
Phalanges hind: more than 20 digits not ossified	1	19**	8*	18***
Total skeletal retardations	137	134	137	145

 */**/***: values differ significantly from the corresponding control value (* P<0.05; ** P<0.01; *** P<0.001) using Fisher's exact probability test

Conclusions:

No maternal toxicity occurred and no effects on reproductive performance, nor on embryonic or foetal development including visceral and skeletal examination were noted up to and including a dietary level of 10% isomaltulose equivalent to 6930 mg/kg bw/day.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

The available information comprises an adequate and reliable study (Klimisch score 2) from a reference substance with similar structure and intrinsic properties. Read-across is justified based on common functional group(s), common breakdown products, similarities in PC/ECO/TOX properties(refer to endpoint discussion for further details).

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

Developmental Toxicity

Justification for read-across

There are no data available on developmental toxicity of Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose. In accordance with Regulation (EC) No 1907/2006, Annex XI, 1.5, read-across from structurally related substances is conducted to fulfill the standard information requirements set out in Regulation (EC) No 1907/2006, Annex X, 8.7.

According to Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met.” In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across) “to avoid the need to test every substance for every endpoint”.

All substances contained in Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose represent mono- or disaccharides which all consist of glucose and/or fructose. Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose is the aqueous solution (syrup) of the reaction mass of isomaltulose (CAS 13718-94-0), trehalulose (CAS 51411-23-5), fructose (CAS 57-48-7), glucose (CAS 50-99-7), sucrose (CAS 57-50-1), isomaltose (CAS 499-40-1) and oligosaccharides.

All ingredients are substances naturally occurring in fruits, vegetables and other crops or honey.

Having regard to the general rules for grouping of substances and read-across approach laid down in Annex XI, Item 1.5, of Regulation (EC) No 1907/2006, whereby physicochemical, toxicological and ecotoxicological properties may be predicted from data for reference substance(s) by interpolation to other substances on the basis of structural similarity, isomaltulose (CAS 13718-94-0) is selected as source substances for assessment of developmental toxicity.

The read-across is based on the presence of common functional groups and common breakdown products via biological processes, which result in structurally similar chemicals. In general, disaccharides like isomaltulose, trehalulose and sucrose are enzymatically hydrolysed at the glycosidic bond between the monosaccharide units to equal parts in glucose and fructose (Cheetham, 1982; Goda and Hosoya, 1983; MacDonald and Daniel, 1983; Yamada et al., 1985; Ziesenitz, 1986; Goda et al., 1991; Würsch, 1991; Günther and Heymann, 1998), which subsequently enter well-characterized carbohydrate metabolic pathways (Lina et al ., 2002) as essential energy substrate or they are converted to storable glycogen (see Toxicokinetics). A detailed analogue approach justification is provided in the technical dossier (see IUCLID Section 13).

Developmental toxicity of isomaltulose was tested in a prenatal developmental toxicity study similar to OECD guideline 414 (Lina et al., 1997). 24 mated Wistar rats were exposed to dietary concentrations of 2.5, 5 and 10% isomaltulose continously from Day 0 – 21 of gestation (corresponding to 1 760, 3 520 and 6 930 mg isomaltulose/kg bw/day). Control animals received plain diet. On day 21, dams were sacrificed and foetuses were examined for abnormalities. No mortality occurred over the study period and no clinical signs attributable to the treatment were determined in dams. Further, body weight gain, food and water consumption did not reveal any treatment-related differences. Additionally, gross pathology of the maternal organs and tissues did not reveal severe macroscopic abnormalities. Placental weight, foetal weight and length were comparable between treatment and control groups.
Examinations of foetal soft tissues did not reveal treatment related changes and no visceral malformations were noted. Few visceral anomalies or visceral variations were spread about equally among the treatment and control groups or occurred occasionally in one or a few foetuses. No significant differences between treated and control animals were found at examination of the placentas during foetal screening or on foetal skeletal malformations. Among skeletal ossification a statistically significant increased incidence of foetuses showing incomplete ossification of the skull bones was found in the low-dose group and was considered to be accidental, since it was not confirmed in the higher dose groups. A delayed ossification of the phalanges was more prominent in the treatment groups without any dose-response relationship and hence was not considered as treatment-related. No significant differences in fertility, gestation indicex, number of corpura lutea and implantation, live and dead foetuses, foetal sex ratios and early and late resorptions were noted. There were no differences in ovaries, gravid and empty uterus weights in any of the dose groups compared to control animals. Therefore, isomaltulose did not induce any maternal, embryonic or foetal adverse effects at dietary concentrations of up to 10% (approximately 7 000 mg isomaltulose/kg body weight/day); thus, a NOAEL ≥ 7 000 mg isomaltulose/kg body weight/day was determined for maternal and developmental toxicity (Lina et al., 2002).

Based on the multigeneration study and the developmental toxicity study, sucrose and isomaltulose are considered as non-hazardous in regard to reproductive or developmental toxicity.

Fructose, glucose and sucrose are not further described in the present dossier as sufficient information is known about the intrinsic properties to consider them as non-hazardous which resulted in inclusion on Annex IV of Regulation (EC) 1907/2006. This has been recently verified by the Comission as reviewed by Blainey et al. (2010). The remainder isomaltose occurs naturally at branch sites within amylopectin in starches and is thus present in commercially available starch hydrolysates and maltodextrines, which are both included in Annex IV.

Based on the available data on the surrogate substance isomaltulose, Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose is considered as safe in regard to developmental toxicity.

 

References not included in IUCLID:

Blainey M, Avila Cd, van der Zandt P. Review of REACH Annex IV--establishing the minimum risk of a substance based on its intrinsic properties. Regul Toxicol Pharmacol. 2010 Feb;56(1):111-20.

Cheetham, P.S.J. 1982. The human sucrase-isomaltase complex: Physiological, biochemical, nutritional and medical aspects. In: Lee, C.K.; Lindley, M.G. (Eds.). Developments in Food Carbohydrate - 3. Disaccharidases. Applied Science Publishers; London, Engl./Englewood, New Jersey, pp. 107-140.

Goda, T.; Hoyosa, N. 1983. Hydrolysis of palatinose by rat intestinal sucrase-isomaltase complex. Nihon Eiyo Shokuryo Gakkaishi 36:169-173. Cited In: Würsch, 1991.

Goda, T.; Takase, S.; Hosoya, N. 1991. Hydrolysis of palatinose condens ates by rat intestinal disaccharidases. Nihon Eiyo Shokuryo Gakkaishi 44(5):395-398.

Günther, S.; Heymann, H. 1998. Di- and oligosaccharide substrate specificities and subsite binding engergies of pig intestinal glycoamylase-maltase.Arch Biochem Biophys 354(1):111-116.

Lina, B.A.R.; Jonker, D.; Kozianowski, G. 2002.Isomaltulose (Palatinose®): A review of biological and toxicological studies. Food Chem Toxicol 40(10):1375-1381

MacDonald, I.; Daniel, J.W. 1983. The bioavailability of isomaltulose in man and rat. Nutr Rep Int 28(5):1083-1090.

Würsch, P. 1991. Metabolism and tolerance of sugarless sweeteners. In: Rugg-Gunn, A.J.(Ed.). Sugarless: The Way Forward. Else vier Applied Science; New York, pp. 32-51.

Yamada, K.; Shinohara, H.; Hosoya, N. 1985. Hydrolysis of 1-O-α-D-glucopyranosyl-D-fructofuranose (Trehalulose) by rat intestinal surcrase-isomaltase complex.Nutrition Reports International 32 (5): 1211 - 1220

Ziesenitz, S.C. 1986. Carbohydrasen aus jejunalmucosa des Menschen = [Carbohydrases from the human jejunal mucosa]. Z Ernährungswiss 25(4):253-258. Cited In: Würsch, 1991.

Justification for selection of Effect on developmental toxicity: via oral route:

Hazard assessment is conducted by means of a read-across from a structural surrogate. The selected study is an adequate and reliable study based on the identified similarities in structure and intrinsic properties between source and target substance and overall assessment of quality, duration and dose descriptor level (refer to endpoint discussion for further details).

Justification for selection of Effect on developmental toxicity: via inhalation route:

No study required since exposure of humans via inhalation is unlikely taking into account the physicochemical properties of the substances and the lack of exposure to aerosols, particles or droplets of inhalable size under normal conditions of use.

Justification for selection of Effect on developmental toxicity: via dermal route:

No study required since sufficient weight of evidence is available to exclude that Reaction mass of 1-O-α-D-glucopyranosyl-D-fructose and 6-O-α-D-glucopyranosyl-D-fructose and fructose and glucose and sucrose is toxic after acute dermal exposure considering the available data on read-across to the main components, which have been shown not to be toxic after acute or repeated oral exposure. Moreover, sufficient information is known for the ingredients glucose, fructose and sucrose to consider them as non-hazardous and to include them in Annex IV of Regulation (EC) No. 987/2008.  Therefore, in accordance with Annex XI, Section 1.2 of Regulation (EC) 1907/2006 further testing on vertebrate animals for that property shall be omitted and further testing not involving vertebrate animals may be omitted.  

Justification for classification or non-classification

Based on read-across, the available data on toxicity to reproduction do not meet the classification criteria according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.