Registration Dossier

Diss Factsheets

Administrative data

Description of key information

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
other: migrated information: read-across from supporting substance (structural analogue or surrogate) (obsolete)deactivated phrase
Adequacy of study:
key study
Study period:
7 Nov - 13 Dec 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP-Guideline study, tested with the source substance Isomaltulose (Cas# 13718-94-0). According to the ECHA guidance document "Practical guide 6: How to report read-across and categories (March 2010)", the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
Qualifier:
according to guideline
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
(BAYERISCHES LANDESAMT FÜR GESUNDHEIT UND LEBENSMITTELSICHERHEIT, LANDESINSTITUT FÜR ARBEITSSCHUTZ UND PRODUKTSICHERHEIT, München, Germany)
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
other: CBA/CaOlaHsD
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann, Borchen, Germany
- Age at study initiation: 8 - 9 weeks
- Weight at study initiation: 16 - 23 g
- Housing: 5 animals per cage in IVC cages, type II L, polysulphone cages on Altromin saw fibre bedding (pre-screen test: lot no. 010812, main study: lot no. 160812)
- Diet: Altromin 1324 maintenance diet for rats and mice, ad libitum
- Water: tap water, sulphur acidified to a pH value of approx. 2.8, ad libitum
- Acclimation period: at least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 10
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12 / 12

Vehicle:
dimethyl sulphoxide
Concentration:
6.25, 12.5 and 25% (w/v)
No. of animals per dose:
5 (main study), 3 (pre-screen test including 2 test and 1 control animal)
Details on study design:
RANGE FINDING TESTS:
- Compound solubility: The solubility test revealed a maximum technically applicable concentration of 25% (w/v) of the test item in the vehicle DMSO
- Irritation:
2 mice were treated with 25% test item solution. 1 animal treated with the vehicle DMSO served as negative control. All animals were observed for any clinical signs of systemic toxicity or local irritation at the application site. Body weights were recorded prior to the first treatment (Day 1) and prior to termination (Day 6). Both ears of each mouse were observed for erythema/eschar formation and scored. Measurement of ear thickness was performed on Day 1 (pre-dose), Day 3 (approximately 48 hours after the first dose) and Day 6.
No mortality or any signs of systemic toxicity were observed. No significant signs of irritation were observed as indicated by an erythema score ≥ 3 and/or an increased ear thickness of ≥ 25% on any day of measurement (25%: 0.19 mm on day 1, 0.17 mm on day 3, 0.18 on day 6; vehicle: 0.18 mm on day 1, 0.185 on day 3 and 0.19 0n day 6). According to this, the maximum technically applicable concentration of 25 % (w/v) was selected as the highest test concentration in the main test.
- Lymph node proliferation response: Lymph node proliferation was not assessed in the range-finding test.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Name of test method: 3H-methyl thymidine incorporation determined by ß-scintillation
- Criteria used to consider a positive response: The test item was considered as a skin sensitizer, if exposure to at least one concentration of the test item resulted in an at least 3-fold or greater stimulation index in test animals compared to control mice (SI ≥ 3).
- Validation criteria: The positive control substance phenylenediamine (1%) should produce a positive LLNA response shown by a SI > 3 over the negative control group without causing excessive skin irritation (i.e. SI > 20) or systemic toxicity.

TREATMENT PREPARATION AND ADMINISTRATION:
25 µL of the test item was applied on the entire dorsal surface of each ear of each dose. The application was repeated once daily over three consecutive days. Local irritation reactions were assessed. On Day 6, an injection of 250 µL phosphate buffered saline containing 20 µCi of 3H-methyl thymidine was made into the tail vein of each experimental mouse. Approx. 5 h later, animals were sacrificed and the draining auricular lymph node of each ear was excised. A single cell suspension of lymph node cells, individually pooled for each animal, was prepared. After repeated washings, cells were precipitated with 5% trichloracetic acid at 4 °C overnight. The 3H-methyl thymidine-incorporation was measured in a ß-counter and expressed as the number of disintegrations per minute (DPM) individually for each animal.

CLINICAL OBSERVATIONS:
All animals were observed prior to the application and once a day thereafter to detect signs of toxicity. Irritation at the site of application was monitored by erythema scoring according to the scoring scale defined in OECD GL 429 and recorded for each animal individually.
Positive control substance(s):
other: phenylenediamine (1% in DMSO)
Positive control results:
The positive control substance induced a significant lymphoproliferative response as shown by a stimulation index value of 13.7.
Parameter:
SI
Remarks on result:
other: No significant (SI ≥ 3) lymphoproliferative response was noted for the test substance at the tested concentrations. The observed stimulation index values were 1.0, 1.0 and 0.9 at test item concentrations of 6.25%, 12.5% and 25%, respectively.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Remark
Remarks:
No significant or dose-dependent increase in the DPM values was observed in the test groups compared to the controls. The determined mean DPM values per lymph node were 858.5 ± 261.9, 860.4 ± 305.6, 895.9 ± 92.9 and 744.1 ± 216.5 for the control, low- mid- and high-dose animals, respectively.

No significant effects on the body weight were observed in any treatment group. Furthermore, no mortality or symptoms of systemic toxicity were noted. No signs of dermal irritation were noted at the application site in any test animals.

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
CLP: not classified
DSD: not classified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

Sensitisation

Justification for read-across

There are no data available on skin sensitisation 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 VII, 8.3.

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 skin sensitisation.

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).

 

Skin sensitisation

Sensitisation of isomaltulose (CAS 13718-94-0) was assessed in a GLP-conducted local lymph node assay according to OECD 429 in female CBA/CaOlaHsD mice (Lütkenhaus, 2013b). In regard to the results of a preliminary test, test animals were exposed to 6.25, 12.5 and 25% (w/v) concentrated test solutions suspended in DMSO. According to OECD 429, the test substance was applied topically to the entire surface of each ear of each mouse once daily for three consecutive days. The proliferative response of lymphocytes in the draining auricular lymph nodes was determined via 3H-methyl thymidine incorporation and the stimulation indices relative to the sham treated controls were calculated. No mortality, clinical signs of toxicity, alterations in body weight or local effects at the application site were observed until the end of the study.Further, no dose-related lymphoproliferative response exceeding the threshold level (SI ≥ 3) was noted for the test substance at the tested concentrations. In detail, the observed stimulation index values were 1.0, 1.0 and 0.9 at test item concentrations of 6.25, 12.5 and 25%, respectively. Thus, isomaltulose did not induce skin sensitisation in the conducted study.

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 sensitising potential of the read across surrogate 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 not considered to exhibit sensitising properties.

 

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.

Migrated from Short description of key information:

Skin sensitisation (OECD 429): not sensitising

Justification for selection of skin sensitisation endpoint:

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 quality assessment (refer to the endpoint discussion for further details).

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:

Justification for selection of respiratory sensitisation endpoint:

Study not required according to Annex VII-X of Regulation (EC) No. 1907/2006.

Justification for classification or non-classification

Based on read-across, the available data on skin sensitisation 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.

Categories Display