Sulfuric acid, mono-C12-14-alkyl esters, magnesium salts

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

22 September - 30 October 1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1988

Report date:

1988

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

not stated

Deviations:

yes

Remarks:

lack of details on test substance

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

A non-LLNA test is available that was performed prior to the current data requirements, stipulated in Regulation (EC) No 1907/2006. In accordance with the same Regulation, the data were included to avoid unnecessary testing.

Test material

Test material information

Constituent 1

Reference substance name:

Sulfuric acid, mono-C12-14-alkyl esters, magnesium salts

EC Number:

292-222-1

EC Name:

Sulfuric acid, mono-C12-14-alkyl esters, magnesium salts

Cas Number:

90583-23-6

Molecular formula:

Not applicable, UVCB substance

IUPAC Name:

Sulfuric acid, mono-C12-14 (even numbered)-alkyl esters, magnesium salts

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: Pirbright white

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Winkelmann, Borchen, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Weight at study initiation (pre-tests): 359.2 g, 362.7 g and 342.4 g
- Weight at study initiation ( main test): 335.6 g (control group) and 346.2 g (test group)
- Housing: Makrolon cage, type IV; 5 animals per cage in the pre-test; 4 animals per cage in the main test
- Diet: Altromin-Haltungsdiät 3032 DG, Altromin GmbH, Lage, Germany, ad libitum
- Water: tap water, ad libitum
- Acclimation period: 6 days under test conditions
- Indication of any skin lesions: only healthy animals were used

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 50 - 60
- Air changes (per hr): not specified
- Photoperiod (hrs dark / hrs light): 12 / 12

Study design: in vivo (non-LLNA)

Induction

Route:

intradermal and epicutaneous

Vehicle:

water

Concentration / amount:

1st induction (intradermal injection): 0.1% in water / 0.1 mL
2nd induction (topical application): 2.5% in water / 1 mL

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

water

Concentration / amount:

1% in water / 0.2 mL

No. of animals per dose:

5 (dose finding for intradermal induction), 5 (dose finding for topical induction), 5 (dose finding for challenge), 20 (test group), 20 (control group)

Details on study design:

RANGE FINDING TESTS:
- Dose finding for intradermal induction: The concentrations of 0.1, 0.5, 1.0 and 2.0% in water were applied to determine concentrations of the test substance suitable for induction of irritating effects.
- Dose finding for topical induction: The concentrations of 5.0 and 10% in water were applied to determine concentrations of the test substance suitable for induction of irritating effects.
- Dose finding for topical challenge: The concentrations of 0.5 and 1.0% in water were applied to determine concentrations of the test substance suitable for induction of sensitising effects.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:

Intradermal (3 paris of injections):
Injection 1: 0.1 mL of a 1:1 mixture (v/v) FCA/0.9% NaCl solution
Injection 2: 0.1 mL of a 0.1% aquesous solution of test substance
Injection 3: 0.1 mL of a 1:1 mixture (v/v) of 0.2% aqueous solution of test substance/FCA

Epicutaneous: 2.5% aqueous solution of test substance

- Control group:

Intradermal (3 pairs of injections)
Injection 1: 0.1 mL of a 1:1 mixture (v/v) FCA/0.9% NaCl solution
Injection 2: not applicable since aqua dest. was used as solvent
Injection 3: 0.1 mL of a 1:1 mixture (v/v) of aqua dest./FCA

Epicutaneous: not applicable since aqua dest. was used as vehicle

- Site: left and right flank
- Frequency of applications: 1 intradermal injection (day 1), 1 topical application (day 8)
- Concentrations: 0.1% in water for intradermal injection and 2.5% in water for topical application

B. CHALLENGE EXPOSURE
- No. of exposures: 1 (challenge)
- Day of challenge: 14 days after epicutaneous induction
- Exposure period: 24 h
- Test groups: test substance in water
- Control group: test substance in water
- Site: right flank (test group), left flank (control group)
- Concentrations: 1% in water
- Evaluation (hr after challenge): 24 and 48 h

Challenge controls:

The control group is actually a challenge control. Animals were challenged on the flank treated during induction and on the other flank as challenge control.

Positive control substance(s):

not specified

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

1%

No. with + reactions:

0

Total no. in group:

20

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

1%

No. with + reactions:

0

Total no. in group:

20

Results 3

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

1%

No. with + reactions:

0

Total no. in group:

20

Results 4

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

1%

No. with + reactions:

2

Total no. in group:

20

Clinical observations:

slight edema

Results 5

Group:

positive control

Remarks on result:

other: no information on positive control in study report available

Any other information on results incl. tables

Clinical signs / local reactions

No mortalities occured in the course of the investigation. After the first (intradermal injection) induction typical signs of irritating effects of Freund's Complete Adjuvans (FCA) were observed in all animals. The test substance induced only slight to moderate irritation following the intradermal injection. After removal of the patch of the 2nd (topical) induction, necrosis finally leading to scabby patches at the sites of the intradermal injection of the test substance were noted. 24 h after removal of the patch of the (topical) challenge exposure, no skin reactions were observed in either control group nor test group animals. However, 48 h after patch removal, 2/20 test animals exhibited a slight edema. Since skin reactions occured only in 2/20 test animals and 0/20 control animals, no tabular representation of the study results is considered necessary.

Body weight development

Animals in the test group exhibited a lower weight gain when compared to control group animals. The reduced weight gain was attributed to the fact that test animals underwent the 48 h 2nd (topical) induction treatment under occlusive conditions which prevented usual food intake. The animals of the control group did not undergo this 48 h treatment and hence were able to gain further weight. Details of the body weight development of test and control animals are summarised in the following tables.

Table 1: Body weight development of control group animals

Animal no.	Weight prior to treatment [g]	Weight after 1 week [g]	Weight after 3 weeks [g]	Weight gain [g]
1	326.4	354.9	455.5	129.1
2	378.4	436.6	558.2	179.8
3	351.9	375.7	440.3	88.4
4	336.0	377.4	433.6	97.6
5	337.6	386.9	449.7	112.1
6	359.5	413.0	466.0	106.5
7	363.8	464.6	558.9	195.1
8	328.7	382.7	459.0	130.3
9	335.9	382.1	467.7	131.8
10	366.5	435.0	498.9	132.4
11	354.3	409.6	505.5	151.2
12	317.2	335.7	402.1	84.9
13	333.5	390.4	470.7	137.2
14	361.6	403.0	477.9	116.3
15	348.2	412.9	486.1	137.9
16	357.7	424.5	512.1	154.4
17	330.3	378.2	432.4	102.1
18	347.3	422.1	539.0	191.7
19	364.4	391.2	453.2	88.8
20	325.6	349.9	410.4	84.8
Average	346.2	396.3	473.9	127.6

Table 2: Body weight development of test group animals

Animal no.	Weight prior to treatment [g]	Weight after 1 week [g]	Weight after 3 weeks [g]	Weight gain [g]
1	343.3	383.6	439.3	96.0
2	336.3	393.5	459.2	122.9
3	350.4	401.6	446.2	95.8
4	357.8	409.7	451.6	93.8
5	331.9	395.2	425.3	93.4
6	338.8	386.5	439.2	100.4
7	327.6	402.1	461.6	134.0
8	296.0	346.7	396.9	100.9
9	356.3	387.4	460.7	104.4
10	321.3	403.3	452.1	130.8
11	310.8	37.7	421.3	110.5
12	382.5	453.1	523.4	140.9
13	352.2	409.4	434.8	82.6
14	366.3	424.7	459.8	93.5
15	322.3	393.8	417.6	95.3
16	296.4	345.9	392.0	95.6
17	358.5	417.1	479.9	121.4
18	326.5	372.3	454.2	127.7
19	321.4	375.9	422.6	101.2
20	316.2	372.1	443.3	127.1
Average	335.6	392.5	444.1	108.4

Applicant's summary and conclusion

Interpretation of results:

other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008.