Fatty acids, C16-18 (even numbered, C18 unsaturated), 2-ethylhexyl esters, epoxidized

Dermal absorption

Administrative data

Endpoint:

dermal absorption in vivo

Type of information:

experimental study

Remarks:

Performed with metabolic degradation product

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

Critical study for SIDS endpoint

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Principles of method if other than guideline:

AME (absorption, metabolism, excretion) following dermal application of [14C]-2-EH. Excretion balance studies were conducted with 2-ethylhexanol (2-EH) in female Fischer 344 rats following single high (500 mg/kg) and low (50 mg/kg) oral doses of [14C]2-El, following repeated oral dosing with unlabelled 2-EH at the low level, following dermal exposure for 6 h with a 1 g/kg applied dose of [14C]2-EH, and following a 1 mg/kg i.v. dose of [14C]2 -EH.

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-ethylhexan-1-ol

EC Number:

203-234-3

EC Name:

2-ethylhexan-1-ol

Cas Number:

104-76-7

Molecular formula:

C8H18O

IUPAC Name:

2-ethylhexan-1-ol

Specific details on test material used for the study:

The test item 2-ethylhexan-1-ol is a metabolic degradation product of the substance to be registered.

Radiolabelling:

yes

Test animals

Species:

rat

Strain:

Fischer 344

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Inc., Kingston, NY, USA
- Age at study initiation: 9-11 weeks
- Weight at study initiation: 125-150g.
- Housing: suspended, stainless-steel mesh cages prior to study, and were transferred to the study room to allow acclimatization for at least 1 day prior to dosing.
- Individual metabolism cages: yes/no
- Diet: certified rodent diet (Agway‘” Prolab RMH 3000 or Agway Prolab RMH 3200 meal) adlibitum except for a 4-h period immediately after dosing.
- Water: Domestic tap water was available ad libitum
- Acclimation period: isolation for at least 5 days prior to use

Administration / exposure

Type of coverage:

other: Pyrex glass cylinders

Vehicle:

unchanged (no vehicle)

Duration of exposure:

96 hours

Doses:

1 g/kg bw

Details on study design:

TEST SITE
- Preparation of test site: shaved dorsal skin
- Area of exposure: 2.27 mm² (inner diameter of the Pyrex glass cylinder)
- Type of cover / wrap if used: adhesive

SITE PROTECTION / USE OF RESTRAINERS FOR PREVENTING INGESTION:
adhesive

REMOVAL OF TEST SUBSTANCE
- Washing procedures and type of cleaning agent: aspiration of remaining test material, repeated washing with a 40% Hisoderm soap solution
- Time after start of exposure: 6 hrs

SAMPLE COLLECTION
- Collection of blood: from the tail vein; 5 samples within the first 10 min; then at 25, 50, 75, and 95 min.
- Collection of urine and faeces: separately, at intervals 0-8; 8-24; 24-48; 48-96 hrs
- Collection of expired air: (1) in sodium hydroxide traps and (2) in silica gel traps; time intervals as above
- Terminal procedure: no data
- Analysis of organs: no

SAMPLE PREPARATION
- Storage procedure: freezing until use (urine, faeces); blood: heparinised
- Preparation details: combustion (total radiaoactivity in urine and faeces); one-step digestant-scintillant treatment (blood); enzymatic treatment of urine for metabolite identification (ß-glucuronidase, sulfatase, acid) follwoed by filtering

ANALYSIS
- Method type(s) for identification: GC-MS, HPLC-MS-MS, Liquid scintillation counting, TLC

Results and discussion

Signs and symptoms of toxicity:

not specified

Dermal irritation:

not specified

Absorption in different matrices:

- Non-occlusive cover + enclosure rinse: 11.43%
- Skin wash: 38.95%
- Skin test site: 34.99%
- Urine: 3.32%
- Cage wash: 0.87%
- Faeces: 0.57%
- Expired air: 1.84%

Total recovery:

- Total recovery: 91.97%
- Recovery of applied dose acceptable: yes
- Results adjusted for incomplete recovery of the applied dose: no

Percutaneous absorption

Percutaneous absorption 1

Time point:

96 h

Dose:

1 g/kg bw

Parameter:

percentage

Absorption:

ca. 7 %

Percutaneous absorption 2

Time point:

96 h

Dose:

1 g/ kg bw

Parameter:

rate

Absorption:

0.57 mg cm-2 h-1

Any other information on results incl. tables

Table 4. Recovery of radioactivity from a 6-h, 1^.0 g/kg dermal application of neat [¹⁴C]2-ethylhexanol to a 2^.27 cm²area on the clipped backs of female Fischer 344 rat

		Collection period (h)
Sample	0-8	8-24	24-48	48-96	Total
Urine	1.41±0.47	1.66 ±0.25	0.19±0.03	0.07±0.02	3.32±0.72
Faeces	0.03±0^.02	0.39±0.066	0.15±0.06	0.01±0.01	0.57±0.09
Cage wash (water)	0.58±0.13	0.24±0.03	0.04±0.01	0.03±0.01	0.87±0
Silica gel breath traps	0.94±0.83	0.40±0.16	0.04±0.01	0.03±0.01	1.41±0.92
Sodium hydroxide breath traps	0.24±0.04	0.13±0.07	0.03±0.01	0.03± 0.01	0.43±0.06
Dose recovered from the exposure site at 6h					34^.99±16.61
Washing recovery from the exposure site at 6h					38.95±9.78
Dose on cell cover at 6h					11.43±5.17
Total recovery	3.19 ±0.77	2.81±0.38	0.44±0.11	0.17±0.03	91.97±5.75

Values are the mean per cent of dose SSD recovered from four animals.

Table 5. Recovery of radioactivity following a 1 mg/kg i.v. (tail vein) dose of [¹⁴C]2-ethylhexanol (in saline) to female Fischer 344 rats.

		Collection period (h)
Sample	0-8	8-24	24-48	48-96	Total
Urine	35.79±1.89	14.50+2.99	1.59±0.28	1.40±0.26	53.28±3.70
Faeces	0.19±0.30	2.89±0.95	0.55±0.15	0.22±0.05	3.84±1.19
Cage wash (water)	15.94±3.36	3.66±1.74	0.66+0.28	0.92±0.44	21.17±2.92
Silica gel breath	0.22 ± 0.03	0.15±0.04	0.04 ±0.01	0.03 ± 0.01	0.44± 0.07
traps Sodium hydroxide	19.12 ± 1.02	2.15±0.18	0.94±0.11	0.77±0.07	22.97±1.23
breath traps Total recovery	71.24±3.42	23.35+3.01	3.77±0.67	3.33±0.42	101.69±1.11

Values are the mean per cent of dose +/- SD recovered from four animals

Table 6. Quantitation of urinary metabolites of 2-ethylhexanol in female Fischer 344 rats.

				Per cent of dose per peak
	Single low oral dose		Single high oral dose		Repeated low oral dose		Dermal dose
Peak ID	Free	Glucuronide	Free	Glucuronide	Free	Glucuronide	Free	Glucuronide
5-OH-EHA	1.11±0-64	1.28±1.26	3.06±0.88	1.18±0.77	0.73±0.21	2.93±2.27	<0.01	0.30±0.37
2-Ethyladipate + 5-OH-EHA	5.55±2.27	24.12±6.84	4.13+0.95	8.17+4.09	2.37+1.62	25.30±2.19	<0.01	1.61±0.37
6-OH-EHA	1.56±1.10	6.81+1.97	0.80+0.71	6.26±0.55	1.43±0.87	6.89±1.65	<0.01	0.57±0.32
Lactones of 5-OH-EHA	0.44±0.17	0.19±0.25	0.28±0.05	0.01	0.56±0.41	0.82±1.54	<0.01	0.01
2-Ethyl-5-hexenoic acid	<0.01	0.16±0.11	0.01	0.10±0.13	0.06±0.12	0.20±0.11	<0.01	0.01
EHA	0.54±0.68	6.30±1.21	4.77±3.69	20.14±3.25	1.27±1.08	7.45+2.42	<0.01	0.52±0.23
2-Ethylhexanol	0.03+0.05	1.53±0.51	0.01	0.34±0.31	0.22±0.40	1.19±0.15	<0.01	0.06±0.07

Values represent the mean±SD of four animals over the 0-24-h collection period. Quantitation is based on hplc separation and radiochemical detection of metabolites. The structural assignments are based on hplc and glc-mass selective detection analysis of samples and authentic standards both before and after enzymic hydrolysis.

Applicant's summary and conclusion

Conclusions:

Bioavailability of 2-EH via the dermal route is low. Metabolism and excretion of absorbed material is fast, with no difference to the oral route.