1-Propanaminium, 2-hydroxy-N-(2-hydroxypropyl)-N,N-dimethyl-, esters with C18-unsatd. fatty acids, Me sulfates (salts)

Dermal absorption

Administrative data

Endpoint:

dermal absorption in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1993-01-26-1993-01-29

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:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 427 (Skin Absorption: In Vivo Method)

Deviations:

yes

Remarks:

Two dose levels should be used in single dose studies. (Only one dose was tested)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Ethanaminium, 2-hydroxy-N-(2-hydroxyethyl)-N,N-dimethyl-, esters with C16-18 and C18-unsatd. fatty acids, chlorides

Cas Number:

1079184-43-2

Molecular formula:

n.a. (UVCB)

IUPAC Name:

Ethanaminium, 2-hydroxy-N-(2-hydroxyethyl)-N,N-dimethyl-, esters with C16-18 and C18-unsatd. fatty acids, chlorides

Details on test material:

- Name of test material (as cited in study report): N-Bis-(Talloyloxyethyl)-N-Dimethyl ammonium chloride (FV-Base)
- radiolabeled position [N-methyl C14 radiolabelled]
- Molecular formula (if other than submission substance): N/A
- Molecular weight (if other than submission substance): no data
- Smiles notation (if other than submission substance): N/A
- InChl (if other than submission substance): N/A
- Structural formula attached as image file (if other than submission substance): N/A
- Substance type:
- Physical state: solid

Radiolabelling:

yes

Remarks:

C14-labeled; 10 µCi/0.1 g (116 mg/ml)

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Labs
- Age at study initiation: N/A
- Weight at study initiation: 175-225 g
- Fasting period before study: Overnight before dosing and for four hours after dosing.
- Housing: For 72 hours after dosing, housed in coated metabolism cages designed for separation of urine, faeces and expired CO2.
- Individual metabolism cages: yes
- Diet (e.g. ad libitum): Purina Rat Chow ad libitum throughout study
- Water (e.g. ad libitum): ad libitum
- Acclimation period: At least 4 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data


IN-LIFE DATES: From: 1/26/93 To: 1/29/93

Administration / exposure

Type of coverage:

other: Permabond 102 adhesive was used to glue an contoured 3.0 cm diameter glass ring to the middle of the back of each animal. A porous glass disk fitted into the top of the ring to permit air circulation over the test site and to prevent loss of the test mate

Vehicle:

ethanol

Duration of exposure:

single dose, duration of exposure is not further specified.

Doses:

- Actual doses: 1.62 mg/cm2 (62.7 mg/kg bw) dermal dose of radio lablled test substance was administered. (0.1 g of dosing solution applied with a syringe and spread over the skin inside a glass ring.)
- Actual doses calculated as follows: Dose of active was 1.62 mg/cm2 (2.4 µmoles/cm2) or 62.7 mg/kg bw (92.6 µmoles/kg).
- Dose volume: effective surface area = 7.63 cm2 = 0.1 g of dosing solution.
- Rationale for dose selection: N/A

No. of animals per group:

4 male rats ( - one rat orally ingested the material and was excluded from analyses).

Control animals:

no

Details on study design:

DOSE PREPARATION
- Method for preparation of dose suspensions: Approximately 2 grams of the dose solution was supplied. Dose solution was prepared by mixing 16 mg of the radiolabeled test substance (208.8 µCi) with 214 mg of unlabeled test substance in 1.8 g of absolute ethanol and warming until dissolved. The amount of dose solution delivered was determined by measuring the difference between the weight of the dosing apparatus plus solution before and after dosing.
- Method of storage: room temperature


APPLICATION OF DOSE: The hair was clipped from the back of the animal using a small animal clipper and only animals with skin which appeared normal was used. Permabond 102 adhesive was used to glue a 3.0 cm diameter glass ring to the middle of the back of each animal. 0.1 g of the dose solution was applied with a syringe and spread over the skin inside the ring.

VEHICLE
- Justification for use and choice of vehicle (if other than water): N/A
- Amount(s) applied (volume or weight with unit):0.1 ml/animal dosing solution
- Concentration (if solution): absolute ethanol (100%)
- Lot/batch no. (if required): N/A
- Purity: N/A


TEST SITE
- Preparation of test site: The hair was clipped from the back of the animal using a small animal clipper and only animals with skin which appeared normal were used.
- Area of exposure: 7.63 cm2
- % coverage: N/A
- Type of cover / wrap if used: Permabond 102 adhesive was used to glue a 3.0 cm diameter glass ring to the middle of the back of each animal. 0.1 g of the dose solution was applied with a syringe and spread over the skin inside the ring.
- Time intervals for shavings or clipplings: Animals were clipped prior to dosing.


SITE PROTECTION / USE OF RESTRAINERS FOR PREVENTING INGESTION: no


REMOVAL OF TEST SUBSTANCE
- Removal of protecting device: N/A
- Washing procedures and type of cleansing agent: N/A
- Time after start of exposure: N/A


SAMPLE COLLECTION
- Collection of blood: Blood was collected from the inferior vena cava in a heparinized syringe. A portion of the sample was submitted for analysis. The plasma fraction from the remainder of the blood was prepared by centrifugation and submitted for analysis.
- Collection of urine and faeces: The male rats were fitted with faecal cups and placed in individual metabolism cages. The accumulated faeces and urine were collected at 24, 48, and 72 hours and frozen. At the end of the 24-, 48- and 72 hour collection period, cages were washed with alcohol followed by distilled water and the washes were collected. If urinary bladder contained urine at necropsy, it was rinsed into the 48 -72 hour urine collection.
- Collection of expired air: CO2 was collected at 24, 48, and 72 hours. The CO2 safety trap (one per rat) was collected at the 72 hour test period.
- Terminal procedure: Animals were observed at least once daily for general health status and observations were recorded. At the end of the 72 hours test period, rats were sacrificed with an overdose of carbon dioxide.
- Analysis of organs: The following were sampled for radiochemistry: Urine, faeces, CO2, blood, plasma, liver (entire), kidneys, testes, heart, lung (entire), spleen, pancreas, brain, bone marrow, muscle (hind limb, right), bone (femur, both), adipose, skin (test site), skin (adjacent), GI Tract, GI Tract wash, carcass, cage wash, cell wash (from blood samples).


SAMPLE PREPARATION
- Storage procedure: samples were frozen as appropriate.

- Preparation details: The test site skin and annular strip of skin from the perimeter of the test site were taken. After removing the other organs and tissues, they were rinsed with water and blotted in paper towel. The fat and connective tissues were removed from the organs and they were placed in tared sample jars. The organs with internal cavities were cut open and rinsed with water. Bone samples from both femurs were taken after bone marrow had been removed. Bones were rinsed with water, washings were discarded. The adipose tissue was sampled from an area of the psoas muscle. The carcass was frozen before grinding in a Wiley mill. Any animals that died were discarded without necropsy and without taking samples for radioassay.


ANALYSIS
- Method type(s) for identification (e.g. GC-FID, GC-MS, HPLC-DAD, HPLC-MS-MS, HPLC-UV, Liquid scintillation counting, NMR, TLC) Liquid scintillation counting.
- Liquid scintillation counting results (cpm) converted to dpm as follows: Not discussed.
- Validation of analytical procedure: Not discussed.
- Limits of detection and quantification: Not discussed.


OTHER:

Details on in vitro test system (if applicable):

N/A

Results and discussion

Signs and symptoms of toxicity:

not specified

Dermal irritation:

not specified

Absorption in different matrices:

Mean +/- SE:

- Non-occlusive cover + enclosure rinse: N/A
- Skin wash: N/A
- Skin test site: 110. +/- 4.3 % of dose
- Skin, untreated site: N/A
- Blood: <0.13 +/- 0.026 µg/g dose
- Carcass: 0.68 +/- 0.59 % of dose
- Urine: 3.4 +/- 2.4 % of dose
- Cage wash + cage wipe: <0.19 +/- 0.098 % of dose
- Faeces: 0.35 +/- 0.18 % of dose
- Expired air (if applicable): <0.18+/- 0.005
- Serial non-detects in excreta at termination: N/A
- Receptor fluid, receptor chamber, donor chamber (in vitro test system): N/A
- Skin preparation (in vitro test system):N/A
- Stratum corneum (in vitro test system): (i.e tape strips) N/A

Total recovery:

- Total recovery: The total percent recovery of the dosed radioactivity was 117 +/- 0.88 % (mean +/- SE; n=3). All subsequent radioactivity recovery data reported for this study were normalized for relative radioactivity.
- Recovery of applied dose acceptable: N/A
- Results adjusted for incomplete recovery of the applied dose: The total percent recovery of the dosed ratioactivity was 117 +/- 0.88 % (mean +/- SE; n=3). All subsequent radioactivity recovery data reported for this study were normalized for relative radioactivity.
- Limit of detection (LOD): Not discussed. The limit of the LSC calibration curves is no better than +/- 5 %, precluding reporting data to more than 2 significant figues.
- Quantification of values below LOD or LOQ: N/A

Percutaneous absorption

Time point:

72 h

Dose:

1.62 mg/cm2

Parameter:

percentage

Absorption:

> 0 - <= 1.4 %

Remarks on result:

other: 72 hours

Remarks:

Extremely low absorption through skin. Most of the test substance (98.6%) remained on the skin

Conversion factor human vs. animal skin:

Not examined

Any other information on results incl. tables

The analytical results from animal #846 were indicative of oral ingestion of the test material. This was suggested by the relatively high radioactive content in the skin adjacent to the test site, urine and tissues compared to the other test animals. Therefore, radiochemical data from this animal were omitted from statistical analysis and subsequent interpretation of results in this study report.

The total percent recovery of the dosed radioactivity was 117 + 0.88 % (mean + SE; n=3). All subsequent radioactivity data reported for this study were normalized for relative radioactivity. At the end of the 72-hour period, 1.03 + 0.89 % of the dosed test substance radioactivity was recovered in the urine plus cage wash, 0.16 + 0.02 % in the expired carbon dioxide, 0.13 + 0.06 % in the tissues plus carcass and 0.05 + 0.01 % in the faeces plus GI tract wash. The balance of the dosed radioactivity remained on the test skin site, adjacent skin and dose cell wash. There was no discernible pattern to the elimination of absorbed radioactivity. Low amounts of radioactivity were detected in the urine, and sporadically, in faeces, expired CO2, and cage wash throughout the study.

Inspection of individual tissue radioactivity distribution at 72 hours shows the presence of low counts of residual radioactivity in all tissues. The liver contained the highest radioactive content, 3 times background (whole blood radioactive content). All other tissues were less than three times background.

The extent of radioactivity absorption following dermal administration of the test substance at 1.62 mg/cm2 (62.7 mg/kg bw) in a vehicle of absolute ethanol to fasted, male Sprague Dawley rats was estimated to be < 1.4 % over the 72 hours test period. Essentially most of the test substance was not absorbed through the skin. Assessment for biliary elimination of absorbed test substance was not determined. Residual radioactivity was detected in all tissues and carcass at 72 hours and accounted for ~ 9 % of absorbed radioactivity. The principal route of radioactivity elimination of dermally administered test substance is urine. Low amounts of radioactivity were sporadically detected in expired carbon dioxide and feces.

Applicant's summary and conclusion

Conclusions:

A total of < 1.4 % (normalised for 100 % recovery) of the administered dose was absorbed over the 72-hour test period. Most of the test substance remained on the skin. Following dermal administration of radiolabelled test substance, the principal route for the elimination of radioactivity was via the urine. Low amounts of radioactivity were sporadically detected in expired carbon dioxide and faeces.

Executive summary:

In a metabolism study comparable to OECD Guideline 427, MDEA-Esterquat C16-18 and C18 unsatd. (> 99 % a.i.), Methyl C¹⁴radio labelled was administered to 4 male Sprague-Dawley rats by the dermal route at a single dose of 1.62 mg/cm²(62.7 mg/kg bw).

Radiochemical data from one animal were omitted from the statistical analysis and subsequent interpretation of the results due to oral ingestion of the test material during the study.

A total of 117 ± 0.88 % of the administered radio labelled test substance was recovered. A total of < 1.4 % (normalised for 100 % recovery) of the administered dose was absorbed over the 72-hour test period. Most of the test substance remained on the skin. About 1.03 % was recovered in urine/cage wash, ~0.16 % in expired CO₂, ~0.13 % in tissue, and ~0.05 % in faeces/GI tract. Of the ~0.13 % recovered in the tissues/carcass, the liver exhibited the highest radioactive content (3 times background).

Following dermal administration of radiolabelled test substance, the principal route for the elimination of radioactivity was via the urine. Low amounts of radioactivity were sporadically detected in expired carbon dioxide and faeces.