1-nitroguanidine

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

September 1984 - January 1985

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1986

Report date:

1986

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: EPA 560/6-82-001

Deviations:

no

Remarks:

slit lamp exam. was added; colour photographic docum. was not performed; 8-hour period with altered temperatur and humidity range; diet was fed at 150 g/d and not ad libitum > none of these dev. had a negative impact on study performance and validity

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1-nitroguanidine

EC Number:

209-143-5

EC Name:

1-nitroguanidine

Cas Number:

556-88-7

Molecular formula:

CH4N4O2

IUPAC Name:

N-nitroguanidine

impurity 1

Reference substance name:

Sodium sulphate

EC Number:

231-820-9

EC Name:

Sodium sulphate

Cas Number:

7757-82-6

Molecular formula:

Na2O4S

IUPAC Name:

disodium sulfate

impurity 2

Reference substance name:

Sodium nitrate

EC Number:

231-554-3

EC Name:

Sodium nitrate

Cas Number:

7631-99-4

Molecular formula:

HNO3.Na

IUPAC Name:

sodium nitrate

impurity 3

Reference substance name:

4,6-diamino-1,3,5-triazin-2(1H)-one

EC Number:

211-455-1

EC Name:

4,6-diamino-1,3,5-triazin-2(1H)-one

Cas Number:

645-92-1

Molecular formula:

C3H5N5O

IUPAC Name:

4,6-diamino-1,3,5-triazin-2(1H)-one

impurity 4

Reference substance name:

6-amino-1,3,5-triazine-2,4(1H,3H)-dione

EC Number:

211-456-7

EC Name:

6-amino-1,3,5-triazine-2,4(1H,3H)-dione

Cas Number:

645-93-2

Molecular formula:

C3H4N4O2

IUPAC Name:

6-amino-1,3,5-triazine-2,4(1H,3H)-dione

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Test substance supplier Sunflower AAP

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Elkhorn Rabbitry, 5265 Starr Way, Watsonville, CA, U.S.A.)
- Age at study initiation: young adults
- Sex: males
- Weight at study initiation: 2.4-3.5 kg
- Housing: individually in stainless steel, screen-bottomed, battery-type cages with automatically flushing dump tanks
- Diet: approx. 150 g/day of Certified Purina Chow Diet 5322 (Ralston Purina Company)
- Water: continous drip from a central line
- Pretest conditioning: 1. quarantine from 6 Sept- 19 Sept 1984
2. animal eyes were examined 24 hours before dosing using slit lamp, fluorescein dye and ultraviolet light
- Acclimation period: at least five days after quarantine


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 14.4-17.8; exception: 8-hour period on 22 Sep 84 when the temperature fluctuated between 12.8 and 26.7 °C
- Humidity (%): 40-70%
- Photoperiod (hrs dark/hrs light): 12/12


IN-LIFE DATES:
Group1 > From: 25 Sep 1984 To: 28 Sep 1984
Group2 > From: 2 Oct 1984 To: 5 Oct 1984

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: untreated eye served as control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.025 g (=0.1 ml)
- Administration: the lower lid was pulled away from the conjunctival cul-de-sac to form a cup into which the compound was dropped. Upper and lower lids were then held gently together for one second to prevent loss of material.

Duration of treatment / exposure:

single application

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

Three animals per dose group (two dose groups).
Group one was dosed on 25 Sep 84 and group 2 was dosed on 2 Oct 84.

Details on study design:

TEST PROCEDURE:
- prior to study initiation animals were examined for any pre-existing abnormalities.
- For each animal, the eye with the nearest normal appearance was designated for treatment; the contralateral eye served as control.
- On the next day, 0.025 g Nitroguanidine was placed in the treated eye of each rabbit in this group (group 1: 25 Sep 1984; group 2: 2 Oct 1984)


SCORING SYSTEM: Draize-type ocular grading


TOOL USED TO ASSESS SCORE:
- slit lamp (24, 48, and 72-hour-observations); additionally any areas appearing grossly abnormal were examined under high magnification
- fluorescein: introduced onto the eye which was then observed under unltraviolet light. Any corneal areas reacting with the dye (a sign of discontinuity of the corneal epithelium) were described with respect to area and intensity of fluorescence
- Examination and grading of ocular reactions were performed in this fashion at 1, 4, 24, 48, and 72 h after dosing.
- Fluorescein staining was omitted from the 1 and 4 hour-observations, as was slit lamp examination.
- Due to an almost lack of reaction during the 72 hours after dosing, the study was terminated, according to protocol, after this observation. Therfore no scoring or observations were performed at 7, 14, or 21 days.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

other: 1 and 4 h after dosing

Score:

0

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

iris score

Basis:

mean

Time point:

other: 1 and 4 h after dosing

Score:

0

Max. score:

2

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

other: 1 and 4 h after dosing

Score:

1

Max. score:

3

Reversibility:

fully reversible within: 1 day

Remarks on result:

other: slight conjunctival injection

Results 6

Irritation parameter:

conjunctivae score

Remarks:

redness

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

3

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 7

Irritation parameter:

chemosis score

Remarks:

conjunctival swelling

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Results 8

Irritation parameter:

chemosis score

Remarks:

conjunctival swelling

Basis:

mean

Time point:

other: 1 and 4 h after dosing

Score:

0

Max. score:

4

Reversibility:

fully reversible

Remarks on result:

no indication of irritation

Any other information on results incl. tables

Control Eyes:

- At no time during the study did no contralateral, untreated eyes exhibit any changes from the pre-dosing examination.

- Fine diffuse stippling was present before and after dosing in the control eye of one rabbit.

- All other untreated controls were normal throughout the entire study.

Nitroguanidine produced no positive reactions. Slight conjunctival redness, indicating mild inflammation, was the only response observed. This reaction, although scorable, did not achieve sufficient severity to warrant consideration as a “positive response”. A likely explanation for this slight redness was the observed presence in the lower conjunctival cul-de-sac (the site of gradable redness) of small amounts of the test material. Most of the eyes with gradable redness exhibited undissolved nitroguanidine. Therefore physical irritation, not pharmacological activity, may have been responsible for the slight response observed. Since human exposure would be closely followed with thorough washing, even this slight response may not occur.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In the present study, nitroguanidine produced no response indicative of a potential to cause occular irritation upon direct contact with the eye.

Executive summary:

In an acute eye irritation study using a modified Draize method (US EPA 560/6-82-001), 0.1 ml (corresponding to 0.025 g) of Nitroguanidine was instilled into the conjunctival sac of the eye of six male albino rabbits (New Zealand White strain). Eyes were not washed afterwards. Animals then were observed for 3 days. Irritation was scored by the method of US EPA 560/6-82-001.

Under the experimental conditions employed, Nitroguanidine produced no positive reactions. Slight conjunctival redness, indicating mild inflammation, was the only response observed. This reaction, although scorable, did not achieve sufficient severity to warrant consideration as a “positive response”. A likely explanation for this slight redness was the observed presence in the lower conjunctival cul-de-sac (the site of gradable redness) of small amounts of the test material. Most of the eyes with gradable redness exhibited undissolved nitro guanidine. Therefore physical irritation, not pharmacological activity, may have been responsible for the slight response observed. Since human exposure would be closely followed with thorough washing, even this slight response may not occur.

In the present study, nitroguanidine produced no response indicative of a potential to cause occular irritation upon direct contact with the eye.