Reaction mass of bisisopropyl peroxydicarbonate and bis-sec-butyl peroxydicarbonate and isopropyl-secbutylperoxydicarbonate

Endpoint summary

Description of key information

There is in vivo data for the reaction mass for skin irritation (similar to OECD 404, 1980) based on this the multi-constituent substance is classified as irritating to skin category 2. This data is supported by available data from the constituents, ), these data were not included in as robust summaries since there is data for the multi-constituent substance, (section 13 QSAR doc appendix 1) and QSARs (section 13 QSAR doc Appendix 2).

The stabilizer is not irritating to eyes. SBP is irritating to the eyes and IPP is irritating to the eyes based on read-across to 16066-38-9, there is a corrosive to the eyes result for IPP but this study is not valid (K3 in vivo data). A BCOP assay is available for the reaction mass. The results of the BCOP show that the multi-constituent substance is not corrosive to the eyes but there is indication of eye irritation.This data is supported by available data from the constituents (section 13 QSAR doc appendix 1), data for SBP included in IUCLID as supporting, and QSARs (section 13 QSAR doc Appendix 2). Based on these results further testing In vivo of the reaction mass is not performed and the reaction mass is classified as irritating to the eyes category 2.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

February 21, 1980 - March 3, 1980

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

No CoA, method used equivalent to OECD 404: 24 hour exposure under occlusive conditions. Intact skin and abraded skin. Skin observed: 24, 72 and 96 hours following test article application and once daily thereafter for a total of 14 days.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

GLP compliance:

yes

Remarks:

FDA GLP 1979

Specific details on test material used for the study:

The test article was received, from the Noury Chemical Corporation, Burt, New York on February 14, 1980. It was identified as "Trigonox ADC Peroxydicarbonate liquid" and was received as a clear liquid.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

SOURCE ANIMAL
- Source: Langshaw Farms, Augusta, Michigan (IRDC Purchase Order #3341)
- Sex: male and female
- Age at study initiation (in days): young adults
- Weight at study initiation: 2204 to 2678 grams
- Housing: individually housed in hanging wire-mesh cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 14 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-24
- Humidity (%): 39-48
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: February 21, 1980 - March 3, 1980

Type of coverage:

occlusive

Preparation of test site:

other: shaved and abraded

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.5 ml

Duration of treatment / exposure:

24 hours

Observation period:

14 days

Number of animals:

3 males and 3 females

Details on study design:

TEST SITE
- Area of exposure: back
- % coverage: 6 cm2
- Type of wrap if used: test site of each animal was then covered with gauze bandaging and Saran Wrap® and overwrapped with several layers of Conform® adhesive tape.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): no
- Time after start of exposure: 24 hours

OBSERVATION TIME POINTS
(indicate if minutes, hours or days) Each test site was observed for skin irritation at 24, 72 and 96 hours following test article application and once daily thereafter for a total of 14 days.

SCORING SYSTEM:
- Method of calculation: Skin Reaction Code, Principles and Procedures for Evaluating the Toxicity of Household Substances, National Academy of Sciences, 1977, page 112; modified (inclusion of 0.5 scores) at International Research and Development Corporation.

Irritation parameter:

erythema score

Basis:

other:

Remarks:

all animals

Time point:

other: 0h, 48h, 72h

Score:

>= 1 - <= 3

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks:

in one animal

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Basis:

other: all animals

Time point:

other: 0h, 48h, 72h

Score:

>= 0 - <= 3

Max. score:

4

Reversibility:

not fully reversible within: 14 days

Remarks:

in one animal

Remarks on result:

no indication of irritation

Irritant / corrosive response data:

See table below:

Erythema

Animal/score at:	24h* (0h)	72h* (48h)	96h* (72h)	mean
1m	2.5	1.5	1.5	1.8 a, b
2m	3	2	2	2.3 a
3m**	2	2	2	2
1f	3	3	1.5	2.5 a
2f	2.5	2.5	1	2 a, b
3f	3.0	3	1.5	2.5 a, b

 

Oedema

Animal/score at:	24h* (0h)	72h* (48h)	96h* (72h)	mean
1m	2.5	1.5	0.5	1.5
2m	3	1.5	1	1.8
3m**	2.5	2.0	2	2.2
1f	2.5	1.5	0.5	1.5
2f	3	0.5	0	1.2
3f	2.5	1	1.5	1.7

 

* following test article application, instead of24, 48 and 72 hours after patch removal.

** very slight oedema and erythema observed after 14 days

a Desquamation present at day 14.

b Fissuring present at day 14

None of the animals died, 2 males and 2 females had diarrhea (M 2( 1), 1( 2), 2(3), 1(4), 2(5), 1(6 -8), 2(11 -14)) and F (2(11-12), 3(13-14)). One of these males had nasal discharge 1(6 -14) and soft stool 1(1). The animals had a decreased body weight at study termination.

Interpretation of results:

Category 2 (irritant) based on GHS criteria

Conclusions:

CLP criteria 1 for classifying Catergory 2 is met:

(1) Mean value of ≥ 2,3 - ≤ 4,0 for erythema/ eschar or for oedema in at least 2 of 3 tested animals from gradings at 24, 48 and 72 hours after patch removal or, if reactions are delayed, from grades on 3 consecutive days after the onset of skin reactions; or
(2) Inflammation that persists to the end of the observation period normally 14 days in at least 2 animals, particularly taking into account alopecia (limited area), hyperkeratosis, hyperplasia, and scaling; or
(3) In some cases where there is pronounced variability of response among animals, with very definite positive effects related to chemical exposure in a single animal but less than the criteria above.

Executive summary:

CLP criteria 1 for classifying Catergory 2 is met

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1970

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

No CoA, pre-GLP, limited information on animals and conditions, 72 hours observation, no data for individual animals

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

GLP compliance:

no

Remarks:

performed pre-GLP

Specific details on test material used for the study:

The test materials were received from the Lucidol Division, Pennwalt Corporation, Buffalo, New York, on June 12, 1970. It was received as a clear liquid packed in dry ice.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: no data
- Age at study initiation: no data
- Weight at study initiation: 1858 - 2447 grams
- Housing: individually in hanging metal cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
Maintained in temperature and humidity controlled quarters throughout the test
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): no data

IN-LIFE DATES: From: To: no data

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

0.1 ml

Duration of treatment / exposure:

A volume of 0.1 ml of the test material was instilled into the cupped conjunctival sac of the right eye of each rabbit.

Observation period (in vivo):

72 hours

Number of animals or in vitro replicates:

3 males and 3 females

Details on study design:

REMOVAL OF TEST SUBSTANCE: no

SCORING SYSTEM: Draize, J. H., Appraisal of the Safety of Chemicals in Foods, Drugs, and
Cosmetics, Assoc. Food and Drugs Officials of the U. S., Austin, Texas, 1959., p. 51,
Modified according to revision in 1964. Edited by A. J. Lehman.

TOOL USED TO ASSESS SCORE: Examinations were made for ocular irritation at 24, 48 and 72 hours. At the 72 hour examination, sodium fluorescein and ultraviolet light were used to aid in revealing possible corneal injury.

Irritation parameter:

cornea opacity score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

mean

Time point:

24/48/72 h

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

24 h

Score:

12.6

Remarks on result:

positive indication of irritation

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

48 h

Score:

10.2

Remarks on result:

positive indication of irritation

Irritation parameter:

conjunctivae score

Basis:

mean

Time point:

72 h

Score:

8.2

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

At each observation interval, conjunctival irritation in varying degrees was observed in each of the 6 animals tested. The irritation noted consisted of slight to moderate redness, slight to marked chemosis and very slight to slight discharge. Seventy-two hours following instillation of the test material, each eye was examined with ultraviolet light and sodium fluorescein.
No corneal damage was observed.
Based upon the results obtained Lupersol 225M would be considered an eye irritant.

 

Conjunctivae /score at:	24h*	48h*	72h*
Redness	2.2 (2.0-2.5)	2.1 (1. 0- 2.5)	1.8 (1. 0- 3.0)
Chemosis	3.8 (3.0-4.0)	3.6 (3.0-4.0)	3.7 (3.0- 4.0)
Discharge	1.3 (0.5-2.5)	1.1 (0.5- 2.0)	1.0 (0.5-2.0)

* average score (range)

 

Score of 0 for Cornea and Iris.

 

No Comments can be made on reversibility of these effects since the observation period was only 72 hours.

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

Eye irritation was observed and based on these results the substance is classifed as irritating to the eyes.

Executive summary:

Eye irritation was observed and based on these results the substance is classifed as irritating to the eyes. Since the animals were only observed for 72 hours no comments can be made on reversibility and individual scores are not available.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-04-26 to 2017-07-11

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: OECD Guideline for the Testing of Chemicals, no. 437 (adopted: 26 July 2013)

Deviations:

yes

Remarks:

Concerning: Calibration of the Opacitometer (see Test System)

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Munich, Germany

Specific details on test material used for the study:

Name: Isopropyl sec-butyl peroxy dicarbonate (multi constituent)
Product Name: Trigonox ADC-NS30
Chemical Name: Reaction mass of bisisopropyl peroxydicarbonate and bis-sec-butyl peroxydicarbonate and isopropylsecbutylperoxydicarbonate
Composition: 71.5% w/w Diethylene glycol bis(allyl carbonate) [142-22-3]
28.0% w/w Isopropyl sec-butyl peroxy dicarbonate [79350-78-4],
Di-sec-butyl peroxydicarbonate [19910-65-7], and
Diisopropyl peroxydicarbonate [105-64-6]
Batch No.: 16041B0206
Physical State: liquid
Colour: colorless
Density: 1.1 g/mL
Storage Conditions: ≤-20°C, protected from light
Expiry Date: 08 August 2017
Safety Precautions: The routine hygienic procedures will be sufficient to assure personnel health and safety.
Peroxide handling
on the lab: The peroxide - water mixture is a 2 liquids phase system. Intensive mixing is required to get the right organic – water ratio in the sample applied for tox testing:
• place the polymer support on the bottom of the other beaker (to fix the inner beaker)
• fill the other glass beaker for 1/3 with ice
• place an inner beaker, with solvent and a magnetic stirring rod, in the outer beaker
• add the peroxide during stirring, apply a stirring speed of
• 200 rpm for peroxide - IPA mixtures, or 500 rpm for peroxide – water (or salt water) mixtures
• maximum handling time: 60 minutes

Vehicle:

physiological saline

Amount / concentration applied:

The test item was used as provided by the sponsor. Handling of the peroxide – water mixture before administration is described above . 750 µL of the test substance or the control substance physiological saline 0.9% NaCl (B. Braun Melsungen, lot no. 17031412, expiry date: 12/2019) was introduced into the anterior chamber.

Duration of treatment / exposure:

After 10 minutes incubation at 32 +/- 1 °C either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red).

Observation period (in vivo):

1 mL of a 4 mg/mL sodium fluorescein solution was added to the anterior chamber and the corneas were incubated for 90 minutes at 32 +/- 1 °C.

Details on study design:

Test System

Preparation of the Corneas:
The assay uses isolated corneas obtained as a by-product from animals freshly slaughtered at the abattoir A. Moksel AG, Buchloe, Germany.
On the test day, fresh eyes were collected from the slaughterhouse and were transported in HBSS containing Pen/Strep on ice to the laboratories. Immediately after arrival of the eyes, cornea preparation was initiated.
The eyes were carefully examined for defects and any defective eyes were discarded.
The tissue surrounding the eyeball was carefully pulled away and the cornea was excised leaving a 2 to 3 mm rim of sclera. The isolated corneas were stored in a petri dish containing HBSS. Before the corneas were mounted in corneal holders (Duratec GmbH) with the endothelial side against the O-ring of the posterior chamber, they had been visually examined for defects and any defective cornea had been discarded. The anterior chamber was then positioned on top of the cornea and tightened with screws. The chambers of the corneal holder were then filled with RPMI (without phenol red) containing 1% FBS and 2 mM L-glutamine (complete RPMI). The posterior chamber was always filled first. The corneas were incubated for one hour at 32 ± 1 °C.


Calibration of the Opacitometer:
The opacitometer (BASF-OP3.0, Duratec GmbH) was switched on at least 15 min before starting the calibration procedure. The filter holder was placed into the opacitometer and the readout was adjusted to
1000 lux ± 10 lux using the “Calibrate”-turning knob. For calibration the glass filter F2 was introduced into the filter holder. The readout lay in the range between 540-560 lux. To test the linearity of the measurement, two additional calibration filters, glass filter F3 and glass filter F4, were measured. For these glass filters, the opacitometer displayed values between 300-310 lux and between 95-105 lux. The calibration procedure was performed before each test, once before the first illuminance measurement and once before the second illuminance measurement and is documented in the raw data.

Treatment of the Corneas:
After the equilibration period, the medium was removed from both chambers and replaced with fresh complete RPMI. An initial measurement was performed on each of the corneas using the opacitometer. Three corneas with illuminance readings approximately equivalent to the median illuminance of all corneas were selected as negative-control corneas. The illuminance of each cornea was read and recorded. Only corneas that had an initial illuminance reading I > I0/1.1651 lux were used for the assay. The medium was removed from the anterior chamber and replaced with the test item or control.
750 ± L of the test substance or the control substance was introduced into the anterior chamber. After 10 minutes incubation at 32 ± 1 °C either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red). Once the medium was free of test substance, the cornea was finally rinsed with complete RPMI (without phenol red). The anterior chamber was refilled with complete RPMI and an illuminance measurement was performed after 2 hours incubation at 32 ± 1 °C. Also, each cornea was observed visually and pertinent observations were recorded.
After the illuminance measurement was performed, the medium was removed from both chambers of the holder. The posterior chamber was refilled with fresh complete RPMI. 1 mL of a 4 mg/mL sodium fluorescein solution was added to the anterior chamber and the corneas were incubated for 90 minutes at 32 ± 1 °C. Then the medium from the posterior chamber was removed and its optical density at 490 nm (OD490) was determined, using a spectrophotometer (Jenway 6405 UV/VIS).




Test Groups:
3 corneas for the test item
3 corneas as negative controls treated with physiological saline 0.9% NaCl
3 corneas as positive controls treated with ethanol 100%
The BCOP assay is considered to be valid if the in vitro irritation score obtained with the positive control falls within the two standard deviations of the current historical mean.
The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.


Evaluation of Results:
The following formula was used to calculate the opacity, whereas the values a and b are equipment-specific variables empirically determined by the manufacturer:
Opacity = (I0/I - b)/a

with a = 0.025 and b = 0.9894

The value I0 = I(zero) is the illuminance through a holder without cornea, but with windows and liquid. This value is determined by taking the mean for a set of cornea holders and is reevaluated periodically.
The change in opacity for each cornea was calculated by subtracting the initial opacity reading from the final opacity reading. These values were corrected by subtracting from each the average change in opacity observed for the negative-control corneas. The mean opacity value for each treatment was calculated by averaging the corrected opacity values of each cornea for a given treatment.
The mean OD490 for the blank cuvettes was calculated. The mean blank OD490 was subtracted from the OD490 of each cuvette (corrected OD490). Any dilutions that were made to bring the OD490 values into the linear range of the spectrophotometer (OD490 should be less than 1.500), were taken into account by multiplying the OD490 value of the dilution by the dilution factor. The final-corrected OD490 of the test article and the positive control were calculated by subtracting the average-corrected OD490 of the negative-control corneas from the corrected OD490 value of each treated cornea:
Final-corrected OD490 = (OD490 – mean blank OD490) – average-corrected negative control OD490
The mean OD490 value of each treatment group was calculated by averaging the final corrected OD490 values of the treated corneas for that treatment condition.
The following formula was used to determine the in vitro irritation score (IVIS):
IVIS = mean opacity value + (15 x mean permeability OD490 value)
The IVIS cut-off values for identifying test substances as inducing serious eye damage (UN GHS Category 1) and test substances not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are given in the table below.

An identification of test substances that should be classified as irritating to eyes (UN GHS Category 2 or Category 2A) or test substances that should be classified as mildly irritating to eyes (UN GHS Category 2B) cannot be made.
For this purpose further testing with another suitable method is required

Irritation parameter:

in vitro irritation score

Run / experiment:

mean

Value:

3.65

Irritant / corrosive response data:

The eye irritancy potential of Isopropyl sec-butyl peroxy dicarbonate (multi constituent) was investigated in the bovine corneal opacity and permeability assay.
The test item was used as provided by the sponsor. Handling of the peroxide – water mixture before administration is described above.
All 3 corneas treated with Isopropyl sec-butyl peroxy dicarbonate (multi constituent) showed slight opacity of the tissue.
The following mean in vitro irritation score was calculated:
3.65

Interpretation of results:

study cannot be used for classification

Conclusions:

No prediction can be made regarding the classification of the test substance Isopropyl sec-butyl peroxy dicarbonate (multi constituent) according to the evaluation criteria. Further testing in another suitable method is required.

Executive summary:

Summary Results

The eye irritancy potential of Isopropyl sec-butyl peroxy dicarbonate (multi constituent) was investigated in the bovine corneal opacity and permeability assay.

Preparation of the test item:                      tested as provided by the sponsor

Visual Observation after treatment:         All 3 corneas treated with Isopropyl sec-butyl peroxy dicarbonate (multi constituent) showed slight opacity of the tissue.

Meanin vitroirritation score:                     3.65

Classification:     

 

X	No prediction can be made

The in vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid.

The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

There is in vivo data for the reaction mass for skin irritation (similar to OECD 404, 1980) based on this the multi-constituent substance is classified as irritating to skin category 2.

The stabilizer is not irritating to eyes. SBP is irritating to the eyes and IPP is corrosive to the eyes based on K3 in vivo data. A BCOP assay is available for the reaction mass. The results of the BCOP show that the multi-constituent substance is not corrosive to the eyes but there is indication of eye irritation.This data is supported by available data from the constituents (section 13 QSAR doc appendix 1), data for SBP included in IUCLID as supporting, and QSARs (section 13 QSAR doc Appendix 2). Based on these results further testing In vivo of the reaction mass is not performed and the reaction mass is classified as irritating to the eyes category 2.