Fast Pyrolysis Bio-oil

Endpoint summary

Description of key information

The potential of Fast Pyrolysis Bio-oil to induce skin corrosion was analyzed by using the GLP compliant guideline study “In Vitro Membrane Barrier Test (CORROSITEX™ Assay)”, comprising a Chemical Detection System covered by a bio-barrier membrane. In the study, the test substance showed corrosive effects. The test substance is classified as “CORROSIVE”, subcategory 1C according to the UN GHS, as the mean time to activate the CDS was >30-60 min (48.75 ± 6.60 min.) (category 2). The test substance is therefore assigned to Packing Group 3 and to EU Risk-Phrase (R 34). Under the regulation 1272/2008/EC [CLP] the corresponding hazard statement is H314 "Causes severe skin burns and eye damage".

 

The potential of Fast Pyrolysis Bio-oil (the test item BIOTOX-21) to induce skin irritation / corrosion was evaluated in rabbits according to OECD (No. 404) and EC (2004/73/EC B.4) guidelines and in compliance with GLP. The test item was first applied for periods of 3 minutes, 1 hour and 4 hours to a single male New Zealand White rabbit. As the test item showed corrosive properties on first animal, the study was considered complete and the test item was not evaluated on other animals. Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing. The mean values of the scores for erythema and oedema were calculated. The test item BIOTOX-21 is corrosive when applied topically to rabbits and according to the classification criteria laid down in 67/548/EEC, the test item BIOTOX-21 should be classified as corrosive and assigned the symbol C, the indication of danger "Corrosive" and the risk phrase R34 "Causes burns". Under the Regulation EC No 1272/2008 (CLP) the corresponding hazard statement is H314.

 

According to the two key studies, fast pyrolysis bio-oil is classified as corrosive, subcategory 1C with hazard statement H314 "Causes severe skin burns and eye damage".

 

Due to classification as corrosive, no study for eye irritation was needed.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Reference 1

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

08 March 2005 - 11 March 2005

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions (final report not dated nor signed by the study director)

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

yes

Remarks:

the relative humidity recorded in the animal room was sometimes outside of the target ranges specified in the study plan

Qualifier:

according to guideline

Guideline:

other: Directive 2004/73/EC B.4, 29th April 2004

GLP compliance:

yes (incl. QA statement)

Remarks:

Agence Francaise de Securite Sanitaire des Produits de Sante

Species:

rabbit

Strain:

New Zealand White

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Controls:

no

Duration of treatment / exposure:

The durations of exposure were 3 minutes, 1 hour and 4 hours.

Observation period:

The skin was examined approximately 1 hour, 24, 48 and 72 hours after removal of the dressing.

Number of animals:

1

Irritation parameter:

edema score

Remarks:

4-hour exposure, mean of scores

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.7

Irritation parameter:

edema score

Remarks:

4-hour exposure

Basis:

animal #1

Time point:

other: 48/72 h

Score:

1

Remarks on result:

other: a very slight edema

Irritation parameter:

edema score

Remarks:

4-hour exposure

Basis:

animal #1

Time point:

other: 1/24 h

Score:

0

Irritation parameter:

erythema score

Remarks:

4-hour exposure

Basis:

animal #1

Time point:

other: 1/24/48 h

Score:

2

Remarks on result:

other: a well-defined erythema

Irritation parameter:

erythema score

Remarks:

4-hour exposure

Basis:

animal #1

Time point:

72 h

Score:

3

Remarks on result:

other: a marked ertythema

Irritation parameter:

erythema score

Remarks:

4-hour exposure, mean of scores

Basis:

animal #1

Time point:

24/48/72 h

Score:

2.3

Irritant / corrosive response data:

The test item Biotox-1 is corrosive under these experimental conditions when applied topically to rabbit skin.

3 -minute exposure - Cutaneous examinations and mean values of the scores recorded for the animal (24, 48 and 72 hours)

Rabbit number	Dermal Irritation	Scores				Mean irritation score (1)
		1h	24h	48h	72h
		D1	D2	D3	D4
19	Erythema	0	1	1	0	0.7
	Oedema	0	0	0	0	0.0
	Other	C	C	C	0

(1) mean of scores on days 2, 3 and 4

h = hour

D = day

C = beige coloration of the skin

1-hour exposure - Cutaneous examinations and mean values of the scores recorded for the animal (24, 48 and 72 hours)

Rabbit number	Dermal Irritation	Scores				Mean irritation score (1)
		1h	24h	48h	72h
		D1	D2	D3	D4
19	Erythema	2	2	2	4	2.7
	Oedema	0	2	2	2	2.0
	Other	C	C	C	ZB

(1) mean of scores on days 2, 3 and 4

h = hour

D = day

C = beige coloration of the skin

ZB = brownish area similar to a severe burn of the skin

4-hour exposure - Cutaneous examinations and mean values of the scores recorded for the animal (24, 48 and 72 hours)

Rabbit number	Dermal Irritation	Scores				Mean irritation score (1)	Interpretation (+) (-)
		1h	24h	48h	72h
		D1	D2	D3	D4
19	Erythema	2	2	2	3	2.3	(+)
	Oedema	0	0	1	1	0.7	(-)
	Other	C	C	C	C/S

(1) mean of scores on days 2, 3 and 4

h = hour

D = day

(+) = irritant according to E.E.C. criteria

(-) = non-irritant according to E.E.C. criteria

S = dryness of the skin

C = beige coloration of the skin

Interpretation of results:

corrosive

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item BIOTOX-21 is corrosive when applied topically to rabbits. According to the classification criteria laid down in 67/548/EEC the test item should be classified as corrosive and assigned the symbol C, the indication of danger "Corrosive" and the risk phrase R34 "Causes burns". Under the Regulation EC No 1272/2008 (CLP) the corresponding hazard statement is H314 "Causes severe skin burns and eye damage".

Executive summary:

The potential of the test item BIOTOX-21 to induce skin irritation was evaluated in rabbits according to OECD (No. 404) and EC (2004/73/EC B.4) guidelines and in compliance with GLP. The test item was first applied for periods of 3 minutes, 1 hour and 4 hours to a single male New Zealand White rabbit. As the test item showed corrosive properties on first animal, the study was considered complete and the test item was not evaluated on other animals. Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing. The mean values of the scores for erythema and oedema were calculated. The interpretation of results was carried out according to the classification criteria laid down in 67/548/EEC.

Reference 2

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-05-13 to 2013-05-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 435 (In Vitro Membrane Barrier Test Method for Skin Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und Lebensmittelsicherheit, Germany

Amount / concentration applied:

TEST SYSTEM
- Bio-barrier membrane

TEST MATERIAL
- Amount applied: 500 µL

Observation period:

4 hours

Details on study design:

This assay is composed of three different steps:

1. Qualification
This step ensures that the sample is compatible with the CORROSITEX™ system. 150 µL or 150 mg of the test substance is added to the Qualify test tube. Solids are shaken to dissolve, if necessary. For immiscible liquids and insoluble solids the vial is shaken and let stand for one minute. If the colour or consistency of the CDS changes at the sample/testing fluid interface, the test material is qualified for the assay. If no reaction is observed within five minutes, the sample is not qualified for the CORROSITEX™ Assay.

2. Categorisation
This step establishes the category for cut-off times for the sample. 150 µL or 150 mg of the test substance is added to the tubes labeled Tube A and Tube B. After shaking a colour change in either of the tubes is matched to the corresponding colour charts on the CORROSITEX™ Testing Protocol Poster. Test materials having high acid/alkaline reserves are defined as Category 1 materials, while those with low acid/alkaline reserves are defined as Category 2 materials. If no colour change is observed in either tube, CONFIRM reagent is added to Tube B. After shaking, the resulting colour is matched to the colour chart on the CORROSITEX™ Testing Protocol Poster.
If the test item has strong colouring or shows other characteristics impairing a clear categorization according to the colour chart, the pH value can be measured in the tubes A and B and is used to confirm/determine the category of the test item

3. Classification
This step determines the appropriate Packing Group for the test sample. The CDS vials are warmed to room temperature (17 - 25˚C) before using. Vials 1 - 4 are utilized for sample replicate testing. Controls are set-up in parallel to the test item in order to confirm the validity of the test: positive control, negative control and one colour control. The Bio-barrier membranes are added on top of the vials. 500 µL oder 500 mg of the test item are applied evenly on the top of the BIOBARRIER disc and break-through time is recorded.

Irritation / corrosion parameter:

penetration time (in minutes)

Value:

ca. 49

Qualification Test / Categorization Test

The test substance was compatible with the CORROSITEX™ Assay, as assessed in the qualification step.As the sample was intensely coloured and an indistinct color change was observed in the categorization tubes A and B, an additional pH measurement was performed to confirm the category of the test item. Since the pH of the 10% solution of the test item was < 7 (3.1) and the pH in tube A was > 5 (6.6), the test item was classified into category 2.

Classification Test

	CORROSITEX™ time [min]	Colour change	Consistency change
Replicate 1	56.00	yes	no
Replicate 2	49.00	yes	no
Replicate 3	40.00	yes	no
Replicate 4	50.00	yes	no
Mean +/- SD	48.75 +/- 6.60
Positive Control	1.40	yes	no
Negative Control	83.00	yes	no

The mean time, required to activate the CDS was 48.75 +/- 6.60 min.

 

Interpretation of results:

corrosive

Remarks:

UN GHS Subcategory 1C, Packing Group: 3, EU Risk-Phrase: R 34) Criteria used for interpretation of results: other: OECD 435

Conclusions:

In this study under the given conditions the test substance showed corrosive effects. The test substance is classified as “CORROSIVE”, subcategory 1C according to the UN GHS, as the mean time to activate the CDS was >30-60 min (category 2). The test substance is therefore assigned to Packing Group 3 and to EU Risk-Phrase (R 34). Under the regulation 1272/2008/EC [CLP] the corresponding hazard statement is H314.

Executive summary:

The potential of the test substance to induce skin corrosion was analysed by using the “In Vitro Membrane Barrier Test (CORROSITEX™ Assay)”, comprising a Chemical Detection System covered by a bio-barrier membrane.

The test substance Fast Pyrolysis Bio-oil proved its ability to activate the CDS and was subsequently subjected to the timescale category test. The test substance was applied undiluted atop the bio-barrier membrane and the time required to activate the CDS was measured in four replicates. The mean time was compared to the given thresholds.

 

As the sample was intensely coloured and an indistinct color change was observed in the categorization assay, an additional pH measurement was performed to confirm the category of the test item. According to this pH-measurement the test item has been classified into category 2.

In this study under the given conditions the test substance showed corrosive effects. The test substance is classified as “CORROSIVE”, subcategory 1C according to the UN GHS, as the mean time to activate the CDS was >30-60 min (48.75 ± 6.60 min.) (category 2). The test substance is therefore assigned to Packing Group 3 and to EU Risk-Phrase (R 34). Under the regulation 1272/2008/EC [CLP] the corresponding hazard statement is H314.

 

The controls confirmed the validity of the study. The positive control activated the CDS within three minutes (1.40 min.), the negative control did not activate the CDS before 60 min. (83.00 min.).

This study satisfies the requirement for Test Guideline OECD 435 for in vitro Membrane Barrier Test Method for Skin Corrosion.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vitro / ex vivo

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

the study does not need to be conducted because the substance is classified as skin corrosion, leading to classification as serious eye damage (Category 1)

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Justification for selection of skin irritation / corrosion endpoint:
GLP Guideline study

Justification for selection of eye irritation endpoint:
No testing needed because the substance is classified as a skin corrosive

Effects on skin irritation/corrosion: corrosive

Effects on eye irritation: highly corrosive

Justification for classification or non-classification

In the in vitro skin corrosivity study under the given conditions the test substance showed corrosive effects. The test substance is classified as “CORROSIVE”, subcategory 1C according to the UN GHS, as the mean time to activate the CDS was >30-60 min (category 2). The test substance is therefore assigned to Packing Group 3 and to EU Risk-Phrase (R 34). Under the Regulation EC No 1272/2008 (CLP) the corresponding hazard statement is H314 "Causes severe skin burns and eye damage".

 

In addition, the test item BIOTOX-21 was found corrosive when applied topically to rabbits in a skin irritation / corrosion study. Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing. According to to the classification criteria laid down in 67/548/EEC, the test item BIOTOX-21 should be classified as corrosive and assigned the symbol C, the indication of danger "Corrosive" and the risk phrase R34 "Causes burns". Under the Regulation EC No 1272/2008 (CLP) the corresponding hazard statement is H314

 

According to Guidance on the application of Regulation EC No 1272/1008 (CLP) (ECHA 2009) in general, a classification for corrosivity in considered to implicitly cover the potential to cause respiratory tract irritation and so the additional category 3 classification is considered to be superfluous.