Spent liquor from alkaline pulping and bleaching containing spent inorganic process chemicals and dissolved organic substances originating from the cellulosic raw material.

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March-April 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-Study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Spent liquor from alkaline pulping and bleaching containing spent inorganic process chemicals and dissolved organic substances originating from the cellulosic raw material.

EC Number:

931-584-3

Cas Number:

66071-92-9

Molecular formula:

UVCB substance, not available. View remarks field.

IUPAC Name:

Spent liquor from alkaline pulping and bleaching containing spent inorganic process chemicals and dissolved organic substances originating from the cellulosic raw material.

Test material form:

liquid

Details on test material:

- Name of test material (as cited in study report): Black liquor- Substance type: UVCB- Physical state: liquid

Specific details on test material used for the study:

Name: "Black Liquor".Chemical name: Sulfite liquors and Cooking liquors, spent.Molecular formula: UVCB.Batch No.:Not stated.CAS No.: 66071-92-9.EC No.: 266-111-3.Appearance: Black and viscous liquid. The tested sample contained 47.3 % water and 52.7% dry solids.Solubility: In water: The substance is a water solution.pH: ~ over 13.Conditions of storage: Room temperature. Storage in the dark but may be used under light.Stability at conditions of storage: Not available.Expiry date: Not available.

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

Species, strain: Rats, Crl:CD(SD).Supplier: Charles River Deutschland GmbH, D-97633 Sulzfeld.Number and sex: 12 females.Age: Between 8 and 12 weeks at the time of the administration.Health conditions: A health inspection was performed prior to the commencement of treatment to ensure that the animals were in a good state of health. Hygiene: Optimal hygienic conditions.Room number: EI1-11.Room temperature: Ranges from 19.93 to 21.13 °C (continuous control and recording).Relative humidity: Ranges from 43.51 to 58.59 % (continuous control and recording).Air exchange: 12 per hour.Light: Artificial light from 6 a.m. to 6 p.m.Cages: Single caging in Makrolon cages type III (37.5 cm x 21.5 cm bottom area, 18 cm height). Wire mesh lids. Sanitation of cages once a week.Bedding material: Aspen wood chips, Fa. ABEDD Dominik Mayr KEG, A-8580 Köflach, autoclaved. Random samples of the bedding material are analysed for contaminants by the supplier. Changes 1 / week.Environmental enrichment: Nibbling wood bricks (10 cm x 2 cm x 2 cm) and nesting material, both from the same material and source as the bedding material, were offered to the animals once a week.Feed: Ssniff R/M-H maintenance diet for rats and mice (item V1534-300) ad libitum, supplied by Ssniff Spezialdiäten GmbH, 59494 Soest, Germany. Analysis of the feed for ingredients and contaminants is performed randomly by Ssniff.Exception: The feed was withdrawn the evening before the administration of the test substance and was offered again about three hours afterwards.Water: Tap water from an automatic watering system, ad libitum. Random samples of the water are analysed by the "AGES", A-1226 Vienna, to check, if the water fulfils the requirements for drinking water for humans.Identification: Labelling with felt-tipped pen on the tail and on the cage.Acclimatisation: At least 7 days.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

A peroral administration was performed once in the morning by stomach intubation using a metal gavage.Vehicle: deionised water.Dose volume: 10 mL per kg body weight.Justification: The test substance was sufficiently soluble in water and water shall be used preferably, according to the guidelines.The solutions were prepared freshly before administration and were administered within 30 minutes after the preparation.As no prior information on the toxicity of the test substance was available, a starting dose of 300 mg of the test substance per kg body weight was chosen. The further proceeding was in accordance with the guideline/directive:•Step 1: 300 mg (wwt.) per kg body weight.•Step 2: 300 mg (wwt.) per kg body weight.•Step 3: 2000 mg (wwt.) per kg body weight.•Step 4: 2000 mg (wwt.) per kg body weight.

Doses:

300 and 2000 mg/kg body weight (substance wet weight)

No. of animals per sex per dose:

3 per step (6 females per dose)

Control animals:

no

Details on study design:

Observations were performed within the periods 0 - 0.5, 0.5 - 1, 1 - 2, 2 - 4 and 4 - 6 hours after administration (p.a.) of the test substance and then at least once a day for a total of 14 days. Observations included but were not limited to changes in skin, fur, eyes, the occurrence of secretions and excretions, autonomic activity, changes in gait, posture and the presence of convulsions.Body weights were determined•before administration.•7 days p.a.•14 days p.a.Body weight gain was calculated for each week of the study, i.e. between •0 and 7 days p.a.•7 and 14 days p.a.The animals were sacrificed by inhalation of 80 % CO2 + 20 % air 14 days p.a. and subjected to a necropsy including a gross pathological examination.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 5 000 mg/kg bw

Based on:

test mat.

Remarks:

wet weight (52.7% dry solids, 47.3 % water))

Mortality:

All animals survived until the scheduled termination of the study.

Clinical signs:

other: All animals did not show any clinical signs during the entire observation period.

Gross pathology:

No abnormal findings were made in all animals at the necropsy 14 d p.a.

Any other information on results incl. tables

Table:  Body weights and body weight gain

           Individual data, means and standard deviations SD.

Dose	Animal	Body weight (g)			Body weight gain (g)
mg/kg (Step No.)	No.	before administr.	7 days p.a.	14 days p.a.	death	0-7 days p.a.	7-14 days p.a.
300	11	215	260	263	-	45	3
(1)	12	210	250	254	-	40	4
	13	222	261	269	-	39	8
	mean	215.7	257.0	262.0	-	41.3	5.0
	SD	6.0	6.1	7.5	-	3.2	2.6
300	14	213	251	264	-	38	13
(2)	15	219	255	264	-	36	9
	16	219	255	264	-	36	9
	mean	217.0	253.7	264.0	-	36.7	10.3
	SD	3.5	2.3	0.0	-	1.2	2.3
2000	21	215	236	242	-	21	6
(3)	22	222	251	271	-	29	20
	23	212	240	244	-	28	4
	mean	216.3	242.3	252.3	-	26.0	10.0
	SD	5.1	7.8	16.2	-	4.4	8.7
2000	24	204	229	227	-	25	-2
(4)	25	200	220	225	-	20	5
	26	218	242	252	-	24	10
	mean	207.3	230.3	234.7	-	23.0	4.3
	SD	9.5	11.1	15.0	-	2.6	6.0

Table:    Observations in life

Findings	Dose (mg/kg), Step No.	Animal Nos.	Observation time (p.a.) first    last		Maximum grade of severity
no clinical signs	300, 1	11, 12, 13	0 h / 14 d	-
	300, 2	14, 15, 16	0 h / 14 d	-
	2000, 3	21, 22, 23	0 h / 14 d	-
	2000, 4	24, 25, 26	0 h / 14 d	-

Table:    Necropsy findings

             Number of animals examined: 12 females.

SYSTEM Organ, finding	Dose (mg/kg)	Step No.	Animal Nos.
no abnormal findings	300	1	11, 12, 13
	300	2	14, 15, 16
	2000	3	21, 22, 23
	2000	4	24, 25, 26

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

According to Commission Directive 2001/59/EC "Black Liquor" does not require classification for acute oral toxicity.

Executive summary:

The aim of the study was to investigate acute toxic effects of the test substance after a single peroral administration to rats.

Methods and investigations were performed in conformance with the OECD-Guideline 423, 17 December 2001 and theCouncil Regulation (EC) No 440/2008, Method B.1 tris.

"Black Liquor" was administered once orally via gavage as a solution in deionised water to female Crl:CD(SD) rats.

The dosing was performed sequentially to groups of 3 animals per step using a starting dose of 300 mg (wwt.) per kg body weight and 2000 mg (wwt.) per kg body weight as the second dose. The dose volume was 10 mL per kg body weight for all groups.

Investigations

·      Body weights: before administration, 7 and 14 days after the administration (p.a.).

·      Clinical observations: at least once per day.

·      Necropsy: The animals were sacrificed and necropsied 14 days p.a.

Results

Dose (mg/kg)	Step No.	No. of animals			Prominent findings
		exposed	affected	deceased	in life	post mortem
300	1	3	0	0	none	none
300	2	3	0	0	none	none
2000	3	3	0	0	none	none
2000	4	3	0	0	none	none

 

Presence of signs in life	no signs
Full recovery of the survivors	not applicable
Body weights	all but 1/6 of the high dosed animals gained weight in both weeks p.a.
Findings in life and post mortem indicate	no toxic effects present
LD50, oralaccording to OECD	> 2635 mg/kg (substance dry wt.) body weight, (> 5000 mg/kg (substance wet wt.) body weight

Conclusion

No toxic effects of the test substance were noted by signs in life and post mortem at a dose of 2000 mg test substance per kg body weight. No mortality occurred.

As no animals died, the LD_{50, oral}was determined to be > 2000 mg/kg (wwt.) body weight. The tested Balck liquor sample contained 47.3 % water and 52.7% dry solids.

Based on the guidance given in OECD guideline 423 (annex 2c), the LD_{50, oral}can be assessed to be > 5000 mg/kg (wwt.) body weight, that is equal to > 2635 mg/kg (substance dry weight)/ body weight.

According to CLP regulation "Black Liquor" does not require classification for acute oral toxicity.