Polyphosphoric acids, ammonium salts

Acute Toxicity: inhalation

Administrative data

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 29 October 2014 to 09 December 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was performed in accordance with an accepted guideline (OECD TG 436) and under the conditions of GLP. No deficiencies are noted and the results are considered to be relaible.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 436 (Acute Inhalation Toxicity: Acute Toxic Class Method)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EU Method B.52 (Acute Inhalation Toxicity - Acute Toxic Class Method, 2014)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Date of Inspection: 12 to 14 March 2014 Date of Signature on Certificate: 12 May 2014

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Polyphosphoric acids, ammonium salts

EC Number:

269-789-9

EC Name:

Polyphosphoric acids, ammonium salts

Cas Number:

68333-79-9

Molecular formula:

[NH4PO3]n

IUPAC Name:

undecaammonium bis(phosphonatooxy)phosphinate dihydrogen phosphate hydrogen (phosphonatooxy)phosphonate hydrogen phosphate

Test material form:

gas under pressure: refrigerated liquefied gas

Details on test material:

- Name of test material (as cited in study report): Polyphosphoric acids, ammonium salts
- Physical state: Green liquid
- Composition of test material, percentage of components: P205 total 37.1 w%, P205 ortho 13.6 w%, N-NH4 11.0 w%
- Lot/batch No.: Synthese 11 - 06/10/2014
- Expiration date of the lot/batch: 14 October 2015
- Storage condition of test material: Approximately 4 °C in the dark over silica gel

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Ltd., Oxon, UK
- Age at study initiation: Eight to twelve weeks
- Weight at study initiation: 200-350 g
- Fasting period before study: None
- Housing: Solid-floor polypropylene cages with stainless steel lids, furnished with softwood flakes (Datesand Ltd., Cheshire , UK) and provided with environmental enrichment items: wooden chew blocks and cardboard "fun tunnels" (DatesandLtd., Cheshire)
- Diet (e.g. ad libitum): Harlan 2014C Rodent Diet, Harlan Laboratories, UK Ltd., Oxon, UK provided ad libitum
- Water (e.g. ad libitum): Mains drinking water provided ad libitum
- Acclimation period: At least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-25
- Humidity (%): 30-70
- Air changes (per hr): At least 15 per hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark/12 hrs light (06:00 to 18:00)

Administration / exposure

Route of administration:

inhalation: aerosol

Type of inhalation exposure:

nose only

Vehicle:

air

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: A glass concentric jet nebuliser (Radleys, Saffron Walden, Essex, UK) was located at the top of exposure chamber. The nebuliser was connected to a plastic syringe attached to an infusion pump, which provided a continuous supply of test item under pressure, and to a metered compressed air supply.
- Exposure chamber volume: Approximately 30 litres (dimensions: 28 cm diameter x 50 cm high)
- Method of holding animals in test chamber: Each rat was individually held in a tapered, polycarbonate resin restraining tube fitted onto a single tier of the exposure chamber and sealed by means of a rubber 'O' ring.
- Source and rate of air: Compressed air was supplied by means of an oil free compressor; the chamber flow rate was maintained at 60 L/min providing 120 air changes per hour
- Method of conditioning air: Passed through a water trap and respiratory quality filters
- System of generating particulates/aerosols: Nebuliser
- Method of particle size determination:
The particle size of the generated atmosphere inside the exposure chamber was determined three times during the exposure period during a Marple Personal Cascade Impactor (Westech IS Ltd, Beds., UK). This device consisted of six impactor stages (8.4, 7.3, 3.6, 1.3, 0.94, and 0.43 µm cut points) with stainless steel collection substrates and a backup glass fibre filter, housed in an aluminium sampler. The sampler was temporarily sealed in a sampling port in the animals' breathing zone and a suitable, known volume of exposure chamber air was drawn through it using a vacuum pump. The collection substrates and backup filter were weighed before and after sampling and the weight of test item, collected at each stage, calculated by difference. The mean amount for each stage was used to determine the cumulative amount below each cut-off point size. In this way, the proportion (%) of aerosol less than 8.4, 7.3, 3.6, 1.3, 0.94 and 0.43 µm was calculated.
- Treatment of exhaust air: The extract from the exposure chamber passed through a 'scrubber' trap and was connected with a high efficiency filter to a metered exhaust system.
- Temperature, humidity, pressure in air chamber: The tempreature and relative humidity inside the exposure chamber was measured by an electronic thermometer/humidity meter (Hanna Instruments Ltd., Beds., UK) located in a vacant port in the animals' breathing zone of the chamber and recorded every thirty minutes throughout the four-hour exposure period.
- Oxgen concentration in air chamber: Oxygen levels were measured by an electronic oxygen analyser (Servomex (UK) Ltd, Crowborough, East Sussex) located in a port in the animals breathing zone during the four-hour exposure period. The test atmosphere was generated to contain at least 19% oxygen.

TEST ATMOSPHERE
- Brief description of analytical method used: During the characterisation phase of the study the test atmosphere was sampled twice and filter samples were then submitted for chemical analysis to determine if the original test item was similar to the composition of the airborne test item. A standard and samples were analysed spectrophotometrically using the following conditions:

Spectrophotometer: Camspec M550
Wavelength: 190-700 mm
Cell path length: 1 cm
Reference medium: water

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

Target concentrated of 5.0 mg/L. Mean achieved atmosphere concentration: 4.85 mg/L - 97% of target.

No. of animals per sex per dose:

3 animals per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Observations for clinical signs were made at hourly intervals during exposure, immediately on removal from the restraining tubes at the end of exposure, one hour after termination of exppsure and subsequently once daily for up to fourteen days.
Individual body weights were recorded on arrival, prior to treatment on the day of exposure and on Days 1,3, 7 and 14 or at death.
- Necropsy of survivors performed: Yes

Results and discussion

Effect levels

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 4.85 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

0/3 males died and 1/3 females died.

Clinical signs:

other: Individual clinical observations are given in Table 1 and 2. Signs of hunched posture and pilo-erection are commonly seen in animals for short periods on removal from the chamber following 4-hour inhalation studies. Wet fur is commonly recorded both duri

Body weight:

Individual body weights, together with body weight changes, are given in Table 3.

All animals exhibited body weight losses on the first day post-exposure. Two males and two female animals exhibited further body weight losses from Days 1 to 3 post-exposure. Body weight gains were noted in all surviving aimals during the remainder of the recovery period.

Gross pathology:

Individual necropsy findings are given in Table 4.

No macroscopic abnormalities were detected amongst animals that survived until the end of the recovery period at necropsy.

The following macroscopic abnormalities were detected at necropsy in the female animal that was humanely killed during the course of the study.

Liver - pale and accentuated lobular pattern
Stomach - gaseous distension
Small intestine - gaseous distension
Large intestine - gaseous distension

Any other information on results incl. tables

Key to clinical observations

Da = distended abdomen

Dh = dehydration

H = hunched posture

P = pilo-erection

Rd = decreased respiratory rate

Rg = gasping respiration

Ri = increased respiratory rate

Rl = labored respiration

Rn = noisy respiration

Wf = wet fur

X* = animal killed in extremis

0 = no abnormalities detected

Table 1: Individual clinical observations (day of exposure)

Mean achieved atmosphere concentration (mg/L)	Animal number and sex	Hours During Exposure			On removal from chamber	One hour post-exposure
		1	2	3
4.85	1 Male	Wf	Wf	Wf	Wf H P Ri	Wf H P Ri
	2 Male	Wf	Wf	Wf	Wf H P Ri	Wf H P Ri
	3 Male	Wf	Wf	Wf	Wf H P Ri	Wf H P Ri
	4 Female	Wf	Wf	Wf	Wf H P Ri	Wf H P Ri
	5 Female	Wf	Wf	Wf	Wf H P Ri Rn Rg	Wf H P Ri Rn Rg
	6 Female	Wf	Wf	Wf	Wf H P Ri	Wf H P Ri

Table 2: Individual clinical observations (recovery period)

Mean achieved atmosphere concentration (mg/L)	Animal number and sex	Days Post Exposure
		1	2	3	4	5	6	7	8-14
4.85	1 Male	H P Ri	H P Ri	H P Ri	H Ri	H Ri	H	0	0
	2 Male	H P Ri	H P Ri	H	H	H	H	0	0
	3 Male	H P Ri	H P Ri	H P Ri	H Ri	H Ri	H	0	0
	4 Female	H P Ri	H P Ri	H Ri	H Ri	H Ri	Ri	0	0
	5 Female	H P Rd Rl Rn Rg	H P Ri Rn	H P Rd Rl Rn Rg Da Dh X*
	6 Female	H P Ri	H P Ri	H Ri	H Ri	H Ri	Ri	0	0

Table 3 Individual Body Weights

Mean Achieved Atmosphere Concentration (mg/L)	Animal Number and Sex	Body Weight (g) on Day:							Increment (g) During Days:
		-20	0	1	3	7	14	At Death	-20-0	0-1	1-3	3-7	7-14
0.54	1 Male	200	296	270	254	262	281		96	-26	-16	8	19
	2 Male	206	298	273	284	300	349		92	-25	11	16	49
	3 Male	204	299	267	254	270	307		95	-32	-13	16	37
	4 Female	208	219	202	201	208	222		11	-17	-1	7	14
	5 Female	206	238	212	187	-	-	187	32	-26	-25	-	-
	6 Female	207	247	217	220	241	244		40	-30	3	21	3

Table 4 Individual Necropsy Findings

Mean Achieved Atmosphere Concentration (mg/L)	Macroscopic Observations	Animal Number and Sex
		1 Male	2 Male	3 Male	4 Female	5* Female	6 Female
4.85	Liver: Pale           Accentuated                                                   lobular pattern					P P
	Stomach: Gaseous distension					P
	Small intestine: Gaseous distension					P
	Large intestine: Gaseous distension					P
		N	N	N	N		N

P = finding present

N = no abnormalities detected

* = Animal humanely killed during study

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

One death occurred in a group of six rats exposed to a mean achieved atmosphere concentration of 4.85 mg/L for four hours. It was therefore considered that the acute inhalation median lethal concentration ( 4 hr LC50) of polyphosphoric acids, ammonium salts (liquid) in the RccHan™: WIST strain rat, was greater than 4.85 mg/L. In accordance with Regulation (EC) No. 1272/2008 (EU CLP) ammonium polyphosphate is not considered to be classified as acutely toxic via the inhalation route.

The test item did not meet the criteria for classification according to Regulation (EC) No. 1272/2008 on the Classification, Labelling and Packaging of Substances and Mixtures.

According to the criteria of classification of GHS (Globally Harmonised Classification System) the substance is classified in Acute Toxicity Inhalation Category 5 due to the mortality and clinical signs observed in one animal.

Executive summary:

A study was performed to assess the acute inhalation toxicity of the test item. The method used was designed to be compatible with that described in the OECD Guideline for Testing of Chemicals (2009) No. 436 "Acute Inhalation Toxicity - Acute Toxic Class Method" and Method B.52 Acute Inhalation Toxicity - Acute Toxic Class Method, 2014, of Commission Regulation (EC) No. 440/2008.

A group of six RccHan™: WIST strain rats (three males and three females) was exposed to an aerosol atmosphere. The animals were exposed for four hours using a nose only exposure system, followed by a fourteen day observation period.

The mean achieved atmospheric concentration was as follows:

Atmosphere Concentration
Mean Achieved (mg/L)	Standard deviation	Nominal (mg/L)
4.85	0.48	42.9

The characteristics of the achieved atmosphere were as follows:

Mean Achieved Atmosphere Concentration (mg/L)	Mass Median Aerodynamic Diameter (µm)	Inhalable Fraction (% < 4µm)	Geometric Standard Deviation
4.85	2.48	72.7	2.22

The mortality data were summarised as follows:

Mean achieved atmosphere concentration (mg/L)	Deaths
	Male	Female	Total
4.85	0/3	1/3	1/6

Clinical observations

Common abnormalities noted during the study included increased respiratory rate, hunched posture, pilo-erection and wet fur. One animal also exhibited decreased respiratory rate, labored respiration, gasped respiration, noisy respiration, dehydration and a distended abdomen. Surviving animals recovered to appear normal on Day 7 post-exposure.

Body weight

All animals exhibited body weight losses on the first day post-exposure. Two males and two female animals exhibited further body weight losses from Days 1 to 3 post-exposure. Body weight gains were noted in all surviving animals during the remainder of the recovery period.

Necropsy

No macroscopic abnormalities were detected amongst animals that survived until the end of the recovery period at necropsy.

The following macroscopic abnormalities were detected at necropsy in the female animal that was humanely killed during the course of the study:

Liver - pale and accentuated lobular pattern;

Stomach - gaseous distension;

Small intestine - gaseous distension;

Large intestine - gaseous distension.

The 4 hr LC₅₀ was considered to be greater than 4.85 mg/L.