Diammonium dihydrogenpyrophosphate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

No data available.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 December 2001 - 24 May 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or
(3) a constant pattern in the changing of the potency of the properties across the category

The proposed source chemical (diammonium hydrogenorthophosphate) is highly soluble in water (> 10000 mg/L). In aqueous media soluble inorganic orthophosphates will dissociate to their ionic constituents; in this case ammonium and orthophosphate ions. Diammonium dihydrogenpyrophosphate will dissociate to ammonium cations and pyrophosphate anions. The pyrophosphate anions are unstable in aqueous solutions with the degree of instability varying according to pH. In distilled water they will hydrolyse slowly via abiotic mechanisms to orthophosphate. In natural waters a number of different processes can occur; abiotic hydrolysis, biotic degradation (as a result of the action of phosphatases which cleave pyrophosphates into orthophosphate subunits) and assimilation by organisms in the water. Thus, the target substance (diammonium dihydrogenpyrophosphate) and the source substance (diammonium hydrogenorthophosphate) will be primarily absorbed as the same inorganic ions: ammonium and orthophosphate and are expected to behave in a similar manner under test conditions.
All (bio) transformation products of the source chemical are common to the target chemical and as such the data is considered to be adequate and reliable for use in the assessment of diammonium dihydrogenpyrophosphate for the toxicity hazard assessment.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report attached.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report attached.

4. DATA MATRIX
See read-across justification report attached.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent, England.
- Age at study initiation: 9-10 weeks
- Weight at study initiation: 298 to 386 g for males and 191 to 263 g for females
- Fasting period before study: not applicable
- Housing:Unless paired for mating, the animals were singly housed in RB3 modified cages consisiting of high density polypropylene bodies with lids and floors of stainless steel grid. These cages were suspended in batteries over trays lined with absorbent paper. RB3 modified cages were used
throughout the study with the exception of reproductive subgroup females from Day 17 after mating, where RB3 solid-bottomed cages were used.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 16 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23°C, except on 3 occasions during which the temperature was minimally 16 or 18°C and the target temperature range was
re-established within 24 hours.
- Humidity (%): 40-70%, except on 1 occasion during which the humidity was minimally 16% and the target temperature range was re-established on
the next day.
- Air changes (per hr): approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 12 December 2001 To: 10 February 2002

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The formulation for Group 4 (150 mg/ml) was mixed by adding the required volume of vehicle (water purified by reverse osmosis) to the required
weight of DAP and magnetically stirring for up to 1 hour until a visibly homogenous brown suspension was formed. Formulation for Groups 3 and 2
were prepared by direct dilution of this suspension.

VEHICLE
- Concentration in vehicle: 25 - 75 - 150 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: up to 2 weeks (although all animals were mated and were seperated within 1 week)
- Proof of pregnancy: sperm in vaginal smear or at least three copulation plugs referred to as day 0 of pregnancy
- No unsuccessful pairing occurred.
- Further matings after two unsuccessful attempts: no, not applicable.
- After successful mating each pregnant female was caged (how): singly housed in RB3 solid-bottomed cages.
- Any other deviations from standard protocol: no.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Samples of each formulation prepared for administration during Weeks 1, 4 and 6 of the study were analysed using spectrophotometry for test
material content and found to be satisfactory.

Duration of treatment / exposure:

Animals were divided between two subgroups (toxicity and reproductive subgroups).
Males of both subgroups and females of the toxicity subgroup were treated until termination during week 6 of treatment. Doses were administered to the reproductive subgroup females for two weeks prior to pairing, throughout pairing and gestation until Day 3 of lactation. Animals that were
in parturition at the time of dosing were not dosed that day. Control animals received the vehicle over the same treatment period.
Animals were not dosed on their scheduled day of necropsy.

Frequency of treatment:

Daily

Details on study schedule:

- Age at mating of the reproductive subgroup animals: 11-12 weeks

Remarks:

Doses / Concentrations:
250 mg/kg/day
Basis:
actual ingested

Remarks:

Doses / Concentrations:
750 mg/kg/day
Basis:
actual ingested

Remarks:

Doses / Concentrations:
1500 mg/kg/day
Basis:
actual ingested

No. of animals per sex per dose:

The toxicity subgroup consisted of 5 males and 5 females per group and the reproductive subgroup consisted of 10 females and
5 males per group.

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: As no treatment-related effects were seen on parameters assessed in a preliminary study at 1000 mg/kg/day, the high
dosage of 1500 mg/kg/day was selected by the Sponsor to attempt to elicit an overt toxic response and establish a reference level for toxicity.
The concept of a 'Limit Dose' is accepted by the OECD and is generally set in the region of 1000 mg/kg/day; therefore, it was not considered
necessary to investigate a dosage above 1500 mg/kg/day.

- Rationale for animal assignment (if not random): The animal numbers were assigned to groups in non-sequential manner due to the need for the
functional observation battery and detailed clinical signs to be performed without knowledge of the treatment groups to which animals were
assigned.

- Section schedule rationale (if not random): The sequence in which the animals of the toxicity subgroup were killed after completion of the
observation period was selected to allow satisfactory inter-group comparison. Reproductive subgroup males were killed after the toxicity
subgroup animals and reproductive subgroup females were killed on Day 4 of lactation.

Positive control:

No.

Parental animals: Observations and examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least twice daily
- Cage side observations: evidence of reaction to treatment or ill-health

DETAILED PHYSICAL OBSERVATIONS: Yes
- Time schedule: weekly
- A more detailed physical examination was performed on each animal by an observer. After removal from the home cage, animals were
assessed for physical condition and behaviour during handling and after being placed in a standard arena. During these examinations particular
attention was paid to possible signs of neurotoxicity such as convulsions, tremor and abnormalities of gait or behaviour.

ADDITIONAL DETAILED OBSERVATIONS: Yes
- Time schedule: during Week 1 daily according to the following frequency:
1. Pre-dose observation, 2.As each animal was returned to its home cage, 3.At the end of dosing each group, 4.Between 1 and 2 hours after
completion of dosing all groups and 5.As late as possible in the working day.
From Week 2 to termination daily observations were limited to 1 and 4.

BODY WEIGHT: Yes
- Time schedule for examinations: Each male and toxicity subgroup female: on the day that treatment commenced, weekly thereafter, and at necropsy. Reproductive subgroup females: on the first day of treatment, weekly until pairing and on Days 0, 7, 14, 17 and 20 after mating, and Days 1 and 4 of
lactation, and at necropsy.

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Time schedule for observations: All males and toxicity subgroup females: weekly throughout the study with the exception of the males whilst in
pairing with the reproductive subgroup females. Reproductive subgroup females: weekly before pairing then on Days 0,7,14, 17 and 20 after mating
and Days 1 and 4 of lactation. No food consumption was recorded for toxicity subgroup males or reproductive subgroup animals during pairing.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OTHER:
Mating performance and fertility:
All 10 males in each group (toxicity and reproductive subgroups) were mated with the 10 reproductive subgroup females after all animals had
received 2 weeks of treatment. Pairing was on a one-to-one basis within treatment groups for up to 2 weeks, although all animals mated and were
separated within 1 week. Each morning, following pairing, the trays beneath the cages were checked for ejected copulation plugs and a wet vaginal
smear was prepared from each female and examined for the presence of spermatozoa. The day on which a sperm positive vaginal smear or at least
three copulation plugs were found was designated Day 0 of gestation. Once mating had been confirmed, males and females were separated and the
males were returned to their normal group housing.

Parturition observations and gestation length:
From Day 20 after mating reproductive subgroup animals were checked 3 times daily for evidence of parturition. The females were permitted to
deliver their young naturally and rear their own offspring until Day 4 of lactation. Numbers of live and dead offspring were recorded during the
parturition process.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight
gain data: Yes

HAEMATOLOGY: Yes
- Time schedule for collection of blood: During week 5, following FOB examinations
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes, overnight
- How many animals: toxicity subgroup animals (5 males + 5 females per dose group)
- Parameters checked: Haematocrit (Hct), Haemoglobin (Hb), Red blood cell count (RBC), Mean cell haemoglobin (MCH), Mean cell haemoglobin
concentration (MCHC), Mean cell volume (MCV), Total white cell count (WBC), Differential WBC count, Neutrophils (N), Lymphocytes (L), Eosinophils (E), Basophils (B), Monocytes (M), Large unstained cells (LUC), Platelet count (Pit), Reticulocyte count (Retic), Blood film for abnormal morphology and
unusual cell types, including normoblasts (The most common morphological changes, anisocytosis (aniscyto), micro/macrocytosis (Microcyto/
Macrocyto), hypo/hyperchromasia (Hypochrom/Hyperchrom), Platelet clumping (PIt. Clump)), Prothrombin time (PT) and Activated partial
thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: During week 5, following FOB examinations
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes, overnight
- How many animals: toxicity subgroup animals (5 males + 5 females per dose group)
- Parameters checked: Alkaline phosphatase (ALP), Alanine aminotransferase (ALT), Aspartate aminotransferase (AST),
Gamma-glutamyl transpeptidase (gGT), Total Bilirubin (Bili), Urea, Creatinine (Creat), Glucose (Gluc), Total cholesterol (Chol), Sodium (Na),
Potassium (K), Chloride (Cl), Calcium (Ca), Inorganic Phosphorus (Phos), Magnesium (Mg), Total protein (Total Prot), Albumin (Alb),
Albumin/globulin ratio (A/G Ratio)

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: After 4 weeks of treatment
- Dose groups that were examined: Toxicity subgroup animals (5 males + 5 females per dose group)
- Battery of functions tested: Approach response, Touch response, Startle reflex (auditory), Tail pinch response, Grip strength: forelimb and
hindlimb and motor activity.

Oestrous cyclicity (parental animals):

No

Sperm parameters (parental animals):

testis weight, epididymis weight, histopathological examinations (see 'Postmorten examinations (Parental animals))

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: no

PARAMETERS EXAMINED
The following parameters were examined in offspring:
number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, weight gain, physical or behavioural abnormalities.

GROSS EXAMINATION OF DEAD PUPS:
yes, for external and internal abnormalities and assessment of the stomach for milk content.

Postmortem examinations (parental animals):

SACRIFICE
- Toxicity subgroup animals: animals were killed during Week 6 of treatment by CO2 inhalation.
- Reproductive subgroup animals: Males were killed after the toxicity subgroups and females were killed on Day 4 of lactation by CO2 inhalation.

GROSS NECROPSY / ORGAN WEIGHTS
- Gross necropsy consisted of detailed examination of the external features and orifices, the neck and associated tissues and the cranial, thoracic,
abdominal and pelvic cavities and their viscera.
- Toxicity subgroup animals:
The requisite organs were weighed after being dissected free of adjacent fat and other contiguous tissue. External and cut surfaces of the organs and tissues were examined as appropriate, abnormalities and interactions were noted and the required tissue samples and any abnormalities
preserved in appropriate fixative.
The following tissues were retained: Adrenals (left and right)*, Aorta - thoracic, Brain*, Caecum, Colon, Duodenum, Epididymides (left and right)*,
Eyes, Heart*, Ileum, Jejunum, Kidneys (left and right)*, Liver*, Lungs (including bronchi), Lymph nodes - mandibular, Lymph nodes - mesenteric,
Lymph nodes - regional to masses, Mammary area, Oesophagus, Optic nerves, Ovaries (left and right)*, Pancreas, Pituitary*, Prostate *, Rectum,
Salivary glands, Sciatic nerves, Seminal vesicles*, Skin, Spinal cord, Spleen*, Sternum, Stomach, Testes (left and right)*, Thymus*, Thyroids with
parathyroids**, Trachea, Urinary bladder, Uterus with cervix*, Vagina
* Organ weight recorded / * * Organ weight recorded after fixation.

- Reproductive subgroup animals:
Males were killed after the toxicity subgroup animals and the testes, epididymides, seminal vesicles with coagulating gland and prostate were retained for each animal. Females were killed on Day 4 of lactation, the number of implantation sites was recorded and the ovaries, uterus with oviducts and
cervix, vagina, pituitary and mammary tissue retained in appropriate fixative. Abnormal tissues were also retained in appropriate fixative.


HISTOPATHOLOGY
- Toxicity subgroup animals:
Organs (including abnormalities) preserved at macroscopic necropsy were processed for animals from the Control and high dosage groups
(Groups 1 and 4). Following treatment related findings in Group 4, the stomachs and kidneys of intermediate and low dosage animals were also
processed and examined. Tissue samples were dehydrated, embedded in paraffin wax and sectioned at approximately four to five micron thickness. Staining was with haematoxylin and eosin, except testes, which were stained with periodic acid/schiff (PAS).
The following tissues were scheduled for histopathology: Abnormalities, Adrenals (cortex and medulla), Aorta (thoracic), Brain (cerebrum,
cerebellum, midbrain), Caecum, Colon, Duodenum, Epididymides (longitudinal section to include the caput and cauda epididymides), Eyes
(longitudinal section of each (both examined)), Heart (including auricular and ventricular regions), Ileum, Jejunum, Kidneys (including cortex,
medulla and papilla regions), Liver (section from all main lobes), Lungs (section from two major lobes, to include bronchi), Lymph nodes,
Mammary area, Oesophagus, Ovaries, Pancreas, Pituitary, Prostate, Rectum, Salivary glands, Sciatic nerves (only one examined), Seminal vesicles,
Skin, Spinal cord (transverse and longitudinal sections at the cervical, thoracic and lumbar levels), Spleen, Sternum (with bone marrow),
Stomach (keratinised, glandular and antrum), Tissue (region to be examined (where appropriate)), Testes, Thymus, Thyroid (included parathyroid
in section, where possible), Trachea, Urinary bladder, Uterus (uterus section separate from cervix section (both examined)), Vagina.
For bilateral organs sections of both the left and right organs were examined. A separate section was prepared from each of the remaining tissues.
In addition to the general histopathological examination of all testicular elements, including tubular and interstitial compartments,
spermatogenesis was assessed taking into consideration the stages of the spermatogenic cycle. Where no disturbances were detected, the testes
were recorded as normal.

- Reproductive subgroup animals:
Histology for reproductive subgroup animals was restricted to the retained reproductive organs and abnormalities observed at macroscopic
necropsy.

Postmortem examinations (offspring):

SACRIFICE
Offspring of were killed by intraperitoneal injection of sodium pentobarbitone on Day 4 of age and subjected to a macroscopic necropsy.
Any offspring dying before scheduled termination were subjected to necropsy and assessment of the stomach for milk.

GROSS NECROPSY
Gross necropsy consisted of detailed examination of the external features and orifices, the neck and associated tissues and the cranial, thoracic,
abdominal and pelvic cavities and their viscera.
Abnormal tissues were also retained in appropriate fixative.

HISTOPATHOLOGY / ORGAN WEIGTHS
Abnormalities observed at macroscopic necropsy.

Statistics:

For some parameters, the similarity of the data was such that analyses were not considered to be necessary.
For categorical data, the proportion of animals was analysed using Fisher's Exact test.
For continuous data, Bartlett's test was first applied to test the homogeneity of variance between the groups. Using tests dependent on the outcome
of Bartlett's test, treated groups were then compared with the Control group, incorporating adjustment for multiple comparisons where necessary.
For bodyweight gains and organ weights, whenever Bartlett's test was found to be statistically significant, a Behrens-Fisher test was used to perform
pairwise comparisons, otherwise a Dunnett's test was used.
The following sequence of statistical tests was used for grip strength and motor activity, bodyweight change during gestation and lactation,
bodyweight and bodyweight change for offspring, food consumption and clinical pathology data:
- If 75% of the data were the same value, then a frequency analysis was applied. Treatment groups were compared using a Mantel test for a trend in
proportions and also pairwise Fisher's Exact tests for each dose group against the control.
- If Bartlett's test for variance homogeneity was not significant at the 1 % level, then parametric analysis was applied. If the F 1 test for monotonicity of dose-response was not significant at the 1 % level, Williams' test for a monotonic trend was applied. If the F 1 test was significant, Dunnett's test was
performed instead.
- If Bartlett's test was significant at the 1 % level, then logarithmic and square-root transformations were tried. If Bartlett's test was still significant, the non-parametric tests were applied. If the HI test for monotonicity of dose-response was not significant at the 1 % level, Shirley's test for a monotonic
trend was applied. If the HI test was significant, Steel's test was performed instead.

Significant differences were expressed at the 5% (p<0.05) or 1% (p<0.01) level.

Reproductive indices:

Percentage mating = 100 * (animals with evidence of mating) / (animals paired)
Conception rate = 100 * (animals achieving a pregnancy) / (animals with evidence of mating)
Fertility index = 100 * (animals achieving a pregnancy) / (animals paired)
Gestation index = 100 * (number of live litters born) / (number pregnant)

Offspring viability indices:

Post-implantation survival index = 100 * (total number of offspring born) / (total number of implantation sites)
Live birth index = 100 * (total number of offspring on Day 1 of littering) / (total number of offspring born)
Viability index = 100 * (total number of offspring on Day 4 of littering) / (total number of offspring on Day 1 of littering)

Clinical signs:

effects observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Other effects:

no effects observed

Reproductive function: oestrous cycle:

not examined

Reproductive function: sperm measures:

no effects observed

Reproductive performance:

no effects observed

CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
One toxicity subgroup female receiving 1500 mg/kg/day was found dead with findings that were consistent with a dosing error. This death was
therefore considered to be unrelated to treatment. A dosage-dependant increase in transient post-dosing salivation was apparent, and this was
considered more likely to be due to the palatability of the test formulations than to an effect of the test material. A dosage-dependant increase in
the number of animals with reddening of the extremities was also apparent mainly during the early stages of treatment.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
The body weight gain of males at 1500 mg/kg/day appeared to be suppressed when compared with Control, such that gain between Weeks 0-5 for this group was 78% of Control. Body weight gain appeared to be unaffected for toxicity subgroup females or reproductive subgroup females
before pairing. Bodyweight gain for reproductive subgroup females receiving 1500 mg/kg/day was reduced during the first week of gestation,
after which values returned to Control-comparable levels through to termination at Day 4 of lactation. Food consumption of males at
1500 mg/kg/day appeared to be marginally lower when compared with Control. There were no effects of treatment on the amount of food
consumed by females of both toxicity and reproductive subgroups.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
No effects.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
No effects

ORGAN WEIGHTS (PARENTAL ANIMALS)
Bodyweight relative kidney and liver weights for toxicity subgroup females at 1500 mg/kg/day were increased, but in the absence of a histological correlate for these findings, this was of uncertain toxicological significance.

GROSS PATHOLOGY (PARENTAL ANIMALS)
A number of toxicity subgroup males and females at 750 and 1500 mg/kg/day had thickened stomachs and associated findings. A number of
toxicity subgroup males and females at 750 and 1500 mg/kg/day exhibited horizontal bands on the incisors.

HISTOPATHOLOGY (PARENTAL ANIMALS)
Stomachs of toxicity subgroup animals showed minimal or slight submucosal inflammation at all doses (0/5, 3/5, 4/5 and 2/5 males and 0/5, 2/5, 4/5 and 4/5 females at 0, 250, 750 and 1500 mg/kg bw respectively). It is thought that these findings may have been associated with an irritant
effect of the test formulations rather than systemic toxicity. Histological processing of the teeth that showed horizontal banding failed to detect
any involvement of areas examined suggesting that the banding was probably restricted to the enamel of the teeth.

OTHER FINDINGS (TOXICITY SUBGROUP ANIMALS)

HAEMATOLOGY:
There was a reduction in activated partial thromboplastin time for toxicity subgroup males at 750 and 1500 mg/kg/day (74 and 76% of control,
resp.), although this was not dosage related. No treatment related changes in these parameters were observed for toxicity subgroup females.

CLINICAL CHEMISTRY:
The following changes were recorded among toxcicity subgroup males for which an effect of treatment could not be discounted: a non dosage-
dependant elevation of alkaline phosphatase levels at 750 and 1500 mg/kg/day (132 and 131% of control, resp.); reduced glucose and
phosphorous levels at 1500 mg/kg/day (79 and 82% of control, resp.); a small but dosage-dependant reduction in total protein at 750 and 1500 mg/kg/day (93 and 91% of control, resp.) with a slightly elevated albumin/globulin ratio at the top dosage (117% of control). Changes in toxicity
subgroup females were limited to a decrease in phosphorous levels at 1500 mg/kg/day (81% of control) although a non-significant increase in
alkaline phosphatase levels at 1500 mg/kg/day (122% of control) in particular suggested a similar change to that recorded in males.

NEUROBEHAVIOUR:
No effects.

Dose descriptor:

NOAEL

Effect level:

>= 1 500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects on reproduction/developmental toxicity (highest dose tested)

Remarks on result:

other: Generation: P and F (migrated information)

Clinical signs:

no effects observed

Mortality / viability:

no mortality observed

Body weight and weight changes:

no effects observed

Sexual maturation:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

no effects observed

Histopathological findings:

no effects observed

VIABILITY (OFFSPRING)
No effects

CLINICAL SIGNS (OFFSPRING)
No effects

BODY WEIGHT (OFFSPRING)
No effects.

GROSS PATHOLOGY (OFFSPRING)
No effects.

HISTOPATHOLOGY (OFFSPRING)
No effects.

OTHER FINDINGS (OFFSPRING)
The percentage of males in the litters did not indicate any preferential mortality amongst either sex.

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

>= 1 500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effects on reproduction/developmental toxicity (highest dose tested)

Remarks on result:

other: Generation: P and F (migrated information)

Reproductive effects observed:

not specified

Conclusions:

NOAEL: 1500 mg/kg/day for reproduction/developmental toxicity.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 500 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

One screeing study is available for an analogous substance. This study is considered to be a Klimisch score 2 (conducted to an appropriate method and under GLP). No further testing is considered necessary.

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

 

In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied. 

The similarities may be based on:

(1)  a common functional group

(2)  the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or

(3)  a constant pattern in the changing of the potency of the properties across the category

 

 Read-across from diammonium hydrogenorthophosphate to diammonium dihydrogenpyrophosphate: 

 

 Diammonium pyrophosphate is a soluble inorganic salt consisting of pyrophosphate anions and ammonium cations. Similarities between the source chemical (diammonium hydrogenorthophosphate) and the target chemical (diammonium dihydrogenpyrophosphate) are based on the following:  

 

 (1)  Both substances contain the bio transformation product; ammonium. 

 

 (2)  Both substances contain the bio transformation product: orthophosphate. In vivo, it is considered that soluble pyrophosphates will be metabolised to the orthophosphate form. Predominantly this occurs as a result of intestinal alkaline phosphatase action.  

 

 The target substance (diammonium dihydrogenpyrophosphate) and the source substance (diammonium hydrogenorthophosphate) will be primarily absorbed as the same inorganic ions: ammonium and orthophosphate. NH4+ ions are immediately transformed into urea by the liver and do not exist in the blood in relevant amounts unless in case of liver failure. 

 The effects of the target compound (diammonium dihydrogenpyrophosphate) are expected to be equal to or worse than the effects of the source substance (diammonium hydrogenorthophosphate) for the property under consideration. 

 Short description of key information:    

In an OECD 422 screening study the NOAEL for both the parental and the offspring generations was > 1500 mg/kg bw/day.  

Justification for selection of Effect on fertility via oral route:
Only one relevant study available.

Effects on developmental toxicity

Description of key information

In an OECD 422 screening study the NOAEL for both the parental and the offspring generations was > 1500 mg/kg bw/day.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

12 December 2001 - 24 May 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:
(1) a common functional group
(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or
(3) a constant pattern in the changing of the potency of the properties across the category

The proposed source chemical (diammonium hydrogenorthophosphate) is highly soluble in water (> 10000 mg/L). In aqueous media soluble inorganic orthophosphates will dissociate to their ionic constituents; in this case ammonium and orthophosphate ions. Diammonium dihydrogenpyrophosphate will dissociate to ammonium cations and pyrophosphate anions. The pyrophosphate anions are unstable in aqueous solutions with the degree of instability varying according to pH. In distilled water they will hydrolyse slowly via abiotic mechanisms to orthophosphate. In natural waters a number of different processes can occur; abiotic hydrolysis, biotic degradation (as a result of the action of phosphatases which cleave pyrophosphates into orthophosphate subunits) and assimilation by organisms in the water. Thus, the target substance (diammonium dihydrogenpyrophosphate) and the source substance (diammonium hydrogenorthophosphate) will be primarily absorbed as the same inorganic ions: ammonium and orthophosphate and are expected to behave in a similar manner under test conditions.
All (bio) transformation products of the source chemical are common to the target chemical and as such the data is considered to be adequate and reliable for use in the assessment of diammonium dihydrogenpyrophosphate for the toxicity hazard assessment.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See read-across justification report attached.

3. ANALOGUE APPROACH JUSTIFICATION
See read-across justification report attached.

4. DATA MATRIX
See read-across justification report attached.

Reason / purpose for cross-reference:

reference to same study

Qualifier:

according to guideline

Guideline:

other: OECD 422

Deviations:

no

GLP compliance:

yes

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Limited, Margate, Kent, England.
- Age at study initiation: 9-10 weeks
- Weight at study initiation: 298 to 386 g for males and 191 to 263 g for females
- Fasting period before study: not applicable
- Housing:Unless paired for mating, the animals were singly housed in RB3 modified cages consisiting of high density polypropylene bodies with lids and floors of stainless steel grid. These cages were suspended in batteries over trays lined with absorbent paper. RB3 modified cages were used
throughout the study with the exception of reproductive subgroup females from Day 17 after mating, where RB3 solid-bottomed cages were used.
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 16 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23°C, except on 3 occasions during which the temperature was minimally 16 or 18°C and the target temperature range was
re-established within 24 hours.
- Humidity (%): 40-70%, except on 1 occasion during which the humidity was minimally 16% and the target temperature range was re-established on
the next day.
- Air changes (per hr): approximately 15 air changes per hour
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 12 December 2001 To: 10 February 2002

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
The formulation for Group 4 (150 mg/ml) was mixed by adding the required volume of vehicle (water purified by reverse osmosis) to the required
weight of DAP and magnetically stirring for up to 1 hour until a visibly homogenous brown suspension was formed. Formulation for Groups 3 and 2
were prepared by direct dilution of this suspension.

VEHICLE
- Concentration in vehicle: 25 - 75 - 150 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Samples of each formulation prepared for administration during Weeks 1, 4 and 6 of the study were analysed using spectrophotometry for test
material content and found to be satisfactory.

Details on mating procedure:

- M/F ratio per cage: 1/1
- Length of cohabitation: up to 2 weeks (although all animals were mated and were seperated within 1 week)
- Proof of pregnancy: sperm in vaginal smear or at least three copulation plugs referred to as day 0 of pregnancy
- No unsuccessful pairing occurred.
- Further matings after two unsuccessful attempts: no, not applicable.
- After successful mating each pregnant female was caged (how): singly housed in RB3 solid-bottomed cages.
- Any other deviations from standard protocol: no.

Duration of treatment / exposure:

Animals were divided between two subgroups (toxicity and reproductive subgroups).
Males of both subgroups and females of the toxicity subgroup were treated until termination during week 6 of treatment. Doses were administered to the reproductive subgroup females for two weeks prior to pairing, throughout pairing and gestation until Day 3 of lactation. Animals that were
in parturition at the time of dosing were not dosed that day. Control animals received the vehicle over the same treatment period.
Animals were not dosed on their scheduled day of necropsy.

Frequency of treatment:

Daily

No. of animals per sex per dose:

The toxicity subgroup consisted of 5 males and 5 females per group and the reproductive subgroup consisted of 10 females and
5 males per group.

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: As no treatment-related effects were seen on parameters assessed in a preliminary study at 1000 mg/kg/day, the high
dosage of 1500 mg/kg/day was selected by the Sponsor to attempt to elicit an overt toxic response and establish a reference level for toxicity.
The concept of a 'Limit Dose' is accepted by the OECD and is generally set in the region of 1000 mg/kg/day; therefore, it was not considered
necessary to investigate a dosage above 1500 mg/kg/day.

- Rationale for animal assignment (if not random): The animal numbers were assigned to groups in non-sequential manner due to the need for the
functional observation battery and detailed clinical signs to be performed without knowledge of the treatment groups to which animals were
assigned.

- Section schedule rationale (if not random): The sequence in which the animals of the toxicity subgroup were killed after completion of the
observation period was selected to allow satisfactory inter-group comparison. Reproductive subgroup males were killed after the toxicity
subgroup animals and reproductive subgroup females were killed on Day 4 of lactation.

Maternal examinations:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: At least twice daily
- Cage side observations: evidence of reaction to treatment or ill-health

DETAILED PHYSICAL OBSERVATIONS: Yes
- Time schedule: weekly
- A more detailed physical examination was performed on each animal by an observer. After removal from the home cage, animals were
assessed for physical condition and behaviour during handling and after being placed in a standard arena. During these examinations particular
attention was paid to possible signs of neurotoxicity such as convulsions, tremor and abnormalities of gait or behaviour.

ADDITIONAL DETAILED OBSERVATIONS: Yes
- Time schedule: during Week 1 daily according to the following frequency:
1. Pre-dose observation, 2.As each animal was returned to its home cage, 3.At the end of dosing each group, 4.Between 1 and 2 hours after
completion of dosing all groups and 5.As late as possible in the working day.
From Week 2 to termination daily observations were limited to 1 and 4.

BODY WEIGHT: Yes
- Time schedule for examinations: Each male and toxicity subgroup female: on the day that treatment commenced, weekly thereafter, and at necropsy. Reproductive subgroup females: on the first day of treatment, weekly until pairing and on Days 0, 7, 14, 17 and 20 after mating, and Days 1 and 4 of
lactation, and at necropsy.

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Time schedule for observations: All males and toxicity subgroup females: weekly throughout the study with the exception of the males whilst in
pairing with the reproductive subgroup females. Reproductive subgroup females: weekly before pairing then on Days 0,7,14, 17 and 20 after mating
and Days 1 and 4 of lactation. No food consumption was recorded for toxicity subgroup males or reproductive subgroup animals during pairing.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OTHER:
Mating performance and fertility:
All 10 males in each group (toxicity and reproductive subgroups) were mated with the 10 reproductive subgroup females after all animals had
received 2 weeks of treatment. Pairing was on a one-to-one basis within treatment groups for up to 2 weeks, although all animals mated and were
separated within 1 week. Each morning, following pairing, the trays beneath the cages were checked for ejected copulation plugs and a wet vaginal
smear was prepared from each female and examined for the presence of spermatozoa. The day on which a sperm positive vaginal smear or at least
three copulation plugs were found was designated Day 0 of gestation. Once mating had been confirmed, males and females were separated and the
males were returned to their normal group housing.

Parturition observations and gestation length:
From Day 20 after mating reproductive subgroup animals were checked 3 times daily for evidence of parturition. The females were permitted to
deliver their young naturally and rear their own offspring until Day 4 of lactation. Numbers of live and dead offspring were recorded during the
parturition process.

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight
gain data: Yes

HAEMATOLOGY: Yes
- Time schedule for collection of blood: During week 5, following FOB examinations
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes, overnight
- How many animals: toxicity subgroup animals (5 males + 5 females per dose group)
- Parameters checked: Haematocrit (Hct), Haemoglobin (Hb), Red blood cell count (RBC), Mean cell haemoglobin (MCH), Mean cell haemoglobin
concentration (MCHC), Mean cell volume (MCV), Total white cell count (WBC), Differential WBC count, Neutrophils (N), Lymphocytes (L), Eosinophils (E), Basophils (B), Monocytes (M), Large unstained cells (LUC), Platelet count (Pit), Reticulocyte count (Retic), Blood film for abnormal morphology and
unusual cell types, including normoblasts (The most common morphological changes, anisocytosis (aniscyto), micro/macrocytosis (Microcyto/
Macrocyto), hypo/hyperchromasia (Hypochrom/Hyperchrom), Platelet clumping (PIt. Clump)), Prothrombin time (PT) and Activated partial
thromboplastin time (APTT)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: During week 5, following FOB examinations
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes, overnight
- How many animals: toxicity subgroup animals (5 males + 5 females per dose group)
- Parameters checked: Alkaline phosphatase (ALP), Alanine aminotransferase (ALT), Aspartate aminotransferase (AST),
Gamma-glutamyl transpeptidase (gGT), Total Bilirubin (Bili), Urea, Creatinine (Creat), Glucose (Gluc), Total cholesterol (Chol), Sodium (Na),
Potassium (K), Chloride (Cl), Calcium (Ca), Inorganic Phosphorus (Phos), Magnesium (Mg), Total protein (Total Prot), Albumin (Alb),
Albumin/globulin ratio (A/G Ratio)

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: After 4 weeks of treatment
- Dose groups that were examined: Toxicity subgroup animals (5 males + 5 females per dose group)
- Battery of functions tested: Approach response, Touch response, Startle reflex (auditory), Tail pinch response, Grip strength: forelimb and
hindlimb and motor activity.

Ovaries and uterine content:

See section 7.8.1.

Fetal examinations:

See section 7.8.1.

Statistics:

For some parameters, the similarity of the data was such that analyses were not considered to be necessary.
For categorical data, the proportion of animals was analysed using Fisher's Exact test.
For continuous data, Bartlett's test was first applied to test the homogeneity of variance between the groups. Using tests dependent on the outcome
of Bartlett's test, treated groups were then compared with the Control group, incorporating adjustment for multiple comparisons where necessary.
For bodyweight gains and organ weights, whenever Bartlett's test was found to be statistically significant, a Behrens-Fisher test was used to perform
pairwise comparisons, otherwise a Dunnett's test was used.
The following sequence of statistical tests was used for grip strength and motor activity, bodyweight change during gestation and lactation,
bodyweight and bodyweight change for offspring, food consumption and clinical pathology data:
- If 75% of the data were the same value, then a frequency analysis was applied. Treatment groups were compared using a Mantel test for a trend in
proportions and also pairwise Fisher's Exact tests for each dose group against the control.
- If Bartlett's test for variance homogeneity was not significant at the 1 % level, then parametric analysis was applied. If the F 1 test for monotonicity of dose-response was not significant at the 1 % level, Williams' test for a monotonic trend was applied. If the F 1 test was significant, Dunnett's test was
performed instead.
- If Bartlett's test was significant at the 1 % level, then logarithmic and square-root transformations were tried. If Bartlett's test was still significant, the non-parametric tests were applied. If the HI test for monotonicity of dose-response was not significant at the 1 % level, Shirley's test for a monotonic
trend was applied. If the HI test was significant, Steel's test was performed instead.

Significant differences were expressed at the 5% (p<0.05) or 1% (p<0.01) level.

Indices:

See section 7.8.1.

Historical control data:

See section 7.8.1.

Details on maternal toxic effects:

Maternal toxic effects:yes

Details on maternal toxic effects:
CLINICAL SIGNS AND MORTALITY (PARENTAL ANIMALS)
One toxicity subgroup female receiving 1500 mg/kg/day was found dead with findings that were consistent with a dosing error. This death was
therefore considered to be unrelated to treatment. A dosage-dependant increase in transient post-dosing salivation was apparent, and this was
considered more likely to be due to the palatability of the test formulations than to an effect of the test material. A dosage-dependant increase in
the number of animals with reddening of the extremities was also apparent mainly during the early stages of treatment.

BODY WEIGHT AND FOOD CONSUMPTION (PARENTAL ANIMALS)
The body weight gain of males at 1500 mg/kg/day appeared to be suppressed when compared with Control, such that gain between Weeks 0-5 for this group was 78% of Control. Body weight gain appeared to be unaffected for toxicity subgroup females or reproductive subgroup females
before pairing. Bodyweight gain for reproductive subgroup females receiving 1500 mg/kg/day was reduced during the first week of gestation,
after which values returned to Control-comparable levels through to termination at Day 4 of lactation. Food consumption of males at
1500 mg/kg/day appeared to be marginally lower when compared with Control. There were no effects of treatment on the amount of food
consumed by females of both toxicity and reproductive subgroups.

REPRODUCTIVE FUNCTION: SPERM MEASURES (PARENTAL ANIMALS)
No effects.

REPRODUCTIVE PERFORMANCE (PARENTAL ANIMALS)
No effects

ORGAN WEIGHTS (PARENTAL ANIMALS)
Bodyweight relative kidney and liver weights for toxicity subgroup females at 1500 mg/kg/day were increased, but in the absence of a histological correlate for these findings, this was of uncertain toxicological significance.

GROSS PATHOLOGY (PARENTAL ANIMALS)
A number of toxicity subgroup males and females at 750 and 1500 mg/kg/day had thickened stomachs and associated findings. A number of
toxicity subgroup males and females at 750 and 1500 mg/kg/day exhibited horizontal bands on the incisors.

HISTOPATHOLOGY (PARENTAL ANIMALS)
Stomachs of toxicity subgroup animals showed minimal or slight submucosal inflammation at all doses (0/5, 3/5, 4/5 and 2/5 males and 0/5, 2/5, 4/5 and 4/5 females at 0, 250, 750 and 1500 mg/kg bw respectively). It is thought that these findings may have been associated with an irritant
effect of the test formulations rather than systemic toxicity. Histological processing of the teeth that showed horizontal banding failed to detect
any involvement of areas examined suggesting that the banding was probably restricted to the enamel of the teeth.

OTHER FINDINGS (TOXICITY SUBGROUP ANIMALS)

HAEMATOLOGY:
There was a reduction in activated partial thromboplastin time for toxicity subgroup males at 750 and 1500 mg/kg/day (74 and 76% of control,
resp.), although this was not dosage related. No treatment related changes in these parameters were observed for toxicity subgroup females.

CLINICAL CHEMISTRY:
The following changes were recorded among toxcicity subgroup males for which an effect of treatment could not be discounted: a non dosage-
dependant elevation of alkaline phosphatase levels at 750 and 1500 mg/kg/day (132 and 131% of control, resp.); reduced glucose and
phosphorous levels at 1500 mg/kg/day (79 and 82% of control, resp.); a small but dosage-dependant reduction in total protein at 750 and 1500 mg/kg/day (93 and 91% of control, resp.) with a slightly elevated albumin/globulin ratio at the top dosage (117% of control). Changes in toxicity
subgroup females were limited to a decrease in phosphorous levels at 1500 mg/kg/day (81% of control) although a non-significant increase in
alkaline phosphatase levels at 1500 mg/kg/day (122% of control) in particular suggested a similar change to that recorded in males.

NEUROBEHAVIOUR:
No effects.

Dose descriptor:

NOAEL

Effect level:

>= 1 500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: developmental toxicity

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
VIABILITY (OFFSPRING)
No effects

CLINICAL SIGNS (OFFSPRING)
No effects

BODY WEIGHT (OFFSPRING)
No effects.

GROSS PATHOLOGY (OFFSPRING)
No effects.

HISTOPATHOLOGY (OFFSPRING)
No effects.

OTHER FINDINGS (OFFSPRING)
The percentage of males in the litters did not indicate any preferential mortality amongst either sex.

Dose descriptor:

NOAEL

Effect level:

>= 1 500 mg/kg bw/day (actual dose received)

Based on:

test mat.

Basis for effect level:

other: no adverse effects on reproduction/developmental toxicity (highest dose tested)

Abnormalities:

not specified

Developmental effects observed:

not specified

Conclusions:

NOAEL: 1500 mg/kg/day for reproduction/developmental toxicity.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Quality of whole database:

One screening study is available for an analogous substance. This study is considered to be a Klimisch score 2 (conducted to an appropriate method and under GLP) with no deficiencies. The additional studies that form the weight of evidence are performed on sodium pyrophosphates, these studies are performed to an old method with some deficiencies but are considered to be reliable as part of a weight of evidence approach.

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

 

In accordance with REACH Annex XI, Section 1.5, of Regulation (EC) No. 1907/2006 (REACH) the standard testing regime may be adapted in cases where a grouping or read-across approach has been applied.

The similarities may be based on:

(1) a common functional group

(2) the common precursors and/or the likelihood of common breakdown products via physical or biological processes, which result in structurally similar chemicals; or

(3) a constant pattern in the changing of the potency of the properties across the category

 

 Read-across from sodium acid pyrophosphate or tetrasodium pyrophosphate to diammonium dihydrogenpyrophosphate: 

 

 Sodium acid pyrophosphate is a soluble inorganic salt consisting of pyrophosphate anions and ammonium cations. Similarities between the source chemicals (sodium acid pyrophosphate / tetrasodium pyrophosphate) and the target chemical (diammonium dihydrogenpyrophosphate) are based on the following: 

 

 (1) The source chemicals (sodium acid pyrophosphate / tetrasodium pyrophosphate) and the target substance (diammonium dihydrogenpyrophosphate) are structurally similar substances. Both are soluble inorganic phosphate salts. In vivo both substances will be (bio)transformed into their respective ionic forms; phosphate anions (either diphosphate or orthophosphate depending on degree of metabolism) and cations; either ammonium or sodium. Exposure to the non-common compound, the sodium ion, will not result in toxicological effects at the dose levels tested. Similarly the effects of different phosphate moieties are not considered to be toxicologically significant due to in vivo (bio) transformation. 

 

 (2) Both substances contain the bio transformation product: orthophosphate. Pyrophosphate will ultimately be converted to orthophosphate in vivo. 

 

 The key moiety for the assessment of diammonium pyrophosphate is the ammonium cation and as such the effects of the target compound (diammonium dihydrogenpyrophosphate) are expected to be equal to or worse than the effects of the source substances (sodium acid pyrophosphate / tetrasodium pyrophosphate) for the property under consideration. 

 

   

   

 Justification for selection of Effect on developmental toxicity: via oral route:    

Study is considered to be the most relevant study for assessing the developmental toxicity of diammonium dihydrogenpyrophosphate

 

 

 

Justification for classification or non-classification

There are no data to suggest that diammonium dihydrogenpyrophosphate is a reproductive or developmental toxicant and as such in accordance with Regulation (EC) No. 1272/2008 (EU CLP) no classification is proposed.