Phosphoric acid, dodecyl ester, potassium salt

Endpoint summary

Description of key information

The oral gavage administration of the test item at doses of 50, 250 and 1000 mg/kg bw/d to rats for 90 consecutive days indicated the following:

There were no mortalities and clinical signs at all dose levels. The body weights were were unaffected by the treatment, while the food consumption was lower at 1000 mg/kg bw/d in both sexes. There were no ocular and neurological abnormalities at all the dose levels. Hematological inverstigations revealed increased total leucocyte count, neutrophils, lymphocytes and monocytes in females at 250 mg/kg bw/d and 1000 mg/kg bw/d in both sexes which was reversible after 28 days recovery period. In addition, at 1000 mg/kg bw/d the reticulocyte counts were decreased. Coagulation and urine parameters were unaffected at all doses. the clinical chemistry evaluation revealed increased ALP activity in females at 205 mg/kg bw/d and increased ALP, AST, triglcerides concentration, decreased total protein and globuline in both sexes at 1000 mg/kg bw/d. The changes in clinical chemistry were reversible after 28 days recovery period. At 1000 mg/kg bw/d the terminal fasting body weight in females was lower, relative weight of liver was increased in both sexes, the absolute weight of epididymides was decreased and hypertrophy of the zona glomerulosa layer in adrenals in both sexes of 1000 mg/kg bw/d which was reversible.

The NOAEL is set at 1000 mg/kg bw/d.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Deviations:

yes

Remarks:

On Day 20, the examination for general clinical signs was done prior to the test item administration for all rats and post dose was carried out 5 to 6 hours after the dose administration instead of 0.5 to 3 hours after dose administartion.

GLP compliance:

yes

Specific details on test material used for the study:

Study Sponsor
(Name and address): Clariant India Limited, Product Stewardship – India, BU ADD, 9th floor, Reliable Tech Park, Thane–Belapur Road,Airoli, Navi Mumbai 400 708, India

Batch Manufactured By: Clariant

Supplied By (Name and Address): Dr. Sunil Deval, Product Stewardship – India, BU ADD, 9th floor, Reliable Tech Park, Thane–Belapur Road, Airoli, Navi Mumbai 400 708, India, Phone: [phone], Mobile: [phone], Email:[e-mail]

Test item: Leomin PN pa

Chemical name (IUPAC): Reaction mass of potassium didodecylphosphate and dipotassium dodecylphosphate and water

CAS NO: CAS 39322-78-6

Sample No.: DEH2 040297

Manufactured date: 23.10.2015

Expiry date: 22.10.2017

Purity as per Certificate of Analysis : 100.0 % (w/w)

Physical appearance: Viscous white paste

Storage conditions: Refrigeration (+2 to + 8°C)

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Vivo Bio Tech Ltd, Sy # 349/A, Pregnapur-502311,Gajwel Mandal, Medak District, Telangana

- Age at study initiation: 7-8 weeks
- Weight at study initiation: Males: 210.29 g to 235.98 g; Females: 141.66 g to 175.47 g
- Fasting period before study: Not applicable
- Housing: Housed two rats/sex/cage except for the last cage of recovery groups wherein one rat/cage was housed
- Diet (e.g. ad libitum): Teklad certified (2014C) global 14% protein rodent maintenance diet - pellet (certified) manufactured by Envigo,
PO Box 44220, Madison, WI 53744-4220, were provided ad libitum to rats.
- Water (e.g. ad libitum): Deep bore-well water passed through activated charcoal filter and exposed to UV rays in Aquaguard on-line
water filter-cum-purifier manufactured by Eureka Forbes Ltd., Mumbai - 400 001, India was provided.
- Acclimation period: 5 days

DETAILS OF FOOD AND WATER QUALITY: There were no known contaminants in the food or water that were expected to interfere with the
results of this study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 to 24 °C
- Humidity (%): 48 to 67%
- Air changes (per hr): 12 - 15 air changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark cycle

IN-LIFE DATES: From: 01 September 2016 To: 02 January 2017

Route of administration:

oral: gavage

Details on route of administration:

Route of test item administration was through oral gavage. This route has been chosen because it is the potential route of human exposure.

Vehicle:

other: Milli-Q water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: Dose formulations were prepared at intervals of 3 to 4 days and used within the established
stability period of 4 days.
Required quantities of the test item was weighed in a beaker (previously pre-calibrated to desired
volume) for each dose levels separately and small volume of vehicle - Milli-Q water (hot Milli-Q water
maintained at approximately 80 - 86.7 °C) was added and mixed by using glass rod till a uniform
formulation was obtained. The resulting pre-mix was made up to the pre-calibrated mark using the
vehicle to get the final desired concentration and mixed well.
The homogeneity of the dosing formulations during administration/sampling was maintained by constant
stirring using magnetic stirrer.


DIET:
Teklad Certified (2014C) Global 14% Protein Rodent Maintenance Diet – pellet
(Certified) manufactured by Envigo, PO Box 44220, Madison, WI 53744-4220,
were provided ad libitum to rats.

VEHICLE: Milli-Q water
- Justification for use and choice of vehicle (if other than water):
As per the information provided by the sponsor, water was used as a vehicle
in the dose range finding study, hence Milli-Q water was used as vehicle for
dose formulation preparation.
- Concentration in vehicle: Not applicable
- Amount of vehicle (if gavage): 10 mL/kg Body weight per day
- Lot/batch no. (if required): Not applicable
- Purity: Not applicable

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Leomin PN pa in the dose formulations was determined using Liquid Chromatograph with Mass Spectrometer (LC-MS/MS)

Duration of treatment / exposure:

The dose formulations were administered by oral gavage to rats of the respective groups once daily at approximately the same time each day (varied by ± 3 hours) for 90 consecutive days. Similarly, vehicle was administered by oral gavage to rats in vehicle control groups for 90 consecutive days. The vehicle or the dose formulations were not administered to the rats in the recovery groups for 28 Days following 90-day treatment period.

The dose volume administered to each rat was 10 mL/kg body weight throughout the study. The dose volume was calculated for individual animal on the first day of the treatment and was adjusted according to the most recent body weights recorded at different intervals of the study.

Frequency of treatment:

Once daily

Dose / conc.:

0 mg/kg bw/day (actual dose received)

Dose / conc.:

50 mg/kg bw/day (actual dose received)

Dose / conc.:

250 mg/kg bw/day (actual dose received)

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

10 males and 10 females for main groups
5 males and 5 females for recovery groups (control recovery and high dose recovery)

Control animals:

yes

Details on study design:

- Dose selection rationale: The dose levels of 50, 250 and 1000 mg/kg bwt/day have been selected in consultation with the sponsor based on
a previously performed dose range finding study along with a concurrent control group.
- Rationale for animal assignment (if not random): Selected randomly as per body weight stratification
- Rationale for selecting recovery groups: control and high dose recovery
- Post-exposure recovery period in recovery groups: 28 days

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Daily twice (pre and post dose)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Weekly

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION: Yes
- Time schedule: Weekly

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: At termination
- Dose groups that were examined: Control and High dose groups

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At termination
- Anaesthetic used for blood collection: Yes
- Animals fasted: Yes
- How many animals: 100
- Parameters checked in table [No.1, 2, 3, 4 (Pathology report)] were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At terminatin
- Animals fasted: Yes
- How many animals: 100
- Parameters checked in table [No.5, 6, 7, 8 (Pathology report)] were examined.

URINALYSIS: Yes
- Time schedule for collection of urine: At termination
- Animals fasted: Yes
- Parameters checked in table [No. 9, 10, 11, 12 (Pathology report)] were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Dose groups that were examined: At prior to terminatin of main and recovery groups
- Battery of functions tested: sensory activity / grip strength / motor activity / :

IMMUNOLOGY: No

OTHER:

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see table 25 (Pathology report) )

HISTOPATHOLOGY: Yes (see table 26 (Pathology report) )

Statistics:

Data captured using ProvantisTM: Parameters such as body weight, body weight gains (derived data), food consumption (derived data), terminal fasting bodyweight, organ weights and their ratios data (derived data), laboratory investigations - Haematology (except coagulation parameters PT and APTT which was entered retrospectively in ProvantisTM and analysed) and Clinical Chemistry was analysed using ProvantisTM built-in statistical tests.

Data captured outside of ProvantisTM: The statistical analysis of the experimental data was carried out using the licensed copies of SYSTAT Statistical
package Ver.12.0. All quantitative variables like neurological examinations (neuromuscular parameters, body weight and body temperature) was tested for
normality (Shapiro-Wilk test) and homogeneity of variances (Levene’s test) within the group before performing a one-factor ANOVA modeling by treatment
groups. Non-optimal (non-normal or heteroschedastic) data was transformed, before ANOVA was performed. Comparison of means between treatment groups and vehicle control group was done using Dunnett’s test when the overall treatment, ‘F’ test was found to be significant.

The data from the vehicle control group (G1) was compared with the data from the treatment groups (G2, G3 & G4).

In the case of recovery groups, data was analysed using the methods stated above. Comparison of mean between treatment recovery group(s) and control
recovery group was performed. When analysis was done outside provantis the comparison of means between the control recovery and high dose recovery group was done using two sampled ‘t’ test.

All analyses and comparisons were evaluated at the 5% (P < 0.05) level. Statistically significant differences (P < 0.05), indicated by the aforementioned tests were designated by the following symbols throughout the report:
+/-: Significantly higher (+) /lower (-) than the vehicle control group
a+/a-: Significantly higher/lower than the vehicle control recovery group

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

Few statistical significant changes in body weights and body weight gains observed are provided below:
- Decrease in the mean body weights in female rats at 1000 mg/kg/day (Days 36, 43 and 50) which were less than 10%.
- Decrease in the body weight gains in male rats (Days 15-22, 36-43) and female rats (Days 1-8, 22-29) at 1000 mg/kg/day.
- Increase in the body weight gains in male rats at 50 and 250 mg/kg/day (Days 64-71)
The above few sporadic changes in mean body weights and gains which were randomly observed during the treatment period were considered incidental and not related to test item. Hence, the body weights and body weight gains were unaffected at all the doses tested.

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

• Decreased absolute and relative reticulocyte counts at 1000 mg/kg/day dose in both sexes
• Increased total leucocyte count, neutrophils, lymphocytes and monocytes at 250 mg/kg/day dose group females and at 1000 mg/kg/day
dose in both sexes.
• All the above test item-related changes were reversible after 28 days recovery period.

There were no test item-related changes in coagulation parameters at all the doses tested.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

• Increased ALP and AST activity at1000 mg/kg/day dose in both sexes and increased ALP activity at 250 mg/kg/day dose females.
• Increased triglyceride concentration at 1000 mg/kg/day dose in both sexes.
• At 1000 mg/kg/day dose, decreased total protein and globulin in both sexes, decreased albumin concentration in females and increased
albumin globulin ratio in males. These changes were associated with decreased food intake.
• All the above test item-related changes were reversible after 28 days recovery period.

Urinalysis findings:

no effects observed

Behaviour (functional findings):

effects observed, non-treatment-related

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

• Decreased terminal fasting body weight at 1000 mg/kg/day dose females.
• Increased relative (body weight ratios) weight of liver at 1000 mg/kg/day dose in both sexes. This change was microscopically associated
with hepatocellular hypertrophy.
• Decreased absolute weight of epididymides at 1000 mg/kg/day dose in males.
• All the above test item-related changes were reversible after 28 days recovery period.

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

• Hepatocellular hypertrophy of liver observed at 1000 mg/kg/day dose in both sexes was mainly seen in centrilobular area and was
considered as test item-related adaptive change. Hepatocellular hypertrophy correlated with increase in relative weight of liver.
This change was reversible after 28 days recovery period.
• Hypertrophy of the zona glomerulosa layer of adrenals was observed at 1000 mg/kg/day dose groups and was considered as
test item–related reversible change.

Histopathological findings: neoplastic:

no effects observed

Other effects:

no effects observed

Details on results:

In view of the results observed, the evaluated “No Observed Adverse Effect Level (NOAEL)” for the test item Leomin PN pa when administered orally for 90 consecutive days to Wistar rats is 1000 mg/kg body weight under the test conditions and doses employed.

Key result

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Complete reversal of observed changes post recovery of 28 days

Key result

Critical effects observed:

no

Conclusions:

The oral gavage administration of the test item, Leomin PN pa at the doses of 50, 250 and 1000 mg/kg body weight/day to Wistar rats for 90 consecutive days indicated the following:

There were no mortalities and clinical signs at 50, 250 and 1000 mg/kg/day doses. There were no ocular and neurological abnormalities at all the dose levels. The body weights were unaffected by treatment, while the food consumption was lower at 1000 mg/kg/day dose in both sexes. Hematological investigations revelaed increased total leucocyte count, neutrophils, lymphocytes and monocytes in females at 250 mg/kg/day dose and 1000 mg/kg/day dose in both sexes which was reversible after 28 days recovery period. In addition at 1000 mg/kg/day dose, the reticulocyte counts were decreased. Coagulation and urine parameters were unaffected at all the doses tested. The clinical chemistry evaluation revealed, increased ALP activity in females at 250 mg/kg/day dose and increased ALP, AST, triglycerides concentration, decreased total protein, and globulin in both sexes at 1000 mg/kg/day dose. The changes in clinical chemistry parameters were reversible after 28 days recovery period. At 1000 mg/kg/day dose, the terminal fasting body weight in females was lower, relative weight of liver was increased in both sexes, the absolute weight of epididymides was decreased, and all these changes were reversible. Microscopic changes included hepatocellular hypertrophy in liver and hypertrophy of the zona glomerulosa layer in adrenals in both sexes at 1000 mg/kg/day dose which was reversible.

In view of the above results observed, the evaluated “No Observed Adverse Effect Level (NOAEL)” for the test item Leomin PN pa when administered orally for 90 consecutive days to Wistar rats is 1000 mg/kg body weight under the test conditions and doses employed.

Executive summary:

The objective of this repeat dose (90-day) oral toxicity study was to assess the systemic toxicity potential of the test item, Leomin PN pa when administered orally by gavage to Wistar rats for a period of 90 consecutive days and to assess the reversibility of any effects following 28 days recovery period. This study may provided information on major toxic effects, target organs and an estimation of a No Observed Effect Level (NOEL) / No Observed Adverse Effect Level (NOAEL).

The test item, Leomin PN pa was suspended in vehicle [Milli-Q water] and administered to Wistar rats at the doses of 50 (low dose - G2), 250 (Mid dose - G3) and 1000 mg/kg/day (high dose - G4/G4R) at an equivolume of 10 mL/kg/day. The rats in vehicle control groups (G1/G1R) received Milli-Q water alone at an equivolume of 10 mL/kg/day. The vehicle or dose formulations were not administered to the recovery groups (G1R and G4R) for 28 days following the 90-day dosing period. The main toxicity groups consisted of 10 rats/sex/group and recovery groups consisted of 5 rats/sex/group.

The identity of the test item was provided by the study Sponsor by Report of Analysis and the test item was not authenticated at the test facility. The Leomin PN pa formulations at 2 and 125 mg/mL concentration levels in the vehicle were found to be stable for 4 days at room temperature. In this study, the dose formulations were analysed three times. The overall mean concentration of Leomin PN pa from all formulations performed were found to be within ± 15% of the claimed concentration and the relative standard deviation (%RSD) was less than 10%. Vehicle sample showed an interference peak at the retention time of the analyte and the same response was also noticed in the Diluent (blank). Based on the complexity of the analysis, the carry over is considered minimum and hence acceptable.

In this study, assessments included clinical signs, body weights, food consumption, ophthalmological examinations, neurological examinations, clinical laboratory investigations of blood and urine, gross pathology, organ weights and histopathological evaluation. Liver and adrenals were considered as target organs and were examined in the lower dose groups and recovery groups.

The salient findings  of 90-day repeat dose study are as follows:

-       Mortality: No mortalities were observed.

-       Clinical Signs: No clinical signs were observed at any of the dose levels tested in both the sexes.

-       Ophthalmological Examination: No ocular abnormalities in any of the rats in both sexes.

-       Neurological Examinations: No test item-related neurological abnormalities were observed at all the dose levels

tested in both the sexes.

-       Body Weights/Food Consumption: No test item-related variations on the body weights, while food consumption

was significantly lower and is considered treatment-related in both the sexes.

-       Hematology and Coagulations: Following treatment-related changes were observed in the hematology parameters:

•  Decreased absolute and relative reticulocyte counts at 1000 mg/kg/day dose in both sexes

•  Increased total leucocyte count, neutrophils, lymphocytes and monocytes at 250 mg/kg/day dose group females

and at 1000 mg/kg/day dose in both sexes.

•  All the above test item-related changes were reversible after 28 days recovery period.

There were no test item-related changes in coagulation parameters at all the doses tested.

-       Clinical Chemistry: Following treatment-related changes were observed in the clinical chemistry parameters:

•  Increased ALP and AST activity at1000 mg/kg/day dose in both sexes and increased ALP activity

at 250 mg/kg/day dose females.

•  Increased triglyceride concentration at 1000 mg/kg/day dose in both sexes.

•  At 1000 mg/kg/day dose, decreased total protein and globulin in both sexes, decreased albumin concentration

in females and increased albumin globulin ratio in males. These changes were associated with decreased food intake.

•  All the above test item-related changes were reversible after 28 days recovery period.

-       Urinalysis: There were no test item-related changes in urinalysis parameters at all the doses tested.

-       Terminal fasting body weights, Organ weights and their ratios: Following treatment-related changes were observed:

•  Decreased terminal fasting body weight at 1000 mg/kg/day dose females.

•  Increased relative (body weight ratios) weight of liver at 1000 mg/kg/day dose in both sexes. This change

was microscopically associated with hepatocellular hypertrophy.

• Decreased absolute weight of epididymides at 1000 mg/kg/day dose in males.

•  All the above test item-related changes were reversible after 28 days recovery period.

-       Gross Pathology: There were no test item-related gross pathological changes at all the doses tested.

-       Histopathology: Following treatment-related changes were observed:

•  Hepatocellular hypertrophy of liver observed at 1000 mg/kg/day dose in both sexes was mainly seen in

centrilobular area and was considered as test item-related adaptive change. Hepatocellular hypertrophy

correlated with increase in relative weight of liver. This change was reversible after 28 days recovery period.

•  Hypertrophy of the zona glomerulosa layer of adrenals was observed at 1000 mg/kg/day dose groups and

was considered as test item–related reversible change.

In view of the results observed, the evaluated “No Observed Adverse Effect Level (NOAEL)” for the test item Leomin PN pa when administered orally for 90 consecutive days to Wistar rats is 1000 mg/kg body weight under the test conditions and doses employed.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

reliable without restriction

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: inhalation

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: dermal

Data waiving:

study scientifically not necessary / other information available

Justification for data waiving:

other:

Critical effects observed:

not specified

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated dose oral toxicity: 90 days

The oral gavage administration of the test item, Leomin PN pa at the doses of 50, 250 and 1000 mg/kg body weight/day to Wistar rats for 90 consecutive days indicated the following:

 

There were no mortalities and clinical signs at 50, 250 and 1000 mg/kg/day doses. There were no ocular and neurological abnormalities at all the dose levels. The body weights were unaffected by treatment, while the food consumption was lower at 1000 mg/kg/day dose in both sexes. Hematological investigations revelaed increased total leucocyte count, neutrophils, lymphocytes and monocytes in females at 250 mg/kg/day dose and 1000 mg/kg/day dose in both sexes which was reversible after 28 days recovery period. In addition at 1000 mg/kg/day dose, the reticulocyte counts were decreased. Coagulation and urine parameters were unaffected at all the doses tested. The clinical chemistry evaluation revealed, increased ALP activity in females at 250 mg/kg/day dose and increased ALP, AST, triglycerides concentration, decreased total protein, and globulin in both sexes at 1000 mg/kg/day dose. The changes in clinical chemistry parameters were reversible after 28 days recovery period. At 1000 mg/kg/day dose, the terminal fasting body weight in females was lower, relative weight of liver was increased in both sexes, the absolute weight of epididymides was decreased, and all these changes were reversible. Microscopic changes included hepatocellular hypertrophy in liver and hypertrophy of the zona glomerulosa layer in adrenals in both sexes at 1000 mg/kg/day dose which was reversible.

 

In view of the above results observed, the evaluated “No Observed Adverse Effect Level (NOAEL)” for the test item Leomin PN pa when administered orally for 90 consecutive days to Wistar rats is 1000 mg/kg body weight under the test conditions and doses employed.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
The OECD 422 study performed with 1-Dodecanol was selcted as the most relevant available repeated dose toxicity study due to its reliability and since it provides the most sensitive NOAEL.
There is a strong evidence from structural considerations and a metabolism study performed with Phosphoric acid 2-ethylhexyl ester in rats that Phosphoric acid dodecyl ester is metabolised to 1-Dodecanol and Phosphate (please refer to chapter 7.1). Phosphoric acid dodecyl ester is supposed to be metabolised to Phosphate and 1 -Dodecanol after entering the organism. Phosphate is known to be an endogenous compound and food additive without reported toxic properties. Therefore, a read across to the second metabolite, 1 -Dodecanol, was conducted in order to estimate the toxicological properties in a combined repeated dose/reproductive toxicity study.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
In accordance with column 2 of REACH Annexes VIII and IX, the repeated dose toxicity study, as required in section 8.6.1 of Annex VIII and in section 8.6.2 of Annex IX, does not need to use the inhalation route because exposure of human via inhalation, especially in a higher extent than via oral application as performed in the animal studies, is considered unlikely taking into account the vapour pressure of the substance and the physical form.

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:
In accordance with column 2 of REACH Annexes VIII and IX, the repeated dose toxicity study, as required in section 8.6.1 of Annex VIII and in section 8.6.2 of Annex IX, does not need to use the inhalation route because exposure of human via inhalation, especially in a higher extent than via oral application as performed in the animal studies, is considered unlikely taking into account the vapour pressure of the substance and the physical form.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
In accordance with column 2 of REACH Annexes VIII and IX, the repeated dose toxicity study, as required in section 8.6.1 of Annex VIII and in section 8.6.2 of Annex IX, does not need to use the dermal route because
- no systemic effects or other evidence of absorption were observed in skin and eye irritation studies in rabbits as well as in the skin sensitization test in guinea pigs
- due its irritant properties only local skin effects are expected to occur; however, since the edema/erythema were reversible these are considered to have no impact on the absorption of the test substance via skin. Due to the combination of its polar (ionic) character and the long extent of the alcoholic chain it is unlikely that higher amounts than tested in a repeated oral toxicity study will be systemically available via the skin barrier.

Justification for selection of repeated dose toxicity dermal - local effects endpoint:
In accordance with column 2 of REACH Annexes VIII and IX, the repeated dose toxicity study, as required in section 8.6.1 of Annex VIII and in section 8.6.2 of Annex IX, does not need to use the dermal route because
- no systemic effects or other evidence of absorption were observed in skin and eye irritation studies in rabbits as well as in the skin sensitization test in guinea pigs
- due its irritant properties only local skin effects are expected to occur; however, since the edema/erythema were reversible these are considered to have no impact on the absorption of the test substance via skin. Due to the combination of its polar (ionic) character and the long extent of the alcoholic chain it is unlikely that higher amounts than tested in a repeated oral toxicity study will be systemically available via the skin barrier.

Repeated dose toxicity: via oral route - systemic effects (target organ) cardiovascular / hematological: other

Justification for classification or non-classification

Due to the most sensitive NOAEL of 1000 mg/kg bw/day derived by an OECD 408 repeated dose toxicity study in rats with phosphoric acid, dodecyl ester, potassium salt, the registration substance does not have to be classified regarding systemic and target organ toxicity after repeated exposure according to the criteria laid down in the EU Classification Labelling and Packaging Regulation (1272/2008/EC).