Fatty acid chlorides, C8-14 (even numbered), reaction products with glycine

Endpoint summary

Description of key information

The registration substance Hostapon SG is of low subacute toxicity based on the findings in the 28 -day oral toxicity study:

Hostapon SG was investigated for its repeated dose toxicity in rats according to the Guideline OECD 407. The study included recovery animal groups. Animals were treated at doses of 0, 62.5, 250 and 1000 mg/kg/day. No significant treatment related systemic effect was found up to the highest dose level. At dose of 1000 mg/kg/day microscopic changes in the forestach was found. The lesions consisted of acanthosis, parakeratosis, inflammation, ulceration and erosions. After recovery period of 14 days, these changes were completely reversible. Comparable effects were not seen in the animals of low and mid dose groups. NOAEL of 250 mg/kg/day was assigned for the forestomach local effects and NOAEL of 1000 mg/kg/day was assigned for the systemic effects.

The registration substance Hostapon SG is likely of low chronic toxicity based on the read-across approach:

The chronic toxicity of the registration substance Hostapon SG was evaluated based on the read-across approach using the repeated dose toxicity data for sodium N-lauroyl sarcosinate

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compounds are likely exhibit comparable toxicity profiles.

Sodim N-Lauroyl sarcosinate was given to rats  by gavage daily at doses of 0, 30, 100, and 250 mg/kg bw for up to 92 days. Body weight gain decrease and effects on the stomach was found for animals of 100 and 250 mg/kg/day. The NOAEL was 250 mg/kg bw/day.The NOEL was 30 mg/kg/day and the NOAEL 250 mg/kg bw/day.

Sodium N-lauroyl sarcosinate was investigated in rats for its chronic toxicity and reproduction toxicity in a 2 -year feeding study. The doses levels applied were 0, 0.2, 1 and 2% in the diet. Males and females were housed together so that fertility could be assessed. No effect on the fertility was found. Histopathological changes of stomach tissues were evident at dose levels of 1 and 2% in the diet. No significant systemic effect was found up to the highest dose level.

Sodium N-lauroyl sarcosinate were applied to weanling rats by feeding at 2% in diet for 6 months. No effect on weight gain, feeding efficiency, general health, or behavior was observed, and no abnormalities of the internal organs were observed.

Likewise, the registration substance Hostapon SG is likely of low chronic toxicity.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2012

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Japanese Guidelines for Screening Toxicity Testing of Chemicals: Testing Methods for New Substances, enacted July 13, 1974, amended December 5, 1986

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

swissmedic; date of decision: 19-November-2010

Limit test:

no

Specific details on test material used for the study:

The test material contains sodium chloride by ca 20% (w/w).

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Animals: Rat, RccHanTM: WIST(SPF)
Rationale: Recognized by international guidelines as a recommended test system.
Breeder: Harlan Laboratories B.V.,Kreuzelweg 53, 5961 NM Horst / Netherlands
Number of Animals:
Group 1: 10 males and 10 females
Group 2: 5 males and 5 females
Group 3: 5 males and 5 females
Group 4: 10 males and 10 females
Group 10: 1 male and 1 female
Age (at Delivery): Ca. 7 weeks
Body Weight Range
(at Acclimatization):
Males: 199 to 233 g (mean 214 g)
Females: 133 to 154 g (mean 144 g)
Identification:
Acclimatization: Cage card and tail mark (later ear tattoo)
Treatment: Cage card and individual ear tattoo
Randomization: Randomly allocated to groups by body weight.
Acclimatization: Under test conditions after health examination. Only animals without any visible signs of illness were used for the study.


ENVIRONMENTAL CONDITIONS

Standard laboratory conditions. Air-conditioned with 10 - 15 air changes per hour, continuously monitored environmental conditions (temp. range: 22 ± 3 °C; relative humidity range: 30 - 70%). The light cycle was set to 12-hour fluorescent light / 12-hour dark cycle with at least eight hours music during the light period.

Accommodation: In groups of five in Makrolon type-4 cages with wire mesh tops and standard softwood bedding (J. Rettenmaier & Söhne GmbH & Co. KG, 73494 Rosenberg / Germany, imported by Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) including paper enrichment (Enviro-dri from Lillico, Biotechnology, Surrey / UK).

Diet: Pelleted standard Harlan Teklad 2914C (batch no. 73/11) rat / mouse maintenance diet (Provimi Kliba AG, 4303 Kaiseraugst / Switzerland) was available ad libitum. The feed batch was analyzed for contaminants.

Water: Community tap-water from Itingen was available ad libitum in water bottles.

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

Method of administration: Oral, by gavage
Rationale for Method: Administration by gavage is a common and accepted route of exposure for studies of this type.
Frequency of Administration: Daily.
Daily Target Dose Level:
Group 1: 0 mg/kg/day
Group 2: 62.5 mg/kg/day
Group 3: 250 mg/kg/day
Group 4: 1000 mg/kg/day
Rationale for Dose Level Selection: The dose levels were selected based on a previous non-GLP 14-day dose range finding toxicity study in Wistar rats. The animals were treated up to dose of 1000 mg/kg/day. No effect was found upon clinical observation. No effect on the body weight development was found.
Dose Volume: 10 mL/kg body weight
Dose Concentrations:
Group 1: 0 mg/mL/day
Group 2: 6.25 mg/mL/day
Group 3: 25 mg/mL/day
Group 4: 100 mg/mL/day
Duration of Pre-Randomization Phase: 1 day
Duration of Acclimatization Phase: At least 5 days
Duration of Treatment Phase: 28 days
Duration of Recovery Phase: 14 days

PREPARATION OF DOSING SOLUTIONS:
-For the purpose of this study the test material was prepared at the appropriate concentrations
as a solution in highly purified water.

VEHICLE
-Highly purified water

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analysis was performed by Harlan Laboratories Ltd. using a HPLC method provided by the Sponsor.

After experimental start and during week 3, duplicate samples of the control group as well as three samples (top, middle and bottom) of about 2 g of each concentration were taken prior to dosing for analysis of homogeneity and concentration. Duplicate samples of about 2 g of each concentration were taken to confirm stability (4 hour and 8 days).

The samples were delivered to the analytical department (Harlan Laboratories Ltd., Analytics, Zelgliweg 1, 4452 Itingen / Switzerland) and stored there at -20 ± 5 °C until analysis.

The test item was used as analytical standard.

Dose formulation samples (primary samples or duplicates) were discarded upon written confirmation by the study director after acceptance of the draft report.

Duration of treatment / exposure:

Test duration: 28 days

Frequency of treatment:

Dosing regime: 7 days/week

Dose / conc.:

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

Dose / conc.:

62.5 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:

Group 1: 10 males and 10 females at 0 mg/kg bw/day
Group 2: 5 males and 5 females at 62.5 mg/kg bw/day
Group 3: 5 males and 5 females at 250 mg/kg bw/day
Group 4: 10 males and 10 females at 1000 mg/kg bw/day
Group 10: 1 male and 1 female (reserve animals)

Reserve animals were exchanged as needed during the acclimatization period and then removed from the study. Any raw data collected during the acclimatization period on reserve animals were not reported but retained in the raw data.

Control animals:

yes, concurrent vehicle

Details on study design:

Method: Oral, by gavage
Rationale for Method: Administration by gavage is a common and accepted route of exposure for studies of this type.
Frequency of Administration: Daily.
Daily Target Dose Level:
Group 1: 0 mg/kg/day
Group 2: 62.5 mg/kg/day
Group 3: 250 mg/kg/day
Group 4: 1000 mg/kg/day
Rationale for Dose Level Selection: The dose levels were selected based on a previous non-GLP dose range finding toxicity study in Wistar rats, Harlan Laboratories study D43306.
Dose Volume: 10 mL/kg body weight
Dose Concentrations:
Group 1: 0 mg/mL/day
Group 2: 6.25 mg/mL/day
Group 3: 25 mg/mL/day
Group 4: 100 mg/mL/day
Duration of Pre-Randomization Phase: 1 day
Duration of Acclimatization Phase: At least 5 days
Duration of Treatment Phase: 28 days
Duration of Recovery Phase: 14 days

Positive control:

None

Observations and examinations performed and frequency:

-Viability / Mortality
Observations for viability / mortality were recorded twice daily.

-Daily Observations
The animals were observed daily for clinical signs before commencement of administration as well as daily on days 1 - 28 (twice daily during days 1 - 3) during the treatment period, and once daily during days 1 to 14 of the recovery period.

-Weekly Behavioral Observations
The animals were observed in their home cages, outside their home cages in a standard arena and in the hand. These observations were performed once before commencement of administration and once weekly (weeks 1 to 3) thereafter.

-Functional Observational Battery (Screen)
During week 4, relevant parameters from a modified Irwin screen test were evaluated in all animals. The results are present in the summary and individual tables of the weekly detailed clinical observations under week 4.

-Grip Strength
Forelimb and hindlimb grip strength measurements were performed using a push-pull strain gauge (Mecmesin, AFG 25N). The animals were placed with the forepaws inside a triangular grasping ring and with the hind paws outside a triangular grasping ring. Using one hand, the animals were held towards the base of the tail and steadily pulled away or towards the ring until the grip was broken. Each measurement was repeated three times, the means were calculated and recorded.

-Locomotor Activity
Locomotor (decreased or increased) activity was measured quantitatively with AMS Föhr Medical Instruments GmbH (FMI) and DeMeTec GmbH Activity Monitor System. Animals were monitored during the fourth treatment week for a 60-minute period and the total activity of this time period was recorded. Low beams count was reported in 10-minute intervals as well as the total activity of the measuring period.

-Food Consumption
The food consumption was recorded once during the acclimatization period and weekly thereafter, using an on-line electronic recording system consisting of a Mettler balance connected to the Harlan Laboratories computer.

-Body Weights
Body weights were recorded weekly during acclimatization, treatment and recovery periods and before necropsy, using an on-line electronic recording system consisting of a Mettler balance connected to the Harlan Laboratories computer.

-Clinical Laboratory Investigations
Blood and Urine Sampling:
After 4 Weeks: 20-Feb-2012 (allocation A and B)
After 6 Weeks: 05-Mar-2012 (allocation B)

Blood samples were drawn from the retro-orbital plexus from all animals under light isoflurane anesthesia. The animals were fasted in metabolism cages for approximately 18 hours before blood sampling but allowed access to water ad libitum. The samples were collected early in the working day to reduce biological variation caused by circadian rhythms. Blood samples were drawn from the retro-orbital plexus using a micro-hematocrit glass capillary tube.

Urine was collected during the 18 hours fasting period into a specimen vial, using a metabolism cage.

-Hematology
The following hematology parameters were determined:

-Complete Blood Cell Count

Erythrocyte count
Hemoglobin
Hematocrit
Mean corpuscular volume
Red cell volume distribution width
Mean corpuscular hemoglobin
Mean corpuscular hemoglobin concentration
Hemoglobin concentration distribution width
Reticulocyte count
Reticulocyte maturity index (low, medium, high fluorescence)
Leukocyte count, total
Differential leukocyte count: Neutrophils, Eosinophils, Basophils, Lymphocytes, Monocytes, Large unstained cells, Platelet count

-Hemoglobin Derivatives

Methemoglobin Heinz bodies (slides were prepared but not evaluated)
Coagulation

Prothrombin time (= Thromboplastin time) Activated partial Thromboplastin time

-Clinical Biochemistry
The following clinical biochemistry parameters were determined:

Glucose
Urea
Creatinine
Bilirubin, total
Cholesterol, total
Triglycerides
Phospholipids
Aspartate aminotransferase
Alanine aminotransferase
Lactate dehydrogenase
Alkaline phosphatase
Bile acids Gamma-glutamyl-transferase
Creatine kinase
Sodium
Potassium
Chloride
Calcium
Phosphorus
Protein, total
Albumin
Globulin
Albumin/Globulin ratio

-Urinalysis
The following urine parameters were determined:

-Physical Examination

Urine volume (18 hour)
Specific gravity (relative density)
Color
Appearance

-Chemical Examination

pH value
Nitrite
Protein
Glucose
Ketones Urobilinogen
Bilirubin
Erythrocytes
Leukocytes

-Necropsy
Sacrifice:

After 4 Weeks: 20-Feb-2012 (allocation A)
After 6 Weeks: 05-Mar-2012 (allocation B)

All allocation A and B animals were weighed and necropsied. Descriptions of all macroscopical abnormalities were recorded. All animals were anesthetized by intraperitoneal injection of pentobarbitone and killed by exsanguination.

A blood sample was taken from each animal by heart puncture (ca. 2 mL) into appropriate tubes (Vacutainer glass tubes containing SST-Gel and Clot Activator or the equivalent) for serum preparation and then placed on ice or cool plates. Following centrifugation, the serum was divided in 2 aliquots of at least 350 µL and transferred to plastic (polypropylene) tubes. These samples were stored at approximately -80 °C and protected from light in case they were needed for analysis of hormone activity.

-Organ Weights
The organs from allocation A and B animals were weighed before fixation and recorded on the scheduled dates of necropsy. Relative organ weights were calculated on the basis of the body weight and brain weight.

The terminal body weight was recorded immediately prior to necropsy and the organ to terminal body weight ratios as well as organ to brain weight ratios were determined.

-Histotechnique
All organ and tissue samples, as defined under Histopathology , were processed, embedded and cut at an approximate thickness of 2 to 4 micrometers and stained with hematoxylin and eosin.

-Histopathology
Slides of all organs and tissues collected at scheduled sacrifices from all animals of the control and high-dose groups and all gross lesions from all animals were examined by the study pathologist. Liver. Lungs and stomach of animals from the low and middle dose groups were also evaluated. Organ and tissue samples taken from animals which died spontaneously were evaluated similarly to those organs taken from animals of the high-dose group.

A description of all abnormalities is attached (see Appendix VI ). Attempts were made to correlate gross observations with microscopic findings.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Other examinations:

-Viability / Mortality
- Organ Weights

Statistics:

The following statistical methods were used to analyze body weight, grip strength, locomotor activity, clinical laboratory data, organ weights and ratios as well as macroscopic findings:

• The Dunnett-test [see References (1)] (many to one t-test) based on a pooled variance estimate was applied if the variables could be assumed to follow a normal distribution for the comparison of the treated groups and the control groups for each sex.

• The Steel-test [see References (2)] (many-one rank test) was applied instead of the Dunnett-test when the data could not be assumed to follow a normal distribution.

• Fisher's exact-test [see References (3)].

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

only forestomach effects at 1000 mg/kg body weight: acanthosis, parakeratosis, inflammation, ulceration and erosions

Details on results:

-Analysis of Dose Formulations
The linearity of the analytical systems used for sample analyses was demonstrated with a good relationship between peak
areas measured and working standard concentrations. All calibration points used met the acceptance limit of ±20% variation
from the calibration curve derived by linear regression analysis. The regression coefficients (R2) calculated exceeded 0.99.
The Hostapon SG dried peak was assigned in sample chromatograms by comparison to that of working standards. In blank
sample chromatograms, no peak appeared at the retention time of Hostapon SG dried, confirming the absence of the test item
in the vehicle control samples (purified water).
The application formulations investigated during the study were found to comprise Hostapon SG dried in the range of 93.6% to
101.9%, meeting the required content limit of ±20%. The dose formulations with Hostapon SG dried were found to be
homogeneous because single results found did not deviate more than 1.4% (acceptance criterion: <15%) from the
corresponding mean.
In addition, the test item was found to be stable in application formulations when kept four hours or eight days at room
temperature due to recoveries which met the variation limit of 10% from the time-zero (homogeneity) mean.
In conclusion, the results indicate the accurate use of the test item Hostapon SG dried and purified water as vehicle during this
study. Application formulations were found to be homogeneously prepared and sufficiently stable under storage conditions.


-Viability / Mortality
At 1000 mg/kg/day, one male and one female rat were found dead before scheduled necropsy. Males no. 22 was found dead
on day 9 of treatment; female no. 58 on day 11 of treatment. Macroscopical changes were generally commensurate with those
of dose formulation aspiration.
All other rats survived until their respective scheduled necropsies.


-Daily Observations
There were no test item-related clinical signs noted at any dose level during the treatment period and there were no late
affects noted during the recovery period.
Breathing noise (râles) were noted in a single male at 1000 mg/kg/day on treatment day 6 only. All remaining males and all
females were without clinical signs.


-Weekly Behavioral Observations
There were no clinical signs noted at any dose level during the weekly behavioral observations performed during weeks 1, 2 or
3.


-Functional Observational Battery (Screen)
There were no test item-related clinical signs noted at any dose level during the functional observational battery performed
during week 4. Dyspnea was noted in two males treated with 1000 mg/kg/day. There were no signs seen in any other male or
in any female.
-Grip Strength
There were no differences between the respective mean fore- and hind-limb grip strength values at any dose level.
-Locomotor Activity
There were no test item-related differences in the mean locomotor activity values when compared with the respective control
values.
In males treated with 62.5 mg/kg/day, the locomotor activity was significantly elevated (p<0.05) from 40-50 minutes only, and
the males at higher dose levels were not affected. Significantly elevated mean locomotor activity was noted in high-dose
females from 0-10 minutes (p<0.05) when compared with the control values. All other dose groups were similar to the values
noted in the respective controls.


-Food Consumption
The mean daily food consumption was reduced in females treated with the test item at 1000 mg/kg/day when compared with
the controls. A trend for marginally lower food consumption was also noted in the females treated with 250 mg/kg/day. The
males were considered to be unaffected.
In males treated with 1000 mg/kg/day, the mean absolute daily food consumption remained slightly below that of the control
males during all measurement intervals. The differences to the control ranged from -1.9% to -3.9%, but this was considered to
be due to marginally lower body weights, as the relative food consumption values of the high-dose males were nearly identical
to those of the control males. Females treated with 1000 mg/kg/day consumed less feed that the control females (ranging from
-14.3% to -29%). These differences were largely confirmed in the relative food consumption values of these females and
generally similar values were noted in both cages. During the recovery period, the daily food consumption of the males
previously treated with 1000 mg/kg/day exceeded that of the control males, whereas in females, treated and untreated females
were similar.
At 250 mg/kg/day, markedly lower mean absolute daily food consumption values were noted in males, ranging from -6.3% to -
11.8% lower that the control males. As in the high-dose males, however, the mean relative food consumption values were only
slightly lower. Females at 250 mg/kg/day had slightly lower absolute mean daily food consumption values that were confirmed
by similarly reduced mean relative body weights.
The mean absolute and mean relative daily food consumption values of the males treated with 62.5 mg/kg/day were nearly
identical with those of the control males throughout the treatment period. Female rats treated with 62.5 mg/kg/day consumed
less food, but the difference was within the range of typical biological variation. Similar, very minor differences were also seen
in the mean relative food consumption values.


-Body Weights
Although slight variations in body weight development were noted, these changes were not clearly dose-related and were
therefore considered to be unrelated to the treatment with the test item.
At 1000 mg/kg/day, marginally lower mean absolute body weights were noted when compared with the control males. These
differences, however, were not considered to be related to the treatment as the mean body weight gain values compared
favorably at all measurement intervals. Females at this dose level, showed slightly lower mean body weights from day 8
onwards and the mean body weight gain values were significantly lower (p<0.05 on days 8, 22 and 28 or p<0.01 on day 15 of
treatment) when compared with the controls. During the recovery period, males gained significantly more weight on days 8
and 14 (both p<0.05) of recovery when compared with the controls, whereas females ad significantly reduced mean body
weights on day 1 (p<0.01) and day 8 (p<0.05) of recovery.
At 250 mg/kg/day, the males had lower mean absolute body weights from day 1 onwards, but without attaining statistical
significance. The mean body weight gain values of these males were also slightly lower but without statistical significance. In
females, the mean absolute body weights and the mean body weight gain values compared favorably with those of the
controls.
At 62.5 mg/kg/day, the mean absolute body weights and mean body weight gain values of the control males and treated males
were similar. The females treated with 62.5 mg/kg/day had marginally lower mean body weights and body weight gain when
compared with the control females.


-Clinical Laboratory Investigations
-Hematology
There were no test item-related changes in the hematology parameters of males or females at any dose level.
In males treated with 1000 mg/kg/day, there were no differences in the hematology parameter after the end of the treatment
period. The slight but statistically significant increase in the hemoglobin level (p<0.05) noted in the females treated with 1000
mg/kg/day, in the absence of any other relevant findings, was considered to be of no toxicological relevance.
At 250 mg/kg/day, significantly higher mean absolute and mean relative monocyte counts (both p<0.05) were noted in females
that were not dose-dependent and considered to be unrelated to the treatment with the test item. The increased mean
absolute eosinophil count was also statistically significant (p<0.05), but also dose-unrelated and therefore, no toxicological
relevance was associated with this finding.
At 62.5 mg/kg/day, a significant increase in the mean red cell count (p<0.05) was noted in males but in the absence of any
dose-response relationship, considered to be incidental.
After the recovery period, males had a minimal but statistically significant reduction of the mean absolute basophil count that
remained within the range of the historical control data and was considered to be incidental. Females had significantly reduced
(p<0.01) mean absolute leukocyte count when compared with the controls. This difference was primarily due to a significant
reduction of the mean absolute lymphocyte count (p<0.01) and, to a lesser extent, the ‘large unstained cell’ count (p<0.01). In
the case of the former parameter, the control value was considered to be elevated and this was considered to result in the
statistical significances.


-Clinical Biochemistry
There were no test item-related changes noted at any dose level. Minor differences between groups were either unrelated to
dose, seen only in one sex or due to atypical differences in the control values.
At 1000 mg/kg/day, males had significantly higher alanine aminotransferase activity (p<0.01) and significantly lower lactate
dehydrogenase activity (p<0.05) when compared with the controls. Phosphorus levels were reduced with statistical
significance (p<0.05), In females, significantly elevated potassium (p<0.05), chloride (p<0.05) and phosphorus (p<0.01) levels
were noted when compared with the controls.
At 250 mg/kg/day, males had significantly higher alanine aminotransferase activity (p<0.01) when compared with the controls.
Although the increased alanine aminotransferase activity seen in males at 100 and 250 mg/kg/day exceeded the historical
control values, the differences were not dose-dependent and were not seen in females, and were therefore considered to be
without toxicological relevance.
At 62.5 mg/kg/day, there were no differences when compared with the respective control values.
After the recovery period, males showed significantly elevated urea (p<0.05), significantly reduced creatine kinase (p<0.01)
when compared with the control males. Both values remained within the ranges of the respective historical control values,
whereas the second value is not generally associated with systemic toxicity and considered to be incidental.


-Urinalysis
After the treatment period, there were no differences in the urinalysis parameters at any dose level. There were no late effects
at the end of the recovery period.


-Pathology
-Organ Weights
There were a small number of typical background changes in the mean absolute organ weights of females treated with 1000
mg/kg/day. The mean absolute pituitary weight was slightly, but significantly, reduced (p<0.01) when compared with the
control females. The males were not affected and this change was considered to be of slight lower mean terminal body
weights. The mean pituitary-to-body weight ratio was also significantly lower (p<0.05) than that of the control females but the
difference was 0.001 (ie 0.007% in the control females vs 0.006% in the females at 1000 mg/kg/day), and the mean pituitaryto-brain weight was significantly lower (p<0.01) when compared with the controls. However, there were no microscopical
changes that would indicate a test item-related response.
The mean testes-to-body weight and the mean epididymides weights of the males treated with 1000 mg/kg/day were
significantly increased (p<0.05 and p<0.01, respectively) when compared with the control males. These differences were
considered to be due to slightly low control values and there were no microscopical differences. The mean epididymides-tobrain weight was also significantly elevated in males treated with 1000 mg/kg/day (p<0.05) but this difference was nearly
identical to the value recorded in the males treated with 62.5 mg/kg/day, and hence was without a dose-response relationship
and toxicological relevance.
There were no differences in the mean absolute or relative organ weights of the males or females treated with 250 mg/kg/day
or 62.5 mg/kg/day when compared with the controls.
After the recovery period, males previously treated with 1000 mg/kg/day had slightly lower absolute mean adrenal weights that
attained statistical significance (p<0.05) when compared with the controls but were without any correlating microscopical
changes. The mean ovary-to-brain weight ratio of the females treated previously with 1000 mg/kg/day were significantly lower
(p<0.05) when compared with the control females.


-Macroscopic Findings
-Unscheduled Deaths
There were two unscheduled deaths, both at 1000 mg/kg/day. One male rat (no. 22), found dead on day 9 of treatment, had
pulmonary findings generally associated with dosing error and which were considered to be the cause of death, rather than an
unidentified systemic toxic effect. One female rat (no. 58) was found dead on day 11 of treatment and also had pulmonary
changes associated with aspiration of dose formulation reflux.
After 4 Weeks of Treatment
There were no test item-related macroscopical changes in males or females after 4 weeks of treatment. All macroscopic
findings seen in males or females occurred at low incidence or without a clear dose-response relationship and were therefore
considered to be unrelated to the test item.
After 2 Weeks of Recovery
There were no late test item-related macroscopical changes in males or females after 2 weeks of recovery. All macroscopic
findings seen in males or females occurred at low incidence, in control rats or without a clear dose-response relationship and
were therefore considered to be unrelated to the test item.


-Microscopic Findings
At the end of the treatment period, microscopic changes related to the treatment with the test item were observed in the
forestomach of animals at 1000 mg/kg/day.
These lesions consisted of acanthosis, parakeratosis, inflammation, ulceration and erosions, and were not seen in the
stomachs of rats in the low-and mid-dose groups. After the 14 days recovery period, these changes were completely
reversible.
In the lungs of some animals treated with 1000 mg/kg/day, amorphous material in the broncho-alveolar lumen was observed at
the microscopic examination. It was the result of aspiration of the formulation following its reflux from the stomach. This
aspiration was the cause of the death of the male and female rats in the high dose group. The amorphous material in turn
caused inflammation, necrosis, edema and emphysema or collapse of the lung parenchyma. However, these lung changes
were not considered a direct toxic effect of the test item.
The examination of the ovaries and the feminine genital tract did not reveal any difference in estrus cycling between control
and treated animals.

Key result

Dose descriptor:

NOAEL

Remarks:

systemic

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect up to the highest dose level

Key result

Dose descriptor:

NOAEL

Remarks:

local

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

histopathology: non-neoplastic

Key result

Critical effects observed:

no

Histopathological findings in stomach : Incidence and Mean Severity Grade
	Males				Females
Group	Control	62.5 mg/kg/day	250 mg/kg/day	1000 mg/kg/day	Control	62.5 mg/kg/day	250  mg/kg/day	1000 mg/kg/day
Animals Examined	5	5	5	5	5	5	5	5
Acanthosis	-	-	-	5/2.4	-	-	-	5/2.6
Parakeratosis	-	-	-	4/2.3	-	-	-	5/2.6
Inflammation	-	-	-	5/1.8	-	-	-	5/2.8
Forestomach ulceration	-	-	-	1/2.0	-	-	-	2/1.5
Forestomach erosion	-	-	-	-	-	-	-	3/1.0

Conclusions:

Hostapon SG was investigated for its repeated dose toxicity in rats according to the Guideline OECD 407. NOAEL of 250 mg/kg/day was assigned fro the forestomach local effects and NOAEL of 1000 mg/kg/day was assigned for the systemic effects.

Executive summary:

The registration substance Hostapon SG was investigated for its repeated dose toxicity in rats according to the Guideline OECD 407. The study included recovery animal groups. Animals were treated at doses of 0, 62.5, 250 and 1000 mg/kg/day. No significant treatment related systemic effect was found up to the highest dose level. At dose of 1000 mg/kg/day microscopic changes in the forestach was found. The lesions consisted of acanthosis, parakeratosis, inflammation, ulceration and erosions. After recovery period of 14 days, these changes were completely reversible. Comparable effects were not seen in the animals of low and mid dose groups. NOAEL of 250 mg/kg/day was assigned for the forestomach local effects and NOAEL of 1000 mg/kg/day was assigned for the systemic effects.

Reference 2

Endpoint:

sub-chronic toxicity: oral

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

other information

Justification for type of information:

Justification is provided in Chapter 13.

Reason / purpose for cross-reference:

assessment report

Reason / purpose for cross-reference:

assessment report

Reason / purpose for cross-reference:

assessment report

Critical effects observed:

not specified

Conclusions:

The chronic toxicity the registration substance Hostapon SG is derived based on the read-across to sodium N-lauroyl sarcosinate. A low chronic toxicity can be reliably derived.

Executive summary:

The chronic toxicity of the registration substance Hostapon SG was evaluated based on the read-across approach using the chronic toxicity data for sodium N-lauroyl sarcosinate

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compounds are likely exhibit comparable toxicity profiles.

Sodium N-lauroyl sarcosinate was investigated in rats for its chronic toxicity and reproduction toxicity in a 2 -year feeding study. The doses levels applied were 0, 0.2, 1 and 2% in the diet. Males and females were housed together so that fertility could be assessed. No effect on the fertility was found. Histopathological changes of stomach tissues were evident at dose levels of 1 and 2% in the diet. No significant systemic effect was found up to the highest dose level.

Sodium N-lauroyl sarcosinate were applied to weanling rats by feeding at 2% in diet for 6 months. No effect on weight gain, feeding efficiency, general health, or behavior was observed, and no abnormalities of the internal organs were observed.

Likewise, the registration substance Hostapon SG is likely of low chronic toxicity.

Reference 3

Endpoint:

chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Validity of the study assessmed by the CIR Expert Panel

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

other: 2-year feeding toxicity study

Version / remarks:

The study integrates the mating of males and females

Principles of method if other than guideline:

2-year feeding toxicity study

GLP compliance:

not specified

Species:

rat

Strain:

Wistar

Sex:

male/female

Route of administration:

oral: feed

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

2 years

Frequency of treatment:

daily (feeding)

Dose / conc.:

20 000 mg/kg diet

Remarks:

0.05% of the test item in the diet for the first 6 months and 2.0% of the test item in the diet for the remaining 18 months

Dose / conc.:

10 000 mg/kg diet

Remarks:

1.0% of the test item in the diet for 24 months

Dose / conc.:

2 000 mg/kg diet

Remarks:

0.2% of the test item in the diet for 24 months

Dose / conc.:

0 mg/kg diet

Remarks:

Basal diet alone

No. of animals per sex per dose:

25 per sex per group

Control animals:

yes, plain diet

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

no effects observed

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

hyperplasia of squamous epithelium with keratin formation of the mucosa of the stomach at two highest dose groups

Histopathological findings: neoplastic:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

20 000 mg/kg diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no effect up to the highest dose

Critical effects observed:

not specified

Citation:

"...Two hundred albino Sherman Wistar rats (25 rats/sex/group; 100-150 g) were fed Sodium Lauroyl Sarcosinate during a 2-year chronic oral toxicity study. Group 1 was fed 0.05% Sodium Lauroyl Sarcosinate in the daily diet for the first 6 months of the study and 2.0% of dies for the remaining 18 months. For the entire study period, group 2 was fed 0.2% of diet, group 3 was fed 1.0% of diet, and group 4 was fed the basal diet alone. Rats were killed at 1,3,6, and 24 months for necropsy and tissues collected for microscopic examination; organs examined included the liver, spleen, heart, lungs, stomach, large intestine, small intestine, adrenal glands, gonads, pancreases, and brain. Blood samples were drawn and analyzed at 30 days, 90 days, 6 months, and 24 months for red and white blood cell counts, hemoglobin content, and differential count. Males and females were housed together so that ferility could be assessed. At 1,3,6 months, no significant differences were observed in lesions, fertility, mortality, hematology, or body weight gain between rats of the control and treated groups. At 24 months, the only consistent difference that could be attributed to the test article was minor hyperplasia of the stratified squamous epithelium with excess keratin formation of the cardiac mucosa of the stomach in rats receiving the highest exposure to the test article-group1 (2% in the diet after 6months) and group 3 (1% in the diet)...."

Conclusions:

Sodium N-Lauroyl sarcosinate was investigated in rats for its chronic toxicity and reproduction toxicity in a 2 -year feeding study. The doses levels applied were 0, 0.2, 1 and 2% in the diet. Males and females were housed together so that fertility could be assessed. No effect on the fertility was found. Histopathological changes of stomach tissues were evident at dose levels of 1 and 2% in the diet. No significant systemic effect was found up to the highest dose level.

Executive summary:

The subacute toxicity of the registration substance Hostapon SG was evaluated based on the read-across approach using the repeated dose toxicity data for sodium N-lauroyl sarcosinate

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compounds are likely exhibit comparable toxicity profiles.

Sodium N-lauroyl sarcosinate was investigated in rats for its chronic toxicity and reproduction toxicity in a 2 -year feeding study. The doses levels applied were 0, 0.2, 1 and 2% in the diet. Males and females were housed together so that fertility could be assessed. No effect on the fertility was found. Histopathological changes of stomach tissues were evident at dose levels of 1 and 2% in the diet. No significant systemic effect was found up to the highest dose level.

Likewise, the registration substance Hostapon SG is likely of low repeated dose toxicity.

Reference 4

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Validity of the study assessmed by the CIR Expert Panel

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

other: 6 month treatment of weanling rats

GLP compliance:

not specified

Species:

rat

Strain:

Wistar

Details on species / strain selection:

Weanling rats

Sex:

male/female

Route of administration:

oral: feed

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

6 months

Frequency of treatment:

daily (feeding)

Dose / conc.:

20 000 mg/kg diet

Remarks:

0.05% of the test item in the diet for the first 6 months and 2.0% of the test item in the diet for the remaining 18 months

No. of animals per sex per dose:

Not mentioned

Control animals:

yes, plain diet

Key result

Dose descriptor:

NOAEL

Effect level:

20 000 mg/kg diet

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: No effect found at the limit dose

Critical effects observed:

not specified

Citation:

"... Weanling rats were given diet containing 2% Sodium Lauroyl Sarcosinate for 6 months. No effect on weight gain, feeding efficiency, general health, or behavior was observed, and no abnormalities of the internal organs were observed. ....."

Conclusions:

Weanling rats were given diet containing 2% Sodium Lauroyl Sarcosinate for 6 months. No effect on weight gain, feeding efficiency, general health, or behavior was observed, and no abnormalities of the internal organs were observed.

Executive summary:

The subacute toxicity of the registration substance Hostapon SG was evaluated based on the read-across approach using the repeated dose toxicity data for sodium N-lauroyl sarcosinate

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compounds are likely exhibit comparable toxicity profiles.

Weanling rats were given diet containing 2% Sodium Lauroyl Sarcosinate for 6 months. No effect on weight gain, feeding efficiency, general health, or behavior was observed, and no abnormalities of the internal organs were observed.

Likewise, the registration substance Hostapon SG is likely of low repeated dose toxicity.

Reference 5

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Validity of the study assessmed by the CIR Expert Panel

Reason / purpose for cross-reference:

read-across source

Key result

Dose descriptor:

NOEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

gross pathology

histopathology: non-neoplastic

organ weights and organ / body weight ratios

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no adverse effect up to the highest dose

Citation:

"... Groups of 15 male and 15 female Sprague-Dawley albino rats were dosed orally by gavage daily with 0, 30, 100, and 250 mg/kg bw Sodium Lauroyl Sarcosinate in distilled water for 91 or 92 days. Body weight gains were decreased in males of the 100 and 250 mg/kg bw groups; the decrease was statistically significant compared to controls for 5 of 13 wks in the middose group and 8 of 13 wks in the high doses group. Absolute stomach weights (in males), stomach-to-body weight ratios, and stomach-to-brain-weight ratios (in males and females) were statistically significantly increased in the 100 and 250 mg/kg bw dose groups, there was an increase in stomach wall thickness and yellow discoloration of non-glandular gastric mucosa, and histopathology revealed an increase in incidence and severity of squamous cell hyperplasia, hyperkeratosis/parakeratosis, inflammation and edema of the non-glandular gastric mucosa in both male and females of these groups. Weights of several other organs that were statistically significantly different from control values were not considered toxicologically significant. There were no toxicologically significant changes in hematology or clinical chemistry parameters. No test material-related mortality was reported. The no-observed effect level (NOEL), lowest observable adverse effect level (LOAEL), and noobservable adverse effect level (NOAEL) were 30, 100, and 250 mg/kg bw/day, respectively......"

Conclusions:

Sodim N-Lauroyl sarcosinate was given to rats by gavage daily at doses of 0, 30, 100, and 250 mg/kg bw for up to 92 days. Body weight gain decrease and effects on the stomach was found for animals of 100 and 250 mg/kg/day. The NOAEL was 250 mg/kg bw/day.

Executive summary:

The subacute toxicity of the registration substance Hostapon SG was evaluated based on the read-across approach using the repeated dose toxicity data for sodium N-lauroyl sarcosinate

The registration substance and the read-across sources are amides of fatty acids and amino acids and can be characterized as "N-fatty acyl amino acids", of which endogeous occurence and metabolism are known. Based on the comparable chemical structure, comparable phys-chem data and expected comparable metabolism, these compounds are likely exhibit comparable toxicity profiles.

Sodim N-Lauroyl sarcosinate was given to rats  by gavage daily at doses of 0, 30, 100, and 250 mg/kg bw for up to 92 days. Body weight gain decrease and effects on the stomach was found for animals of 100 and 250 mg/kg/day. The NOAEL was 250 mg/kg bw/day. The NOEL was 30 mg/kg/day and the NOAEL 250 mg/kg bw/day.

Likewise, the registration substance Hostapon SG is likely of low repeated dose toxicity.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

Reliable and robust database:
- One 28-day toxicity study on the registration substance, chronic toxicity study on the structurally related substance. Consistent outcomes in all available studies
- Consistent outcomes when compared to the metabolism considerations

Justification for classification or non-classification

The registration substance was investigated for its repeated dose toxicity according to the Guideline OECD 407. No significant systemic effect was found up to the dose of 1000 mg/kg/day and no significant local effect was found up to the dose of 250 mg/kg/day. No classification is warranted.