Fatty acids, tall-oil, magnesium salts

Endpoint summary

Description of key information

The hazard information on the substance is based on read across from Tall oil and 2-ethylhexanoic acid and reveals low toxicity. Furthermore, the source substances contain higher or similar concentrations of the constituents used for read across compared to the target substance. There are no scientific reasons indicating that the constituents of the substance can interact in a way that will influence the toxicological/ecotoxicological properties of the substance. The parental NOEL was considered to be 1000 p.p.m. For reproductive parameters the NOEL was considered to be 5000 p.p.m

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2002-03-02 to 2002-07-02

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Justification for read-across, see attached file.

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

There was no certificate of analysis or detailes of the test item supplied by the sponsor. Given the nature of the test item (distillation products) it was considered that this deviation from the protocol did not affect the intergrity od the study. As the test item was a complex mixture of distillation products the study was conducted on the tst item as recieved by the sponsor (Pine Chemicals Association)

GLP compliance:

yes (incl. QA statement)

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Route of administration:

oral: feed

Vehicle:

acetone

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Diet prepared for Week 1 and Week 4 of treatment was sampled.

Duration of treatment / exposure:

The males were treated for at least 4 weeks overall, starting from 2 weeks prior to mating until termination.
The females were treated for 2 weeks prior to mating, then through mating until termination after Day 4 of lactation

Frequency of treatment:

The animals were dosed continuously via the diet.

Dose / conc.:

0 ppm

Dose / conc.:

1 000 ppm

Dose / conc.:

5 000 ppm

Dose / conc.:

20 000 ppm

No. of animals per sex per dose:

Four groups of 10 male and 10 female

Control animals:

yes, concurrent no treatment

Clinical signs:

no effects observed

Description (incidence and severity):

At 20000 p.p.m: Increased male liver weight, small decreases were noted in adrenal gland weight in both sexes, small decreases in ovary weight in, females; slightly increased spleen weight in males.

At 5000 p.p.m. liver weight in male was increased. Female adrenal gland weight was reduced.

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

At 20000 p.p.m. in-life observations included decreased weight gain.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

At 20000 p.p.m. in-life observations included decreased food consumption in both sexes.

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

not examined

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

At 20000 p.p.m. increases in bilirubin and alkaline phosphatase were noted in both sexes, small decreases in albumin and white blood cell count in females, cholesterol were slightly increased in males.

At 5000 p.p.m alkaline phosphatase in both sexes were increased.

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Other effects:

effects observed, treatment-related

Description (incidence and severity):

The only indication of reproductive toxicity was a marginal decrease in implant sites at 20000 ppm with a corresponding decrease in the mean total number of pups born compared to all other dose groups. However, due to the very slight differences compared to the Control group, there is some doubt as to the reproducibility of this finding.

Details on results:

LINICAL SIGNS AND MORTALITY: There were no clinical signs of toxicity or deaths.


BODY WEIGHT AND WEIGHT GAIN: At 20000 ppm there was a transient decrease in weight gain in both sexes. In males decreased weight gain was most notable for over the first week, although absolute weights were significantly lower over the first 3 weeks of treatment. In females there was a notable decrease throughout the pre-mating phase. The resulting deficit in body weight was never regained in either sex. In pregnant females reduced weight gain was evident over Day 7-20 of gestation, compared to the Control animals. There were no effects on body weight at 5000 and 1000 ppm.


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): At 20000 ppm food consumption in males was reduced for the first 2 weeks of treatment (attaining significance during Week 1) and in Week 4 (not recorded Week 3 as paired for mating). In females, food consumption was significantly decreased during the premating period. Consumption was also reduced during the first half of the gestation period, compared to the Control
animals. There were no effects on food consumption at 5000 and 1000 ppm.


HAEMATOLOGY (See Table 4): At 20000 ppm there was a non-significant decrease in white blood cells in females.


CLINICAL CHEMISTRY (See Table 4): Alkaline phosphatase levels were significantly increased in females at 5000 and 20000 ppm, and in males at 20000 ppm. In males there was a non-significant increase in levels at 5000 ppm and in females at 1000 ppm there was an equivocal increase, but given the small group size it was considered that the difference was too small to reflect an effect of treatment. Total bilirubin was increased in both sexes at 20000 ppm. In addition, at 20000 ppm, cholesterol levels were increased in males; albumin (and consequently total protein) were reduced in females.


ORGAN WEIGHTS (see Table 3): In males, at 20000 ppm there was a decrease in body weights, with liver weights being essentially similar to Controls. At 5000 and 1000 ppm liver weight was slightly greater than Controls. Following covariance analysis there was a dose related increase in liver weights, with the increases at 5000 and 20000 ppm attaining statistical significance. In females, slight non-significant increases in liver weights following covariance analysis at 5000 and 20000 ppm were too small to attribute to treatment. In males at 20000 ppm spleen weight was notably increased following variance and covariance analysis. Adrenal gland and thymus weights were slightly but significantly decreased. Following covariance analysis adrenal gland weight was still significantly decreased, but for the thymus there was no significant difference from Controls. In females, ovary, adrenal gland and kidney weights were significantly reduced at 5000 and 20000 ppm, with pituitary gland weight reduced at 20000 ppm. Following covariance analysis, kidney and pituitary gland weights were essentially similar to Controls, but a decrease in ovary weight at 20000 ppm, and adrenal gland weight at 5000 and 20000 ppm was still evident, but not significant.


GROSS PATHOLOGY: No findings attributable to treatment.


HISTOPATHOLOGY: No findings attributable to treatment. All histology findings were typical of spontaneously arising background findings in rats of this strain and age.

Key result

Dose descriptor:

NOEL

Effect level:

1 000 ppm

Based on:

test mat.

Basis for effect level:

other: Decreased weight gain and food consumption in both sexes at 20000ppm. Changes in liver function in both sexes at 20000ppm. At 5000 ppm there was increased liver weight and alkaline phosphatase in both sexes.

Key result

Dose descriptor:

NOEL

Effect level:

5 000 ppm

Based on:

test mat.

Basis for effect level:

other: Reproductive toxicity

Critical effects observed:

not specified

Conclusions:

In conclusion, under the conditions of this study, toxicity was exhibited at levels of 5000 and 20000 p.p.m., but there were no clear effects of toxicity at 1000 p.p.m.  Therefore the parental NOEL was considered to be 1000 p.p.m. For reproductive parameters the NOEL was considered to be 5000 p.p.m.

Executive summary:

Aim:

The test item, Tall Oil (CAS No. 8002-26-4), is a chemical undergoing testing by the Sponsor. The objective of this study was to provide initial information on possible effects on reproduction and development in rats.

Method:

Four groups of 10 male and 10 female Sprague-Dawley rats received the test item via the diet at concentrations of 0, 1000, 5000 and 20000 p.p.m. Tall Oil. The males were dosed for at least 4 weeks, starting from 2 weeks prior to mating. The females were dosed from 2 weeks prior to mating until at least Day 6 of lactation.

The animals were monitored for clinical signs, body weight, food consumption, mating and litter performance.

Blood samples were taken from 5 males and 5 females per group for laboratory investigations. Males were sampled during Week 5: females were sampled on Day 6 of lactation. All animals were subjected to necropsy, which included weighing of major organs.  Histopathology was conducted on tissues from 5 males from Control and High dose, and 7 females from the Control and 8 females from the High dose.

Results

At 20000 p.p.m. in-life observations included decreased weight gain and food consumption in both sexes. Increased male liver weight following covariance analysis, and increases in bilirubin and alkaline phosphatase were noted in both sexes.  In addition, small decreases were noted in adrenal gland weight in both sexes, and in albumin, white blood cell count and ovary weight in females; spleen weight and cholesterol were slightly increased in males.

At 5000 p.p.m. liver weight in males and alkaline phosphatase in both sexes were increased.  Female adrenal gland weight was reduced. The only indication of reproductive toxicity was a marginal decrease in implant sites at 20000 ppm with a corresponding decrease in the mean total number of pups born compared to all other dose groups.  However, due to the very slight differences compared to the Control group, there is some doubt as to the reproducibility of this finding.

Conclusion

In conclusion, under the conditions of this study, toxicity was exhibited at levels of 5000 and 20000 p.p.m., but there were no clear effects of toxicity at 1000 p.p.m.  Therefore the parental NOEL was considered to be 1000 p.p.m. For reproductive parameters the NOEL was considered to be 5000 p.p.m.

Reference 2

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EPA OTS 795.2600 (Subchronic Oral Toxicity Test)

GLP compliance:

yes

Species:

mouse

Strain:

B6C3F1

Sex:

male/female

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Diets were prepared containing 0.0, 0.1, 0.5 or 1.5% of the test material. The diets were frozen in closed amber glass bottles until used. The test material was stable for at least 35 days in the diets when refrigerated. Stability, homogeneity and concentration analyses were conducted using GC.

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analyzed concentrations (mean ± SD; GC): 0.091 (± 0.004), 0.45 (± 0.02) and 1.45 (± 0.05)%

Duration of treatment / exposure:

91-93 days

Frequency of treatment:

7 days/week

No. of animals per sex per dose:

10

Control animals:

yes, plain diet

Details on study design:

- Dose selection rationale: based on a 14-day feeding study in mice
- Post-exposure recovery period in satellite groups: 28-29 days

Positive control:

Not included

Observations and examinations performed and frequency:

CLINICAL OBSERVATIONS: Yes
- Detailed observations: on the mornings of body weight measurement.
- Cage side observations: every workday afternoon and on mornings on which body weights were not collected. Animals were checked for mortality on weekends.

BODY WEIGHT: Yes
- Time schedule for examinations: days 0, 3, 7, and at least once weekly thereafter.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Feed weights (g) were collected on days 3, 7, and at least once weekly thereafter.

OPHTHALMOSCOPIC EXAMINATION: Yes, using an indirect ophthalmoscope after dilation of the pupils with 1% Mydriacyl.
All mice were examinde prior to the start of the study. During the last week of exposure, five male and five female non-recovery animals from each dose level were examined.

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at necropsy
- Anaesthetic used for blood collection: Yes (CO2 anaesthesia)
- Animals fasted: overnight
- How many animals: five animals per sex, per dose.
- Parameters: hemoglobin concentration, hematocrit, red blood cell count, white blood cell count, differential white blood cell count, platelet count, red blood cell indices, and examination of the blood smears for cellular morphology.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at necropsy
- Animals fasted: overnight
- How many animals: five animals per sex per dose
- Parameters: Non-recovery groups: aspartate aminotransferase, alanine aminotransferase, total bilirubin, total protein, albumin, creatinine, urea nitrogen, glucose, gamma glutamyl transpeptidase, triglycerides, cholesterol, sodium, potassium, chloride, calcium and phosphorus. Recovery groups: alanine aminotransferase, total bilirubin, urea nitrogen, triglycerides, cholesterol.

URINALYSIS: Yes
In the week prior to termination of exposure, five males and five females from each dose group were place in metabolism cages for 24-hour urine collections. Parameters: specific gravity, osmolality, volume, glucose, bilirubin, ketones, blood, protein, urobilinogen, nitrite, leukocytes and pH.

Sacrifice and pathology:

Organ weights: liver, kidneys, adrenal glands, testes, ovaries, brain.
Histopathology: all non-recovery high-dose and control animals: trachea, lungs, heart, aorta, esophagus, stomach, duodenum, jejunum, ileum, cecum, colon, rectum, pancreas, liver, salivary glands, kidneys, urinary bladder, pituitary gland, adrenal glands, thyroid glands, parathyroid glands, thymus, spleen, mesenteric lymph nodes, bone marrow (femoral), brain (including sections of medulla/pons, cerebellar cortex, and cerebral cortex), sciatic nerve, quadriceps femoris, testes, ovaries, vagina, uterus, fallopian tubes, sternum with bone marrow, and gross lesions.
The liver, kidneys, lungs, target organs, and gross lesions for animals from all dose levels were examined.
Because no signs of toxicity or target organ involvement was observed, no histopathology was performed on cervical/mid-thoracic/lumbar spinal cord, epididymides, male accessory sex glands, male mammary gland, female mammary gland, femur (including articular surface), skin, and exorbital lachrymal glands.
For recovery animals, histopathology was performed on the liver, kidneys, lungs, and gross lesions.

Statistics:

Means were calculated for body weight, feed consumption, and organ weights. Numerical data were evaluated using Bartlett's test, one-way ANOVA, and Duncan's multiple range test.

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

A reduced mean body weight was observed in males and females of the high-dose group (5.2% and 13.8% below control weights on day 91, respectively) and in females of the mid-dose group (5.6% below control weights on day 91). At the end of the recovery period, body weight was still 4.7% and 10.1% below control weights in males and females of the high-dose group, respectively.

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

In males of the high-dose group, feed consumption was below control levels at several time points. In females of the high-dose group, this occured once in the main study group, but consumption was almost continuously below control level in the recovery group. At the end of the recovery period, high-dose males consumed slightly more than the control and high-dose females consumed slightly less the the control group, but neither difference was statistically significant.

Food efficiency:

effects observed, treatment-related

Description (incidence and severity):

In high-dose males, bilirubin and triglyceride levels were reduced. Cholesterol was elevated in the mid- and high-dose males and alanine aminotransferase (ALT) only in the high-dose group.
Females of the high-dose group exhibited increased levels of urea nitrogen. Bilirubin and triglyceride levels were below control levels in the mid- and high-dose groups, and cholesterol levels were higher relative to controls.
In the recovery males, ALT, bilirubin and triglycerides were lower than controls for the high-dose group, and cholesterol levels were increased, though not statistically significant. In the recovery females, no effects were observed.

Urinalysis findings:

effects observed, treatment-related

Description (incidence and severity):

Dose-dependent elevations in ketone levels were noted in females, which returned to control levels by the end of the recovery period.

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

- Liver weights were increased in high-dose animals. At the mid-dose level, relative liver (to body) weight was increased in males and females and absolute and relative (to brain) liver weights were increased in mid-dose males only. At the end of the recovery period, liver weight differences had largely disappeared.
- Differences in kidney weight (viz. increased relative kidney/body weight in mid- and high-dose females, increased in relative kidney/brain weight in high-dose females, decreased relative kidney/brain weight in high-dose males at the end of the recovery period) reflect differential growth in body and brain weights rather than any significant differrences in kidney weight. This also applies to the observed decrease in adrenal gland weight in mid- and high-dose females, and to the increase in relative testes weight in the high-dose males after the exposure period.
- Absolute brain weight for the high-dose females was decreased at the end of exposure, but not after the recovery period. In the high-dose males, an increased relative brain/body weight was observed at the end of exposure and after the recovery phase. These differences reflect growth retardation early in the animals' lives.

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Hepatocyte hypertrophy with dose-dependent severitywas observed in all animals of the high-dose group and in males of the mid-dose group. The hypertrophy was characterized by an increase in the cell size with compression of the adjoining sinusoids. Affected hepatocytes were primarily located in the portal area, but in the high-dose group the lesions were diffuse and more eosinophilic. Furthermore, a decreased incidence of small cytoplasmic vacuoles was observed in a dose-dependent manner.
Slight increases in cytoplasmic basophilia of the proximal convoluted tubules were observed in the kidneys of the high-dose group. The same cells contained small numbers of cytoplasmic vacuoles, brush borders were absent and nuclei were slightly enlarged and vesicular with marginated chromatin (4/10 males and females were affected).
Stomach lesions in high-dose males consisted of minimal acanthosis and hyperkeratosis of the non-glandular forestomach (6/10 animals were affected).
At the end of the recovery period, the observed changes were largely reversed. Single animals still showed hepatocyte hypertrophy and decreased severity and amount of vacuolization.

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: other: Reduced feed consumption and decreased rate of body weight gain

Key result

Critical effects observed:

not specified

The liver enlargement was considered to be primarily an adaptive change rather than a toxic effect.

Conclusions:

a NOAEL of ca. 200 mg/kg bw/day based on decreased body weight gain and reduced feed consumption.

Reference 3

Endpoint:

sub-chronic toxicity: oral

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

See attached file for justification for read across

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Key result

Dose descriptor:

NOEL

Effect level:

1 000 ppm

Based on:

test mat.

Basis for effect level:

other: Parental toxicity

Key result

Dose descriptor:

NOEL

Effect level:

5 000 ppm

Based on:

test mat.

Basis for effect level:

other: Reproductive Toxicity

Dose descriptor:

NOAEL

Effect level:

200 mg/kg bw/day (nominal)

Based on:

test mat.

Basis for effect level:

body weight and weight gain

food consumption and compound intake

Critical effects observed:

not specified

Conclusions:

In conclusion, under the conditions of this study, toxicity was exhibited at levels of 5000 and 20000 p.p.m., but there were no clear effects of toxicity at 1000 p.p.m.  Therefore the parental NOEL was considered to be 1000 p.p.m. For reproductive parameters the NOEL was considered to be 5000 p.p.m.

Executive summary:

In the Tall oil sub-chronic study, the parental NOEL was considered to be 1000 p.p.m. For reproductive parameters the NOEL was considered to be 5000 p.p.m.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

200 mg/kg bw/day

Study duration:

chronic

Quality of whole database:

Studies have been carried out according GLP and the recommended OECD guideline

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

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

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

According to the study results on crude tall oil and 2 -Ethylhexanoic acid the substance requires no clasification, because the results are not sufficient for classification.