reaction mass of Calcium chloride and Sodium chloride

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1951

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study was conducted before GLP compliance. The study contains sufficient amount of information for interpretation of the results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1951

Report date:

1951

Materials and methods

Test guideline

Qualifier:

no guideline available

Principles of method if other than guideline:

Non-iodized sodium chloride, hereafter referred to as "salt" was fed to rats in their diets for periods of 90 days. A group of 40 Carworth Farms, Wistar
strain, albino rats was selected after a preliminary observation period. The animals were randomized among the 8 cages so that each cage contained 5 male or 5 female rats. The basic diet was the modified Food Research Laboratory Diet 2C which consisted of dried whole milk, ground whole wheat, inactivatedly yeast, U.S.P liver extract,and iodised salt. The iodized salt was present as 2% of the basic diet. The diets were continuously available in McCollum-type food cups. Groups of five males and 5 females received diets that contained 32, 8, 2, and 0% added salt respectively. In terms of total salt added plus that in the basic diet of these groups the percentages were 34, 10, 4, and 2%.

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

Sodium chloride

EC Number:

231-598-3

EC Name:

Sodium chloride

Cas Number:

7647-14-5

Molecular formula:

ClNa

IUPAC Name:

sodium chloride

Constituent 2

Reference substance name:

231-589-3

IUPAC Name:

231-589-3

Details on test material:

- Name of test material (as cited in study report): Sodium Chloride
- Physical state: Salt
- Lot/batch No.: E 277

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Carwoth Farms

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DIET PREPARATION
- Mixing appropriate amounts with (Type of food):The basfe diet was the modified Food Research Laboratory Diet 2C
which consisted of dried whole milk, ground whole wheat, inactivated yeast, U.S.P. liver extract,and iodised salt. The iodized salt was present as 2% of the basic diet.

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

no data

Duration of treatment / exposure:

90 days

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0.00, 1.33, 5.73, 31.04 gms. added salt/kg/day
Basis:
nominal in diet

Doses / concentrations 2

Remarks:

Doses / Concentrations:
1.30, 2.67, 7.16, 32.92 gms total salt/kg/day
Basis:
actual ingested

No. of animals per sex per dose:

5 males and 5 females per dose.

Control animals:

yes, plain diet

Details on study design:

no data

Positive control:

no data

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: No data
- Time schedule:no data
- Cage side observations checked in table [No.?] were included.: No data

DETAILED CLINICAL OBSERVATIONS: No data
- Time schedule:No data

BODY WEIGHT: Yes


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Other examinations:

None of the 40 rats used in this study died. At the end of 91 days of doses, the rats were weighed and sacrificed, Portions of kidney, liver, and lung were taken for histopathological examination, Livers and kidneys were weighed and the mean organ weight, expressed as percentage of body weight, calculated. These data, as well as those obtained on body weight gain and diet consumption were satatistically analysed for the sexes separately and combined.

Statistics:

No statistical differences were found in total diet eaten (or spilled). The mean weight gain of the combined sex groups dfffered significantly from the
controls only in the 3 s group. These rats gained 82 grams in 91 days while the controls gained 174 grams. However, the rats that received 8% and 2% added salt gained only 130 and 156 grams. The correlation between weight gain and salt content of the diet was -0.969, a significant finding that could occur only 5 times in 1000 by chance. When the body weigt data were examined separately by sex, each group of rats was found to have gained statistically significantly less weight than did the male controls (+92.6, +161.6, +217.6, and +238.8 for the 32, 8, 2, and 0% added salt groups ). The females differed only in the 32% groups. when the weight gain of each group was recalculated as grams gain per 100 grams basal diet eaten, the same statistical differences in growth data were found as when the weight data were calculated as grams weight gain.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Description (incidence and severity):

None of the 40 rats used in this study died.

Mortality:

no mortality observed

Description (incidence):

None of the 40 rats used in this study died.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

When the body weight data were examined separately by sex, each group of male rats was found to have gained statistically significantly less weight than did the male controls (+92.6, +161.6, +217.6, and +238.8 for the 32, 8, 2, and 0% added salt groups )

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

no effects observed

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

Livers and Kidneys were weighed and the mean organ weight, expressed as percentage of body weight, calculated.

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Portion of kidney, liver and lung were taken for histopathological examination.

Histopathological findings: neoplastic:

not specified

Details on results:

CLINICAL SIGNS AND MORTALITY: No mortality was observed in all 40 rats
BODY WEIGHT AND WEIGHT GAIN: Weight gain observed and recorded
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Food consumption and compound intake was recorded.
FOOD EFFICIENCY: increased


ORGAN WEIGHTS: Livers and kidneys were weighed


HISTOPATHOLOGY: Portions of kidney, liver and lung were taken for histopathological examination.

Effect levels

Key result

Dose descriptor:

LOAEL

Effect level:

1.33 other: grams per kg/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

Remarks on result:

other:

Remarks:

only in males and at high doses.

Target system / organ toxicity

Key result

Critical effects observed:

yes

Lowest effective dose / conc.:

1.33 other: g/kg/day

System:

other: Body weight

Organ:

other: No organ affected. Only impact on body weight change.

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

yes

Any other information on results incl. tables

Table are attached in attached background material.

Applicant's summary and conclusion

Conclusions:

The LOAEL for 91 doses of salt in the diet of rats is 2%, 1.33 grams/kg/day of added salt in male rats only. Since this doses is high, no classification is required for repeated toxicity according to UN and EU GHS criteria.

Executive summary:

In this study, a group of 5 males and 5 female rats were fed diets for 91 days that contained 32, 8, 2 and 0% salt in excess of the 2% already present as a normal constituent of basic FRL-2C rat diet.

The salt added to the dry diet in this study was Baker and Adamson crystal reagent Sodium Chloride, code 2232 from General Chemical Division of Allied Chemical and Dye Corp., New York, N.Y. It was identified as their lot E 277 and labeled as having passed A.C.S Specifications. This salt does not contain added potassium iodide as did the salt present as 2% of the basic diet.

Non-iodized sodium chloride, was fed to rats in their diets for periods of 90 days. A group of 40 Carworth Farms, Wistar strains, albino rats was selected after a preliminary observation period. The animals were randomized among the 8 cages so that each cage contained 5 male or 5 female rats. The basic diet was the modified Food Research Laboratory Diet 2C which consisted of dried whole milk, ground whole wheat, inactivated yeast, U.S.P. liver extract and iodized salt. The iodized salt was present as 2% of the basic diet. The diet were continuously available in McCollum-type food cups. Groups of five males and 5 females received diets that contained 32, 8, 2 and 0% added salt respectively. In terms of total salt added plus that in the basic diet of these groups the percentages were 34, 10, 4 and 2%.

None of the 40 rats used in the study died. At the end of 91 days of doses, the rats were weighed and sacrifice. Portions of kidney, liver, and lung were taken for histopathological examination. Livers and kidneys were weighed and the mean organ weight, expressed as percentage of the body weight, calculated. These data, as well as those obtained on body weight gain and diet consumption were statistically analyzed for the sexes separately and combined.

No statistical differences were found in total diet eaten. The mean weight gain of the combined sex groups differed significantly from the controls only in the 32% group. These rats gained 82 grams in 91 days while the controls gained 174 grams. However, the rats that received 8% and 2% added salt gained only 130 and 156 grams. The correlation between weight gain and salt content of the diet was -0.969, a significant finding that could occur only 5 times in 1000 by chance. When the body weight data were examined separately by sex, each group of male rats was found to have gained statistically significantly less weight than did the male controls (+92.6, +161.6, +217.6 and +236.8 for the 32,8,2 and 0% added salt groups.). The females differed only in the 32% group. When the weight gain of each group was recalculated as grams gain per 100 grams basal diet eaten, the same statistical differences in growth data were found as when the weight data were calculated as grams weight gain.

The mean liver weights of the 32% group of male and female rats combined and of the females separately were statistically lower than those of their respective controls. The mean kidney weights of the 32% females were significantly higher than those of the controls.

However, no pathological changes were detected by microscopic examination of these organs.

The LOAEL for 91 days of salt in the diet of rats is 2%, 1.33 grams/kg/day of added salt for male rats body weight decrease.