Fatty acids, rape-oil, mixed esters with 1,4:3,6-dianhydro-d-glucitol, sorbitan and sorbitol

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

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

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions (non-GLP, limited documentation, no opthalmoscopy and neurobehavioural examination, 15 animals/dose group, no NOAEL identified).

Data source

Reference

Reference Type:

publication

Title:

Short-term toxicity study of sorbitan monolaurate (Span 20) in rats

Author:

Cater, B.R. et al.

Year:

1978

Bibliographic source:

Food Cosmet Toxicol 16(6): 519-26

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

yes

Remarks:

(non-GLP, limited documentation, no opthalmoscopy and neurobehavioural examination, 15 animals/dose group)

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Sorbitan laurate

EC Number:

215-663-3

EC Name:

Sorbitan laurate

Cas Number:

1338-39-2

Molecular formula:

C18H34O6

IUPAC Name:

1,4-anhydro-6-O-dodecanoyl-D-glucitol

Details on test material:

- Physical state: oily liquid
- Analytical purity: approx. 95%
- Impurities (identity and concentrations): water (max. 1%), sulphated ash (max. 0.25%), arsenic (max. 3 ppm), lead (max. 10 ppm), copper (max. 50 ppm), zinc (max. 25 ppm)

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: A. Tuck & Son, Rayleigh, Essex
- Housing: 5 per cage

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The animals were fed a diet (ground Spratts Laboratory Diet No.1) containing the test substance.

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

90 days

Frequency of treatment:

continuously (via diet)

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
2100, 4200, 8000 mg/kg bw/day
Basis:
other: mean intake of test compound, males

Doses / concentrations 2

Remarks:

Doses / Concentrations:
2300, 4500, 8400 mg/kg bw/day
Basis:
other: mean intake of test compound, females

Doses / concentrations 3

Remarks:

Doses / Concentrations:
2.5, 5.0, 10.0 %
Basis:
nominal in diet

No. of animals per sex per dose:

15

Control animals:

yes, concurrent no treatment

Details on study design:

The animals were weighed and the food and water intakes of each cage was measured weekly.

Positive control:

no

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: No data

DETAILED CLINICAL OBSERVATIONS: Yes

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE :
- Food consumption for each cage was determined and mean daily diet consumption calculated as g food/kg body weight/day

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 2, 6, and 13 weeks
- Anaesthetic used for blood collection: Yes (at 13 weeks collection): barbiturate anaesthesia
- Animals fasted: no
- How many animals: all
- Parameters examined: haemoglobin, packed cell volume, counts of erythrocytes, reticulocytes and total and differential white blood cells

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at 6 and 13 weeks of treatment (females: all parameter after 6 and 13 weeks, males: total protein, albumin only after 13 weeks)
- Animals fasted: no
- How many animals: all
- Parameters examined: total protein, albumin, activity of glutamic-oxalacetic transaminase, glutamic-pyruvic transaminase and lactic dehydrogenase

URINALYSIS: Yes
- Time schedule for collection of urine: 6 h period during the final week
- Animals fasted: no
- Parameters examined: albumin, glucose, ketones, bile salts, blood

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
brain, pituitary, thyroid, heart, liver, spleen, kidneys, adrenal glands, gonads, stomach, small intestine, caecum

HISTOPATHOLOGY: Yes
brain, pituitary, thyroid, heart, liver, spleen, kidneys, adrenal glands, gonads, stomach, small intestine, caecum, lung, salivary gland, aortic arch, thymus, various lymph glands, urinary bladder, colon, rectum, pancreas, uterus, skeletal muscle

Other examinations:

Measurement of specific gravity and volume of urine:
6 h period of water deprivation
2 h period following a water load (25 ml/kg)
4 h period following a 16 h period of water deprivation

Statistics:

Student´s t-test for body and organ weights and White (1952) test for food consumption

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

decreased body weights at 5 and 10%

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

lower food consumption

Food efficiency:

not specified

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

effects observed, treatment-related

Description (incidence and severity):

increase in mid dose group males, decrease in high dose females

Ophthalmological findings:

not specified

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Hb, erythrocyte cound, reticulocyte counts changed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not specified

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

increased liver and kidney weights

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

increase of periportal fat at 5 and 10%

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
No deaths or abnormal behaviour was seen.

BODY WEIGHT AND WEIGHT GAIN
Decreased body weights were seen in the 5 or 10% group after 48 h of start of treatment (-19 and -32%, respectively) and remained less until the end of the study (-19 and -27%, respectively), see Table 1. The effect was detectable, but less marked in the 2.5% group (statistically significant after 6-9 days of treatment, about 10%).

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
The average food intake was lower over the treatment period, related to the dietary concentration of the test substance (see Table 1).
Paired-feeding study:
Body weights of treated animals were lower than those of the pair-fed controls. The differences were less than with the ad. lib. controls.

HAEMATOLOGY
Haemoglobin concentration and packed cell volume was decreased in all groups and both sexes in a dose dependent manner beginning with the second week of the study period (see Table 2). Red blood cell counts were decreased at the lower treatment levels, but increased at the highest treatment level, beginning with week 6 in booth sexes. A tendency for increased reticulocyte counts and decrease in total leucocyte counts was seen in both sexes.

CLINICAL CHEMISTRY
Results were similar in the test and the control animals.

WATER CONSUMPTION AND COMPOUND INTAKE
An increase in mid dose group males and a decrease in high dose females was observed for water consumption over the hole study period.

URINALYSIS
There was a tendency for the treated male rats to produce less urine with a higher specific gravity.

ORGAN WEIGHTS
Absolute weights of brain, heart, parts of gastro-intestinal tract and testes were unchanged or decreased, but relative organ weight was increased in treated groups (see Table 3). Spleen weights were decreased, with unchanged relative organ weight. Absolute and relative liver weights in males and females were increased (10% group). Absolute kidney weights were increased (5, 10% group), relative kidney weights were increased in all dose groups.
Paired-feeding study:
Liver and kidney weights were significantly increased. Relative organ weights of those organs, as well as of heart and small intestine were increased.
HISTOPATHOLOGY: NON-NEOPLASTIC
Periportal vacuolation was observed in the livers in the highest dose group. An increase of periportal fat was determined in females (5, 10% group) and males (10% group). Early nephrosis was observed in test and control animals.

Effect levels

1

Dose descriptor:

NOAEL

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Due to the high dose levels administered, no NOAEL but only a LOAEL can be determined.

Remarks on result:

not determinable

Remarks:

no NOAEL identified

2

Dose descriptor:

LOAEL

Effect level:

2 300 other: mg/kg bw/day: mean intake of test compound

Based on:

test mat.

Sex:

female

Basis for effect level:

other: effects on body weight, food consumption, haematology, organ weights and histopathology

3

Dose descriptor:

LOAEL

Effect level:

2 100 other: mg/kg bw/day: mean intake of test compound

Based on:

test mat.

Sex:

male

Basis for effect level:

other: effects on body weight, food consumption, haematology, organ weights and histopathology

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Table 1: Mean data body weights and on food consumption and resulting intake of test substance

		Body weight (g) at day					Food consumption (g/rat/day) at day
Diet	Dietary level (%)	0#	1	27	55	90	0#	1	27	55	90	Mean food consumption (g/rat/day)	Mean intake of test compound
Males
Control	0	86	93	263	371	432	13.3	14.5	21.3	22	18.9	20.2	0
Test substance	2.5	85	87	238**	332**	389**	10.7	13.3	19.2	19.4	17.5	18.5*	2.1
	5	85	86	214***	294***	350***	9.6	10.1	16.4	18.7	17.8	16.9***	4.2
	10	84	81**	179***	263***	316***	7.5	7.8	16	17.1	15.9	15.4***	8
Females
Control		69	75	176	221	250	10.5	11.7	15.7	16.3	13.7	15.3	0
Test substance	2.5	71	73	162**	205*	233	9.1	9.3	13.1	15.5	14.7	14.7	2.3
	5	71	70*	152***	198**	225**	7.3	6.8	12.9	15.5	14.3	13.6*	4.5
	10	71	68**	141***	172***	191***	4.6	5.9	11.7	12.8	12.3	11.8***	8.4

#: pretreatment value on day 1 of study

*p<0.05, **p<0.01, ***p<0.0001

Table 2: Mean haematological findings after 2, 6 or 13 weeks

						Leucocytes
							Differential (%)
Sex and diet	Dietary level	Hb (g/100 mL)	PCV (%)	RBC (10^6/mm³)	Retics (% of RBC)	Total (10^3/mm³)	N	E	L	M
Week 2
Male
Control	0	14.4	44	6.7	3.4	6.7	11	1	87	1
Test substance	5	13.6*	41*	6.6	1.5**	5.6	12	1	86	1
	10	13**	41*	7.5	4.4	5.5	14	1	84	1
Female
Control	0	14.7	44	7	1.9	5	16	1	82	1
Test substance	5	14.2	41*	6.7	1.1	3.9	11	0	88	1
	10	13***	40**	6.3*	2.7	3.4	11	0	88	1
Week 6
Male
Control	0	15.6	47	7.2	1.2	5.3	11	1	87	1
Test substance	5	14**	42**	6.9	0.8*	5	8	0	91	1
	10	13.2***	41**	8.5*	2.4**	4.4	14	1	84	1
Female
Control	0	15	44	7.1	0.7	5.3	13	1	85	1
Test substance	5	13.9***	41*	6.5*	0.6	4	12	2	85	1
	10	14.2*	41	8*	1	4.4	12	0	88	0
Week 13
Male
Control	0	15.3	45	8	0.3	6	14	1	84	1
Test substance	2.5	15.2	44	7.2*	0.6	4.4**	18	1	80	1
	2.5	14.2**	43**	7.1*	0.7*	4.6*	16	1	81	2
	10	13.7***	41***	8.2	0.9*	4.8*	16	1	83	1
Female
Control	0	14.6	45	7	0.9**	3.7	17	1	81	1
Test substance	2.5	14.8	45	7.2	0.7	3.7	14		84	1
	5	14**	42**	7	0.7	3.6	13	1	85	1
	10	12.3***	39***	8.1**	0.8	4.2	15	1	82	2

*p<0.05, **p<0.01, ***p<0.0001

Hb: Haemoglobin, PCV: Packed cell volume, RBC: Red blood cells, Retics: Reticulocytes, N: Neutrophils, E: Eosinophils, L: Lymphocytes, M: Monocytes

Table 3: Mean relative organ weights (g/100g body weight) of rats feed the test diets after 13 weeks

Sex and diet	Dietary level	Brain	Heart	Liver	Kidneys	Adrenals#	Gonads$	Pituitary#	Thyroid#	Terminal body weight (g)
Male
Control	0	0.46	0.25	2.51	0.56	12.8	0.94	2.2	3.5	418
Test substance	2.5	0.51*	0.27	2.38	0.63***	15.9**	0.98	2.3	3.9	377**
	5	0.55***	0.28	2.54	0.75***	15.7*	1.05	2.3	3.7	338***
	10	0.6***	0.31***	3.67***	0.71***	14.5	1.15***	2.6*	4	305***
Female
Control	0	0.72	0.3	2.5	0.6	25.8	47.1	4.9	4.9	245
Test substance	2.5	0.78*	0.31	2.44	0.69***	26.5	48.5	4.8	4.5	227*
	5	0.79*	0.32	2.67	0.88***	29	47.9	4.5	5.3	221**
	10	0.9***	0.35***	4.39***	0.97***	25.7	50.4	4.3	5.6	190***

*p<0.05, **p<0.01, ***p<0.0001

#: Weights of this organs are express in mg/100 bw

$: Weights of female gonads are expressed in mg/100 g bw