Okra, ext.

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Oct - Nov 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Version / remarks:

December 17, 2001

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

(from the competent authority) Landesamt für Umwelt Baden-Württemberg

Test type:

acute toxic class method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Okra, ext.

EC Number:

294-510-2

EC Name:

Okra, ext.

Cas Number:

91723-07-8

Molecular formula:

not applicable

IUPAC Name:

Hibiscus abelmoschus, ext.

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No.of test material: 0016468911
- Expiration date of the Batch: December 09, 2019
- Purity : 100 % UVCB substance (substance of unknown or variable composition, complex reaction products or biological materials)
- Physical state / color: solid / beige

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: refrigerator
- Stability under test conditions: The stability of the test item in the vehicle will be determined indirectly by concentration control analysis. For this purpose, samples taken were stored at room temperature over the maximum duration of the administration period, subsequently deep-frozen and sent to the sponsor for analysis.

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was prepared for each test group shortly before administration by stirring with a high-speed homogenizer and a magnetic stirrer. The homogeneity of the test item preparation during administration was ensured by stirring with a magnetic stirrer.
- Final preparation of a solid: Aqueous preparation corresponds to the physiological medium; vehicle: deionized water

FORM AS APPLIED IN THE TEST (if different from that of starting material) : suspension

Test animals

Species:

rat

Strain:

Wistar

Remarks:

Crl:WI (Han) SPF

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga GmbH, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: approx. 10 weeks
- Weight at study initiation: animals of comparable weight (+/- 20 % of the mean weight)
- Fasting period before study: Feed was withdrawn from the animals at least 16 hours before administration, but water was available ad libitum.
- Housing: single housing in Makrolon cage, type III
- Diet (e.g. ad libitum): R/M maintenance, low phytoestrogen; Ssniff, Spezialdiäten GmbH (Experimental Animal Diets Inc., 59494 Soest, Germany), ad libitum
- Water (e.g. ad libitum): tap water ad libitum
- Acclimation period: at least 5 days before the beginning of the experimental phase; during the acclimatization period, the animals were accustomed to the environmental conditions of the study and to the diet

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C +/- 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12 h / 12 h

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Remarks:

deionized

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 20 g/100 mL
- Amount of vehicle (if gavage): 10 mL/kg bw
- Justification for choice of vehicle: aqueous preparation corresponds to the physiological medium

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg bw

DOSAGE PREPARATION (if unusual): The test item was prepared for each test group shortly before administration by stirring with a high-speed homogenizer and a magnetic stirrer.

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: By request of the sponsor a starting dose of 2000 mg/kg bw was chosen in the first step with 3 female animals. Because no mortality occurred, a further dose of 2000 mg/kg bw was administered to another group of 3 female animals in the second step. As no animal died, the study was terminated.

Doses:

2000 mg/kg bw

No. of animals per sex per dose:

3

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: Individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation. Clinical signs for each animal were recorded several times on the day of administration and at least once during each workday thereafter.
- Necropsy of survivors performed: yes; necropsy with gross-pathology examination was performed on the last day of the observation period after sacrifice by CO2-inhalation in a chamber with gradually increasing concentrations

Statistics:

Calculations were performed using Microsoft Excel 2010 and checke with a calculator.

Results and discussion

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Remarks on result:

other: no mortality occurred

Mortality:

No mortality occurred in both 2000 mg/kg bw test groups.

Clinical signs:

other: No clinical signs were observed during clinical examination in both 2000 mg/kg bw test groups.

Gross pathology:

There were no macroscopic pathological findings in any animal sacrificed at the end of the observation period (6 females in both 2000 mg/kg bw test groups).

Any other information on results incl. tables

Table 1: Mortality

Dose [mg/kg bw]	2000	2000
Sex	Female	Female
Administration	1	2
No. of animals	3	3
Mortality (animals)	No mortality	No mortality

Table 2: Maximum Incidence of Clinical Signs

Dose [mg/kg bw]	2000			2000
Sex	Female			Female
Administration	1			2
No. of animals	3			3
Animal No.	R343	R344	R345	R349	R350	R351
Abnormalities	-	-	-	-	-	-

Table 3: Body weights

Dose [mg/kg bw]	2000					2000
Administration	1					2
Animal No.	R343	R344	R345	Mean weight	Standard deviation	R349	R350	R351	Mean weight	Standard deviation
Body weight at study day [g] 0 7 14	175 200 207	175 196 202	175 197 204	175.0 197.7 204.3	0.00 2.08 2.52	194 222 230	184 204 209	186 207 215	188.0 211.0 218.0	5.29 9.64 10.82

Table 4: Gross pathology

Dose [mg/kg bw]	2000			2000
Administration	1			2
No. of animals	3			3
Animal No.	R343	R344	R345	R349	R350	R351
Macroscopic pathologic abnormalities	-	-	-	-	-	-

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of this study the median lethal dose of the test substance after oral administration was found to be greater than 2000 mg/kg bw in rats.

Executive summary:

In an acute oral toxicity study performed according to the Acute Toxic Class Method, a dose of 2000 mg/kg bw of the test item (preparations in deionized water) was administered by gavage to two test groups of three fasted Wistar rats each (6 females).

2000 mg/kg bw (first and second test group):

- No mortality occurred

- No clinical signs were observed

- All animals gained weight in a normal range throughout the study period.

- There were no macroscopic pathological findings in any animal sacrificed at the end of the observation period (6 females).

The acute oral LD50 was calculated to be greater than 2000 mg/kg bw in rats.