Resinoid of Boswellia serrata (Burseraceae) obtained from exudate by hexane extraction

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2020

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2020

Report date:

2020

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Resinoid of Boswellia serrata (Burseraceae) obtained from exudate by hexane extraction

EC Number:

948-256-0

IUPAC Name:

Resinoid of Boswellia serrata (Burseraceae) obtained from exudate by hexane extraction

Specific details on test material used for the study:

Name: Resinoid of Boswellia serrata (Burseraceae) obtained from exudate by hexane extraction
EC number: 948-256-0
Batch number: 06/1234
Appearance: Amber crystal
Purity: 100% UVCB
Expiry date: 31 May 2020
Storage conditions: Refrigerated (2-8°C), protected from light and humidity (store in a tightly closed container)
Safety precautions: Routine safety precautions (gloves, goggles, face mask, lab coat) for unknown materials were applied to assure personnel health and safety.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Samples were taken at the test concentration as well as from the control, at the beginning and at the end of the renewal periods.

Test solutions

Vehicle:

yes

Details on test solutions:

Reconstituted water (ISO medium, according to OECD 202) was used as dilution water for both the range finding and definitive tests. The same composition of reconstituted water was used for the tests and for breeding the test animals.
OECD medium was constitued of stock solution 1 (macro nutrients), stock solution 2 (iron), stock solution 3 (trace elements) and stock solution 4 (bicarbonate).

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

Species and strain:
Daphnia magna
Source:
István Szent University,
2100 Gödöllő, Páter Károly u. 1, Hungary
Breeding
The Daphnia are bred in Ecotoxicological Laboratory of Charles River Laboratories Hungary Kft.
The health of the stock animals is continuously monitored by visual daily checking. Abnormal behaviour or significant decrease of population is recorded.
Justification of strain:
Daphnia magna is the standard species of the acute immobilisation test.
Number of animals:
There were 20 animals in test group and control group respectively, divided into 4 replicates (5 animals / replicate)
Age of the animals:
They were less than 24 h old at the beginning of the test
Acclimatization:
There was no acclimatization because the water used was similar to the culture water.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

The immobility of the Daphnia was determined by visual observation 24 and 48 hours after the start of the test. Those animals not able to swim within 15 seconds after gentle agitation of the test beaker are considered to be immobile.
The number of immobilised animals and the percentage of immobility were determined at 24 and 48 hours.

Test conditions

Hardness:

The reconstituted water (ISO medium) had a total hardness of 249 mg/L (as CaCO3).

Test temperature:

The water temperature was measured at the start and at the end of the renewal periods in one control test vessel. The test temperature was in the range of 20.6 – 20.8°C measured in the test vessels.
The additionally measured temperature in the climate chamber was between 20.4 – 21.0°C.

pH:

The pH of the test solution was not adjusted and not varied by more than 1.5 units in any one test. The pH was measured at the start and at the end of the renewal periods in one test vessel both in the control and test item group and was in the range of 7.02 – 7.26.

Dissolved oxygen:

The dissolved oxygen concentration was measured in the control and in the test concentration in one replicate at the start and at the end of the renewal periods and was in the range of 7.4 – 8.6 mg/L.

Nominal and measured concentrations:

The concentration levels used (48 h) of the Preliminary Range-Finding Test are summarised below:
Nominal concentrations (mg/L nominal loading rate WAF): untreated control, 0.1, 1.0, 10.0, 100.0.


Because significant inhibition was not observed at the highest concentration level during the preliminary range-finding test, only one test concentration in the test medium (100 mg/L nominal loading rate WAF) and one control group were tested in a Limit Test.

Details on test conditions:

Light: The test was carried out in 16-hour light and 8-hour dark cycle.

PERFORMANCE OF THE TEST
The test duration was 48 hours. Twenty animals, divided into four groups (glass beaker with a volume of 50 mL) of five animals each (~40 mL test solution/flask; 8 mL test solution/animal) were used at the test concentration and for the control in a semi-static system. The animals were not fed during the test.
Based on the analytical method validation, the test was performed under semi-static conditions. The frequency of the water renewal periods was 24 hours.
The choice of the test concentration was done on the basis of the results of a preliminary range-finding test.

-Preliminary range-finding test
A concentration range-finding test was conducted to determine the approximate toxicity of the test item so that appropriate test concentrations could be selected for use in the definitive test. Ten daphnids (divided into two replicates) in each test concentration and control were exposed for 48 hours under semi-static conditions (in absence of stability data).
During the formulation procedure the stock solution was prepared by the same method described above.
The concentration levels used (48 h) of the Preliminary Range-Finding Test are summarised below:
Nominal concentrations (mg/L nominal loading rate WAF): untreated control, 0.1, 1.0, 10.0, 100.0.

-test item concentration in the definitive test
Because no significant toxic response was observed during the preliminary concentration range-finding test, a limit test was carried out using only one test concentration at 100 mg/L nominal loading rate WAF and one control group in a semi-static system.
The biological results are based on the nominal loading rate (WAF).


OBSERVATIONS
The immobility of the Daphnia was determined by visual observation 24 and 48 hours after the start of the test. Those animals not able to swim within 15 seconds after gentle agitation of the test beaker are considered to be immobile.
The number of immobilised animals and the percentage of immobility were determined at 24 and 48 hours.
The water temperature, the oxygen concentrations and pH of the control and the test solution were measured at the beginning and at the end of the renewal periods.

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Effect concentrations 2

Key result

Duration:

24 h

Dose descriptor:

EL50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Effect concentrations 3

Key result

Duration:

48 h

Dose descriptor:

EL100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Effect concentrations 4

Key result

Duration:

48 h

Dose descriptor:

NOELR

Effect conc.:

ca. 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat. (dissolved fraction)

Basis for effect:

mobility

Results with reference substance (positive control):

There were no immobilized animals in the control group and the dissolved oxygen concentration at the end of the test in the control and test vessels was more than 3 mg/L.
All validity criteria were within acceptable limits and therefore the study can be considered as valid.

Reported statistics and error estimates:

No statistical analysis was performed because of the lack of toxic effects.
The EL50, NOELR, LOELR and EL100 values were determined directly from the raw data.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Executive summary:

Acute toxicity of Resinoid of Boswellia serrata (Burseraceae) obtained from exudate by hexane extraction was assessed with Acute immobilisation test on Daphnia magna, over an exposure period of 48 hours in a semi-static system.

 

Under the conditions of this Daphnia magna acute immobilisation study the observed endpoints for the effect of Resinoid of Boswellia serrata (Burseraceae) obtained from exudate by hexane extraction were the following:

24h and 48h EL50>100mg/L

48NOELR=100mg/L