Registration Dossier

Diss Factsheets

Ecotoxicological information

Toxicity to aquatic algae and cyanobacteria

Administrative data

Endpoint:
toxicity to aquatic algae and cyanobacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08.02.2018-18.05.2018
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 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)
GLP compliance:
yes

Test material

Constituent 1
Reference substance name:
Cyperus scariosus, ext.
EC Number:
294-955-2
EC Name:
Cyperus scariosus, ext.
Cas Number:
91771-62-9
IUPAC Name:
Cyperus scariosus, ext.
Test material form:
liquid
Specific details on test material used for the study:
Batch manufactured and supplied by : Jagat Aroma Oils DistIllery
Kannauj-209725
Uttar Pradesh, India

Test item : Caprilo (Cyperus Scariosus ext. Oil)

CAS No. : 91771-62-9/68916-60-9

Physical appearance : Amber coloured or Light Brown Liquid

Purity as per certificate of analysis : 100% Naturally pure

Batch No. : Lot # 11/CYP/2016

Manufactured date : November 2016

Expiry date : October 2018

Recommended storage condition : Ambient (+15 to +25 °C)


Notes: a) Date of receipt of test item at test facility: 05 December 2017
b) Test Item code by test facility: J09-01

Sampling and analysis

Analytical monitoring:
not specified
Details on sampling:
not specified

Test solutions

Vehicle:
no
Details on test solutions:
Test medium was algal medium as described in the OECD test guideline 201 (OECD, 2006) and the preparation details are given in Appendix 5.

Chemical Batch No. Manufacturer
MgCl2. 6H2O MF6M 561553 Merck Specialities Private Limited, Mumbai, India
ZnCl2 B0755316248
CaCl2. 2H2O D14A/1314/0804/31 s.d. fine chem. Limited
Mumbai, India
NH4Cl D14A/2314/2103/31
MgSO4. 7H2O H14A/1813/1707/31
Na2EDTA. 2H2O J08W/2908/2209/31
MnCl2. 4H2O E13A/1613/1204/13
CoCl2. 6H2O H09Z/1709/2203/31
CuCl2. 2H2O G09Z/2109/0106/31
Na2MoO4. 2H2O L15A/3315/0912/31
H3BO3 A16A/1115/0705/31
NaHCO3 D14A/1414/0702/31
KH2PO4 0000136538 Himedia Laboratories Private Limited, Mumbai, India
FeCl3. 6H2O 161215 Central Drug House (P) Limited, New Delhi, India

Test organisms

Test organisms (species):
Pseudokirchneriella subcapitata (previous names: Raphidocelis subcapitata, Selenastrum capricornutum)
Details on test organisms:
Pseudokirchneriella subcapitata ATCC 22662

This algal species was used as the test system as it is a recommended algal species in the guideline.
6.1.2.1 Source
American Type Culture Collection
P.O.Box 1549
Manassas, VA 20108
USA
6.1.2.2 Culture Maintenance
Algal cells were maintained at the test facility through sub-culturing once in 5 to 10 days from the continuous culture into fresh medium in a conical flask. These conical flasks were incubated between 21 and 25°C (±2°C) under a continuous illumination of 8000±1600 Lux.

Study design

Test type:
static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
72 h

Test conditions

Hardness:
not specififed
Test temperature:
22.1 to 22.4 C
pH:
7.80 to 7.83
Dissolved oxygen:
not specififed
Salinity:
not specififed
Conductivity:
not specififed
Nominal and measured concentrations:
not specififed
Details on test conditions:
Algal cells with test concentrations were incubated in the following conditions during range finding and the definitive test.

Parameters Range finding test Definitive test
Duration of the test 72 ± 1 hours 72 ± 1 hours
Test containers 250 mL conical flask 250 mL conical flask
pH of test concentration 7.78 to 7.83 7.80 to 7.83
Temperature of growth chamber (C) 22.1 to 22.4 22.1 to 22.4
Mean continuous light intensity (Lux) 7138 to 7263 7138 to 7263
Reference substance (positive control):
yes

Results and discussion

Effect concentrations
Key result
Duration:
72 h
Dose descriptor:
EC50
Effect conc.:
ca. 83.69 mg/L
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
growth rate

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The EC values calculated for growth rate and yield on the basis of the nominal concentration of the test item are presented below:

At 72 hours EC values (mg/L)
ErC50 83.69
ErC20 2.8756
ErC10 0.4937
EyC50 1.425
EyC20 0.0677
EyC10 0.0138

Note: ErC refers to growth rate and EyC refers to yield.
Executive summary:

The effect of Caprilo (Cyperus Scariosus ext. Oil) was tested on the growth of freshwater single cell green algaPseudokirchneriella subcapitata.

 

Range finding test was carried out to select the test concentrations for the definitive test. In the range finding test, the tested concentration was 0.00001, 0.0001, 0.001, 0.01, 0.1, 1, 10 and 100 mg/L with single replicate. The test item exhibited 4.73, 5.41, 15.88, 29.05, 43.92, 46.62, 85.14 and 88.18 % reduction in cell biomass at the tested concentrations of 0.00001, 0.0001, 0.001, 0.01, 0.1, 1, 10 and 100 mg/L at 72 h.

 

The alga was exposed to the test item at the nominal concentrations of 0.25, 0.8, 2.56, 8.19, 26.21 and 83.87 mg/L (factor of about 3.2) along with a negative control and a reference substance (Potassium dichromate) at 0.9 mg/L. The cell growth was measured at 24, 48 and 72 hours after the initiation of the test.

 

The concentration/effect relationship was determined using two factors namely growth rate and yield at the end of the test (72 hours). The per cent inhibition of growth rate was 11.45, 13.5, 16.23, 19.09, 41.77 and 55.34 % and the percent yield inhibition was 39.39, 44.65, 50.94, 56.78, 84.56 and 92.00 % at the nominal concentrations of 0.25, 0.8, 2.56, 8.19, 26.21 and 83.87 mg/L, respectively, when compared to the negative control. There was 23.98 % and 65.24 % inhibition of average growth rate and yield of alga, respectively, in the reference substance group at 0.9 mg/L. Observed cells were found morphologically normal.

 

The EC values calculated for growth rate and yield on the basis of the nominal concentration of the test item are presented below:

 

At 72 hours

EC values (mg/L)

ErC50

83.69

ErC20

2.8756

ErC10

0.4937

EyC50

1.425

EyC20

0.0677

EyC10

0.0138

 

Note: ErCrefers to growth rate and EyC refers to yield.