Hydroxychloroquine

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

Study undertaken to GLP certification, based on OECD guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 201 (Freshwater Alga and Cyanobacteria, Growth Inhibition Test)

Deviations:

no

Principles of method if other than guideline:

Guideline study

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Hydroxychloroquine

EC Number:

204-249-8

EC Name:

Hydroxychloroquine

Cas Number:

118-42-3

Molecular formula:

C18H26ClN3O

IUPAC Name:

2-({4-[(7-chloroquinolin-4-yl)amino]pentyl}(ethyl)amino)ethan-1-ol

Test material form:

solid

Remarks:

white solid powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch number of test material:
HYDROXYCHLOROQUINE (CAS N°118-42-3) – Batch 5U054 (Synonym: PLAQUENIL)

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
Nominal concentrations were 0.625, 1.25. 2.5, 5, 10 mg/l
Geometric mean concentrations were 0.403, 0.768, 2.064, 4.832, 9.553 mg/l

- Sampling method:
A Hydroxychloroquine quantification was performed according to the validated analytical method to determine the concentration of test item at T0 and T72h in the test for all tested concentrations

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
A stock solution of test item was prepared by dissolution of the test item in the test media.

Test organisms

Test organisms (species):

Raphidocelis subcapitata (previous names: Pseudokirchneriella subcapitata, Selenastrum capricornutum)

Details on test organisms:

TEST ORGANISM
- Common name: Green algae
- Source (laboratory, culture collection) Culture lab:
- Age of inoculum (at test initiation): 4 days

Study design

Test type:

static

Water media type:

freshwater

Remarks:

OECD TG 201 medium

Limit test:

no

Total exposure duration:

72 h

Remarks on exposure duration:

None

Post exposure observation period:

None

Test conditions

Hardness:

No data (OECD TG 201 medium)

Test temperature:

23°C ± 2°C

pH:

7.7 to 8.7

Dissolved oxygen:

No data (OECD TG 201 medium)

Salinity:

Not applicable

Conductivity:

No data (OECD TG 201 medium)

Nominal and measured concentrations:

Nominal concentrations were 0.625, 1.25. 2.5, 5, 10 mg/l
Geometric mean concentrations were 0.403, 0.768, 2.064, 4.832, 9.553 mg/l

Details on test conditions:

TEST SYSTEM
- Test vessel: glass beaker of 250mL filled with 100ml of test solution
- Type (delete if not applicable): closed with synthetic wadding
- Initial cells density: 5.18 * 10^3 cells/ml
- Control end cells density: 1.058 * 10^6 cells/ml
- No. of vessels per concentration (replicates): 3
- No. of vessels per control (replicates): 6

GROWTH MEDIUM
- Standard medium used: yes (OECD TG 201)

OTHER TEST CONDITIONS
- Photoperiod:constant illumination

Reference substance (positive control):

yes

Remarks:

Potassium dichromate (K2Cr2O7)

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

3.11 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 2

Duration:

72 h

Dose descriptor:

EC50

Effect conc.:

1.83 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks:

yield

Effect concentrations 3

Key result

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

1.83 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 4

Duration:

72 h

Dose descriptor:

EC10

Effect conc.:

0.84 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks:

yield

Effect concentrations 5

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

0.768 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 6

Duration:

72 h

Dose descriptor:

NOEC

Effect conc.:

0.768 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks:

yield

Effect concentrations 7

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

2.06 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

growth rate

Effect concentrations 8

Duration:

72 h

Dose descriptor:

LOEC

Effect conc.:

2.06 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

biomass

Remarks:

yield

Details on results:

- Exponential growth in the control (for algal test): yes

Results with reference substance (positive control):

- Results with reference substance valid? Yes
Potassium dichromate (K2Cr2O7) concentration 0.92 - 1.46 mg/l
72 hour EC50: 1.01 mg/l

Reported statistics and error estimates:

Endpoints and confidence limits were obtained by calculation of ECx according to Regtox® Macro (“Vindimian”) using the Hill model. NOEC and LOEC were estimated by statistic comparison using Statgraphics® software. Data homoscedasticity was checked by Levene test and data were compared by ANOVA.
Tukey HSD test (Honestly Significant Difference. with p < 0.05) was used for pairwise comparisons of several treatment groups.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The reliable (Klimisch 1) OECD 201 Guideline study on algal growth inhibition reported a 72-hr EC50 of 3.11 mg/l and an EC10 of 1.83 mg/L based on growth of Pseudokirchneriella subcapitata (measured geometric mean).

Executive summary:

The reliable OECD 201 Guideline algal growth inhibition study was undertaken to GLP standards. Ecotoxicity test results were based on geometric mean of measured concentrations

Results according to OECD 201 test guideline:

	HYDROXYCHLOROQUINE
Growth inhibition	BPL15-0032.001
ErC50-72h Confidence limits 95%	3.11 mg/L  (2.96 – 3.27 mg/L)
ErC10-72h Confidence limits 95%	1.83 mg/L (1.70 – 1.98 mg/L)
NOEC	0.768 mg/L
LOEC	2.06 mg/L
Inhibition of yield
EyC50-72h Confidence limits 95%	1.83 mg/L (1.45 – 2.26 mg/L)
EyC10-72h Confidence limits 95%	0.840 mg/L (0.489 – 1.59 mg/L)
NOEC	0.768 mg/L
LOEC	2.06 mg/L

 

ErC50-xh: Effective Concentration corresponding to 50% of growth inhibition.

ErC10-xh: Effective Concentration corresponding to 10% of growth inhibition.

EyC50-xh: Effective Concentration corresponding to 50% of yield inhibition.

EyC10-xh: Effective Concentration corresponding to 10% of yield inhibition.

NOEC: No Observed Effect Concentration.

LOEC: Lowest Observed Effect Concentration.