Nicotine

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1989

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

2 October 2012

Deviations:

yes

Remarks:

Exposure duration was 16 days instead of 21 days

Qualifier:

according to guideline

Guideline:

EPA OPPTS 850.1300 (Daphnid Chronic Toxicity Test)

Version / remarks:

1982

Deviations:

not specified

Qualifier:

according to guideline

Guideline:

other: American Society for Testing Materials (1980) Standard practice for conducting acute toxicity tests with fish, macroinvertebrates, and amphibians. E729-80, American Society for Testing and Materials, Philadelphia, Pennsylvania

Principles of method if other than guideline:

-Exposure duration was 16 days instead of 21 days
-Species was Daphnia pulex instead of Daphnia magna STRAUS

GLP compliance:

not specified

Specific details on test material used for the study:

purity > 97%

Analytical monitoring:

yes

Details on sampling:

Set A: Chemicals in water --> determination of losses due to physical characteristics such as volatility or adhesion to glass
Set B: Chemicals in test medium, including algae and daphnids --> determination of additional losses from uptake by organisms

Chemical concentrations were analyzed at 1 hr, 48 hr, and 72 hr after preparation to simulate exposure in test media at the beginning and end of the renewal cycle (2-3 d)

Vehicle:

yes

Remarks:

water

Test organisms (species):

Daphnia pulex

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

16 d

Hardness:

160-200 mg/L as CaCO3

Test temperature:

20°C

pH:

not specified

Dissolved oxygen:

8-9 mg/L

Salinity:

not specified

Conductivity:

not specified

Nominal and measured concentrations:

Nominal: 0.02, 0.07, 0.12, 0.18, 0.24 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: 1L beakers, containing five uniquely labeled, glass test tubes (3 cm diameter)
- Fill volume: 700 mL + 1 mL chemical stock
- Renewal rate of test solution (frequency/flow rate): new culture media were provided for each treatment three times a week (Monday, Wednesday, Friday)
- No. of vessels per concentration (replicates): two replicate chambers per concentration, each containing five D. pulex in individual test tubes


TEST MEDIUM / WATER PARAMETERS
- Alkalinity: 100-125 mg/L as CaCO3


OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod:
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : Fecundity and growth

Reference substance (positive control):

no

Key result

Duration:

16 d

Dose descriptor:

LOEC

Effect conc.:

0.18 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Key result

Duration:

16 d

Dose descriptor:

LOEC

Effect conc.:

0.07 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Key result

Duration:

16 d

Dose descriptor:

NOEC

Effect conc.:

0.02 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Key result

Duration:

16 d

Dose descriptor:

NOEC

Effect conc.:

0.12 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

Recovery rates from the gas chromatography analysis at 1 hr after preparation in water only was relatively low (about 50% of expected) for nicotine. At 48 and 72 hr after preparation, concentrations had dropped substantially (<25% of expected). Uptake by algae and daphnids from the test medium did not add to chemical loss 1 hr after preparation but further reduced nicotine to negligible concentrations (<5% of expected) after 48 hr. The estimated LOEC was based on nominal concentrations instead of actual concentrations and therefore indicate upper limits. The actual concentrations of nicotine were much lower than the nominal concentrations, possibly due to adsorption to glassware, the volatility of these contaminants, and uptake by organisms.

Nicotine significantly reduced growth and fecundity of daphnids at nominal concentrations from 0.02 to 0.24 mg/L. The LOEC for length was 0.07 mg/L (NOEC=0.02 mg/L). The LOEC for fecundity was 0.18 mg/L (NOEC=0.12 mg/L). Mortality also increased at higher concentrations (about 70%).

Reported statistics and error estimates:

- Analysis of variance (SAS Institute Inc. 1982)
- A set of orthoganol contrasts was used to compare control with treatments, to determine the first concentration that produced significant differences (p<0.05) in fecundity or growth, designated as the lowest-observable effect concentration (LOEC)

Mortality (%, mean and range) of Daphnia pulex by day 16. Two replicates at each concentration were noted.

mg/L	Mortality [%], mean and range
0.0	10	0 -20
0.02	6	0 -13
0.07	4	0 -7
0.12	10	0 -20
0 .18	20	13 -27
0.24	66	33 -100

Recovery rates (mean % of nominal concentrations, standard error in parentheses) of nicotine through time after preparation in water only and in test media.

Substance	% Recovery in water only			% Recovery in test media
	1 hr	48 hr	72 hr	1 hr	48 hr	72 hr
Nicotine	57 (3.0)	24 (2.0)	9 (4.5)	89 (6.0)	3 (3.0)	0 (0)

Validity criteria fulfilled:

yes

Remarks:

mortality in controls <20% by day 16

Conclusions:

The toxic effect of the test item (S)-Nicotine to Daphnia pulex was assessed in a semi-static concentration-response test. The 16-day LOEC was determined to be 0.07 mg/L for growth and 0.18 mg/L for reproduction. The concentrations were verified in the analytical part. The estimated LOEC is based on nominal concentrations instead of actual concentrations and therefore indicate upper limits. The actual concentrations of nicotine were much lower than the nominal concentrations, possibly due to adsorption to glassware, the volatility, and uptake by organisms.

Executive summary:

A semi-static long-term study was performed using five nicotine concentrations of 0.02, 0.07, 0.12, 0.18, 0.24 mg/L (nominal). Exposure duration was 16 days to test the effect of nicotine on growth and reproduction of Daphnia pulex. One Daphnia neonate was placed in each test tube in each chamber (15 per treatment, 90 per test). The cumulative number of neonates produced by daphnids over 16 d was used to measure fecundity. The test was considered valid since mortality in the controls was <20% by day 16. Nicotine concentrations were quantified by GC, equipped with a photo ionization detector. Nicotine significantly reduced growth and fecundity of daphnids at nominal concentrations from 0.02 to 0.24 mg/L. The LOEC for length (growth) was 0.07 mg/L, the LOEC for fecundity (reproduction) was 0.18 mg/L.

Description of key information

One published long-term study available in Daphnia pulex. The overall quality of this study is considered appropriate for risk assessment (Klimisch 2).

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

0.02 mg/L