Salicylaldehyde

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

16 March 1981 - 20 March 1981

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Remarks:

The results of this study on the registered substance are a part of a whole battery of fish acute toxicity tests on 198 substances. Even though the test was not performed according to guideline and GLP, the study follows main recommendations of OECD 203 TG and US EPA 850-1075 about the method and fulfil validity criteria: - the mortality did not exceed 10% in the control at the end of the exposure - analytical measurement of test concentration (+/-85%) - dissolved oxygen concentration was maintained between 60 and 100 % for most of tests except in 12 tests where limits were exceed. The minor vulnerabilities are: - No accurate details about dissolved oxygen concentration was for the registered substance study - Reported exposure are the mean of the before and after exchange values: Raw data are not available

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

not applicable

Qualifier:

equivalent or similar to guideline

Guideline:

EPA OPPTS 850.1075 (Freshwater and Saltwater Fish Acute Toxicity Test)

Deviations:

not applicable

Principles of method if other than guideline:

Although this study was not performed according to guidelines, the study is well detailled and follow similar recommendations to guidelines.

GLP compliance:

no

Analytical monitoring:

yes

Details on sampling:

All test exposure chambers were sampled at approximately mid-depth at 0 and 96 hr and one of each duplicate exposure chamber at 24, 48 and 72 hr.

Vehicle:

no

Test organisms (species):

Pimephales promelas

Details on test organisms:

Test Organism:
a) Supplier: Test organism was cultured from brood stock provided by the U.S. EPA Environmental Research Laboratory-Duluth.
b) Age: 29 and 32 days old.
c) Pretreatment: Test organism was held at 25 °C in flowing water with a 16 hour light-controlled photoperiod. Adults fish were fed frozen adult brine shrimp (Artemia sp.), and fry were fed freshly hatched brine shrimp nauplii to excess three times daily until 24 hour before a test (twice on most weekend days) and a dry flake ration (Tetramin) during their first week. Only groups of healthy fish were used for the test.

Fish sizes:
mean length: 16.8 mm
SD length: 1.958 mm
Loading: 0.653 g/L
Mean weight : 0.081 g
SD Weight: 0.0236 g

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

Total hardness was determined at least once for each test. The compleximetric or EDTA (ethylenediaminetetraacetic acid) method (APHA, 1980) was used to determine hardness. Total hardness concentrations (mg/L as CaCO3) were usually in the mid-forty range.

Laboratory water: total hardness 41.3-58.6 mg/L,
Lake Superior water: total hardness 44.0-53.0 mg/L.

Test temperature:

Water temperature was determined using a partial immersion mercury thermometer. Measurements were made in each exposure chamber daily.

Water Temperature expected: 25 +/- 1 °C
Mean measured Temperature: 24.8 +/- 1 °C

pH:

Laboratory water: pH 6.5-7.8
Lake Superior water: pH 7.4-8.2

Dissolved oxygen:

Dissolved oxygen was determined in high, medium, low and control exposure chambers at least three times (0, 24 and 96 hr) during a test if surviving fish existed in chamber. Determinations were made with an oxygen sensitive electrode (Yellow Springs Instrument, Yellow Springs, OH45387, Model 54 polarograph) which was calibrated weekly using the azide modification of the winkler method (APHA, 1980). Among 198 substances tested, test mean concentration were maintained between 60 and 100% of water saturation except in 12 tests where the limits were exceed. In 7 tests, the dissolved oxygen concentration was between 50 and 60% on at least one day, and in 5 tests, the dissolved oxygen concentration slightly exceed 100% of saturation on one or more occasions. Results of these tests were considered to be unaffected by the dissolved oxygen concentrations and have been included in data set.
Dissolved Oygen: 8.0 +/- 0.19 mg/L

Conductivity:

Total alkalinity was determined by a tetrametric method (APHA, 1980).
Alkalinity: 33.0 mg/L CaCO3

Nominal and measured concentrations:

Nominal Concentrations: control, 0.95, 1.6, 2.6, 4.4 and 7.3 mg/L

Details on test conditions:

Test Conditions:
a) Dilution Water Source: Dechlorinated laboratory water or Lake Superior water.
b) Dilution Water Chemistry: (Laboratory water) pH 6.5-7.8, total hardness 41.3-58.6 mg/L, (Lake Superior water) pH 7.4-8.2, total hardness 44.0-53.0 mg/L.
c) Exposure Vessel Type: 28.0 L test solution in a 3.1-L test tank.
d) Loading Rate: 0.653 g / L
e) Nominal Concentrations: control, 0.95, 1.6, 2.6, 4.4 and 7.3 mg/L
f) Vehicle/Solvent and Concentration: Not used.
g) Stock Solution Preparation: Stock solution (concentration of 567 mg/L) was prepared. Test solutions were prepared by diluting this stock solution with dilution water.
h) Number of Replicates: 2
i) Fish per Replicate: 25
j) Renewal Rate of Test Water: Daily; Test tank were also exchanged daily because of microbial growth.
k) Water Temperature: 25 +/- 1 °C
l) Light Condition: 16:8 hours, light-darkness cycle
m) Feeding: None
During test period, the fish were routinely observed for behavioural responses (effects) and deaths. Dead fish were removed and recorded at 3, 6, 12, 24, 48, 72 and 96 hour after initial exposure.
At the termination of tests, control fish were weighted (wet) and measured (standard length).

-Statistical Method:
a) Data Analysis: trimmed Spearman-Karber method (Hamilton et al., 1977)
b) Method of Calculating Mean Measured Concentrations: Arithmetic mean. Average values were corrected by recovery rate (85.5%).

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.3 mg/L

95% CI:

>= 2.2 - <= 2.5

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

Water Chemistry:
pH: 7.05 +/- 0.07
Dissolved Oygen: 8.0 +/- 0.19 mg/L
Water Temperature: 24.1 +/- 0.19 °C
Hardness: 49.6 mg/L CaCO3
Alkalinity: 33.0 mg/L CaCO3
-Size of the control fish at the end of the test:
Length: 16.8 +/- 1.958 cm, Wet weight: 0.081 +/- 0.0236 g
-Effect Data (mortality):
LC50 (96hr) 2.3 mg/L (mc)
95% Confidence interval: 2.2 – 2.5 mg/L.
mc: based on measured concentrations
-Effect Data (effect): EC50 (96hr): 2.3 mg/L (mc)
95% Confidence interval: 2.2 – 2.5 mg/L.
- Cumulative Mortality: None of test organisms were killed during exposure period at control and concentrations of 0.95 mg/L (nominal).

Sublethal observations / clinical signs:

Table1. Concentration of the Test Substance in Test Water.

Nominal Conc. (mg/L)	Replicate	Measured concentrations (mg/L)					Average*(mg/L)	Corrected Average**(mg/L)
		03/16/81	03/17/81	03/18/81	03/19/81	03/20/81
Control	1	0	-	0	-	0	0	0
	2	-	0	-	0	-	0	0
0.95	1	0.44	0.68	-	0.67	0.76	0.64	0.75
	2	0.39	-	0.67	-	0.7	0.59	0.69
1.6	1	0.89	1.1	-	1.3	1.3	1.1	1.3
	2	0.81	-	1.3	-	1.4	1.2	1.4
2.6	1	1.5	1.7	-	1.8	1.9	1.7	2
	2	1.6	-	1.9	-	1.9	1.8	2.1
4.4	1	2.9	-	-	-	-	2.9	3.4
	2	2.8	-	-	-	-	2.8	3.3
7.3	1	5.1	-	-	-	-	5.1	6
	2	5.4	-	-	-	-	5.4	6.3

*: Arithmetic Mean.

**: Corrected by recovery rate, 85.5%.

Reported concentrations are the means of the before and after exchange values.

Table 2. The Cumulative Numbers of Dead Fish.

Nominal Conc. (mg/L)	Replicate	Cumulative Number of Dead
		3 hr	4.5 hr	6 hr	12 hr	24 hr	48 hr	72 hr	74 hr	96 hr
Control	1	0	0	0	0	0	0	0	0	0
	2	0	0	0	0	0	0	0	0	0
0.95	1	0	0	0	0	0	0	0	0	0
	2	0	0	0	0	0	0	0	0	0
1.6	1	0	0	0	0	0	0	0	0	0
	2	0	0	0	0	0	0	0	0	1
2.6	1	0	0	0	1	1	2	6	7	8
	2	0	0	0	2	3	3	4	5	7
4.4	1	5	25	25	25	25	25	25	25	25
	2	3	25	25	25	25	25	25	25	25
7.3	1	25	25	25	25	25	25	25	25	25
	2	25	25	25	25	25	25	25	25	25

Affected fish stopped schooling and some fish lost equilibrium prior to death.

Validity criteria fulfilled:

not applicable

Conclusions:

Based on measured concentration of the test substance, the 96 h-LC50 was 2.3 mg/L (95% Cl: 2.2 to 2.5 mg/L).

Executive summary:

This study was performed to assess the 96h acute toxicity of the registered substance to fathead minnow (Pimephales promelas) under semi-static conditions in conditions similar to OECD TG 203 and EPA OPPTS 850.1075 without GLP compliance. Although this study was not performed according to guidelines and GLP compliance, the study is well detailled and follow similar recommendations to guidelines: analytical monitoring, cumulative mortality of control at 0% at the end of the test, monitoring of dissolved oxygen concentration. Minor vulnerabilities are reported but this study can be considered as reliable with a klimish score 2.

The nominal test substance concentrations tested were 0.95, 1.6, 2.6, 4.4 and 7.3 mg/L. A blank control (test water without test substance) was selected in parallel to the test concentration. The analytical monitoring was performed by Gas-liquid chromatography in all test exposure chambers sampled at approximately mid-depth at 0 and 96 hr and one of each duplicate exposure chamber at 24, 48 and 72 hr. Twenty five fish per group were used at the test concentrations and in the control. One replicate was perfomed for each tested concentration and control.

The measured concentrations of test substance in blank control were all lower than the limit of detection. The mean corrected measured concentrations of 0.95, 1.6, 2.6, 4.4 and 7.3 mg/L test groups ranged from 0.69 to 6.3 mg/L considering a percentage of recovery of 85.5 (+-1.9). The analytically measurement confirmed nominal concentrations.

The cumulative mortality of blank control was 0% at the end of the test, which can be considered as a criterion for the validity of the test. 

Based on measured concentration of the test substance, the 96 h-LC50 was 2.3 mg/L (95% Cl: 2.2 to 2.5 mg/L). The highest nominal concentration causing 0% mortality at 96 h (LC0) was 0.95 mg/L and the lowest nominal concentration causing 100% mortality at 96h (LC100) was 4.4 mg/L.

Description of key information

96h-LC50 (Pimephales promelas) = 2.3 mg/L; Similar to OECD TG 203 and EPA OPPTS Guideline, Brooke et al. (1984)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

2.3 mg/L