Reaction products of sodium glucoheptonate with iron sulfate and ammonium hydroxide

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: OECD guideline study; GLP compliance; read-across

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

not specified

Remarks:

the study reviewed in the report was stated to be GLP-compliant

Specific details on test material used for the study:

No details on properties are given.

Analytical monitoring:

yes

Details on sampling:

No details given.

Vehicle:

not specified

Details on test solutions:

No details given.

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Age at study initiation (mean and range, SD): 4 months
- Length at study initiation (length definition, mean, range and SD): 2.9 ± 0.1 cm
- Weight at study initiation (mean and range, SD): 0.22 ± 0.03 g

ACCLIMATION
- Acclimation period: Fish were acclimated for 7 days before test. No food was fed before 1 day and during test.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not reported.

Hardness:

34 mg/L as CaCO3

Test temperature:

23.7 ± 0.3 °C

pH:

3.50 – 7.42

Dissolved oxygen:

65 – 97 %

Salinity:

Not applicable.

Nominal and measured concentrations:

Measured concentrations were 11.7, 16.5, 24.0, 34.4, 62.6 and 99.2 mg/L (nominal concentrations at 10, 15, 24, 39, 63 and 100 mg/L).

Details on test conditions:

TEST SYSTEM
- Test vessel: 8.7 L aquarium with 5 L test solution
- No. of organisms per vessel: 7


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Tap water passed through activated carbon and membrane filter (1 μm)
- Alkalinity: 24 mg/L as CaCO3

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16/8 (light/dark)
- Light intensity: 890 – 1,030 Lux

TEST CONCENTRATIONS
- Test concentrations: 10, 15, 24, 39, 63 and 100 mg/L

Reference substance (positive control):

not required

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

46.6 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: 95% CL: 36.1 – 61.5 mg/L

Details on results:

- Mortality of control: No dead fish.

Results with reference substance (positive control):

Not reported.

Reported statistics and error estimates:

LC50 value and 95 % confidence limit were calculated by Probit method (EPA/600/4-85/13, 1985).

Sublethal observations / clinical signs:

Table 1. Cumulative mortality of Oryzias latipes

Nominal concentrations (mg/L)	Measured concentrations (mg/L)	Number of organisms tested	Cumulative number of dead fish
			24 hrs	48 hrs	72 hrs	96 hrs
Control	Control	7	0	0	0	0
10	11.7	7	0	0	0	0
15	16.5	7	0	0	0	0
24	24.0	7	0	0	0	0
39	34.4	7	0	0	0	1
63	62.6	7	0	0	1	6
100	99.2	7	2	4	7	7

Table 2. pH of test solutions

Nominal concentrations (mg/L)	Measured concentrations (mg/L)	0 hr	24 hrs	48 hrs	72 hrs	96 hrs
Control	Control	7.42	7.15	7.17	7.25	7.27
10	11.7	6.75	7.00	7.07	7.09	7.17
15	16.5	6.58	6.82	6.97	7.01	7.07
24	24.0	6.36	6.49	6.65	6.67	6.80
39	34.4	6.11	5.89	5.34	4.11	4.19
63	62.6	5.93	5.61	4.72	3.65	3.50
100	99.2	5.71	5.53	4.21	-	-

Table 3. The results of acute toxicity test with pH adjustment

Nominal concentrations (mg/L)	pH	Number of organisms tested	Cumulative number of dead fish
			24 hrs	48 hrs	72 hrs	96 hrs
100	3.50	5	5	5	5	5
100	4.00	5	0	0	0	0
100	4.50	5	0	0	0	0
100	5.00	5	0	0	0	0
100	5.50	5	0	0	0	0
100	6.00	5	0	0	0	0

When iron dichloride was dissolved in water, the test solution became acidic. So, a preliminary test was conducted to evaluate the effect of pH on fish. 100 mg/L of iron dichloride was dissolved and pH of the test solutions were adjusted to 3.5, 4.0, 4.5, 5.0, 5.5 and 6.0. A group of 5 fish was used without duplication. The mortality was affected at pH 3.5 only. Therefore, mortality increase in 62.6 mg/L at 96 hours was due to the low pH.

Validity criteria fulfilled:

yes

Remarks:

Validity criteria (e.g. mortality in the control, constant conditions, dissolved oxygen concentration, concentration of the test substance) were met.

Conclusions:

LC50 (96 h) = 46.6 mg/L (based on measured concentration)

Executive summary:

The acute toxicity of the read-across substance Iron dichloride (CAS 7758-94-3) towards fish was determined according to OECD Guideline 203 in compliance with GLP. The following test concentrations have been used: 0 (control), 10, 15, 24, 39, 63 and 100 mg/L (based on nominal concentrations). Measured concentrations were 11.7, 16.5, 24.0, 34.4, 62.6 and 99.2 mg/L (Method applied: ICP-AES). Oryzias latipes was chosen as test organism which was exposed for 96 h to the test substance. 7 fish were tested for each concentration (incl. control). At 39 mg/L test concentration, one dead fish was observed after 96 hours. At 63 mg/L, 6 dead fish were observed after 96 hours. At 100 mg/L, all fish died after 96 hours.

When iron dichloride was dissolved in water, the test solution became acidic. The influence of the pH value on the mortality of the fish has, thus, been evaluated as well. The mortality was affected at pH 3.5 only. Therefore, mortality increase in 63 mg/L at 96 hours was due to the low pH. The LC50 (96 h) value was determined to be 46.6 mg/L (based on measured concentration). LC50 (96 h) with 95 % confidence limit was 36.1 – 61.5 mg/L.

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: study conducted according to OECD guidelines; quality assurance and GLP certificates; read-across

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

No details given.

Analytical monitoring:

yes

Details on sampling:

No details given.

Vehicle:

not specified

Details on test solutions:

No details given.

Test organisms (species):

Oryzias latipes

Details on test organisms:

No details given.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

No details provided.

Hardness:

No details provided.

Test temperature:

24 +/- 1°C

pH:

min: 7.1 - max : 7.6

Dissolved oxygen:

Dissolved oxygen concentrations were over 60% of the saturation value (8.25 mg/L at 24.0°C)

Salinity:

Not applicable.

Nominal and measured concentrations:

Measured concentrations of the test substance were within +/- 20% of the nominal concentrations.

Details on test conditions:

TEST SYSTEM
- No. of organisms per vessel: 5 (range finding test)
- Biomass loading rate: 10 fish/concentration (definitive test)

OTHER TEST CONDITIONS
- Photoperiod: fluorescent light; 16 hours light/8 hours dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): toxicological symptoms and deaths

TEST CONCENTRATIONS
- Range finding study
- Results used to determine the conditions for the definitive study: yes

Reference substance (positive control):

not required

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality and toxicological symptoms

Duration:

96 h

Dose descriptor:

LC0

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: mortality and toxicological symptoms

Details on results:

No details provided.

Results with reference substance (positive control):

No data.

Reported statistics and error estimates:

None reported.

Sublethal observations / clinical signs:

Measured concentrations of the test substance were within +/- 20% of the nominal concentrations. Results were based on the nominal concentrations.

No toxicological symptoms nor any death were observed at 100 mg/L (limit test).

Validity criteria fulfilled:

yes

Remarks:

Validity criteria (e.g. mortality in the control, constant conditions, dissolved oxygen concentration, concentration of the test substance) were met.

Conclusions:

NOEC/LC0 (96 h) > 100 mg/L

Executive summary:

The acute toxicity of the read-across substance Sodium gluconate (CAS 527-07-1) towards fish was determined according to OECD Guideline 203 in compliance with GLP. A range finding test (5 fish/vessel/concentration) was conducted before the definitive test. Based on the results of the range finding test, only two concentrations were tested in the definitive test: 0 and 100 mg/L (limit test). Oryzias latipes served as test organism.

10 fish/concentration were used for the definitive test. Neither toxicological symptoms nor any death were observed at 100 mg/L. Conclusively, the NOEC/LC0 (96 h) was set to > 100 mg/L based on the nominal concentration.

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: well documented publication; method meets basic scientific principles; read-across

Qualifier:

no guideline followed

Principles of method if other than guideline:

10 fish/concentration tested (3 replicates/concentration); tested concentrations are not clarified; duration of 96 hours; static test

GLP compliance:

no

Specific details on test material used for the study:

No details are provided.

Analytical monitoring:

yes

Details on sampling:

No details reported.

Vehicle:

no

Details on test solutions:

Chemically pure chloride compounds of metals were dissolved in deionized water for the preparation of desired stock solutions.

Test organisms (species):

other: Labeo rohita

Details on test organisms:

TEST ORGANISM
- Age at study initiation (mean and range, SD): 30-, 60- and 90-days

ACCLIMATION
- Acclimation period: acclimatized for two weeks prior to test

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Post exposure observation period:

Not reported.

Hardness:

Age group 30-day: 99.91 ± 0.32
Age group 60-day: 100.02 ± 0.31
Age group 90-day: 105.15 ± 0.40

Test temperature:

Age group 30-day: 30.24 ± 0.02
Age group 60-day: 30.23 ± 0.03
Age group 90-day: 30.25 ± 0.01

pH:

Age group 30-day: 7.04 ± 0.01
Age group 60-day: 7.05 ± 0.02
Age group 90-day: 7.00 ± 0.02

Dissolved oxygen:

Age group 30-day: 5.11 ± 0.52
Age group 60-day: 5.00 ± 0.20
Age group 90-day: 4.77 ± 0.17

Salinity:

not applicable

Nominal and measured concentrations:

The data obtained from analyses confirmed that the determined iron, zinc, lead, nickel and manganese concentrations coincided with the estimated data.

Details on test conditions:

TEST SYSTEM
- Test vessel: 70 L aquariums
- Material, size, headspace, fill volume: glassware
- Aeration: During all the trails constant air was supplied to all the test mediums with an air pump through capillary system.
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 3

TEST MEDIUM / WATER PARAMETERS
- Ammonia (mg/L):
Age group 30-day: 1.05 ± 0.05
Age group 60-day: 1.40 ± 3.00
Age group: 90-day: 1.70 ± 3.11

-Sodium (mg/L):
Age group 30-day: 240.33 ± 22.35
Age group 60-day: 380.31 ± 16.17
Age group: 90-day: 221.01 ± 16.45

-Potassium (mg/L):
Age group 30-day: 9.74 ± 2.56
Age group 60-day: 5.88 ± 1.06
Age group: 90-day: 7.25 ± 2.00

-CO2 (mg/L):
Age group 30-day: 2.12 ± 0.16
Age group 60-day: 2.19 ± 0.22
Age group: 90-day: 2.16 ± 0.15

Reference substance (positive control):

not specified

Remarks:

Not reported.

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

49.75 mg/L

Nominal / measured:

not specified

Conc. based on:

element

Remarks:

Fe

Basis for effect:

mortality (fish)

Remarks on result:

other: ± 2.78 mg/L; 30-days fish

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

53.18 mg/L

Nominal / measured:

not specified

Conc. based on:

element

Remarks:

Fe

Basis for effect:

mortality (fish)

Remarks on result:

other: ± 2.74 mg/L; 60-days fish

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

58.18 mg/L

Nominal / measured:

not specified

Conc. based on:

element

Remarks:

Fe

Basis for effect:

mortality (fish)

Remarks on result:

other: ± 2.74; 90-days fish

Details on results:

The fish had the lowest mean iron LC50 concentration of 49.75 ± 2.78 mg/L for 30-days fish, followed by that of 60- and 90-days that had the average values of 53.18 ± 2.74 and 58.18 ± 2.74 mg/L, respectively. The differences among three fish age groups for LC50 values were statistically significant. Lethal concentrations of 30-, 60- and 90-days fish varied significantly as 83.45 ± 5.14, 85.53 ± 5.37 and 90.53 ± 5.37 mg/L, respectively. However, the differences between 30- and 60-days age groups were statistically non-significant.

Results with reference substance (positive control):

Not applicable.

Reported statistics and error estimates:

The 96-h LC50, lethal values and their 95% confidence intervals were estimated by using Probit analysis.

Sublethal observations / clinical signs:

No mortality was observed among control fish.

Validity criteria fulfilled:

not applicable

Conclusions:

LC50 (96 h) = 49.75 mg Fe/L

Executive summary:

The acute toxicity of iron dichloride (CAS 7758-94-3) to fish (Labeo rohita) has been studied in a static test including the determination of 96-h LC50 values. No guideline was followed and GLP compliance was not fulfilled. The test was performed at constant temperature (30°C), pH (7) and hardness (100 mg/L). Three fish age groups (30-, 60- and 90-days) were tested for their sensitivity. Physico-chemical variables such as water temperature, pH, total hardness, dissolved oxygen, total ammonia, sodium, potassium and carbon dioxide were also studied during the experiment. 10 fish/concentration were tested (3 replicates/concentration) for an exposure duration of 96 hours. Specific test concentrations are not clarified. In order to avoid the sudden stress to fish, the concentrations of metals in aquariums were increased gradually, 50% test concentration being reached in three and half hours and full toxicant concentration in seven hours. The data obtained from analytical analyses (methods described in S.M.E.W.W., 1989) confirmed that the determined iron concentrations coincided with the estimated data.

The fish had the lowest mean iron LC50 concentration of 49.75 ± 2.78 mg/L for 30-days fish, followed by that of 60- and 90-days that had the average values of 53.18 ± 2.74 and 58.18 ± 2.74 mg/L, respectively. The differences among three fish age groups for LC50 values were statistically significant. No mortality was observed among control fish.

Description of key information

There is no data available for the target substance iron glucoheptonate (CAS 23351 -51 -1) on acute toxicity towards fish. However, there is data available for the read-across substances sodium gluconate (CAS 527 -07 -01) and iron dichloride (CAS 7758-94-3). This data is used within a frame of a weight-of-evidence approach to assess the toxicity of iron glucoheptonate.

The LC50 (96 h) value of 46.6 mg/L reported in a study with iron dichloride on Oryzias latipes (OECD SIDS, 2004) was converted to the target substance FeGHA. The outcome is an LC50 (96 h) of 145.33 mg FeGHA/L.

The NOEC (96 h) reported for Sodium gluconate was > 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

145.33 mg/L

Additional information

There is no data available for the target substance Iron glucoheptonate (CAS 23351-51-1) on acute toxicity towards fish. However, there is data available for the read-across substances Sodium gluconate (CAS 527-07-01) and Iron dichloride (CAS 7758-94-3).

The acute toxicity of the read-across substance Sodium gluconate (CAS 527-07-1) towards fish was determined according to OECD Guideline 203 in compliance with GLP (OECD SIDS, 2004). A range finding test (5 fish/vessel/concentration) was conducted before the definitive test. Based on the results of the range finding test, only two concentrations were tested in the definitive test: 0 and 100 mg/L (limit test).Oryzias latipes served as test organism. 10 fish/concentration were used for the definitive test. Neither toxicological symptoms nor any death were observed at 100 mg/L. Conclusively, the NOEC/LC0 (96 h) was set to > 100 mg/L based on the nominal concentration.

The acute toxicity of the read-across substance Iron dichloride (CAS 7758-94-3) towards fish was determined according to OECD Guideline 203 in compliance with GLP (OECD SIDS, 2004). The following test concentrations have been used: 0 (control), 10, 15, 24, 39, 63 and 100 mg/L (based on nominal concentrations). Measured concentrations were 11.7, 16.5, 24.0, 34.4, 62.6 and 99.2 mg/L (Method applied: ICP-AES).Oryzias latipes was chosen as test organism which was exposed for 96 h to the test substance. 7 fish were tested for each concentration (incl. control). At 39 mg/L test concentration, one dead fish was observed after 96 hours. At 63 mg/L, 6 dead fish were observed after 96 hours. At 100 mg/L, all fish died after 96 hours. When iron dichloride was dissolved in water, the test solution became acidic. The influence of the pH value on the mortality of the fish has, thus, been evaluated as well. The mortality was affected at pH 3.5 only. Therefore, mortality increase in 63 mg/L at 96 hours was due to the low pH. The LC50 (96 h) value was determined to be 46.6 mg/L (based on measured concentration). LC50 (96 h) with 95 % confidence limit was 36.1 – 61.5 mg/L.

In addition, the acute toxicity of Iron dichloride (CAS 7758-94-3) to fish (Labeo rohita) has been studied in a static test including the determination of 96-h LC50 values. No guideline was followed and GLP compliance was not fulfilled. The test was performed at constant temperature (30°C), pH (7) and hardness (100 mg/L). Three fish age groups (30-, 60- and 90-days) were tested for their sensitivity. Physico-chemical variables such as water temperature, pH, total hardness, dissolved oxygen, total ammonia, sodium, potassium and carbon dioxide were also studied during the experiment. 10 fish/concentration were tested (3 replicates/concentration) for an exposure duration of 96 hours. Specific test concentrations are not clarified. In order to avoid the sudden stress to fish, the concentrations of metals in aquariums were increased gradually, 50% test concentration being reached in three and half hours and full toxicant concentration in seven hours. The data obtained from analytical analyses (methods described in S.M.E.W.W., 1989) confirmed that the determined iron concentrations coincided with the estimated data. The fish had the lowest mean iron LC50 concentration of 49.75 ± 2.78 mg/L for 30-days fish, followed by that of 60- and 90-days that had the average values of 53.18 ± 2.74 and 58.18 ± 2.74 mg/L, respectively. The differences among three fish age groups for LC50 values were statistically significant. No mortality was observed among control fish.

Derivation of (non)-effective concentrations for Iron glucoheptonate (CAS 23351-51-1)

There are studies available for two different read-across substances (Sodium gluconate; Iron dichloride). Since it is expected that toxicity of the target substance is rather triggered by the iron cation, data on the metal salt are considered for the Chemical Safety Assessment.

 

A NOEC (96 h) of > 100 mg/L was determined for the substance Sodium gluconate (OECD SIDS, 2004). By consideration of the molecular weights of the read-across substance Sodium gluconate (MW = 218.14 g/mol) and the target substance Iron glucoheptonate (MW = 354.8 g/mol; trihydrated form), the NOEC value for the read-across substance can be converted in order to account for the target substance.

100 mg/L x 354.8 g/mol / 218.14 g/mol = 162.65 mg/L

The calculated NOEC (96 h) value of 162.65 mg/L for the target substance will be used for the Chemical Safety Assessment.

 

The LC50 (96 h) value for Iron dichloride was determined to be 46.6 mg/L (OECD SIDS, 2004). Since this value refers to the compound itself and not only to the metal ion, a conversion is necessary. By consideration of the molecular weights of Iron dichloride (126.751 g/ mol) and Iron (55.85 g/mol), the LC50 value for Iron is calculated as follows:

46.6 mg/L x 55.85 g/mol / 126.751 g/mol = 20.5 mg Fe/L

The LC50 of the target substance FeGHA is calculated with regard to the molecular weight as follows

MW Fe in FeGHA = 55.84 g/mol

MW FeGHA = 300.8 g/mol

LC50 (FeGHA) in target = LC50 (Fe) in source / 55.84 x 300.8

LC50 (Fe) in source = 20.5 mg/L

LC50 (FeGHA) = 20.5 / 55.84 x 300.8 = 110.45 mg/L

Purity of FeGHA: 76 %

16.57 / 0.76 = 145.33 mg/L

The calculated LC50 (96 h) value of 145.33 mg/L for the target substance will be used for the Chemical Safety Assessment.