(4-amino-1-hydroxy-1-phosphonobutyl)phosphonic acid;hydrate

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2003-08-04 to 2003-08-08

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2003

Report date:

2003

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

sodium;(4-amino-1-hydroxy-1-phosphonobutyl)-hydroxyphosphinate

Cas Number:

121268-17-5

Molecular formula:

C4H12NaNO7P2 · 3 H2O

IUPAC Name:

sodium;(4-amino-1-hydroxy-1-phosphonobutyl)-hydroxyphosphinate

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot No.of test material: Sponsor and 005Y030

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Stability under test conditions: Stable in the vehicle
- Solubility and stability of the test substance in the solvent/vehicle: Soluble in the vehicle up toi 10,000 mg/L following sonofication

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: Solutions of 10 and 100 mg of test ingredient / L vehicle were prepared as intial stock solutions
- Final dilution of a dissolved solid, stock liquid or gel: 0.010, 0.10,1.0,10, and 100 mg test ingreidient / L vehicle for preliminary test
6.3, 13, 25, 50, and 100 mg test ingredient / L vehicle for main test

FORM AS APPLIED IN THE TEST (if different from that of starting material)
0.010, 0.10,1.0,10, and 100 mg test ingreidient / L vehicle for preliminary test
6.3, 13, 25, 50, and 100 mg test ingredient / L vehicle for main test

Sampling and analysis

Analytical monitoring:

yes

Remarks:

Measurements of test substance concentration, water temperature, light intensity,

Details on sampling:

- Concentrations: 0.010, 0.10,1.0,10, and 100 mg test ingreidient / L vehicle for preliminary test
6.3, 13, 25, 50, and 100 mg test ingredient / L vehicle for main test

- Sampling method: Water quality (i.e., temperature, pH and dissolved oxygen concentration) was measured daily in each replicate. The diurnal range of the water bath temperature was continuously monitored using a minimum/maximum thermometer and recorded daily. Specific conductivity, total alkalinity and total hardness of the dilution water were measured at test initiation. Specific conductivity was measured using a Corning Model Check-mite CD-30 digital conductivity meter. Water hardness and alkalinity were determined by EDTA and potentiometric titration, respectively (APHA et al., 1992). Dissolved oxygen concentrations and testchamber temperatures were determined using a YSI Model 58 oxygen meter utilizing a membrane electrode. The pH was measured with an Oakton pH Testr2 Waterproof meter. Test substance concentrations were determined in the control and test treatments at test initiation and termination. Water samples were collected from midway in the water column of each chamber using volumetric pipets. At test initiation,
approximately 10 mL was removed from each replicate test chamber and quantitatively pooled for analysis. At test termination, 10 mL was again removed from each replicate test chamber and quantitatively composited.

- Sample storage: Samples were analyzed immediately after sampling.

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Controls: Negative vehicle control
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): organic solvent, water
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)): 0.010, 0.10,1.0,10, and 100 mg test ingreidient / L vehicle for preliminary test. 0, 6.3, 13, 25, 50, and 100 mg test ingredient / L vehicle for main test

Test organisms

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Juvenile fathead minnow
- Source: Toxikon Corporation
- Age at study initiation (mean and range, SD): 7 days
- Length at study initiation (length definition, mean, range and SD): from 1.0 to 1.4centimeters (cm) total length (mean and standard deviation = 1.1 ± 0.16 cm)
- Weight at study initiation (mean and range, SD): 0.0049 to 0.0234 grams (g) wet weight (mean and standard deviation= 0.0108 ± 0.0065 g)

ACCLIMATION
- Acclimation period: 7 days
- Acclimation conditions (same as test or not): Yes
- Health during acclimation (any mortality observed): None

FEEDING DURING TEST
None

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

None

Test conditions

Hardness:

68 mg/Las calcium carbonate

Test temperature:

23.5 to 24.2°C

pH:

6.3 to 7.6

Dissolved oxygen:

5.8 to 7.3 mg/L

Salinity:

17 mg/Las CaCO3

Conductivity:

414 microSiemens (µS).

Nominal and measured concentrations:

Nominal concentrations: 0.010, 0.10,1.0,10, and 100 mg test ingreidient / L vehicle for preliminary test. 0, 6.3, 13, 25, 50, and 100 mg test ingredient / L vehicle for main test

The mean measured concentration of the test treatments were 6.24, 11.5, 22.8, 44.3 and 90.7 mg ALENDRONATE SODIUM /L, which were 99.0, 88.5, 91.2, 88.6 and 90.7% of the nominal concentrations respectively.

Details on test conditions:

TEST SYSTEM
- Test vessel: 1 L glass beakers
- Type (delete if not applicable): covered to prevent evaportation.
- Material, size, headspace, fill volume: Contained 0.80 L of test solution or dilution water
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 2
- No. of vessels per control (replicates): 0
- No. of vessels per vehicle control (replicates): 2
- Biomass loading rate: 0.095 g fo fish tissue per liter of test solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Town of Jupiter
- Total organic carbon: <1.00 ug/L
- Particulate matter:
- Metals: 3.1 mg/L
- Pesticides: <0.040 ug/L
- Chlorine: 87 mg/L
- Alkalinity: 17 mg/Las CaCO3
- Ca/mg ratio: 19 mg/L
- Culture medium different from test medium: No
- Intervals of water quality measurement: At test initiation

OTHER TEST CONDITIONS
- Adjustment of pH: None
- Photoperiod: 16 hours light and 8 hours darkness
- Light intensity: 5.1 and 6.7 micromols per square meter per second

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Survival of fathead minnow was monitored daily. The fish were also monitored for any abnormalities in their behavior or appearance.

TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study: 0.010, 0.10,1.0,10, and 100 mg test ingreidient / L vehicle for preliminary test.
- Test concentrations: 0, 6.3, 13, 25, 50, and 100 mg test ingredient / L vehicle for main test
- Results used to determine the conditions for the definitive study: After 96 hours of exposure, there was zero percent mortality in the control and in the 0.010, 0.10, 1.0 and 10 mg ALENDRONATE SODIUM /L test concentrations. There was 100% mortality in the 100 mg ALENDRONATE SODIUM /L test concentration. Fish utilized in the range-finding test averaged 1.1 cm in total length and0.0080 gain net weight as measured from the control fish at testtermination.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

ca. 27 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 2

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

ca. 25 mg/L

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Effect concentrations 3

Duration:

72 h

Dose descriptor:

LC50

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

not determinable

Details on results:

- Behavioural abnormalities: None
- Mortality of control: None
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: A film formed on the 50 mg and 100 mg/ L concentrations in the main test

Reported statistics and error estimates:

Based on the results of the test, the 96-hour LC50 value was calculated to be 27 mg ALENDRONATE SODIUM /L with 95 percent confidence limits of 22 .and 33 mg ALENDRONATE SODIUM / L.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

This study validly determined the 96 hour LC50 and NOEC for the test substance according to the guideline. The 96-hour LC50 was calculated to be 27 mg ALENDRONATE SODIUM /L. The 72 -hour LC50 could not be calculated. The 96-hour NOEC was 25 mg ALENDRONATE SODIUM /L based on a lack of statistically significant mortality at this test concentrati