N-cyclohexylcyclohexanamine 2-[1-[[[(1R)-1-[3-[(1E)-2-(7-chloroquinolin-2-yl)ethenyl]phenyl]-3-[2-(2-hydroxypropan-2-yl)phenyl]propyl]sulfanyl]methyl]cyclopropyl]acetate

Short-term toxicity to fish

Administrative data

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

22 to 26th September 2003

Reliability:

1 (reliable without restriction)

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)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-[1-[[1-[3-[2-[(7-Chloro-2-quinolyl)]vinyl]phenyl]-3-[2-(1-hydroxy-1-methyl-ethyl)phenyl]-propyl]sulfanylmethyl]cyclopropyl]acetic acid sodium salt

EC Number:

604-813-7

Cas Number:

151767-02-1

IUPAC Name:

2-[1-[[1-[3-[2-[(7-Chloro-2-quinolyl)]vinyl]phenyl]-3-[2-(1-hydroxy-1-methyl-ethyl)phenyl]-propyl]sulfanylmethyl]cyclopropyl]acetic acid sodium salt

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Montelukast sodium; Purity as is 98.8%, fine white powder
Received in Amber glass bottles, dessicated, stored at room temperature and was stored in the dark at room temperature

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Prior to initiation of the definitive test, water samples were collected from the exposure tanks to verify test concentrations and proper chemical delivery system function. During the test, water samples (SO mL in volume from each replicate) were collected from the controls and test solution and composited prior to analysis to verify actual exposure concentrations. The primary DMF stock utilized during the definitive test was also sampled and analyzed at test initiation

Test solutions

Vehicle:

yes

Remarks:

DMF

Details on test solutions:

A single primary stock was prepared for the limit test at a nominal concentration of 10,000 mg Montelukast Sodium Salt in DMF by diluting 1.0123 g of test substance in 10.0 mL DMF (Mallinckrodt Lot# 5356 N26437). The measured stock concentration was 10,700 mg. During the test, a total volume of 42 µL of stock was injected into the chemical mixing box (total volume of 4,160 mL) during each diluter cycle to provide a high nominal test concentration

Test organisms

Test organisms (species):

Pimephales promelas

Details on test organisms:

Juvenile fathead minnow, Pimephales promelas, were received on September 10, 2003 at Toxikon Corporation from Aquatox Inc., Hot Springs, Arkansas. P. promelas were maintained in filtered laboratory freshwater for a 12-day period prior to test initiation. P. promelas appeared to be in good physical condition at test initiation. Mortality during the 7-day period prior to test initiation was zero percent.
P. promelas used for the definitive test ranged from 1.0 to 1.4 centimeters (cm) total length (mean and standard deviation: 1.2 ± 0.1 cm) and from 0.0051 to 0.0141 grams (g) wet weight (mean and standard deviation: 0.0093 ± 0.0031 g) as measured from the control fish at test termination. Loading was calculated to be 0.0062 g of fish tissue per liter of test solution.

Study design

Test type:

flow-through

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Post exposure observation period:

Survival was monitored daily and fish were also monitored for any abnormal behaviour or appearances

Test conditions

Hardness:

Filtered freshwater with an initial hardness of 60 mg/L as CaC03

Test temperature:

Measure daily and ranged between 21.9 - 23 degree

pH:

measured daily and ranged between 7.1-7.4

Dissolved oxygen:

measured daily and ranged from 6.2 to 7 mg/L

Salinity:

initial alkalinity of 11 mg/L

Conductivity:

Initial conductivity of 350 µS

Nominal and measured concentrations:

Nominal Concentration 100 µg/L
Measured Concentration 77.8 µg/L

Details on test conditions:

Studies were previously conducted with the test substance in saltwater, which gave recoveries at around 100 µg Montelukast Sodium Salt /L the limit of aqueous solubility (Toxikon Study numbers 02J0038b, 02J0038c). The highest concentration tested in the freshwater studies was therefore 100 µg Montelukast Sodium Salt./L.

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Key result

Duration:

96 h

Dose descriptor:

NOEC

Effect conc.:

> 0.778 mg/L

Nominal / measured:

meas. (initial)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: limit of solubility

Details on results:

The diluter functioned properly during the entire definitive test based upon daily calculations and observations of test substance delivery. The mean measured concentration of the single test treatment was 77.8 µg Montelukast Sodium Salt /L, which was 77.8% of the nominal concentration. Chemical recovery ranged between 74.8 and 82.3 µg Montelukast Sodium Salt /L during the 96-hour exposure. The solution appeared clear and colorless and the concentration remained stable throughout the test.
Mortality of fathead minnow exposed for 96 hours to Montelukast Sodium Salt was zero percent in the control, solvent control and in the single test concentration (Table 2). The 96-hour LCso could not be calculated due to a lack of significant mortality during the exposure period. The 96-hour NOEC can be stated to be the limit of solubility, which was the highest test concentration used, or 77.8 µg Montelukast Sodium Salt/L

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

The 96-hour LC50 could not be calculated due to the lack of significant mortality at the specified time, and therefore can be stated to be greater than the limjt of solubility or> 77.8 µg SINGULAIR /L. The no-observable-effect ¥oncentration (NOEC) was 77.8 µg SINGULAIR /L based on the lack of significant mortality at this test concentration.

Executive summary:

A flow-through freshwater toxicity test was conducted at Toxikon Corporation, Jupiter, Florida, to determine the acute toxicity of Montelukast sodium salt to fathead minnow, Pimephales promelas, in accordance with OECD guideline 203.  The criterion for effect was death. Results of the test are expressed as a 96-hour median lethal concentration (LC50),  the concentration of Montelukast sodium salt

calculated to result in death to SO percent of the test population at the specified time.