Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
fish, juvenile growth test
Type of information:
experimental study
Adequacy of study:
key study
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 215 (Fish, Juvenile Growth Test)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Reference substance name:
.beta.-Cyclodextrin, 2-hydroxypropyl cycloheptaamylose
IUPAC Name:
.beta.-Cyclodextrin, 2-hydroxypropyl cycloheptaamylose
Constituent 2
Chemical structure
Reference substance name:
-
EC Number:
420-920-1
EC Name:
-
Cas Number:
128446-35-5
Molecular formula:
Hill formula: (C42H70-nO35)(C3H7O)n; n(mittel)=5,25
IUPAC Name:
2-Hydroxypropyl-.beta.-cyclodextrine ethers
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Molecular weight: 1380-1500 g/mol
Description: white powder
Batch: 74T001
Purity: >95%
Test substance storage: at room temperature in the dark
Stability under storage conditions: stable
Expiry date: 31 January 2005
Stability in water: at least 96h

Sampling and analysis

Analytical monitoring:
no

Test solutions

Vehicle:
yes

Test organisms

Test organisms (species):
Cyprinus carpio
Details on test organisms:
TEST ORGANISM
- Common name: Carp
- Strain: Teleostei, Cyprinidae
- Source: Zodiac, proefacc, "De Haar Vissen", L.U., Wageningen, the Netherlands
- Age at study initiation (mean and range, SD): 2.6cm, 0.4 SD
- Weight at study initiation (mean and range, SD): mean weight: 0.63g, range: >= 0.58g, =< 0.73g (day0), >= 1.07g, =< 1.35g (day28)

FEEDING DURING TEST
- Food type: Trouvit
- Amount: a rate of 4 % of initial body weight per vessel
- Frequency: Daily
- general: The daily ration was generally divided into two equal portions and given to the fish in two feeds per day. After 14 days of exposure the ration was recalculated based on the fish weights then determined. Fish were not fed 24 hours prior to weighing.

Study design

Test type:
flow-through
Water media type:
freshwater
Limit test:
no
Total exposure duration:
28 d

Test conditions

Hardness:
Hardness: 2.2 mmol/l
Nitrate (expr. as N): 0.82 mg/l
Nitrite (expr. as N): < 0.030 mg/l
Test temperature:
20.0-22.3 °C
pH:
7.6-8.3
Dissolved oxygen:
> 6 mg O2/L
Nominal and measured concentrations:
10, 18, 32, 56 and 100 mg/L
Controls: Test medium without hydroxypropylated .beta.-cyclodextrin or other additives (0 mg/L; group code 1)
Details on test conditions:
TEST SYSTEM
- Test vessel: Day 0-13: 10 litres (ca. 44x15x20 cm); day 13-28: 30 litres (50x30x25 cm) both consisting of glass plates sealed with a
thin film of silicone and covered by a removable glass plate
- Aeration: test solutions were not aerated during exposure period
- Renewal rate of test solution (flow rate): 6 L/h
- No. of organisms per vessel: 16
- No. of vessels per concentration (replicates): 5
- No. of vessels per control (replicates): 1
- No. of vessels per vehicle control (replicates): 5

TEST MEDIUM / WATER PARAMETERS
- Total organic carbon: 3.0-51.2 mg/L
- Metals: Selenium <2.0 µg/l, Arsenic < 4.0 µg/l, Cadmium < 0.40 µg/l, Chromium < 1.0 µg/l, Copper 14 µg/l, Mercury < 0.050 µg/l, Lead < 5.0 µg/l, Zinc 13 µg/l, Manganese < 0.010 mg/l, Calcium 75 mg/l, Magnesium 8.8 mg/l, Iron < 0.050 mg/l
- Chlorine: was not presented

OTHER TEST CONDITIONS
- Adjustment of test system: from day 13 onwards
- Photoperiod: 16h daily

VEHICLE CONTROL PERFORMED: yes
Reference substance (positive control):
yes
Remarks:
pentachlorophenol

Results and discussion

Effect concentrationsopen allclose all
Duration:
28 d
Dose descriptor:
LC50
Effect conc.:
> 100 mg/L
Conc. based on:
test mat.
Duration:
28 d
Dose descriptor:
LC10
Effect conc.:
> 100 mg/L
Conc. based on:
test mat.
Duration:
28 d
Dose descriptor:
other: EC50 juvenile growth
Effect conc.:
> 100 mg/L
Conc. based on:
test mat.
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
100 mg/L
Conc. based on:
test mat.
Details on results:
No specific analytical method was available for measuring hydroxypropylated .beta.-cyclodextrin concentrations. As a consequence TOC-analysis was performed. The TOC content of a hydroxypropylated .beta.-cyclodextrin stock solution of 2 g/I averaged 1039 mg/I. This value was based on 4 measurements performed on stock solutions prepared on days 0 and 7. Hence, the nominal TOC content of hydroxypropylated .beta.-cyclodextrin was expected to correspond to 52%. TOC-analysis of the samples taken during the 28-day test period from the target concentrations of 32 mg/I and higher revealed that recoveries for organic carbon were at more than 800/0 compared to the
expected carbon content. Results further showed that measured TOC concentrations were maintained within 80-120% of the mean measured values at these target concentrations.
The TOC-results at the target concentrations of 10 and 18 mg/I were difficult to interpret. This was a consequence of the limited accuracy at TOC levels below 10 mg/I, i.e. close to the limit of detection of the equipment, which resulted in a high variation rate.
No valid results were obtained on day 21. However, as the flow-through system functioned properly and results of the other sampling days were reliable it was considered that actual concentrations on day 21 did not differ from those obtained on the other sampling days.
The overall conclusion was that measured TOC-concentrations were in agreement with what was expected based on the nominal carbon content of hydroxypropylated .beta.-cyclodextrin. Further results will therefore be based on the target concentrations.
Results with reference substance (positive control):
Under the conditions of the present test PENTACHLOROPHENOL induced no lethal effects in carp at or below 0.15 mg/1. The 96h-LC50 for carp exposed to pep was 0.21 mg/I (95 % confidence interval between 0.19 and 0.27 mg/I). The 24h-LC50 was 0.24 mg/I (95% confidence interval between 0.21 and 0.30 mg/I), and remained unchanged until 72h. The range of the 96h-LC50 for carp is generally between 0.10 and 0.46 mg/I based on historical data of reference tests performed approximately every 3 months from April 1988 until the end 2000, and annually since then. The response observed in carp originating from the present batch falls within this range.

Concentraions mg/L: 0.06 (0% mortality), 0.10 (0% mortality), 0.15 (0% mortality), 0.22 (60% mortality), 0.32 (100% mortality)

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
This prolonged toxicity test investigated the possible effects of hydroxypropylated .beta.-cyclodextrin on survival and growth of rainbow trout exposed to various concentrations for a period of 28 days in a flow-through system.
Hydroxypropylated .beta.-cyclodextrin induced no visible effects in carp at concentrations up to and including 100 mg/L. Exposure to hydroxypropylated .beta.-cyclodextrin concentrations up to and including 100 mg/I did not result in reduction of juvenile growth.
Executive summary:

Prolonged (28 days) juvenile growth test in Carp with hydroxypropylated .beta.-cyclodextrin.

The study procedures described in this report were based on the OECD guidelines for Testing of Chemicals 215: "Fish, juvenile growth test - 28 days", accepted January 21, 2000.

The main test was performed with carp exposed to a range of hydroxypropylated .beta.-cyclodextrin target concentrations of 10, 18, 32, 56 and 100 mg/L,in a flow-through system. Sixteen fish were exposed per concentration and to untreated test medium (blank-control). Stock solutions were dosed via a computer-controlled system and entered a mixing chamber separately from the tap water supply (flow-rates 0.3 to 6 L/h). In addition, all test solutions were refreshed four times during the first 13 days of exposure to ensure optimal quality and cleanness of the test solutions. The functioning of the system was checked daily during the 28-day test period.

Fish were fed daily with Trouvit at 4% of the body weight per vessel. After 14 days of exposure the ration was recalculated based on the fish weights then determined. Sampling for analysis was performed weekly in all test solutions. No specific analytical method was available for measuring hydroxypropylated .beta.-cyclodextrin concentrations. Consequently, the Total Organic Carbon (TOC) content was analyzed as a measure for the actual exposure concentrations. Fish were daily observed for possible mortality, while three times a week fish were observed for clinical signs. Specific growth rates were based on the body weights measured at the start of the study and after 14 and 28 days of exposure.

TOC-analysis of the samples taken during the 28-day test period from the target concentrations of 32 mg/Land higher revealed that recoveries for organic carbon were at more than 80% compared to the expected carbon content. Results further showed that measured TOC concentrations were maintained within 80-120% of the mean measured values at these target concentrations. The TOC-results at the target concentrations of 10 and 18 mg/L were difficult to interpret. This was a consequence of the limited accuracy at TOC levels below 10 mg/L, i.e. close to the limit of detection of the equipment, which resulted in a high variation rate.

The overall conclusion was that measured TOC-concentrations were in agreement with what was expected based on the nominal carbon content of hydroxypropylated .beta.-cyclodextrin. Further results will therefore be based on the target concentrations.

The mean weight of the fish in the control groups had increased by an average of > 150% above their initial weight over 28 days. Oxygen concentration was generally found to be > 6 mg O2/L for all measurements. The few occasions with lower oxygen concentrations were only short lasting and did not affect the fish. During the study, temperature was maintained between 20.0 and 22.3°C and variation between the different test chambers was always within 1°C at anyone time during the test.

TOC-analyses showed that measured TOC-concentrations were in agreement with what was expected based on the nominal carbon content of hydroxypropylated .beta.-cyclodextrin. A reference test with pentachlorophenol showed normal sensitivity of the supplied carp.

The L(E)C-values, the threshold levels of effect and the no observed effect concentration (NOEC) were:

Parameter

hydroxypropylated .beta.-cyclodextrin

28-day LC50

>100

28-day LC0

100

28-day LC10

>100

28-day EC50 for juvenile growth

>100

Threshold Level for juvenile growth

>100

NOEC for juvenile growth

100