2-Hydroxypropyl-β-cyclodextrine ethers

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

.beta.-Cyclodextrin, 2-hydroxypropyl cycloheptaamylose

IUPAC Name:

.beta.-Cyclodextrin, 2-hydroxypropyl cycloheptaamylose

Constituent 2

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:

5,10,15,25-tetrakis(hydroxymethyl)-40,44,47,49-tetrakis(2-hydroxypropoxy)-20,30,35-tris[(2-hydroxypropoxy)methyl]-2,4,7,9,12,14,17,19,22,24,27,29,32,34-tetradecaoxaoctacyclo[31.2.2.2³,⁶.2⁸,¹¹.2¹³,¹⁶.2¹⁸,²¹.2²³,²⁶.2²⁸,³¹]nonatetracontane-36,37,38,39,41,42,43,45,46,48-decol

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): hydroxypropylated .beta.-cyclodextrin
- Physical state: White powder
- Analytical purity: >95%
- Lot/batch No.: 74B008
- Expiration date of the lot/batch: 31 December 2002
- Stability in water: At least 96 h
- Stability under storage conditions: Stable
- Storage condition of test material: At room temperature in the dark

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

no

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Source: In-house culture
- Age at study initiation (mean and range, SD): <24h
- Valve height at study initiation, for shell deposition study (mean and range, SD):
- Method of breeding: daphnids were cultured in a temperature-controlled area at 20 +/- 2°C.

Since the culturing and testing parameters of temperature, dilution water, and lighting were the same, no acclimation period was necessary.

Study design

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Test conditions

Hardness:

146-150 mg/L as CaCO3

Test temperature:

20.0 to 22.8°C

pH:

8.32-8.53

Dissolved oxygen:

"new solutions": 7.5 to 8.4 mg/L
"old solutions": 6.2 to 8.8 mg/L

Nominal and measured concentrations:

Nominal Test Concentrations: Control, 62.5. 125, 250. 500, and 1000 mg/L
Mean Measured Concentrations: Control, 62, 127, 251, 513, and 1074 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel: quadruplicate 32 oz
- Type (delete if not applicable): closed: covered with loose-fItting petri dish covers
- Material, size, headspace, fill volume: clear glass jars, dimensions approx. 8.7 cm inside diameter by 17.0 cm height, ~400mL
Solution depth was approximately 6.8 cm
- Aeration: pipets were used to gently bubble air into each solution via a hole in the center
- Renewal rate of test solution (frequency/flow rate): every Monday, Wednesday, and Friday throughout the defInitive test
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- Biomass loading rate: - 1 daphnid per 40 mL of dilution water

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: combination of well water and reverse osmosis water blended to a hardness of 130 to 160 mg/L as CaCO3. The water was filtered with a sediment filter and exposed to ultraviolet light before use.
- Total organic carbon: <1.00 mg/L
- Metals: Aluminium 0.053, Cadmium <0.00008, Chromium 0.00087, Cobalt <0.0055, Copper <0.0038, Iron 0.0086, Lead <0.0012, Mercury 0.000051, Nickel <0.0054, Silver <0.00018, Zinc 0.0171
- Chlorine: <0.020 mg/L
- Alkalinity: 160-166 mg/L as CaCO3
- Conductivity: 338-380 µmhos/cm
- Intervals of water quality measurement: temperature, dissolved oxygen concentration, and pH were measured in "new" control dilution water and parent test solutions on day 0, 2, 4, 7, 9, 11, 14, 16, and 18 prior to daphnid and food addition, and "old" control and test solution replicates (A, B, C, and D) on days 0,2,4,7,9,11, 14, 16, and 18 after daphnid removal. Water quality parameters of temperature, dissolved oxygen concentration, pH, hardness, alkalinity and conductivity were measured on the dilution water daily.
OTHER TEST CONDITIONS
- Photoperiod: 16-hour light and 8-hour dark
- Light intensity: 47 to 52 footcandles

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Range finding study: yes

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

127 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

behaviour

Effect concentrations 2

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

251 mg/L

Nominal / measured:

meas. (not specified)

Conc. based on:

test mat.

Basis for effect:

behaviour

Details on results:

Range-Finding Test
A 14-day static renewal Daphnia magna toxicity test with hydroxypropylated .beta.-cyclodextrin was completed on June 13, 1997. The nominal test concentrations of hydroxypropylated .beta.-cyclodextrin tested were: 10, 100, and 1000 mg/L. Throughout the range-finding test, there was no observed mortality. The number of total young data collected showed 1127, 1154, 1206, and 990 young produced in the control, 10, 100, and 1000 mg/L test solutions, respectively. The percent of total young produced as compared to the control was 102, 107, and 88% for the 10, 100, and 1000 mg/L test solutions, respectively. Temperature and DO measurements for each test solution on day 14 of the range-finding test showed the temperature ranged from 20.3 to 20.5°C and the DO ranged from 2.8 to 6.5 mg/L. The percent DO saturation in the control, 10, and 100 mg/L test solutions was > 60%, while the percent DO saturation in the 1000 mg/L test solutions was <40%. The definitive test concentrations were agreed upon between the sponsor and study director based on the results of this test.
Definitive Study
Analytical Measurements:
The mean measured concentrations of hydroxypropylated .beta.-cyclodextrin collected on days 0, 7, 14, and 21 were 62, 127, 251, 513, and 1074 mg/L representing 99, 102, 100, 103, and 107% of the 62.5. 125, 250, 500, and 1000 mg/L nominal test concentrations. Quality control fortifications were prepared at nominal concentrations ranging from 37.3 to 41.6 mg/L (low spikes) and 932 and 1040 mg/L (high spikes) on each sample day. The recoveries for the quality control fortifications averaged 92% of nominal for the low spikes and 103% of the nominal for the high spikes. Based on analytical recovery, hydroxypropylated .beta.-cyclodextrin appeared to be stable in the exposure system. All results were based on the mean measured test concentration.
Biological Results:
The definitive toxicity test was initiated on July 21, 1997, and lasted 21 days before it was terminated on August II, 1997. All results are presented in this section as mean measured test concentrations of hydroxypropylated .beta.-cyclodextrin.
The mean measured test concentrations of 62, 127, 251, 513, and 1074 mg/L had mean percent survivals of 100 +/- 0.0, 100 +/- 0.0, 95.0 +/- 5.8, 82.5 +/- 12.6, 25 +/- 50.0, and 25.0 +/- 43.6%, respectively. Survival of Daphnia magna was shown to be affected statistically in the 251, 513, and 1074 mg/L mean measured concentrations as compared to the controls. The 7-, 14-, and 21-day EC50 values were estimated to be > 1074, > 1074, and 446 mg/L, respectively.
There were no test concentrations shown to be significantly affected for time to first brood when compared to the control. The mean measured concentrations of 62, 127, 251, 513, and 1074 mg/L each had mean times to first brood of 7.0 +/- 0.0 days.
There were no test concentrations shown to be significantly affected for YAD when compared to the control. The mean measured concentrations of 62, 127, 251, 513, and 1074 mg/L had mean YAD values of 8.56 +/- 0.37, 9.54 +/- 0.28, 9.91 +/- 0.37, 10.24 +/- 0.91, 5.55 +/- 3.7, and 6.87 +/- 2.43, respectively. Throughout the study, all young daphnids produced appeared to be normal.
There were no test concentrations shown to be significantly affected for length when compared to the control. The mean measured concentrations of 62, 127, 251, 513, and 1074 mg/L had mean daphnid lengths of 4.83 +/- 0.02, 4.89 +/- 0.02, 4.87 +/- 0,09, 4.87 +/- 0.00, and 4.69 +/- 0.13 mm, respectively.
There were no test concentrations shown to be significantly affected for weight when compared to the control. The nominal concentrations of 62, 127,251, 513, and 1074 mg/L had mean daphnid weights of 0.93 +/- 0.07, 0.90 +/- 0.04, 0.93 +/- 0.09, 0.93 +/- 0.11, 1.04 +/- 0.00, and 1.15 +/- 0.21 mg, respectively.
Water quality parameters of temperature dissolved oxygen concentration, and pH were measured in the "new" parent control and test solutions on study days 0, 2. 4. 7. 9. 11, 14, 16, and 18, and in each replicate of the "old" control and test solutions on study days
2, 4, 7, 9, 11, 14, 18, and 21. The temperature of the "new" control and test solutions ranged from 20.7 to 22.8°C and the temperature of the "old" control and test solutions ranged from 20.0 to 20.9°C. The pH values of the "new" control and test solutions ranged from 8.1 to 8.5 and the pH values of the "old" control and test solutions ranged from 7.9 to 8.6. Dissolved oxygen concentration of the control and test solutions ranged from 7.5 to 8.4 mg/L in the "new" solutions and ranged from 6.2 to 8.8 mg/L in the "old" solutions. These dissolved oxygen values represented 91 and 100% saturation at 23°C and 22°C, respectively, for the "new" solutions, and 73 and 101% saturation at 21°C and 20°C, respectively, for "old" solutions. The dissolved oxygen saturation (100%) at temperatures of 20, 21, 22, and 23°C, corrected for local altitudinal pressure, are 8.7, 8.5, 8.4, 8.2 mg/L, respectively. All water quality parameters were considered to be within acceptable limits.
All solutions approximately 24 hours old or older had been observed to be free of precipitate and surface film with the exception of some 513 and 1074 mg/L test replicates beginning on day 14. On days 14, 15, and 16, the 513 mg/L replicates A, B, and D, and the 1074 mg/L replicates A, B, and C, were cloudy. On days 17, 18, and 19, the 513 mg/L replicates B and D, and the 1074 mg/L replicates B and C were cloudy. On days 20 and 21, the 513 mg/L B and D replicates, and the 1074 mg/L B replicate were cloudy.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

Based on the statistical endpoint of adult mean survival from this 21-day Daphnia magna chronic toxicity test, MATC limits were determined to be 251 mg/L (LOEC) and 127 mg/L (NOEC).
The geometric mean of the MATC limits is 179 mg/L. The day-7, -14, and -21 ECso values based on immobility/mortality were estimated to be > 1074, > 1074, and 446 mg/L, respectively. The results of the definitive test represent the best estimate of the toxicity of hydroxypropylated .beta.-cyclodextrin to Daphnia magna.

Executive summary:

The primary objective of this study was to evaluate the chronic toxicity of hydroxypropylated .beta.-cyclodextrin to Daphnia magna under static-renewal according to OECD Guideline 202. The test was designed to yield a no-observed effect concentration (NOEC) and a lowest-observed effect concentration (LOEC) based on the statistical analysis of survival, growth, and reproduction.

An estimate of the maximum acceptable toxicant concentration (MATC) was calculated using the NOEC and LOEC. If possible, a 7-, 14-, and 21-day EC50 based on adult immobilization and 95 % confidence limits were to be calculated.

The definitive study was initiated on July 21, 1997, and lasted 21 days before it was terminated on August 11, 1997. The nominal exposure concentrations were 62.5, 125, 250, 500, and 1000 mg/L of hydroxypropylated .beta.-cyclodextrinalong with a control. The test was conducted in 32 oz. clear glass containers containing 400 mL of the test solution. There were four replicates (A, B, C, and D) per test concentration and each initially contained 10 Daphnia magna (<24 hours old). The daphnids were exposed to newly prepared test (and control) solutions every Monday, Wednesday, and Friday, throughout the definitive test. The daphnids were fed and observed daily. After 21 days, survival, time to first brood, young per adult reproduction day, length, and weight were statistically compared to the control to evaluate the toxicity of hydroxypropylated .beta.-cyclodextrin to Daphnia magna.

The terms "new" and "old" will appear throughout this report. The term "new" is defined as newly prepared parent test solutions and control having contained no daphnids or food. The term "old" is defined as approximately 48- or 72-hour old test solutions and control that had contained daphnids and/or food.

Water quality parameters of temperature, dissolved oxygen concentration and pH were measured in "new" and "old" solutions. The temperature of the "new" control and test solutions ranged from 20.7 to 22.8°C and the temperature of the "old" control and test solutions ranged from 20.0 to 20.9°C. The pH values of the "new" control and test solutions ranged from 8.1 to 8.5 and the pH values of the "old" control and test solutions ranged from 7.9 to 8.6. Dissolved oxygen concentrations for the control and test solutions ranged from 7.5 to 8.4 mg/L in the "new" solutions and ranged from 6.2 to 8.8 mg/L in the "old" solutions. These dissolved oxygen values represented 91 and 100% saturation at 23°C and 22°C, respectively, for the "new" solutions, and 73 and 101% saturation at 21°C and 20°C, respectively, for "old" solutions. Each solution during the definitive test was gently aerated. The dissolved oxygen saturation (100 %) at temperatures of 20, 21, 22. and 23 °C corrected for local altitudinal pressure. are 8.7. 8.5, 8.4, 8.2 mg/L, respectively. All water quality parameters were considered to be within acceptable limits.

The parameters of survival, time to first brood, young per adult reproduction day (YAD), and growth (length and weight) were statistically compared to the control. The mean measured test concentrations of 251, 513 and 1074 mg/L were shown to be significantly different(p< 0.05) as compared to the control for the survival parameter. For all other parameters no exposure concentration was shown to be significantly different as compared to the control.

The NOEC and LOEC were determined to be 127 and 251 mg/L, respectively, as indicated by the statistical analysis. The MATC was based on these concentrations and was calculated to be 179 mg/L forhydroxypropylated .beta.-cyclodextrin to Daphnia magna.The day-7, -14, and -21 EC50 values based on immobility/mortality estimated to be >1074, > 1074, and 446 mg/L, respectively.