Santicizer P1400

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OTS 797.1330 (Daphnid Chronic Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
Test Material: P1400 (synonym Santicizer Platinum P-1400)
CAS No.: 1200806-67-2; Lot No.: 945)
Received on: 30 January 2017
Stotage: room temperature
Appereance: colorless to light yellow liquid
Expiration Date: 30 January 2018.

Analytical monitoring:

yes

Details on sampling:

A primary stock solution of P1400 was prepared on 08 March 2017 by weighing 23.1 mg of P1400 into a 25-mL volumetric flask and bringing the flask to volume with cyclohexane for a
concentration of 0.924 mg/mL. Subsequent dilutions of the primary standard in cyclohexane were used as analytical standard solutions.
A primary stock solution of P1400 was prepared on 8 March 2017 by weighing 106.9 mg of P1400 into a 25-mL volumetric flask and bringing the flask to volume with methanol for a concentration of 4.28 mg/mL. This solution and subsequent dilutions in methanol were used as matrix spiking solutions for P1400 method validation samples and quality control (QC) fortification samples during the definitive test.

ll solutions were stored refrigerated when not in use.

Vehicle:

yes

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna neonates (<24-hours old)
- Source: Obtained from an in-house daphnid culture
- Age of parental stock (mean and range, SD): NFrom a continuous breeding stock
- Feeding and Food type: fed a suspension of the algal species Pseudokirchneriella subcapitata (formerly Selenastrum capricornutum) at least once a day supplemented by a prepared artificial diet consisting of a yeast, wheat grass (cereal leaves), and
salmon starter (trout feed) suspension (termed YCT)
- Parents selection: Approximately one day prior to neonate selection, adult daphnids were isolated by transferring the adults to a fresh culture with a water/food suspension. The adults were considered acceptable with the lack of the following confirmed: signs of stress, presence of males, documented delay in first brood production, ephippia present, disease or physical damage
Mortality: No adult mortality occurred during the 48-hour period immediately prior to production of neonates used to initiate the definitive test. The adults producing the young for the definitive test were approximately 28-days old.
Acclimatation: Since the culturing and testing environmental parameters were equivalent (i.e., temperature, dilution water, and lighting), no acclimation period was necessary.
- Culture conditions: temperature-controlled area at approximately 20°C


QUARANTINE (wild caught)
- Duration: not applicable
- Health/mortality: not applicable

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Post exposure observation period:

no

Hardness:

Hardness (mg CaCO3/L)
Fresh Solutions: 146 – 150
Spent Solutions: 140 - 150

Test temperature:

Temperature (ºC)
Fresh Solutions: 19.9 – 20.4
Spent Solutions: 19.6 – 20.2

pH:

pH
Fresh Solutions: 8.4 – 8.6
Spent Solutions: 8.2 – 8.6

Dissolved oxygen:

Dissolved Oxygen (mg/L)
Fresh Solutions: 7.8 to 8.7 mg/L (90 to 100% sat)
Spent Solutions: 5.9 to 8.7 mg/L (68 to 100% sat)

Salinity:

Conductivity was measured with a WTW Conductivity & Salinity Meter Model 3300i.

Conductivity:

Conductivity was measured with a WTW Conductivity & Salinity Meter Model 3300i.

Nominal and measured concentrations:

Nominal Concentrations: 0 (control), 0 (vehicle control), 50, 100, 200, 400, and 800 µg/L
Measured Concentrations:

Details on test conditions:

EXPOSURE SYSTEM

The definitive test was conducted using 125-mL glass jars as test chambers. Test chambers contained approximately 80 mL of control, vehicle control, or test substance solution, and were covered with clear plastic Petri dishes. All test chambers were labeled with the study number, treatment, and replicate. The test chambers were randomly positioned in a temperature-controlled water bath using a computer-generated randomization table, and maintained at 20 ± 1°C. The control, vehicle control, and all test substance treatments were replicated 10 times. Fluorescent lighting was maintained on a 16-hour daylight photoperiod with two 30-minute simulated dawn and dusk periods. The light intensity during the definitive test was 542 lux as measured on Day 0 with a LI-COR Model LI-250A light meter equipped with a photometric sensor.

RANGE FINDING TEST

A 14-day static-renewal range-finding test was initiated at nominal concentrations of 0 (control), 0 (vehicle control), 0.00080, 0.0080, 0.080, and 0.80 mg/L. The control, vehicle control, and all test substance treatments were replicated five times (A through E, 1 daphnid each). After 14 days of exposure there was 80, 100, 100, 100, 80, and 100% survival in the 0 (control), 0 (vehicle control), 0.00080, 0.0080, 0.080, and 0.80 mg/L treatments, respectively. The total number of live young produced during the 14-day exposure was 346, 469, 443, 534, 376, and 66 neonates in the control, vehicle control, 0.00080, 0.0080, 0.080, and 0.80 mg/L treatments, respectively. The mean number of live young produced per living adult after 14 days of exposure were 83, 94, 89, 107, 92, and 9 neonates in the control, vehicle control, 0.00080, 0.0080, 0.080,
and 0.80 mg/L, respectively.

DEFINITIVE TEST

The definitive test was performed with target nominal concentrations of 0 (control), 0 (vehicle control), 50, 100, 200, 400, and 800 µg/L. The primary stock solutions were prepared at a concentration of 8.0 mg/mL by dissolving approximately 0.0400 g of P1400 in 5.0 mL of dimethylformamide (DMF).

Key result

Duration:

21 d

Dose descriptor:

NOEC

Remarks:

Adult Survival

Effect conc.:

673 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

83.8 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Key result

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 673 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

mortality

Key result

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

173 µg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

SURVIVAL
After 21 days of exposure, survival of D. magna exposed to P1400 was 100, 100, 90, 100, 100, 100, and 100% in the 0 (control), 0 (vehicle control), 44.5, 83.8, 173, 346, and 673 µg/L time weighted mean measured treatments, respectively


BEHAVIORAL CHANGES
No morphological or behavioral changes were observed.


EGGS OBSERVATION
Unhatched eggs were observed in one replicate of the 346 mg/L test treatment at day-18. Unhatched eggs were observed in one replicate of the 673 mg/L test treatment at day-12, four replicates on day-14, six replicates on day-16, all replicates on day-18, three replicates on day-20, and seven replicates on day-21.


REPRODUCTION OBSERVATIONS
The first broods were observed after 7 days of exposure in the control, and after 8 days in the vehicle control, 44.5, 83.8, 173, 346, and 673 µg/L treatments. The mean time to first brood was 8, 8, 8, 8, 8, 8, and 9 days in the 0 (control), 0 (vehicle control), 44.5, 83.8, 173, 346, and 673 µg/L treatments, respectively. There were no ephippia observed in any control or test substance treatment chambers during the exposure. There was a significant increase in the mean time to first brood in the 673 µg/L test treatment compared to the pooled control. Based on the time weighted mean measured concentrations, the NOEC, LOEC, and MATC values for time to first brood were 346, 673, and 483 µg/L, respectively. The estimated EC 10 , EC 20 , and EC 50 values for the time to first brood, based on time weighted mean concentrations, were all >673 µg/L, the highest concentration tested.

The first brood production rate was 0.135, 0.133, 0.131, 0.133, 0.133, 0.133, and 0.121 in the 0 (control), 0 (vehicle control), 44.5, 83.8, 173, 346, and 673 µg/L treatments, respectively. There was a significant reduction in mean first brood production rate in the 673 µg/L test treatment compared to the pooled control. Based on the time weighted mean measured concentrations, the NOEC, LOEC, and MATC values for first brood production rate were 346, 673, and 483 µg/L, respectively. The estimated EC 10 , EC 20 , and EC 50 values for the first brood production rate, based on time weighted mean concentrations, were 671 µg/L (95% confidence limits of 646 to 696 µg/L), >673 µg/L, and >673 µg/L, the highest concentration tested.


LIVE YOUNG PRODUCED PER SURVIVING ADULT
The mean number of live young produced per surviving adult during the course of the test was 201, 205, 204, 205, 193, 158, and 16 in the 0 (control), 0 (vehicle control), 44.5, 83.8, 173, 346, and 673 µg/L treatments, respectively (Table 7). There was a significant reduction in mean number of live young produced per surviving adult in the 173, 346, and 673 µg/L test treatments compared to the pooled control. Based on the time weighted mean measured concentrations, the NOEC, LOEC, and MATC values for the number of live young produced per surviving adult were 83.8, 173, and 120 µg/L, respectively. The estimated EC 10 , EC 20 , and EC 50 values for the number of live young produced per surviving adult, based on time weighted mean concentrations, were 293 µg/L (95% confidence limits of 288 to 299 µg/L), 339 µg/L (95% confidence limits of 334 to 344 µg/L), and 433 µg/L (95% confidence limits of 428 to 438 µg/L), respectively.


LIVE YOUNG PRODUCED PER ADULT REPRODUCTIVE DAY
The mean number of live young per adult reproductive day was 14, 15, 15, 15, 14, 11, and 1 in the 0 (control), 0 (vehicle control), 44.5, 83.8, 173, 346, and 673 µg/L treatments, respectively. There was a statistically significant reduction in mean number of live young per adult reproductive day in the 173, 346, and 673 µg/L test treatments as compared to the pooled control. Based on the time weighted mean measured concentrations, the NOEC, LOEC, and MATC values for mean number of live young per adult reproductive day were 83.8, 173. and 120 µg/L, respectively. The estimated EC 10 , EC 20 , and EC 50 values for the mean number of live young per adult reproductive day, based on time weighted mean concentrations, were 294 µg/L (95% confidence limits of 288 to 300 µg/L), 339 µg/L (95% confidence limits of 334 to 345 µg/L), and 433 µg/L (95% confidence limits of 428 to 439 µg/L), respectively.


MEAN LENGTH
Mean length of the daphnids were 4.59, 4.54, 4.56, 4.57, 4.55, 4.42, and 4.08 mm for the 0 (control), 0 (vehicle control), 44.5, 83.8, 173, 346, and 673 µg/L treatments, respectively. There was a significant reduction in individual lengths in the 346 and 673 µg/L test substance treatment as compared to the pooled control (i.e. 4.57 mm). Based on length, the 21-day NOEC, LOEC, and MATC values were 173, 346, and 245 µg/L, respectively. The 21-day EC 10 , EC 20 , and EC 50 based on length and time weighted mean measured concentrations was estimated as 646 µg/L (95% confidence limits 616 to 676 µg/L), >673 µg/L, and >673 µg/L, the highest concentration tested, respectively.


MEAN DRY WEIGHT
Mean dry weights of the daphnids were 0.870, 0.833, 0.879, 0.911, 0.922, 0.890, and 0.605 mg for the 0 (control), 0 (vehicle control), 44.5, 83.8, 173, 346, and 673 µg/L treatments, respectively. There were significant reductions in individual dry weights in the 673 µg/L test substance treatment as compared to the pooled control (i.e. 0.852 mg). Based on dry weight, the 21-day NOEC, LOEC, and MATC values were 346, 673, and 483 µg/L, respectively. The 21-day EC 10 and EC 20 values, based on dry weights and time weighted mean measured concentrations, could not be estimated due to a lack of convergence in the data. The EC 50 value, based on dry weights and time weighted mean measured concentrations was estimated as >673 µg/L, the highest concentration tested.

See attachement for further infomation

Results with reference substance (positive control):

- Results with reference substance valid? yes

Reported statistics and error estimates:

All statistical analyses were performed using SAS software (version 9.3).

See attachement for further infomation

Validity criteria fulfilled:

yes

Conclusions:

The mortality of parent animals in the control did not exceed 20% at the end of the test and no ephippia were produced by the control animals. The mean number of live young produced per surviving control adult daphnid at the end of the test was ≥60. This study is classified as acceptable and satisfies the guideline requirement for a reproduction test with Daphnia magna. Mean number of live young per adult reproductive day and mean number of live young per live adult were the most sensitive biological parameters.
The NOEC Adult Survival and Reproduction were established at 673 µg/L and 83.8 µg/L, respectively.

Executive summary:

A 21-day static-renewal chronic toxicity test with Daphnia magna was performed with the test material P1400. The aim of this study was to evaluate the effect of the test substance on the survival, growth, and reproductive output of Daphnia magna. 

The study was conducted according to the U.S. EPA OCSPP 850.1300 and the OECD Guideline 211.

The test acceptability criteria were met for this study. The mortality of parent animals in the control did not exceed 20% at the end of the test and no ephippia were produced by the control animals. The mean number of live young produced per surviving control adult daphnid at the end of the test was ≥60. This study is classified as acceptable and satisfies the guideline requirement for a reproduction test with Daphnia magna. Mean number of live young per adult reproductive day and mean number of live young per live adult were the most sensitive biological parameters.

According to the results of this study, the NOEC for Adult Survival and Reproduction (Reproduction expressed as number of live young per surviving adult daphnid) were determined to be 673 µg/L and 83.3 µg/L, respectively.

Description of key information

A 21-day static-renewal chronic toxicity test with Daphnia magna exposed to P1400 was conducted to evaluate the effect of the test substance on the survival, growth, and reproductive output of Daphnia magna. This was achieved by measuring the effects of exposure to the test substance on survival, young per surviving adult, length, and dry weight as determined by comparing control and exposure treatments.

The test acceptability criteria were met for this study. The mortality of parent animals in the control did not exceed 20% at the end of the test and no ephippia were produced by the control animals. The mean number of live young produced per surviving control adult daphnid at the end of the test was ≥60. This study is classified as acceptable and satisfies the guideline requirement for a reproduction test with Daphnia magna. Mean number of live young per adult reproductive day and mean number of live young per live adult were the most sensitive biological parameters.

According to the results of this study, the NOEC for Adult Survival and Reproduction (Reproduction expressed as number of live young per surviving adult daphnid) were determined to be 673 µg/L and 83.3 µg/L, respectively.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

NOEC

Remarks:

Reproduction

Effect concentration:

83.3 µg/L