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EC number: 931-597-4 | CAS number: -
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Short-term toxicity to fish
Administrative data
Link to relevant study record(s)
- Endpoint:
- short-term toxicity to fish
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- February 2002
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Non-GLP (no certificate) study conducted according a proposed guideline. Study is reported as a public company report.
- Qualifier:
- according to guideline
- Guideline:
- other: OECD Guideline for testing of chemicals, Draft proposal for a new guideline, Fish Embryo Toxicity (FET) Test (2006)
- Deviations:
- not specified
- GLP compliance:
- not specified
- Analytical monitoring:
- not specified
- Details on sampling:
- Observed with loupe.
- Vehicle:
- no
- Details on test solutions:
- PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Preparation of water extract of ashes (WAF):
- Method: SS-EN 14735:2005/AC:2006
- Concentration: L/S 10
- Shaking time: 24 h
- Filtration: 0.45 µm - Test organisms (species):
- Danio rerio (previous name: Brachydanio rerio)
- Details on test organisms:
- TEST ORGANISM
- Common name: Zebra fish
- Age at study initiation (mean and range, SD): 0 - Test type:
- static
- Water media type:
- freshwater
- Total exposure duration:
- 144 h
- Post exposure observation period:
- Not reported.
- pH:
- See Table 4.
- Salinity:
- See Table 4.
- Nominal and measured concentrations:
- Ash A: 6.25-100%, Ash C: 0.16-25% and Ash F: 6.25-100%
- Details on test conditions:
- TEST SYSTEM
- Embryo cups (if used, type/material, size, fill volume): 250 µl
- Test vessel: 96 multiplate
- Renewal rate of test solution (frequency/flow rate): None
- No. of fertilized eggs/embryos per vessel: 1
- No. of vessels per concentration (replicates): 16
TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: ISO (12890:1999)
- Salinity was adjusted to correspond the salinity in each test concentration
- Chemical parameters: See Table 3
OTHER TEST CONDITIONS
- Adjustment of pH: adjusted to pH 7-8
EFFECT PARAMETERS MEASURED
- Observation intervals: 24, 48 and 144 hours
- Observed effects: deaths, teratogenic and sublethal effects, e.g. malformations in eyes or tails, pigmentation, spontaneous movement, oedema, heart beat, delay in development time
VEHICLE CONTROL PERFORMED: yes - Reference substance (positive control):
- yes
- Remarks:
- Not reported
- Duration:
- 48 h
- Dose descriptor:
- NOEC
- Effect conc.:
- ca. 3.2 other: %
- Conc. based on:
- other: WAF
- Remarks on result:
- other: Ash A
- Duration:
- 48 h
- Dose descriptor:
- NOEC
- Effect conc.:
- ca. 0.78 other: %
- Conc. based on:
- other: WAF
- Remarks on result:
- other: Ash C
- Duration:
- 48 h
- Dose descriptor:
- NOEC
- Effect conc.:
- ca. 25 other: %
- Conc. based on:
- other: WAF
- Remarks on result:
- other: Ash F
- Details on results:
- Significant delay (p<0.001) in hatching time compared to control was observed for ashes A and C (Figure 1 and 2).
- Reported statistics and error estimates:
- See Fig. 1 and 2.
- Sublethal observations / clinical signs:
Table 2. Total content of the elements in the ashes. (TS= dry substance, LOI= Loss on ignition, TOC= Total Organic Carbon, Ntot= total amount of nitrogen)
Metal
Ash A
Ash C
Ash F
Al
mg/kg TS
53500
39400
30000
Ca
mg/kg TS
69000
210000
144000
Fe
mg/kg TS
34300
11500
11300
K
mg/kg TS
14400
38400
56400
Mg
mg/kg TS
9890
12000
15000
Mn
mg/kg TS
594
677
5440
Na
mg/kg TS
36100
49200
11500
P
mg/kg TS
3420
5020
8420
Si
mg/kg TS
284000
65000
220000
Ti
mg/kg TS
4390
8150
1000
As
mg/kg TS
48,8
98,3
< 3
Ba
mg/kg TS
898
1300
1270
Be
mg/kg TS
1,22
0,712
< 0,6
Cd
mg/kg TS
7,32
66,1
8,14
Co
mg/kg TS
20,1
19,3
6,2
Cr
mg/kg TS
360
402
30,5
Cu
mg/kg TS
1690
2580
59,9
Hg
mg/kg TS
0,341
6,09
0,228
La
mg/kg TS
20,7
24,6
< 6
Mo
mg/kg TS
< 6
15,9
< 6
Nb
mg/kg TS
< 6
9,3
< 6
Ni
mg/kg TS
62,2
75,1
17,7
Pb
mg/kg TS
802
2000
52,5
S
mg/kg TS
5280
30900
24800
Sb
mg/kg TS
118
483
1,29
Sc
mg/kg TS
4,25
3,04
< 1
Se
mg/kg TS
2,07
4,24
1,62
Sn
mg/kg TS
242
452
2,19
Sr
mg/kg TS
222
390
492
V
mg/kg TS
33,6
27,7
21,6
W
mg/kg TS
< 60
< 60
< 60
Y
mg/kg TS
12,8
14
9,71
Zn
mg/kg TS
2690
10100
1270
Zr
mg/kg TS
225
120
158
TS
%
87,4
81,8
59,7
LOI
% TS
3,2
13,9
6,6
TOC
% TS
1
1
< 1
N-tot
% av TS
<0,1
<0,1
< 0,1
Table 3. Amounts of metals, chloride, fluoride, sulfur and nitrogen in the leachates of ashes.
Ash A
Ash C
Ash F
Parameter
Unit
Column
Shaked
pH
10
12
11,9
12,4
Conductivity
mS/m
292
5540
1108
1489
Metals
Ca
mg/l
436
4330
95,6
373
Fe
mg/l
0,0102
<0,008
<0.004
0,0095
K
mg/l
63,8
3640
2710
2960
Mg
mg/l
0,344
<0,5
<0.5
<0,2
Na
mg/l
193
4240
395
404
S
mg/l
266
131
1170
1020
Si
mg/l
0,832
0,303
17,2
0,752
Al
μg/l
10100
8,81
848
21,1
As
μg/l
5,3
<20
<3
<1
Ba
μg/l
70,5
2710
64,1
142
Cd
μg/l
0,141
0,444
<0.05
<0,07
Co
μg/l
0,18
<0,1
<0.05
0,157
Cr
μg/l
1,46
32,3
55,9
3,96
Cu
μg/l
300
576
<1
1,66
Hg
μg/l
0,0243
0,0775
<0.02
<0,02
Mn
μg/l
1,1
0,569
0,232
4,04
Mo
μg/l
53,6
155
236
221
Ni
μg/l
3,24
30,1
<0,5
0,51
Pb
μg/l
3,66
7350
<0,2
4,91
Sb
μg/l
30,1
0,225
0,129
0,592
Se
μg/l
1,7
10,1
5,63
6,06
V
μg/l
5,83
9,72
25,8
1,5
Zn
μg/l
5,83
3820
<2
10,4
Anions
Chloride
mg/l
470
16000
569
480
Fluoride
mg/l
<0,20
<12,0
<1,20
<6,00
Sulphate
mg/l
570
330
3080
2800
Other
Ammonium
mg/l
1,4
27
0,024
*****
Nitrate
mg/l
<0,50
<28,0
0,75
*****
Nitrite
mg/l
<0,01
1,14
0,31
*****
TOC/DOC
mg/l
15
2,3
<1
4
Phenol Index
mg/l
0,007
0,009
0,011
<0,005
CODCr
mg/l
33
46
18
*****
BOD7
mg/l
*****
6
<1
*****
Table 4. The validity parameters (pH, oxygen in % and salinity in ‰) for the leachates measured before test start.
Ash A Ash C Ash F pH 9.02 12.55 12.1 Acidity (%) 30 135 93 Salinity (‰ ) 3.4 28.8 5.7 - Conclusions:
- NOEC (144 h) values for three different types of ashes were determined. NOEC for bottom ash formed by burning mainly domestic waste was 3.2 %, NOEC for fresh fly ash formed by burning mainly domestic waste was 0.78 %, and NOEC for fly ash formed by burning mainly biofuels was 25 % of ash WAF in the exposure mixture. Significant delay (p<0.001) in hatching time was reported for bottom ash and for fresh fly ash.
- Executive summary:
A subchronic Fish Embryo Toxicity (FET) test was performed for three different types of ashes according to an OECD Guideline for testing of chemicals, Draft proposal for a new guideline (2006). The used species was Danio rerio. Water Accomodated Fractions (WAFs) of different types of ashes were prepared according to SS-EN 14735:2005/AC:2006 in a ratio L/S 10. Due to delayed toxic responses, observation period used was 144 hours. NOEC for bottom ash formed by burning mainly domestic waste (Ash A) was 3.2 %, NOEC for fresh fly ash formed by burning mainly domestic waste (Ash C) was 0.78 %, and NOEC for fly ash formed by burning mainly biofuels (Ash C) was 25 % of ash WAF in the exposure mixture. Significant delay (p<0.001) in hatching time was reported for bottom ash and for fresh fly ash.
Highest ecotoxicological effects were found for Ash C, for which concentrations of the most toxic heavy metals and salinity were the highest of the studied ashes. The adjusted high salinity affects the complex formation, binding in organic carbon, and bioavailability of metals. The study indicated that the results depend on the preparation method of the ash WAF and therefore, toxicity may be overestimated.
- Endpoint:
- short-term toxicity to fish
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- Not reported.
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: Non-GLP compliant, non-guideline experimental investigation. Study published in scientific, peer reviewed journal.
- Qualifier:
- no guideline available
- Principles of method if other than guideline:
- Oxidative stress and enzyme induction study with freshwater fish Channa punctata and fly ash leachate (24 h).
- GLP compliance:
- no
- Analytical monitoring:
- not specified
- Vehicle:
- no
- Test organisms (species):
- other: Channa punctata
- Details on test organisms:
- TEST ORGANISM
- Common name: Gerai
- Source: obtained from commersial fish suppliers
- Age at study initiation (mean and range, SD): Not reported
- Length at study initiation (length definition, mean, range and SD): 15-17 cm
- Weight at study initiation (mean and range, SD): 70-75 g
- Feeding during test: No
ACCLIMATION
- Acclimation period: 15 d
- Acclimation conditions (same as test or not): glass aquarium; the tank water was kept oxygen saturated by aeration and temperature was maintained at the ambient laboratory temperature (25±2°C)
- Type and amount of food: commercial fish feed
- Feeding frequency: twice a week - Test type:
- static
- Water media type:
- freshwater
- Limit test:
- yes
- Total exposure duration:
- 24 h
- Details on test conditions:
- Ten animals were used both in the test water and in the control.
EFFECT PARAMETERS MEASURED: lipid peroxidation, antioxidant enzyme activity and glutathione reduction at the end of the test
TEST CONCENTRATIONS: Fish were exposed to 1 ml/l of fly ash leachate representing 100 mg/mL of fly ash in the original slurry. This concentration was selected on the basis of range finding study. - Reference substance (positive control):
- no
- Duration:
- 24 h
- Remarks on result:
- not measured/tested
- Remarks:
- Tested parameters included: - induction of lipid peroxidase in liver, kidney and gills - induction of catalase activity in liver and kidney and gills - induction of GST activity in liver and kidney and gills - increase of GSH in liver and kidney and gills. No LC-, EC-, NOEC or LOEC values were determined.
- Details on results:
- Significant differences in the tested parameters compared to control were observed (see Figures).
- Reported statistics and error estimates:
- Statistically significant findings compared to control were:
- induction of lipid peroxidase in liver, kidney and gills (p<0.001)
- induction of catalase activity in liver and kidney (p<0.01) and gills (p<0.001)
- induction of GST activity in liver and kidney (p<0.05) and gills (p<0.01)
- increase of GSH in liver and kidney (p<0.05) and gills (p<0.01) - Sublethal observations / clinical signs:
See illustration.
- Validity criteria fulfilled:
- not specified
- Conclusions:
- Sub-bituminous coal ash was found to induce oxidative stress and enzyme activity in Channa punctata.
- Executive summary:
Oxidative stress induction potential of fly ash leachate (FAL) in Channa punctata was studied in a non-GLP non-guideline 24 h study. The sub-bitominous fly ash was obtained from a thermal power plant dumping site. The leachate was prepared by mixing the ash in water one hour every day for seven days, after which the leachate was filtrated. Amount of fly ash in the studied leachate was 100 mg/mL. After the exposure time, fish were homogenised followed by centrifugation. The supernatant was used for the biochemical analyses such as rate of lipid peroxidation, acitivity of catalase and gluthathione S-transferase enzymes and level of glutathione. Exposure to fly ash leachate induced lipid peroxidation, caused elevated enzyme activities and increased levels of glutathione in all the studied organs. All the effects were statistically significant compared to controls.
- Endpoint:
- short-term toxicity to fish
- Type of information:
- experimental study
- Adequacy of study:
- weight of evidence
- Study period:
- Not reported
- Reliability:
- 3 (not reliable)
- Rationale for reliability incl. deficiencies:
- other: Non-GLP compliant, non-guideline experimental investigation. Study published in scientific, peer reviewed journal.
- Qualifier:
- no guideline available
- Principles of method if other than guideline:
- Cytological and genetic in-vitro study with hepatocytes from freshwater fish Channa punctata.
- GLP compliance:
- no
- Analytical monitoring:
- not specified
- Vehicle:
- no
- Test organisms (species):
- other: Hepatocytes from Channa punctata
- Test type:
- other: in-vitro
- Water media type:
- freshwater
- Limit test:
- no
- Total exposure duration:
- 48 h
- Remarks on exposure duration:
- also 24h
- Duration:
- 48 h
- Remarks on result:
- not measured/tested
- Remarks:
- The studied parameters were apoptosis, DNA fragmentation and laddering, caspases, cytochrome-c, lactate dehydrogenase (LDH). No LC-, EC-, NOEC or LOEC values were determined.
- Sublethal observations / clinical signs:
Apoptosis, DNA fragmentation and DNA laddering
All the studied parameters indicate a proapoptotic effect of fly ash leachates in fish hepatocytes (See figures 1 -2).
Caspases, cytochrome-c and LDH
A concentration-dependent increase in the activity of caspases 3, 7 and 10 was observed in cells exposed to different concentrations of FAL for 48 h. The maximum activity of the caspases was recorded in the cells exposed to the highest concentration of FAL (10%). The increase was significant at FAL concentrations (w/v) of 2% (P < 0.05), 5% (P < 0.01) and 10% (P < 0.001). The increase of caspase-9 was most pronounced of the caspases (P < 0.001). Compared to control, increase of release of cytochrome-c was significant (P<0.05) in all concentrations of FAL except for 1%. LDH activity increased significantly at two highest concentrations (P<0.05 at 5% and P<0.01 at 10%) with high variation in the results. See figures 3A-C.
H2O2, superoxide ions and LPO
FAL exposure also resulted in a significant concentration and time dependent increase in H2O2 production by hepatocytes (Fig. 4A). The maximum increase in production of H2O2was observed at 10% FAL concentration. Also at other FAL concentrations (1%, 2% and 5%) there was significant (P<0.01) increase. However, exposure at low concentration (1% FAL) resulted in a significant increase in 48h analysis (P<0.05). Instead, higher production of superoxide ions was observed at 24h than at 48h. At 10% FAL exposure, there was a significant increase in superoxide ion production at 24 h (P<0.01 and at 48h (P<0.05) compared to control in all concentrations except for 1% at 48h. LPO measurement in C. punctata hepatocytes showed significant increase at all the concentration of FAL (except 1%) at 24 h as well as 48 h (Fig. 4C).
- Validity criteria fulfilled:
- not specified
- Conclusions:
- Sub-bituminous coal fly ash leachate caused significant cytotoxic effects on hepatocytes from Channa punctata. Also, effects in DNA level were obvious.
- Executive summary:
The pro-apoptotic effects of fly ash leachate (FAL) in hepatocytes of Channa punctata was studied in a non-GLP non-guideline in-vitro study. The sub-bitominous fly ash was obtained from a thermal power plant dumping site. The leachate was prepared by mixing the ash in water one hour every day for seven days, after which the leachate was filtrated.
The hepatocytes were isolated with a double perfusion method. The hepatocyte cell density was 4 x 106cells/mL. The medium used was RPMI-1640 medium with 1% FBS, 25 µL/mL gentamycin sulphate and 2.5 µg/mL amphotericin-B. A slurry containing ash was prepared. Cytologic and genetic effects of 10 % sub-bituminous coal fly ash leachate were studied at concentrations of 0, 1, 2, 5 and 10% of FAL from this 10% w/v FAL solution for 24 and 48 h. The studied parameters were apoptosis, DNA fragmentation and laddering, caspases, cytochrome-c, lactate dehydrogenase (LDH)
H2O2, and superoxide ions and lipid peroxidation (LPO). The used techniques were microscopic observation, plate scanning and electrophoresis.
Apoptotic and DNA fragmentation effects of FLA were evident. Significant effects of FAL were observed on caspase activity (P < 0.001), cytochrome-c release (P < 0.05), lactate dehydrogenase activity (P < 0.01). Also the oxidative stress bimarkers showed significant elevation (P < 0.01): H2O2release, superoxide ion production and lipid peroxidation.
Referenceopen allclose all
Description of key information
Short-term toxicity to fish was estimated based on three publications from literature. One was a guideline compliant study in which Dania rerio was exposed to water accomodated fractions (WAFs) of three different types of ashes for 144 h. Statistically significant (p<0.001) delay in hatching time was used as the endpoint and for determining NOEC. The other two studies were non-GLP compliant, non guideline investigations in which Channa punctata was exposed to fly ash lechate for 24 h and 48 h. In the first investigation, oxidative stress induction potential was studied using lipid peroxidation, GST activity, levels of GSH and proteins as endpoints. In the second investigation, the pro-apoptotic effects (apoptosis, DNA fragmentation and laddering, caspases, cytochrome-c, lactate dehydrogenase LDH, H2O2, and superoxide ions and lipid peroxidation) were studied. Highest ecotoxicological effects were found for ash, for which concentrations of the most toxic heavy metals and salinity were highest. Significant differences in parameters indicating oxidative stress potential as well significant cytotoxic effects in hepatocytes and effects in DNA level were observed in exposed fish compared to the control. LC50 was not reported in any of the studies.
Key value for chemical safety assessment
Additional information
NOEC was obtained from a guideline compliant study with zebra fish (Dania rerio) exposed to water accomodated fractions of three different types of ash. NOEC for bottom ash formed by burning mainly domestic waste was 3.2 % ash in WAF, NOEC for fresh fly ash formed by burning domestic waste mainly was 0.78 % ash in WAF, and NOEC for fly ash formed by burning biofuels mainly was 25 % ash in WAF. For 10 % coal ash slurry, sublethal effects in fish were significant. It was suspected that high salinity influenced the outcome of the test by affecting the complex formation, binding in organic carbon, and bioavailability of metals. The study indicates overestimation of the results depending on the preparation method of the ash WAF.
It must be noted that the results represent worst-case scenario since in typical uses of ash, it is not intended to be released to water environment directly. Thus, environmental concentrations of ash and salinity effects in water are lower than in this study.
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