AMMONIUM SULFATE CAS NÂ°: 7783-20-2

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OECD SIDS AMMONIUM SULFATE 5. TOXICITY ID: 7783-20-2 DATE: 18.04.2006 19-MAY-2003 (144) Type: Cytogenetic assay System of testing: V79 cells Concentration: hypertonic solutions with final osmolalities of 500, 750, 1000 or 1500 mOsm/kg H2O (corresponding to about 166.6; 250; 333.3; 500 mM = 22; 33; 44; 66 mg/mL) (converted according to information given by Nowak C (1987), Ter Carc Mut 7, 515) Cytotoxic Concentration: not cytotoxic up to and including the highest tested osmolality (1500 mOsm) Metabolic activation: without Method: other GLP: no data Test substance: other TS: ammonium sulfate, purity 99.5% Remark: Result: The study was designed to investigate the effect of hypertonic solutions on ethylmethane sulfonate induced HPRT mutations and chromosomal aberrations. Treatment of ethyl methanesulfonate-induced cells with hypertonic solutions containing sodium chloride or ammonium sulfate had a clear effect on chromosomal aberration but TGr (6-thioguanine resistance) mutations were only enhanced by ammonium sulfate posttreatment, and not by sodium chloride posttreatment. % ABERRANT METAPHASES (200 metaphases analyzed, 2 independent experiments, fixation time 13 hours): 1.0 %, 1.0%, 2.0%, 4.5% and 8.0% for controls, 500, 750, 1000 and 1500 mOsm ammonium sulfate, respectively. 8.0%, 42.0%, 42.5%, 51% and 43% for EMS, EMS+500, EMS+750, EMS+1000 and EMS+1500 mOsm ammonium sulfate, respectively. % ABERRANT METAPHASES (200 metaphases analyzed, 2 independent experiments, fixation time 16 hours): 1.0 %, 3.5%, 3.5%, 3.0% and 7.5% for controls, 500, 750, 1000 and 1500 mOsm ammonium sulfate, respectively. 8.0%, 8.5%, 19.5%, 30.5% and 26.5% for EMS, EMS+500, EMS+750, EMS+1000 and EMS+1500 mOsm ammonium sulfate, respectively. TG MUTATIONS (per 10e6 survivors, 2 independent experiments: 28.0 %, 48.7%, 55.95%, 49.9% and 65.5% for controls, 500, 750, 1000 and 1500 mOsm ammonium sulfate, respectively. 314.6%, 584.0%, 512.4%, 478.0% and 782.4% for EMS, EMS+500, EMS+750, EMS+1000 and EMS+1500 mOsm ammonium sulfate, respectively. Test condition: 192 It was suggested by the author that hypertonic salt posttreatment leads to conformational changes in the DNA, resulting in an increase in TGr mutations and chromosomal aberrations. YEAR OF STUDY: not reported. TEST SYSTEM: V79 hamster cells were grown at 37 deg C in a humidified atmosphere with 5% CO2 in Earle`s minimal essential medium (MEM), supplemented with 10% fetal calf serum, 100 units penicillin, and 0.128 mg/mL UNEP PUBLICATIONS
OECD SIDS AMMONIUM SULFATE 5. TOXICITY ID: 7783-20-2 DATE: 18.04.2006 dihydrostreptomycin sulfate in 10-cm petri dishes; 48 hr before the treatment 2x10e5 cells from the stock culture were seeded in 10-cm petri dishes with 10 mL of complete medium. MUTAGEN TREATMENT: Before adding the mutagen the culture medium was discarded and replaced by 10 mL of prewarmed MEM. 2 mL of ethyl methanesulfonate (EMS), dissolved in MEM were added to each dish, to give a final concentration of 20 mM. After 1 h of treatment at 37 deg C, the medium was removed and the dishes were washed twice with 10 mL of prewarmed phosphate-buffered saline (PBS). POST-TREATMENT WITH AMMONIUM SULFATE or SODIUM CHLORIDE: The hypertonic treatment was performed immediately after the mutagen treatment. The washing solution was carefully removed, then the cells were covered with 300 uL of PBS to avoid drying. Then 300 uL of ammonium sulfate or sodium chloride solution were added to give a final osmolality of 500, 750, 1000 or 1500 mOsm/kg H2O. Controls and EMS-treated cultures received 600 uL of PBS (300 mOsm/kg H2O). The osmolality of all solutions was measured with a microosmometer. After 30 min at 37 deg C the treatment was stopped by adding 5 mL of PBS. The supernatant of all dishes was collected in centrifuge tubes so that no cells were lost. Then the cells were trypsinized by adding 2 mL of trypsin/EDTA. After 3 min the cells were collected with 2 mL of medium and added to the supernatant. After centrifugation the cells were counted and plated for the chromosomal aberration and TGr mutation assay. CHROMOSOMAL ABERRATIONS: Immediately after the posttreatment with hypertonic salt solutions 10e6 cells were transferred to 6-cm dishes and cultivated for 13 or 16 hours, respectively, in complete medium at 37 deg C, including a 2.5 h colcemid treatment. The cells were fixed in methanol-acetic acid and stained with 5% Giemsa. In some experiments cultures containing BrdUrd were set up in parallel to ensure that only first post-treatment metaphases were scored for chromosomal aberrations. TGr MUTATIONS: 5x10e5 cells were seeded in 10-cm dishes and cultivated for 7 days after the treatment. Then the medium was removed, the cells were washed once with PBS and trypsinized. Then 10e5 cells were seeded in 6-cm dishes (10 per concentration) and grown in 5 mL complete medium including 10.9 ug/mL of 6-thioguanine (TG). Cell survival tests were carried out in parallel to the selection procedure. Colonies were fixed and stained after 8 days of incubation. Reliability: (3) invalid Unsuitable test system (unphysiological, hypertonic test conditions). 12-APR-2006 (145) 5.6 Genetic Toxicity 'in Vivo' Type: Micronucleus assay Species: mouse Sex: male Strain: other: ddY Route of admin.: i.p. UNEP PUBLICATIONS 193
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OECD SIDS AMMONIUM SULFATE FOREWORD
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OECD SIDS AMMONIUM SULFATE 10. Comm
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OECD SIDS AMMONIUM SULFATE Consumer
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OECD SIDS AMMONIUM SULFATE 1.3 Phys
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OECD SIDS AMMONIUM SULFATE - in the
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OECD SIDS AMMONIUM SULFATE The inte
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OECD SIDS AMMONIUM SULFATE 228 µg/
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OECD SIDS AMMONIUM SULFATE also an
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OECD SIDS AMMONIUM SULFATE 3.1.3 Ir
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OECD SIDS AMMONIUM SULFATE diarrhoe
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OECD SIDS AMMONIUM SULFATE Table 3
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OECD SIDS AMMONIUM SULFATE Toxicity
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OECD SIDS AMMONIUM SULFATE Table 7
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OECD SIDS AMMONIUM SULFATE I U C L
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OECD SIDS AMMONIUM SULFATE 1. GENER
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AMMONIUM SULFATE CAS NÂ°: 7783-20-2

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