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EC number: 231-511-9 | CAS number: 7601-89-0
- Life Cycle description
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- Endpoint summary
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- Ecotoxicological Summary
- Aquatic toxicity
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- 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
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- Toxicological Summary
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- Genetic toxicity
- Carcinogenicity
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- Additional toxicological data
Endpoint summary
Administrative data
Description of key information
Ammonium perchlorate was shown to have antithyroid effects, without any other repeat-dose toxic effects, in a Klimisch 2 GLP rat 90-day study at up to 10 mg/kg/day via drinking water. The overall NOAEL was 1 mg/kg/day. All antithyroid effects were demonstrated to be partly to completely reversible within 30 days.
However, the antithyroid effects from a rat 2-generation study were considered to be more reliable for the classification and risk assessment related to repeated exposure, because of this study's higher sensitivity than the 90-day study (40% longer treatment duration: 113-142 days, more individual thyroid data per sex per dose: 56-60 vs. 9-10, 3-fold higher top-dose, presence of highly sensitive populations: pregnant dams and pups) and of the better biological relevance of the dose-relationship (dose-increase, vs. plateauing of effects in the 90-day study). This resulted in a lower NOAEL, reported below, of 0.3 mg/kg/day.
The full RSS of the 2-generation study is under section 7.8.1. In this study conducted at up to 30 mg/kg/day, parental and offspring retained NOAELs for antithyroid effects were both 0.3 mg/kg/day.
Key value for chemical safety assessment
Repeated dose toxicity: via oral route - systemic effects
Link to relevant study records
- Endpoint:
- sub-chronic toxicity: oral
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Study period:
- August 1997 - June 1998
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- Sufficiently compliant to GLP and testing guideline; adequate coherence between data, comments and conclusions. The few deviations have no impact on the reliability and completeness of the conclusions. Histopathological examinations did not distinguish between hypertrophy and hyperplasia, but this was done later by peer-review. Justification for read-across : Mammalian toxicity data of an analogous substance, ammonium perchlorate (CAS no. 7790-98-9), has been used for read across where data gaps for sodium perchlorate exist (repeated toxicity).
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
- Deviations:
- yes
- Remarks:
- Temperature (°C): 18-26 instead of 19-25. Highest dose chosen with the aim to induce antithyroid effects, but not based on dose-limiting toxicity. No histological examination of Peyer’s patches.
- GLP compliance:
- yes
- Remarks:
- No laboratory compliance certificate. No analytical certificate for test item. Perchlorate concentrations were occasionally outside the range validated for stability.
- Limit test:
- no
- Species:
- rat
- Strain:
- Sprague-Dawley
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Laboratories
- Age at study initiation: 6-7 weeks (indicated only in published version of report)
- Weight at study initiation (day -1): 195-197 g (M), 151-153 g (F)
- Fasting period before study: no
- Housing: individual suspended stainless steel wire mesh cages
- Diet (e.g. ad libitum): PMI Certified Rodent Chow, ad libitum
- Water (e.g. ad libitum): reverse osmosis water, ad libitum
- Acclimation period: 11 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-26
- Humidity (%): 30-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 1997-9-9 To: 1998-1-9 - Route of administration:
- oral: drinking water
- Vehicle:
- water
- Details on oral exposure:
- VEHICLE
- Nature: reverse osmosis drinking water
- Justification for use and choice of vehicle: high solubility of the test item in water
- Lot/batch no. (if required): not applicable
- Purity: not applicable
- Analyzed and found to contain no detectable nitrate, a potential interference ion for perchlorate analysis
PREPARATION OF TREATED DRINKING WATER:
- Dilution from a stock solution at 50 mg/mL
- Concentration adapted weekly (using body weight and water consumption) to reach the target dose-levels in mg/kg/day: 0.04-129 µg/mL
- Stirred for 30 min before storage, and again before delivery
- Frequency of preparation: at least every five weeks (stock solution), and every week (diluted solutions)
- Storage : refrigerated (diluted solutions) - Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- Methods:
- validated ion chromatography method
- LOD for perchlorate and nitrate (interference ion): 0.005 µg/mL
Results:
- at 0.05 and 200 µg/mL: concentrations within +/- 10% of nominals
- at 0.05 and 200 µg/mL: stable over 109 days at 21-22°C under light (12h/day)
Comment:
- For the first of 5 concentration measurement dates, group 3 was exposed to lower drinking water perchlorate (nominal and achieved) levels than group 2: it was likely a 10-fold too low nominal concentration setting by error (0.04 µg/mL instead of > 0.4 µg/mL at other preparation dates). Furthermore, this low level was below the minimal concentration demonstrated to be stable (0.05 µg/mL).
- This validation otherwise adequately covers the study's test item use conditions. - Duration of treatment / exposure:
- - 14 days for satellite groups
- 90 days for terminal and recovery groups
Recovery: for 30 days after the 90-day treatment period - Frequency of treatment:
- Continuous (treated water ad libitum)
- Remarks:
- Doses / Concentrations:
0.01, 0.05, 0.2, 1 and 10 mg/kg/day
Basis:
other: nominal dose; achieved dose within +/- 10% of nominal - No. of animals per sex per dose:
- 10M+10F (interim group) + 10M+10F (terminal group): at each dose
+10M+10F (recovery groups): at 0, 0.05, 1 and 10 mg/kg/day (there were thus in total 20M+20F treated for 90 days in these groups) - Control animals:
- yes, concurrent vehicle
- Details on study design:
- - Dose selection rationale: panel of toxicology experts, focused on antithyroid effects, to identify doses with frank effects and a possible NOEL
- Rationale for animal assignment: random
- Rationale for selecting satellite groups: random
- Post-exposure recovery period in satellite groups: 30 days, in "recovery" groups treated for 90 days
- Section schedule rationale: random - Positive control:
- Only for micronucleus interpretation (additional animals treated with cyclophosphamide i.p.)
- Observations and examinations performed and frequency:
- CAGE SIDE OBSERVATIONS: Yes
- Time schedule: once daily
- Mortality and morbidity
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once weekly
BODY WEIGHT: Yes
- Time schedule for examinations: weekly
FOOD CONSUMPTION: Yes
- Food consumption for each animal: weekly
FOOD EFFICIENCY: No
WATER CONSUMPTION AND COMPOUND INTAKE: Yes
- Time schedule for examinations: weekly, individually
- Calculated as mg/kg/day from body weight data
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: day -1, day 85 and day 119.
- Dose groups that were examined: all
HAEMATOLOGY: Yes
- Time schedule for collection of blood: sacrifice (14, 90 or 120 days of study)
- Anaesthetic used for blood collection: No data
- Animals fasted: No
- How many animals: 10 per timepoint
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: sacrifice (14, 90 or 120 days of study)
- Animals fasted: No
- How many animals: 10 per timepoint
URINALYSIS: No
NEUROBEHAVIOURAL EXAMINATION: No (not a deviation as it was done in another study) - Sacrifice and pathology:
- GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
All guideline-required organs and tissues were exmained, except for histological examination of Peyer's patches. - Other examinations:
- - Estrous cycles, all females daily during 3 weeks before sacrifices at 90 and 120 days*
- Sperm analyses, all males at 90 and 120-day sacrifices*
- Bone marrow micronucleus evaluation, rats at 0 and 10 mg/kg/day and positive controls, at 90 and 120-day sacrifices (NB: does not follow requirements of an in vivo micronucleus study)*
*: low-detail sufficient as specific, more sensitive studies cover the endpoints fertility and mutagenicity
- Serum hormone levels: TSH, T3, T4 in all rats of all groups at 14, 90 and 120 days. - Statistics:
- Continuous variables: one-way ANOVA + Tukey-Kramer intergroup comparisons
Chi-square for % females cycled.
All tests at significance level of 5%. - Clinical signs:
- no effects observed
- Mortality:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Food efficiency:
- not examined
- Water consumption and compound intake (if drinking water study):
- no effects observed
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- no effects observed
- Clinical biochemistry findings:
- no effects observed
- Urinalysis findings:
- not examined
- Behaviour (functional findings):
- not examined
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Gross pathological findings:
- effects observed, treatment-related
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Histopathological findings: neoplastic:
- no effects observed
- Details on results:
- MORTALITY:
One female at 0.05 mg/kg/day died during the recovery period. The cause could not be determined, but was not considered to be related to treatment.
BODY WEIGHT GAIN
Slightly lower in males at 10 mg/kg/day over the first week only. Not considered toxicologically relevant.
FOOD CONSUMPTION
Slightly reduced in all treated groups over first 3 weeks only. Not considered toxicologically relevant.
WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study)
Consumption reduced in all treated groups at some occasions during first 3 weeks. Not considered toxicologically relevant.
HAEMATOLOGY
Decreased monocytes at 0.05 and 0.2 mg/kg/day in females at 90 days. Not considered toxicologically relevant.
CLINICAL CHEMISTRY
Some slight changes in 0.01 mg/kg/day males at 90 days. Not considered toxicologically relevant.
ORGAN WEIGHTS
Higher absolute and relative thyroid (weighed with parathyroid) weights at 10 mg/kg/day at 14 days (males only) and 90 days (both sexes). Full recovery in the recovery group.
Some changes in absolute but not relative testes and pituitary weights, without lesions. Not considered toxicologically relevant.
GROSS PATHOLOGY
Reddened thyroids in 3/10 males at 10 mg/kg/day at 90 days.
HISTOPATHOLOGY: NON-NEOPLASTIC
Increased incidence of follicular cell hypertrophy and hyperplasia (based on peer-review) in thyroids of both sexes at 14 and 90 days, at 10 mg/kg/day. Generally minimal.
OTHER FINDINGS
- Estrous cycles: no effect.
- Sperm analyses: no effect.
- Bone marrow micronucleus: neither myelotoxic nor genotoxic effects (NB: cyclophosphamide: genotoxicity, without myelotoxicity)
- Thyroid hormone levels: KEY EFFECTS: dose-related reduction in T3 and T4 and increase in TSH, see table below. They occured at low doses but presented an unusual pattern (plateauing of amplitude + minimal TSH increase for the observed changes in T3/T4), suggesting absence of biological relevance, below 10 mg/kg/day. All effects were reversible. - Dose descriptor:
- NOAEL
- Remarks:
- (histological changes)
- Effect level:
- 1 mg/kg bw/day (nominal)
- Sex:
- male/female
- Basis for effect level:
- other: hypertrophy and hyperplasia of thyroid at 10.0 mg/kg bw/d.
- Critical effects observed:
- not specified
- Conclusions:
- After 90-day oral (drinking water) treatment of rats with ammonium perchlorate, thyroid hormone levels were affected at low doses (T3/T4 decreases, TSH increases) but presented an unusual pattern (plateauing of amplitude + minimal TSH increase for the observed changes in T3/T4), suggesting absence of biological relevance, below 10 mg/kg/day. Non-hormonal adverse effects were only observed at 10 mg/kg/day: thyroid redness, increased thyroid weight and thyroid follicle hypertrophy and minimal hyperplasia.
- Executive summary:
Ammonium perchlorate was provided to groups of 10 male and 10 female rats via drinking water, at dose-levels of 0 (control), 0.01, 0.05, 0.2, 1 and 10 mg/kg/day for 14 and 90 days. Additional 10 rats per sex were treated in the same way at 0, 0.05, 1 and 10 mg/kg/day and kept for a 30-day treatment-free period. Data for the 14-day treatment are ignored here.
- At 10 mg/kg/day, the only toxicologically relevant effects were related to thyroids. Thyroids were reddened at 90 days (3/10 males) and of higher absolute and relative weights than in controls, with follicle hypertrophy and hyperplasia (generally minimal). All these changes were completely reversible in 30 days. Marked thyroid hormone changes were noted in both sexes. Serum levels of T3 and T4 were significantly lower than in controls (-28% to -43%), respectively with almost complete (-9% to -12%) and incomplete (-11% to -31%) recoveries. Serum levels of TSH were significantly higher than in controls (+18% to +21%), with no clear recovery (+9% to +22%).
- At 0.05 to 1 mg/kg/day, poorly dose-related (plateauing) effects on thyroid hormones occured: T3 and T4 reductions (-16% to -32%) and minimal TSH increases (at most +18%), which seems not coherent with the usual high compensation by TSH for moderate changes in T3/T4. The biological relevance seemed therefore low. Recovery was evidenced for T3, T4 and TSH.
- At 0.01 mg/kg/day, T3 and T4 were still significantly lower (-12% to -20%) while TSH was unaffected. Recovery was not assessed.
Reference
Mean thyroid hormone level values in adult rats
At 90 days of treatment | At 120 days (30-day recovery) | |||||||||||
Dose-level (mg/kg/day) | 0 | 0.01 | 0.05 | 0.2 | 1 | 10 | 0 | 0.05 | 1 | 10 | ||
T3 (ng/dL) - males | 180 | 158* | 125*** | 123*** | 122*** | 117*** | 203 | 214 | 212 | 185 | ||
T3 (ng/dL) - females | 170 | 143** | 143** | 137*** | 123*** | 122*** | 224 | 216 | 215 | 197* | ||
T4 (µg/dL) - males | 5.1 | 4.4** | 3.7*** | 3.5*** | 3.5*** | 2.9*** | 4.9 | 4.0*** | 4.0*** | 3.4*** | ||
T4 (µg/dL) - females | 4.4 | 3.5*** | 3.4*** | 3.4*** | 3.2*** | 3.0*** | 3.5 | 3.4 | 3.3 | 3.1 | ||
TSH (ng/mL) - males | 16.2 | 16.9 | 17.8 | 19.0** | 19.1** | 19.1** | 20.9 | 21.3 | 22.2 | 22.8 | ||
TSH (ng/mL) - females | 16.5 | 16.8 | 16.8 | 17.4 | 17.7 | 20.0*** | 13.1 | 15.5** | 15.8** | 16.0*** |
*: p<0.05; **: p<0.01; ***: p<0.001; N= 9-10 rats/sex/dose/timepoint
Incidence of histopathological thyroid follicle lesions in adult rats (peer-reviewed data)
At 90 days of treatment | At 120 days (30-day recovery) | |||||||||
Dose-level (mg/kg/day) | 0 | 0.01 | 0.05 | 0.2 | 1 | 10 | 0 | 0.05 | 1 | 10 |
Hypertrophy - males | 1/10 | 2/10 | 0/10 | 2/10 | 3/10 | 8/10 | 2/10 | 4/10 | 2/10 | 0/10 |
Hypertrophy - females | 0/10 | 0/10 | 3/10 | 2/10 | 1/10 | 5/10 | 0/10 | 0/10 | 1/10 | 0/10 |
Hyperplasia - males | 0/10 | 0/10 | 0/10 | 0/10 | 0/10 | 4/10 | 1/10 | 3/10 | 1/10 | 0/10 |
Hyperplasia - females | 0/10 | 0/10 | 0/10 | 0/10 | 0/10 | 3/10 | 0/10 | 0/10 | 0/10 | 0/10 |
no statistical analysis performed
Endpoint conclusion
- Endpoint conclusion:
- adverse effect observed
- Dose descriptor:
- NOAEL
- 0.3 mg/kg bw/day
- Study duration:
- subchronic
- Species:
- rat
Additional information
- Ammonium perchlorate was provided to groups of 10 male and 10 female rats via drinking water, at dose-levels of 0 (control), 0.01, 0.05, 0.2, 1 and 10 mg/kg/day for 14 and 90 days. Additional 10 rats per sex were treated in the same way at 0, 0.05, 1 and 10 mg/kg/day and kept for a 30-day treatment-free period. Data for the 14-day treatment are ignored. At 10 mg/kg/day, the only toxicologically relevant effects were related to thyroids. Thyroids were reddened at 90 days (3/10 males) and of higher absolute and relative weights than in controls, with follicle hypertrophy and hyperplasia (generally minimal). All these changes were completely reversible in 30 days. Marked thyroid hormone changes were noted in both sexes. Serum levels of T3 and T4 were significantly lower than in controls (-28% to -43%), respectively with almost complete (-9% to -12%) and incomplete (-11% to -31%) recoveries. Serum levels of TSH were significantly higher than in controls (+18% to +21%), with no clear recovery (+9% to +22%). At 0.05 to 1 mg/kg/day, poorly dose-related (plateauing) effects on thyroid hormones occurred: T3 and T4 reductions (-16% to -32%) and minimal TSH increases (at most +18%), which seems not coherent with the usual high compensation by TSH for moderate changes in T3/T4. The biological relevance seemed therefore low. Recovery was evidenced for T3, T4 and TSH. At 0.01 mg/kg/day, T3 and T4 were still significantly lower (-12% to -20%) while TSH was unaffected. Recovery was not assessed. Conclusion: thyroid hormone levels were affected at low doses (T3/T4 decreases, TSH increases) but presented an unusual pattern (plateauing of amplitude + minimal TSH increase for the observed changes in T3/T4), suggesting absence of biological relevance, below 10 mg/kg/day. Non-hormonal adverse effects were only observed at 10 mg/kg/day: thyroid redness, increased thyroid weight and thyroid follicle hypertrophy and minimal hyperplasia. Therefore the NOAEL was the next lower dose of 1 mg/kg bw/day.
- A two-generation study was conducted in rats exposed to ammonium perchlorate at 0, 0.3, 3.0 and 30 mg/kg/day. At 30 mg/kg/day, there was a clear antithyroid effect in P1 and F1 adults and F1 and F2 pups: lower serum T3 (female pups) or T4 levels (male adults), higher serum TSH level (adult P1 males, adult F1 males and females), higher absolute and (when determined) relative thyroid weights (in all conditions), minimal to marked thyroid follicle hypertrophy and hyperplasia (in all conditions), thyroid follicle adenomas in two males. There was no general parental toxicity: the maximal tolerated dose was not reached, limiting sensitivity of the study according to summary author. No effects on reproduction and in particular fertility and pup growth were noted. At 3 mg/kg/day, no relevant hormonal effects were noted; thyroid weight was affected in adults only; however, incidence and severity of thyroid follicle lesions was increased in all conditions (except possibly in F1 adult females were it was not obvious). At 0.3 mg/kg/day, no relevant hormonal effects were noted; thyroid weight was minimally increased in F1 adult females but this was considered not biologically relevant; no relevant effect on thyroid lesions. Conclusion: when taking into account antithyroid effects, the parental NOAEL was 0.3 mg/kg/day. The offspring NOAEL was revised to 0.3 mg/kg/day, as opposed to original report conclusion (3 mg/kg/day), based on anti-thyroid effects at higher doses.
Key repeat-dose study:
Key data for risk assessment come however from a more sensitive 2-generation study as justified in "Short summary":
Repeated dose toxicity: via oral route - systemic effects (target organ) glandular: thyroids
Justification for classification or non-classification
Because of the higher sensitivity (see above) of the 2 -generation study when compared to the 90 -day study, the classification criteria were applied to the 2 -generation study results.
1) Ammonium perchlorate effects qualitatively meet two of the criteria warranting T/Xn R48 or STOT RE classifications:
Type of effect (CLP, § 3.9.2.7.3) |
Associated with AP? |
Comments |
(b) significant functional change in organ systems |
Depends on dose and study |
The toxicologically significant functional effect (in animals) is the impairment of thyroid function defined by hormonal changes in TSH, T3 and T4. They occurred in valid 90-day (showing reversibility) and 2-generation studies in rats. All underlying mechanistic steps (e.g. inhibition of iodine uptake) do not have,per se, any consequence of health before progression into hypothyroidism along increasing exposure. |
(c) consistent significant adverse effects at clinical biochemistry |
Depends on dose and study |
2) The two aforementioned effects did not meet the criteria for non-classification which are described in CLP, § 3.9.2.8.1. Notably, TSH increase can be seen as an adaptative effect, but it only occurs in case of significant effect on T3 and/or T4 synthesis. Therefore, as is the case in therapeutic approach of human subclinical hypothyroidism, it must be considered as adverse.
3) Comparison with repeat-dose toxicity classification thresholds:
- The critical effect occurred at 30 mg/kg/day in a 2-generation study, in particular in F1 male adult rats treated for 125-142 days: a 106% increase (doubling) of TSH with a 26% reduction in T4.
- For this treatment duration of 125-142 days, the mean of 133.5 days was used with Haber’s rule to recalculate (i.e. reduce) the thresholds dose-levels:
Dose-level (mg/kg/day) at which the critical effect occurs |
[0-3.4] |
]3.4-6.8] |
]6.8-34] |
]34-68] |
>68 |
Directive 67/548/EEC |
T, R48/22 |
Xn, R48/22 |
NC |
||
Regulation (EC) No 1272/2008 |
STOT RE 1 |
STOT RE 2 |
NC |
- It was therefore concluded that these effects warranted the following classifications:
- for Directive 67/548/EEC: Xn, R48/22
- for Regulation (EC) No 1272/2008: STOT RE 2, H373(thyroid)
This conclusion to classify for repeated-dose toxic effects can be balanced by the fact that perchlorate has been used for years as a therapy in hyperthyroidism in patients with Grave’s disease: perchlorate’s effect on the thyroid can be seen either as an adverse (presently, in eu- or hypothyroid persons) or as desirable effect (in hyperthyroid persons) depending on the exposed person and the regulatory frame. It can also be recalled that the effect of perchlorate on the thyroid is easy to monitor (follow-up of thyroid hormones), quickly reversible (with even a reversibility during week-ends: see Endpoint summary on human data) and antagonizable (by iodine supplementation).
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