1. Introduction
Ammonium perchlorate (AP) is a strong oxidant widely employed in military and civilian energy materials (propellants). Recently, it has been scrutinized as a persistent environmental pollutant [
1] due to its wide use in fireworks, paints, coatings and other civilian industrial products. The presence of perchlorate in the environment is of both natural [
2,
3,
4], and anthropogenic origins, via improper waste disposal associated with production and use. Exposure of humans to perchlorate via foodstuffs and drinking water has been widely documented [
5]. Additionally, exposure can also derive from inhalation of dust containing AP, especially in workplaces [
6]. The prevalence of trace levels of perchlorate in the environment probably leads to human exposure [
7]. Perchlorate is well known as a potent inhibitor of iodide uptake due to a similar ionic radius as iodide [
8]. Its toxicity is mainly reflected in the inhibition of the sodium iodide symporter (NIS), which results in reduced iodide uptake. Continuous inhibition can produce intracellular iodine deficiency, as that iodine is the important precursor for thyroid hormone synthesis. Long-term exposure to perchlorate may leads to insufficient production of thyroid hormones. Thyroid hormone homeostasis is usually met through upregulation of NIS via the hypothalamus-pituitary-thyroid (H-P-T) feedback mechanism [
9]. Many animal studies have shown that perchlorate can inhibit thyroidal iodide and lead to a decrease of serum thyroid hormone level and increase of serum TSH level in response to iodine insufficiency [
10,
11,
12].
Perchlorate has been used historically in pharmacologic doses to treat hyperthyroidism, and the ability of perchlorate to inhibit thyroid iodide uptake by the sodium-iodide symporter (NIS) has been tested
in vitro [
13]. However, the impacts on thyroid function of long-term low-dose exposures to perchlorate are uncertain and controversial. To our knowledge there is no convincing epidemiological evidence for adverse health effects in human beings through exposure to perchlorate [
14], such that the debate now focuses on the potential toxicity of low level perchlorate exposure on the general population [
15]. Furthermore, a study has pointed out that any discussion about safe levels of perchlorate must be framed in the context of recent study data concerning iodine nutrition status [
1,
16]. Previous research has shown that thyroid dysfunction induced by perchlorate in individuals with low iodine intake was exacerbated [
17].
China was formerly an iodine-deficient country, with 40% of the World’s iodine deficient population. Therefore, the Chinese government implemented a program of distributing iodized salt in 1996, with the aim of eliminating iodine deficiency by 2010. However until now, there have been no reports on the iodine nutrition status associated with occupational exposure to perchlorate in China. China also has large quantities, and increased use of fireworks leaving many workers being exposed to perchlorate. Human studies on perchlorate include cross-sectional epidemiology studies of perchlorate workers with inhalation exposure, and many studies found no significant effects on thyroid homeostasis associated with perchlorate dust exposure [
17,
18]. The possible reason was that there was an adequate supply of iodine in those areas, thus compensating for the perchlorate exposure-related downregulation of iodine uptake.
The present study aimed to examine whether AP dust exposure has an impact on the thyroid homeostasis of occupationally exposed workers. The study was conducted on the workers of the ammonium perchlorate manufacturing company 525 Investment Co., Ltd. Yicheng City, Hubei, which has the second largest manufacturing operation in China. This is the first study to explore the potential biomarkers of occupational AP exposure in Chinese workers, especially those with long-term, low level of AP dust exposure.
4. Discussion
To our knowledge, this is the first report of examining a biomarker of perchlorate exposure for an association with an effect on thyroid homeostasis of Chinese workers. The results of serum chemistry and hematology profiles were within normal limits throughout the study in all participants, which is consistent with the previous findings [
14]. The primary thyroidal parameters (FT
3, FT
4, Log TSH, Tg, and TPO) showed no significant differences between the exposed workers and the controls.
Iodide is required to synthesize hormones critical for fetal and neonatal development. The human thyroid gland contains a huge reserve store of hormone within the intrafollicular thyroglobulin. If iodine intake is sufficient, even a reduction in thyroidal iodide concentration to one-third of its former level may still leave these individuals sufficient iodide to produce thyroid hormone at a normal rate. As shown in
Table 7, the iodine nutritional status of the two groups was sufficient. Further, given that perchlorate has a half-life of about 8–14 h, the plant workers, many of whom worked 12 h shifts, might recover from the inhibitory effects of exposure during off-shift periods.
In this study, no significant difference was observed between the two groups in the main indicators of thyroid function such as FT
3, FT
4 and TSH. Historically, the mode of action analysis suggests that alteration of hormones (T
4, T
3 and TSH) would be the first observed biological effect of perchlorate exposure [
23]. TT
4 was measured in addition to thyroid parameters shown in
Table 4. The mean TT
4 level for the exposed workers (65.6 ± 16.7 ng/mL) was significantly (
p = 0.034) lower than the mean TT4 level for the controls (81.9 ± 46.2 ng/mL). Bruce
et al. [
24] demonstrated that depressed TT4 levels are due to increased nitrate and thiocyanate levels and not to increased perchlorate levels. Neither serum nor urinary levels of nitrate or thiocyanate were measured in this study. A second explanatory hypothesis might be that lower TT
4 levels reflect lower thyroid binding globulin levels, but that also was not measured. Because of the lack of determination of TBG, the exact cause of decrease in total T4 remain unclear. Perchlorate is not known to be associated with binding globulin alterations, so the association observed in this study is not, in fact, in line with the known mode of action of perchlorate. Thus, further studies are needed to clarify these findings.
Concurrent quantification of iodine and perchlorate levels in human urine can provide information on potential risks from perchlorate exposure. Urinary perchlorate is reasonably indicative of human exposure, because 70%–95% of a perchlorate dose is excreted unchanged in the urine with a half-life of 8–14 h [
8,
25], not only urine collection is less invasive than blood collection, but also urinary perchlorate levels tend to be much higher than serum levels due to efficient renal clearance of perchlorate. Urinary perchlorate levels are elevated in the urine because the kidney reabsorbs water from the urine and thus concentrates the urinary solutes. Measuring perchlorate in human urine can assess the combined exposure from all sources [
26]. By comparing the concentration of urinary perchlorate at the end of shift and before the shift could reflect the exposure intensity during the work shift. The geometric mean concentration of urinary perchlorate in the exposure group was markedly higher than that of the control group, which demonstrated a direct association between urinary levels of perchlorate and AP exposure dose in workplace (
Table 4).
Perchlorate is not metabolized in the human body, and is excreted from the urine, with a biological half-life of approximately 8 hours. So the timing of perchlorate exposure is important for individual exposure assessment. The pre-shift urine samples were obtained in the early morning after a day of consecutive work and adequate rest in the evening. The urine perchlorate of the control group was detected, indicating the presence of background exposure. The perchlorate content in the waste water discharged from the plant was also detected, confirming the source of exposure (data not shown). The concentration of urinary perchlorate in the exposure group was markedly higher than that of the control group, which showed that urinary perchlorate is a specific biomarker to distinguish exposed from unexposed individuals.
We found no evidence of an effect on urinary iodine from environmental perchlorate dust exposure in workplace. Iodine nutrition status is crucial to the comparative evaluation of the thyroid function of the two groups. However, iodine levels in spot urine can vary significantly from sample to sample, due to variation in iodine intake throughout the day and from day to day. In the region studied, the main source of iodine intake is iodized salt ingestion. In addition, urinary iodine concentrations can be affected by fluid status, Therefore, creatinine adjustments were used to account for differences in fluid volume status. The median urinary iodine levels of the two groups of workers were close to each other; additionally, the difference was not statistically significant, indicating that the iodine nutrition status of the two groups was almost identical, and had strong comparability.
A previous study has shown that creatinine adjustment for estimation of 24 h excretion from spot samples was not effective for iodine [
27]. However, another study revealed that single-spot urine samples could reflect the participant’s exposure level, providing support for the use of a single sample as an exposure measure in epidemiological studies [
28]. Owing to the slight decrease of median of urinary iodine in the exposed group comparing with that of the control group, our research showed that spot urinary iodine might be used as an indicator of the iodine nutrition status of occupational workers.
The high iodine levels found in the urine of some people (maximum concentration: 595.2 g/g Cr) indicated that effective measures of iodized salt were widely implemented in China. The median urinary iodine concentration in a population provides a good yardstick for its current iodine nutrition. The WHO (2007) has identified median urinary iodine levels ≥ 100 μg/L as an indicator of sufficient iodine intake for a population, while median urinary iodine levels > 300 μg/L indicate excessive iodine intake [
29]. In this study elevated concentrations of iodine (>300 μg/L) were found in five of the urinary samples from the total participants; the concentrations ranged from 320.4 to 480.2 μg/L. This elevated concentrations of urinary iodine indicated that some individuals may have ingested large amounts of iodine from their diet. Furthermore, such high iodide intake may effectively compete with perchlorate binding sites on the NIS, which generally alleviates the effects of perchlorate on thyroid homeostasis.
There have been reported that high dose of ammonium perchlorate dust exposure could lead to pulmonary fibrosis in Chinese workers [
30]. Our results were inconsistent with the previous study, and possible reason lies in the significant decreased dust concentration due to the proper use of protective equipment and the improvement of the production process.
The present study has the general limitations of a cross-sectional analysis. Therefore, the relationship between perchlorate exposure and thyroid function was examined with attention to the potential influences of chance, bias, or confounding. Further, thiocyanate exposure was not actually measured, which was the major source of sodium iodide transporter inhibitors. A larger sample size might help to average such potential kinetic differences. Limitations also include a lack of measurements of specific chemical components (TBG) that may have been responsible for the level of TT4 in serum samples, as well as the limitation of a 2-day monitoring of ammonium perchlorate concentration in the dust in the workplace.