Tippawan Prapamontol

Environmental lead exposure: a public health problem of global dimensions

Lead is the most abundant of the heavy metals in the Earth’s crust. It has been used since prehistoric times, and has become widely distributed and mobilized in the environment. Exposure to and uptake of this non-essential element have consequently increased. Both occupational and environmental exposures to lead remain a serious problem in many developing and industrializing countries, as well as in some developed countries. In most developed countries, however, introduction of lead into the human environment has decreased in recent years, largely due to public health campaigns and a decline in its commercial usage, particularly in petrol. Acute lead poisoning has become rare in such countries, but chronic exposure to low levels of the metal is still a public health issue, especially among some minorities and socioeconomically disadvantaged groups. In developing countries, awareness of the public health impact of exposure to lead is growing but relatively few of these countries have introduced policies and regulations for significantly combating the problem. This article reviews the nature and importance of environmental exposure to lead in developing and developed countries, outlining past actions, and indicating requirements for future policy responses and interventions.

Concentrations of urinary pesticide metabolites in small-scale farmers in Chiang Mai Province, Thailand

Our research goal was to assess exposure to currently used pesticides among small-scale male farmers residing in two topographically different areas in Chiang Mai Province, Thailand. Farmers (N=136) were recruited from Pong Yaeng subdistrict (N=67) and Inthakhin subdistrict (N=69). Each farmer provided a morning urine void for the analysis of 30 urinary metabolites of insecticides, herbicides, and fungicides. Farmers in Pong Yaeng had significantly higher urinary concentrations of metabolites of organophosphorus insecticides and ethylene bisdithiocarbamates, while farmers from Inthakhin had significantly higher concentrations of malathion, 2,4-D, alachlor, and parathion or EPN metabolites. Based upon the metabolites measured in the urine of the farmers, chlorpyrifos and pyrethroid insecticides seemed to be commonly used across both communities; no significant differences in metabolite concentrations of these insecticides were observed between the two farmer groups. The presence of methamidaphos in the urine of farmers suggests that, despite a ban on its use, methamidaphos continues to be used in the communities. A similar finding with metabolites of methyl parathion must be further investigated. Overall, our results suggest that while each community may use different pesticides, Thai farmers are exposed to a wide variety of pesticides with a broad range in exposure magnitude. Furthermore, age, field size, crop production type, and the use of protective equipment were found to be potential factors influencing the degree of exposure.

Urinary pesticide metabolites in school students from northern Thailand

We evaluated exposure to pesticides among secondary school students aged 12-13 years old in Chiang Mai Province, Thailand. Pesticide-specific urinary metabolites were used as biomarkers of exposure for a variety of pesticides, including organophosphorus insecticides, synthetic pyrethroid insecticides and selected herbicides. We employed a simple solid-phase extraction with analysis using isotope dilution high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). A total of 207 urine samples from Thai students were analyzed for 18 specific pesticide metabolites. We found 14 metabolites in the urine samples tested; seven of them were detected with a frequency > or=17%. The most frequently detected metabolites were 2-[(dimethoxyphosphorothioyl) sulfanyl] succinic acid (malathion dicarboxylic acid), para-nitrophenol (PNP), 3,5,6-trichloro-2-pyridinol (TPCY; metabolite of chlorpyrifos), 2,4-dichlorophenoxyacetic acid (2,4-D), cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acids (c-DCCA and t-DCCA; metabolite of permethrin) and 3-phenoxybenzoic acid (3-PBA; metabolite of pyrethroids). The students were classified into 4 groups according to their parental occupations: farmers (N=60), merchants and traders (N=39), government and company employees (N=52), and laborers (N=56). Children of farmers had significantly higher urinary concentrations of pyrethroid insecticide metabolites than did other children (p<0.05). Similarly, children of agricultural families had significantly higher pyrethroid metabolite concentrations. Males had significantly higher values of PNP (Mann-Whitney test, p=0.009); however, no other sex-related differences were observed. Because parental occupation and agricultural activities seemed to have little influence on pesticide levels, dietary sources were the likely contributors to the metabolite levels observed.

A single method for detecting 11 organophosphate pesticides in human plasma and breastmilk using GC-FPD

Organophosphate (OP) pesticides are widely used for crop protection in many countries including Thailand. Aside from causing environmental contamination, they affect human health especially by over-stimulating of the neurotransmission system. OP pesticides, as with other non-persistent pesticides, degrade quickly in the environment as well as are metabolized quite rapidly in humans. Assessing human exposures to these compounds requires analytical methods that are sensitive, robust, and most importantly, suitable for specific laboratory settings. The aim of this study was to develop and validate an analytical method for measuring 11 OP pesticide residues in human plasma and breast milk. Analytes in both plasma and breast milk samples were extracted with acetone and methylene chloride, cleaned-up using aminopropyl solid phase extraction cartridges, and analyzed by gas chromatography with flame photometric detection. The optimized method exhibited good linearity, with the coefficients of determination of 0.996-0.999 and <7% error about the slope. Extraction recoveries from spiked plasma and breast milk samples at low and medium concentrations (0.8-5.0 and 1.6-10ngmL(-1), respectively) ranged from 59.4% (ethion) to 94.0% (chlorpyrifos). Intra-batch and inter-batch precisions ranged from 2.3-18.9% and 5.8-19.5%, respectively. Method detection limits of plasma and breast milk ranged from 0.18-1.36 and 0.09-2.66ngmL(-1), respectively. We analyzed 63 plasma and 30 breastmilk samples collected from farmworkers in Chiang Mai Province to determine the suitability of this method for occupational exposure assessment. Of the 11 pesticides measured, seven were detected in plasma samples and five were detected in breast milk samples. Mass spectrometry was used to confirm results. Overall, this method is rapid and reliable. It offers the laboratories with limited access to mass spectrometry a capacity to investigate levels OP pesticides in plasma and breastmilk in those occupationally exposed for health risk assessment.

Cross validation of gas chromatography-flame photometric detection and gas chromatography–mass spectrometry methods for measuring dialkylphosphate metabolites of organophosphate pesticides in human urine ☆

We report two analytical methods for the measurement of dialkylphosphate (DAP) metabolites of organophosphate pesticides in human urine. These methods were independently developed/modified and implemented in two separate laboratories and cross validated. The aim was to develop simple, cost effective, and reliable methods that could use available resources and sample matrices in Thailand and the United States. While several methods already exist, we found that direct application of these methods required modification of sample preparation and chromatographic conditions to render accurate, reliable data. The problems encountered with existing methods were attributable to urinary matrix interferences, and differences in the pH of urine samples and reagents used during the extraction and derivatization processes. Thus, we provide information on key parameters that require attention during method modification and execution that affect the ruggedness of the methods. The methods presented here employ gas chromatography (GC) coupled with either flame photometric detection (FPD) or electron impact ionization-mass spectrometry (EI-MS) with isotopic dilution quantification. The limits of detection were reported from 0.10ng/mL urine to 2.5ng/mL urine (for GC-FPD), while the limits of quantification were reported from 0.25ng/mL urine to 2.5ng/mL urine (for GC-MS), for all six common DAP metabolites (i.e., dimethylphosphate, dimethylthiophosphate, dimethyldithiophosphate, diethylphosphate, diethylthiophosphate, and diethyldithiophosphate). Each method showed a relative recovery range of 94-119% (for GC-FPD) and 92-103% (for GC-MS), and relative standard deviations (RSD) of less than 20%. Cross-validation was performed on the same set of urine samples (n=46) collected from pregnant women residing in the agricultural areas of northern Thailand. The results from split sample analysis from both laboratories agreed well for each metabolite, suggesting that each method can produce comparable data. In addition, results from analyses of specimens from the German External Quality Assessment Scheme (G-EQUAS) suggested that the GC-FPD method produced accurate results that can be reasonably compared to other studies.

Enhanced sensitivity and selectivity in the detection of polycyclic aromatic hydrocarbons using sequential simplex optimization, the addition of an organic modifier and wavelength programming

Optimized separation of a mixture of 16 priority pollutant polycyclic aromatic hydrocarbons (PAHs) by high performance liquid chromatography (HPLC) using the sequential simplex optimization method was accomplished by varying the starting and ending compositions of acetonitrile and water, linear gradient time, mobile phase flow rate, column temperature and holding time of the final mobile phase composition. Focusing on the two sets of difficult-to-separate pairs (acenaphthene-fluorene and benzo[g,h,i]perylene-indeno[1,2,3-c,d]pyrene), analysis time was reduced by about ten percent through the use of an organic modifier (isopropanol or methanol), under both optimum and near-optimum conditions, while maintaining good separation of the remaining PAHs. High sensitivity for all of the 16 PAHs was achieved by wavelength programming during elution using five wavelengths (224, 235, 254 270 and 296 nm), depending upon the molar absorptivities of the individual compounds. Detection limits (DLs) ranging from 0.002 (benzo[a]pyrene) to 0.140 mug ml(-1) (acenaphthene) were achieved for this set of 16 standard compounds.

Determination of PM10 and its ion composition emitted from biomass burning in the chamber for estimation of open burning emissions

Biomass samples including agricultural waste (rice straw and maize residue) and forest leaf litter were collected from Chiang Mai Province, Thailand for the burning experiment in the self-designed stainless steel chamber to simulate the emissions of PM10. The burning of leaf litter emitted the highest PM10 (1.52±0.65 g kg(-1)). The PM10-bound ions emitted from the burning of rice straw and maize residue showed the same trend, which was K(+)>Cl(-)>SO4(2-)>NH4(+)>NO3(-). However, the emissions from maize residue burning were ~1.5-2.0 times higher than those from the rice straw burning. The ion content emitted from leaf litter burning was almost the same for all ion species. Noticeably, K(+) and Cl(-) concentrations were ~2-4 times lower than those emitted from agricultural waste burning. It can be deduced that K(+) and Cl(-) were highly emitted from agricultural waste burning due to the use of fertilizer and herbicides in the field, respectively. Based on emission values obtained from the chamber, the pollutant emission rate from open burning was calculated. Burned areas in Chiang Mai Province were 3510 and 866 km(2) in 2010 and 2011, respectively. Forest burning was 71-88%, while agricultural land burning accounted for 12-29% (rice field: crop field=1:3) of total burned area. Therefore, emissions of PM10 from open burning in Chiang Mai were 3051 ton (2010) and 705 ton (2011). Major ions emitted from agricultural waste burning were found to be K(+) and Cl(-), while those from forest burning were SO4(2-) and K(+).

Determination of the pyrethroid insecticide metabolite 3-PBA in plasma and urine samples from farmer and consumer groups in northern Thailand

In this study, the enzyme-linked immunosorbent assays (ELISA) were modified to detect 3-PBA in plasma (including the adducted form) and urine among a large group of consumers and farmers in an agricultural area. The samples were collected on the same day in the morning from 100 consumers (50 females, 50 males) and 100 farmers (50 females, 50 males) in the Fang district, Chiang Mai province, northern Thailand. The ELISA was very sensitive having an IC50 value of 26.7 and 15.3 ng/mL, a limit of quantitation of 5 and 2.5 ng/mL and a limit of detection of 1.08 and 1.94 ng/mL for plasma and urine, respectively. These methods had low (< 5%) intra- and inter-assay coefficients of variation. The extraction technique satisfactorily eliminated the matrix effect from samples before ELISA analysis, yielding good recoveries (85.9–99.4% and 87.3–98.0%, respectively). For the volunteer study, the detection rate for plasma 3-PBA was 24% in consumers and 42% in farmers, but the median and range values were similar (median 5.87 ng/mL, range 5.16–8.44 ng/mL in consumers and 6.27 ng/mL, range 4.29–9.57 ng/mL in farmers). The rate of detection in the urine was similar (76% and 69%, in consumers and in farmers), yet the median concentration was significantly higher in farmers (8.86 μg/g creatinine in consumers vs 16.1 μg/g creatinine in farmers) and the range also much wider in farmers (1.62–80.5 μg/g creatinine in consumers and 0.80–256.2 μg/g creatinine in farmers). There was no correlation between plasma 3-PBA and urinary 3-PBA concentrations in the study presumably because plasma 3-PBA is a measure of cumulative exposures while urinary 3-PBA reflects acute exposures. In addition, metabolism and excretion of pyrethroids varies by individual. Nevertheless, this study demonstrated that these volunteers were exposed to pyrethroids. To our knowledge, this is the first report that compared plasma 3-PBA and urinary 3-PBA in a large group of volunteers. The ELISA method provided higher sample throughput with lower cost as compared to the instrumental analysis.

Associations of maternal organophosphate pesticide exposure and PON1 activity with birth outcomes in SAWASDEE birth cohort, Thailand

Prenatal organophosphate (OP) pesticide exposure has been reported to be associated with adverse birth outcomes and neurodevelopment. However, the mechanisms of toxicity of OP pesticides on human fetal development have not yet been elucidated. Our pilot study birth cohort, the Study of Asian Women and Offsprings Development and Environmental Exposures (SAWASDEE cohort) aimed to evaluate environmental chemical exposures and their relation to birth outcomes and infant neurodevelopment in 52 pregnant farmworkers in Fang district, Chiang Mai province, Thailand. A large array of data was collected multiple times during pregnancy including approximately monthly urine samples for evaluation of pesticide exposure, three blood samples for pesticide-related enzyme measurements and questionnaire data. This study investigated the changes in maternal acetylcholinesterase (AChE) and paraoxonase 1 (PON1) activities and their relation to urinary diakylphosphates (DAPs), class-related metabolites of OP pesticides, during pregnancy. Maternal AChE, butyrylcholinesterase (BChE) and PON1 activities were measured three times during pregnancy and urinary DAP concentrations were measured, on average, 8 times from enrollment during pregnancy until delivery. Among the individuals in the group with low maternal PON1 activity (n=23), newborn head circumference was negatively correlated with log10 maternal ∑DEAP and ∑DAP at enrollment (gestational age=12±3 weeks; β=-1.0 cm, p=0.03 and β=-1.8 cm, p<0.01, respectively) and at 32 weeks pregnancy (β=-1.1cm, p=0.04 and β=-2.6 cm, p=0.01, respectively). Furthermore, among these mothers, newborn birthweight was also negatively associated with log10 maternal ∑DEAP and ∑DAP at enrollment (β=-219.7 g, p=0.05 and β=-371.3g, p=0.02, respectively). Associations between maternal DAP levels and newborn outcomes were not observed in the group of participants with high maternal PON1 activity. Our results support previous findings from US birth cohort studies. This is the first study to report the associations between prenatal OP pesticide exposure and birth outcomes in Thailand.

Pilot Study of Pesticide Knowledge, Attitudes, and Practices among Pregnant Women in Northern Thailand

An estimated 200,000 children born in Thailand each year are at risk of prenatal exposure to pesticides and associated neurodevelopmental outcomes because of their mothers’ agricultural occupations. Children born to non-agricultural workers may also be at risk of exposure from other pathways of maternal pesticide exposure, including exposure through home use, diet, and other environmental media. Pesticide exposure in Thailand has been linked to unsafe practices and beliefs about pesticides. However, limited information exists on pesticide knowledge, attitudes, and practices among pregnant women in Thailand or elsewhere. Obtaining this information is essential to understand the factors associated with prenatal pesticide exposure, identify populations potentially at risk, and ultimately protect pregnant women and their children. We administered surveys to 76 pregnant women in northern Thailand and used multivariable logistic regression to evaluate associations among pesticide-related knowledge, pregnancy trimester, and pesticide use behavior. In this pilot study, lower knowledge score and earliest trimester of pregnancy were marginally (p < 0.1) associated with unsafe practices in the home, but not at work. Women who worked in agriculture or applied pesticides before becoming pregnant, or who had a previous child were significantly (p < 0.05) more likely to engage in unsafe behaviors in the home during their current pregnancy. We preliminarily conclude that increasing pesticide-related knowledge among pregnant women may help promote safe practices and reduce prenatal exposure. Knowledge-based interventions may be most effective when implemented early in pregnancy and targeted to agricultural workers and other sub-populations at risk of pesticide exposure.