Geometric Mean Serum Cotinine Concentrations Confirm a Continued Decline in Secondhand Smoke Exposure among U.S. Nonsmokers-NHANES 2003 to 2018.

The objective of this study was to examine long-term trends in serum cotinine (COT) concentrations, as a measure of secondhand smoke (SHS) exposure, in U.S. nonsmokers using data from the National Health and Nutrition Examination Surveys (NHANES) from 2003 to 2018. We analyzed NHANES serum COT results from 8 continuous NHANES 2 year cycles from 2003 to 2018 using a liquid chromatography−tandem mass spectrometry assay that has been maintained continuously at the Centers for Disease Control and Prevention (CDC) since 1992. Serum COT concentrations (based on the geometric means) among nonsmokers in the U.S. decreased by an average of 11.0% (95% confidence interval (CI) [8.8%, 13.1%]; p < 0.0001) every 2 year cycle. From 2003 to 2018, serum COT concentrations in U.S. nonsmokers declined by 55.0%, from 0.065 ng/mL in 2003−2004 to 0.029 ng/mL in 2017−2018 (p < 0.0001). Significant decreases in serum COT concentrations were observed in all demographic groups. While disparities between these groups seems to be shrinking over time, several previously observed disparities in SHS exposure remain in 2017−2018. Serum COT concentrations of the non-Hispanic Black population remained higher than those of non-Hispanic Whites and Mexican Americans (p < 0.0001). Additionally, serum COT concentrations were significantly higher for children aged 3−5 years than other age groups (p ≤ 0.0002), and men continued to have significantly higher serum COT concentrations than women (p = 0.0384). While there is no safe level of exposure to SHS, the decrease in serum COT concentrations in the U.S. population as well as across demographic groupings represents a positive public health outcome and supports the importance of comprehensive smoke-free laws and policies for workplaces, public places, homes, and vehicles to protect nonsmokers from SHS exposure.

Serum Concentrations of Cotinine and Trans-3'-Hydroxycotinine in US Adults: Results From Wave 1 (2013-2014) of the Population Assessment of Tobacco and Health Study.

INTRODUCTION: The Population Assessment of Tobacco and Health (PATH) Study is a nationally representative cohort of tobacco product users and nonusers. The study's main purpose is to obtain longitudinal epidemiologic data on tobacco use and exposure among the US population. AIMS AND METHODS: Nicotine biomarkers-cotinine (COT) and trans-3'-hydroxycotinine (HCT)-were measured in blood samples collected from adult daily tobacco users and nonusers during Wave 1 of the PATH Study (2013-2014; n = 5012; one sample per participant). Participants' tobacco product use and exposure to secondhand smoke were categorized based on questionnaire responses. Nonusers were subdivided into never users and recent former users. Daily tobacco users were classified into seven tobacco product use categories: exclusive users of cigarette, smokeless tobacco, electronic cigarette, cigar, pipe, and hookah, as well as polyusers. We calculated sample-weighted geometric mean (GM) concentrations of cotinine, HCT, and the nicotine metabolite ratio (NMR) and evaluated their associations with tobacco use with adjustment for potential confounders. RESULTS: The GMs (95% confidence intervals) of COT and HCT concentrations for daily tobacco users were 196 (184 to 208) and 72.5 (67.8 to 77.4) ng/mL, and for nonusers they were 0.033 (0.028 to 0.037) and 0.021 (0.018 to 0.023) ng/mL. Exclusive smokeless tobacco users had the highest COT concentrations of all user groups examined. The GM NMR in daily users was 0.339 (95% confidence interval: 0.330 to 0.350). CONCLUSIONS: These nationally representative estimates of serum nicotine biomarkers could be the basis for reference ranges characterizing nicotine exposure for daily tobacco users and nonusers in the US adult population. IMPLICATIONS: This report summarizes the serum nicotine biomarker measurements in Wave 1 of the PATH Study. We are reporting the first estimates of HCT in serum for daily tobacco users and nonusers in the noninstitutionalized, civilian US adult population; the first nationally representative serum COT estimates for daily exclusive users of different tobacco products and daily polyusers; and the first nationally representative estimate of the serum NMR in daily tobacco users by age, race/ethnicity, and sex.

Degradation of Organophosphorus and Pyrethroid Insecticides in Beverages: Implications for Risk Assessment.

Since urinary insecticide metabolites are commonly used as biomarkers of exposure, it is important that we quantify whether insecticides degrade in food and beverages in order to better perform risk assessment. This study was designed to quantify degradation of organophosphorus and pyrethroid insecticides in beverages. Purified water, white grape juice, orange juice, and red wine were fortified with 500 ng/mL diazinon, malathion, chlorpyrifos, permethrin, cyfluthrin, cypermethrin, and deltamethrin, and aliquots were extracted several times over a 15-day storage period at 2.5 °C. Overall, statistically significant loss of at least one insecticide was observed in each matrix, and at least five out of seven insecticides demonstrated a statistically significant loss in all matrices except orange juice. An investigation of an alternative mechanism of insecticide loss-adsorption onto the glass surface of the storage jars-was carried out, which indicated that this mechanism of loss is insignificant. Results of this work suggest that insecticides degrade in these beverages, and this degradation may lead to pre-existing insecticide degradates in the beverages, suggesting that caution should be exercised when using urinary insecticide metabolites to assess exposure and risk.

Predictors of Serum Polybrominated Diphenyl Ether (PBDE) Concentrations among Children Aged 1-5 Years.

Serum concentrations of PBDEs were measured using gas chromatography-tandem mass spectrometry in 80 children aged 15-71 months. Demographic and behavioral data were collected on parental questionnaires; a research nurse recorded anthropometric measures and insurance status. For a subset of children (n = 17), PBDEs were measured in house dust and child handwipes sampled during a home visit. In linear and Tobit regression, log-transformed PBDE congeners were modeled as a function of child characteristics, including neighborhood-level socioeconomic indicators. BDE congeners 47, 99, and 100 were highly correlated and summed for analysis; BDE-153 was examined individually. PBDE serum concentrations were associated with socioeconomic factors; for example, a $20,000 increase in median household income in a child's ZIP code was associated with a 34% decrease (95%CI = 14-49%) in BDE-153 and a 26% decrease (95%CI = 6-42%) in ∑BDE-47,-99,-100. Lower body-mass index (BMI) z-score and household smoking were strong predictors of higher BDE-153 levels. Among children who participated in a home visit, serum PBDE was positively correlated with handwipe PBDE (Spearman r ∑BDE-47, -99, -100 = 0.48, p = 0.09), but not dust PBDE. Results indicate socioeconomic factors and BMI are strong predictors of serum PBDE levels among young children. PBDEs measured on handwipes are more predictive of serum PBDE levels than vacuum-collected dust.

Quantification of Polybrominated and Polychlorinated Biphenyls in Human Matrices by Isotope-Dilution Gas Chromatography-Tandem Mass Spectrometry.

We have developed a highly sensitive and selective analytical method capable of quantifying a total of 15 polybrominated and polychlorinated biphenyls (11 PBBs and 4 PCBs) in human serum. Samples were subjected to liquid-liquid extraction followed by solid-phase extraction prior to measurement using gas chromatography-tandem mass spectrometry in multiple reaction monitoring mode. Quantification was performed using isotope-dilution calibration covering a concentration range of 0.005-12.5 ng/mL. Limits of detection for all target compounds were in the low range (0.7-6.5 pg/mL). The method was validated using in-house pooled human serum fortified at two concentrations (0.5 ng/mL and 1.0 ng/mL), whole semen fortified at one concentration (0.25 ng/mL), and NIST Standard Reference Material (SRM) 1958, which includes five of the target compounds. Method accuracies for all target compounds ranged from 84 to 119% with relative standard deviations (RSDs) of <19%. The measured values for the five target compounds present in the SRM agreed with the certified reference values (89-119% accuracy with RSDs <9%). As this method was developed to support ongoing epidemiologic investigations, we evaluated its suitability by analyzing subsets of serum and whole semen samples from the Michigan PBB Registry cohort. PBB-153, PCB-118, PCB-138, PCB-153 and PCB-180 were detected in all serum samples analyzed, with PBB-77 and PBB-101 detected less frequently in serum. PBB-153, PCB-118, PCB-138, PCB-153 and PCB-180 were detected in at least one whole semen sample.

Production of Insecticide Degradates in Juices: Implications for Risk Assessment.

This study was designed to observe the production of degradates of two organophosphorus insecticides and one pyrethroid insecticide in beverages. Purified water, white grape juice, apple juice, and red grape juice were fortified with 500 ng/g malathion, chlorpyrifos, and permethrin, and aliquots were extracted for malathion dicarboxylic acid (MDA), 3,5,6-trichloro-2-pyridinol (TCPy), and 3-phenoxybenzoic acid (3-PBA) several times over a 15 day period of being stored in the dark at 2.5 °C. Overall, first-order kinetics were observed for production of MDA, and statistically significant production of TCPy was also observed. Statistically significant production of 3-phenoxybenzoic acid was not observed. Results indicate that insecticides degrade in food and beverages, and this degradation may lead to preexisting insecticide metabolites in the beverages. Therefore, it is suggested that caution should be exercised when using urinary insecticide metabolites to assess exposure and risk.

Biological Matrix Effects in Quantitative Tandem Mass Spectrometry-Based Analytical Methods: Advancing Biomonitoring.

The ability to quantify levels of target analytes in biological samples accurately and precisely in biomonitoring involves the use of highly sensitive and selective instrumentation such as tandem mass spectrometers and a thorough understanding of highly variable matrix effects. Typically, matrix effects are caused by co-eluting matrix components that alter the ionization of target analytes as well as the chromatographic response of target analytes, leading to reduced or increased sensitivity of the analysis. Thus, before the desired accuracy and precision standards of laboratory data are achieved, these effects must be characterized and controlled. Here we present our review and observations of matrix effects encountered during the validation and implementation of tandem mass spectrometry-based analytical methods. We also provide systematic, comprehensive laboratory strategies needed to control challenges posed by matrix effects in order to ensure delivery of the most accurate data for biomonitoring studies assessing exposure to environmental toxicants.

Method for the quantification of current use and persistent pesticides in cow milk, human milk and baby formula using gas chromatography tandem mass spectrometry.

The aim of this study was to develop an analytical method for the quantification of organochlorine (OC), organophosphate (OP), carbamate, and pyrethroid insecticide residues in cow milk, human milk, and baby formula. A total of 25 compounds were included in this method. Sample extraction procedures combined liquid-liquid extraction, freezing-lipid filtration, dispersive primary-secondary amine cleanup, and solid-phase extraction together for effective extraction and elimination of matrix interferences. Target compounds were analyzed using gas chromatography with electron impact ionization-tandem mass spectrometry (GC-EI-MS/MS) in the multiple reaction monitoring (MRM) mode. Average extraction recoveries obtained from cow milk samples fortified at two different concentrations (10 ng/mL and 25 ng/mL), ranged from 34% to 102%, with recoveries for the majority of target compounds falling between 60% and 80%. Similar ranges were found for formula fortified at 25 ng/mL. The estimated limits of detection for most target analytes were in the low pg/mL level (range 3-1600 pg/mL). The accuracies and precisions were within the range of 80-120% and less than 15%, respectively. This method was tested for its viability by analyzing 10 human milk samples collected from anonymous donors, 10 cow milk samples and 10 baby formula samples purchased from local grocery stores in the United States. Hexachlorobenzene, p,p-dicofol, o,p-DDE, p,p-DDE, and chlorpyrifos were found in all samples analyzed. We found detectable levels of permethrin, cyfluthrin, and fenvalerate in some of the cow milk samples but not in human milk or baby formula samples. Some of the pesticides, such as azinphos-methyl, heptachlor epoxide, and the pesticide synergist piperonyl butoxide, were detected in some of the cow milk and human milk samples but not in baby formula samples.

HPLC-MS/MS method for the measurement of insecticide degradates in baby food.

A solid phase extraction method was developed to isolate four insecticide degradates from baby food that were measured subsequently using high-performance liquid chromatography-tandem mass spectrometry. The degradates [parent insecticide] measured were malathion dicarboxylic acid [malathion], 3,5,6-trichloro-2-pyridinol [chlorpyrifos, chlorpyrifos methyl] (TCPy), cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid [permethrin, cypermethrin, cyfluthrin], and 3-phenoxybenzoic acid [general pyrethroid]. All degradates produced recoveries between 80 and 120% except TCPy in fruit (122% recovery), and all relative standard deviations were <16%. Use of this method demonstrated that insecticide degradates were found in baby foods frequently purchased in the United States, supporting the need for this method. These data will assist in differentiating whether biomarker levels of insecticide metabolites are the result of exposures to the toxic insecticide or its preformed degradate.

Method for the determination of organophosphorus and pyrethroid pesticides in food via gas chromatography with electron-capture detection.

We have developed a rapid, high-throughput, accurate, multiresidue method for the analysis of selected organophosphorus and pyrethroid pesticides in a variety of food samples suitable for use in public health and epidemiologic investigations of high-use pesticides using modifications of existing methods. The procedure involves a pesticide extraction from the food sample with acetonitrile followed by a salting-out with NaCl and cleanup of the extract with a multilayer solid-phase extraction cartridge composed of a Supelclean ENVI-CARB-II top layer and a primary-secondary amine bottom layer separated by a polyethylene frit. To evaluate the method, we performed fortification studies at 50, 100, and 200 ng/g for 3 organophosphorus and 4 pyrethroid pesticides in 16 different foods. Instrumental analysis was carried out by capillary gas chromatography with electron-capture detection (GC-ECD). Confirmatory analysis was performed by GC coupled with mass spectrometry (MS) in the selected-ion monitoring (SIM) mode. Average recoveries for each fortification level ranged from 49 to 146% with 80% of recoveries between 80 and 120%.

Current and historically used pesticides in residential soil from 11 homes in Atlanta, Georgia, USA.

We used a multiresidue, gas chromatography/mass spectrometry-based method to measure seven pyrethroid, five organophosphorus (OP), and six organochlorine pesticides in soil collected from 11 Atlanta homes in 2006. Our objective was to collect preliminary data for a larger study of pesticide exposures among Atlanta children. The pyrethroid insecticides (cis- and trans-permethrin, bioallethrin) were the most commonly detected analytes, giving evidence of widespread outdoor use among our study homes. Our pyrethroid insecticide detection frequencies were higher than those reported in a recent study of Ohio and North Carolina homes; however, our maximum values were approximately half of those reported. We detected the target OP pesticides in only a few samples, but we found two restricted-use OP pesticides--methyl parathion and terbufos--and thus possible evidence of illegal residential use or environmental persistence in soil. We also detected dichlorodiphenyltrichloroethane (DDT) and dichlorodiphenyldichloroethane (DDE) in samples from six homes. Although our small sample size limits comparison to other studies, our results provide evidence that residential soil is a potential source of human exposure to both current and historically used pesticides.

Pyrethroid and organophosphorus pesticides in composite diet samples from Atlanta, USA adults.

Four pyrethroid (permethrin, cyfluthrin, cypermethrin, deltamethrin) and 3 organophosphorus (chlorpyrifos, diazinon, malathion) pesticides were measured in 4 days of 24 h duplicate diet samples collected from 12 Atlanta adults over two cycles (2005-2006). Samples were composited into 9 categories, by food type, to evaluate their contribution to daily intakes. The resulting 437 samples were analyzed using a multiresidue method using liquid-liquid and solid-phase extraction followed by quantification via gas chromatograph with electron-capture detection. Total daily intakes (mg/kg-d) were calculated by summing the mass of a pesticide in all composites collected that day and dividing by body weight. Chlorpyrifos, diazinon, and cypermethrin in were detected in a range of composite types at frequencies > or =30%, whereas other pesticides were detected at lower frequencies. Concentrations ranged from the detection limits (0.38-0.88 ng/g) to several hundred ng/g, exceeding U.S. tolerances in a few cases. We also detected pesticides in some foods labeled organic. Total daily intakes were below the U.S. Environmental Protection Agency's oral reference doses, except in 6% of cases when the organophosphorus concentrations were summed. Results show frequent dietary exposure of our participants to the target pesticides from a range of food types.