Ralph D. Whitehead

Impact of prenatal chlorpyrifos exposure on neurodevelopment in the first 3 years of life among inner-city children.

OBJECTIVE. The purpose of this study was to investigate the impact of prenatal exposure to chlorpyrifos on 3-year neurodevelopment and behavior in a sample of inner-city minority children. METHODS. As part of an ongoing prospective cohort study in an inner-city minority population, neurotoxicant effects of prenatal exposure to chlorpyrifos were evaluated in 254 children through the first 3 years of life. This report examined cognitive and motor development at 12, 24, and 36 months (measured with the Bayley Scales of Infant Development II) and child behavior at 36 months (measured with the Child Behavior Checklist) as a function of chlorpyrifos levels in umbilical cord plasma. RESULTS. Highly exposed children (chlorpyrifos levels of >6.17 pg/g plasma) scored, on average, 6.5 points lower on the Bayley Psychomotor Development Index and 3.3 points lower on the Bayley Mental Development Index at 3 years of age compared with those with lower levels of exposure. Children exposed to higher, compared with lower, chlorpyrifos levels were also significantly more likely to experience Psychomotor Development Index and Mental Development Index delays, attention problems, attention-deficit/hyperactivity disorder problems, and pervasive developmental disorder problems at 3 years of age. CONCLUSIONS. The adjusted mean 36-month Psychomotor Development Index and Mental Development Index scores of the highly and lower exposed groups differed by only 7.1 and 3.0 points, respectively, but the proportion of delayed children in the high-exposure group, compared with the low-exposure group, was 5 times greater for the Psychomotor Development Index and 2.4 times greater for the Mental Development Index, increasing the number of children possibly needing early intervention services.

Urinary Concentrations of Metabolites of Pyrethroid Insecticides in the General U.S. Population: National Health and Nutrition Examination Survey 1999–2002

Background Pyrethroid insecticides are the most commonly used residential insecticides in the United States. Objectives Our objective was to assess human exposure via biomonitoring to pyrethroid insecticides in a representative sample of the general U.S. population ≥ 6 years of age. Methods By using isotope-dilution high-performance liquid chromatography/electrospray chemical ionization/tandem mass spectrometry, we measured five urinary metabolites of pyrethroid insecticides in 5,046 samples collected as a part of the 1999–2002 National Health and Nutrition Examination Survey (NHANES). Univariate, multivariate, and Pearson correlation analyses were performed using SUDAAN and SAS software, incorporating the appropriate sample weights into the analyses. Multivariate analyses included age, sex, race/ethnicity, creatinine, fasting status, and urine collection time as covariates. Results We detected 3-phenoxybenzoic acid (3PBA), a metabolite common to many pyrethroid insecticides, in more than 70% of the samples. The least-squares geometric mean (LSGM) concentration (corrected for covariates) of 3PBA and the frequency of detection increased from 1999–2000 (0.292 ng/mL) to 2001–2002 (0.318 ng/mL) but not significantly. Non-Hispanic blacks had significantly higher LSGM 3PBA concentrations than did non-Hispanic whites and Mexican Americans in the 2001–2002 survey period and in the combined 4-year survey periods but not in the 1999–2000 survey period. Children had significantly higher LSGM concentrations of 3PBA than did adolescents in both NHANES periods and than adults in NHANES 1999–2000. Cis- and trans-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid were highly correlated with each other and with 3PBA, suggesting that urinary 3PBA was derived primarily from exposure to permethrin, cypermethrin, or their degradates. Conclusions Pyrethroid insecticide exposure in the U.S. population is widespread, and the presence of its metabolites in the urine of U.S. residents indicates that children may have higher exposures than adolescents and adults.

Measurement of dialkyl phosphate metabolites of organophosphorus pesticides in human urine using lyophilization with gas chromatography-tandem mass spectrometry and isotope dilution quantification.

Urinary dialkylphosphate (DAP) metabolites have been used to estimate human exposure to organophosphorus pesticides. We developed a method for quantifying the six DAP urinary metabolites of at least 28 organophosphorus pesticides using lyophilization and chemical derivatization followed by analysis using isotope-dilution gas chromatography–tandem mass spectrometry (GC–MS/MS). Urine samples were spiked with stable isotope analogues of the DAPs and the water was removed from the samples using a lyophilizer. The dried residue was dissolved in acetonitrile and diethyl ether, and the DAPs were chemically derivatized to their respective chloropropyl phosphate esters. The chloropropyl phosphate esters were concentrated, and analyzed using GC–MS/MS. The limits of detection of the method were in the low μg/l (parts per billion) to mid pg/ml range (parts per trillion) with coefficients of variation of 7–14%. The use of stable isotope analogues as internal standards for each of these metabolites allows for sample-specific adjustment for recovery and thus permits a high degree of accuracy and precision. Use of this method with approximately 1100 urine samples collected from pregnant women and children indicate that the low limits of detection allow this method to be used in general population studies.

A multi-analyte method for the quantification of contemporary pesticides in human serum and plasma using high-resolution mass spectrometry

We have developed a sensitive and accurate analytical method for quantifying 29 contemporary pesticides in human serum or plasma. These pesticides include organophosphates, carbamates, chloroacetanilides, and synthetic pyrethroids among others and include pesticides used in agricultural and residential settings. Our method employs a simple solid-phase extraction followed by a highly selective analysis using isotope dilution gas chromatography-high-resolution mass spectrometry. Our method is very accurate, has limits of detection in the low pg/g range and coefficients of variation of typically less than 20% at the low pg/g end of the method linear range. We have used this method to measure plasma pesticide concentrations in females living in an urban area. We found detectable concentrations of carbaryl/naphthalene, propoxur, bendiocarb, chlorpyrifos, diazinon, dicloran, captan and folpet or their metabolites in more than 20% of the plasma samples tested.

An Intervention to Reduce Residential Insecticide Exposure during Pregnancy among an Inner-City Cohort

Background We previously reported widespread insecticide exposure during pregnancy among inner-city women from New York City. Here we report on a pilot intervention using integrated pest management (IPM) to reduce pest infestations and residential insecticide exposures among pregnant New York City African-American and Latina women (25 intervention and 27 control homes). Methods The IPM consisted of professional cleaning, sealing of pest entry points, application of low-toxicity pesticides, and education. Cockroach infestation levels and 2-week integrated indoor air samples were collected at baseline and one month postintervention. The insecticides detected in the indoor air samples were also measured in maternal and umbilical cord blood collected at delivery. Results Cockroach infestations decreased significantly (p = 0.016) after the intervention among intervention cases but not control households. Among the intervention group, levels of piperonyl butoxide (a pyrethroid synergist) were significantly lower in indoor air samples after the intervention (p = 0.016). Insecticides were detected in maternal blood samples collected at delivery from controls but not from the intervention group. The difference was significant for trans-permethrin (p = 0.008) and of borderline significance (p = 0.1) for cis-permethrin and 2-isopropoxyphenol (a propoxur metabolite). Conclusion To our knowledge, this is the first study to use biologic dosimeters of prenatal pesticide exposure for assessing effectiveness of IPM. These pilot data suggest that IPM is an effective strategy for reducing pest infestation levels and the internal dose of insecticides during pregnancy.

Quantification of riboflavin in human urine using high performance liquid chromatography-tandem mass spectrometry.

We developed a selective method to measure riboflavin in human urine. Sample preparation involved solid phase extraction and concentration of the target analyte in urine. The urine concentrate was analyzed using high performance liquid chromatography-tandem mass spectrometry. Riboflavin concentrations were quantified using an isotopically labeled internal standard. The limit of detection was 11 ng/mL, and the linear range was 4.4-20,000 ng/mL. The relative standard deviation at 100, 1000, and 5000 ng/mL was 17%, 17%, and 12%, respectively. The accuracy was 90%. On average, 100 samples, including calibration standards and quality control samples, were prepared per day. Using our method, we measured concentrations of riboflavin in human urine samples that were collected from participants in a study where riboflavin was used as a surrogate chemical to simulate exposure to an environmental toxicant.

High Prevalence of Vitamin B12 Deficiency and No Folate Deficiency in Young Children in Nepal

Many children in low- and middle-income countries may have inadequate intake of vitamin B12 and folate; data confirming these inadequacies are limited. We used biochemical, demographic, behavioral and anthropometric data to describe the folate and vitamin B12 concentrations among six- to 23-month-old Nepalese children. Vitamin B12 (serum B12 < 150 pmol/L) and folate deficiencies (red blood cell (RBC) folate < 226.5 nmol/L) were assessed. We used logistic regression to identify predictors of vitamin B12 deficiency. The vitamin B12 geometric mean was 186 pmol/L; 30.2% of children were deficient. The mean RBC folate concentration was 13,612 nmol/L; there was no deficiency. Factors associated with vitamin B12 deficiency included: (a) age six to 11 months (adjusted odds ratio (aOR) 1.51; 95% confidence interval (CI): 1.18, 1.92) or 12–17 months (aOR 1.38; 95% CI: 1.10, 1.72) compared to 18–23 months; (b) being stunted (aOR 1.24; 95% CI: 1.03, 1.50) compared to not being stunted; (c) and not eating animal-source foods (aOR 1.85; 95% CI: 1.42, 2.41) compared to eating animal-source foods the previous day. There was a high prevalence of vitamin B12 deficiency, but no folate deficiency. Improving early feeding practices, including the consumption of rich sources of vitamin B12, such as animal-source foods and fortified foods, may help decrease deficiency.

Effects of preanalytical factors on hemoglobin measurement: a comparison of two HemoCue® point-of-care analyzers

BACKGROUND In field studies, hemoglobin (Hb) is often measured using a battery-operated, portable HemoCue® hemoglobinometer. METHODS We compared the performance of 2 HemoCue® models (Hb-201+ and Hb-301) and investigated effects of preanalytical factors on Hb results by simulating unfavorable field conditions. RESULTS The Hb-301 produced 2.6% higher results compared to the Hb-201+. Hb had to be measured within 1min of filling the Hb-301 cuvette to avoid artificially elevated concentrations (1.3% per min). The Hb-301 cuvettes withstood elevated temperature (37°C) and humidity (72%) for 3weeks, while the Hb-201+ cuvettes degraded within 10min under those conditions. Both cuvette types withstood elevated temperature for 3weeks. Properly-collected venous and capillary blood produced comparable results. Pooled capillary blood produced comparable results to the second and third but not the fourth drop of blood (3.3% lower). Blood could be stored for ≤4days at 10-30°C before Hb-201+ measurement, but only for 1day at 10-23°C before Hb-301 measurement (≤1% change in Hb). CONCLUSIONS Higher Hb results obtained with the Hb-301 may influence the interpretation of anemia prevalence in health surveys. While the Hb-301 performed better in high humidity conditions, the Hb-201+ provided more user flexibility regarding delayed Hb reading.

Effects of Inflammation on Biomarkers of Vitamin A Status among a Cohort of Bolivian Infants

Globally, vitamin A deficiency (VAD) affects nearly 200 million children with negative health consequences. VAD can be measured by a retinol-binding protein (RBP) and serum retinol concentrations. Their concentrations are not always present in a 1:1 molar ratio and are affected by inflammation. This study sought to quantify VAD and its impact on infant mortality and infectious morbidity during the first 18 months of life in a cohort of mother-infant dyads in El Alto, Bolivia, while accounting for the previously mentioned measurement issues. Healthy mother-infant dyads (n = 461) were enrolled from two hospitals and followed for 12 to 18 months. Three serum samples were collected (at one to two, six to eight, and 12 to 18 months of infant age) and analyzed for RBP, and a random 10% subsample was analyzed for retinol. Linear regression of RBP on retinol was used to generate RBP cut-offs equivalent to retinol <0.7 µmol/L. All measures of RBP and retinol were adjusted for inflammation, which was measured by a C-reactive protein and alpha (1)-acid glycoprotein serum concentrations using linear regression. Infant mortality and morbidity rates were calculated and compared by early VAD status at two months of age. Retinol and RBP were weakly affected by inflammation. This association varied with infant age. Estimated VAD (RBP < 0.7 µmol/L) decreased from 71.0% to 14.8% to 7.7% at two, six to eight, and 12 to 18 months of age. VAD was almost nonexistent in mothers. Early VAD was not significantly associated with infant mortality or morbidity rates. This study confirmed a relationship between inflammation and vitamin A biomarkers for some subsets of the population and suggested that the vitamin A status in early infancy improves with age and may not have significantly affected morbidity in this population of healthy infants.

The Loss of 5-Methyltetrahydrofolate in Human Serum under Suboptimal Preanalytical Conditions Can Only Partially Be Recovered by an Oxidation Product

BACKGROUND Maintaining folate stability during sample handling is important, yet challenging. OBJECTIVE We investigated the effects of suboptimal preanalytical conditions on serum folate stability. METHODS By using an HPLC-tandem MS method we measured folates [5-methyltetrahydrofolate (5-methylTHF), folic acid, MeFox (5-methylTHF oxidation product, pyrazino-s-triazine derivative of 4α-hydroxy-5-methylTHF), and other minor folate forms at or below the limit of detection] in human serum exposed to suboptimal conditions. RESULTS Whole blood samples (n = 21) stored at 32°C for ≤ 3 d (Expt. 1: delayed processing) showed significant decreases in serum total folate (tFOL; sum of folate forms: 11-32%, 5.5-15.9 nmol/L) and 5-methylTHF (36-62%, 14.5-25.1 nmol/L) and a significant increase in MeFox (346-415%, 7.17-8.63 nmol/L). Serum samples (n = 21) stored at 11°C for 7-14 d (Expt. 2: delayed freezing) also showed significant decreases in tFOL (4.6-10.4%, 2.3-5.1 nmol/L) and 5-methylTHF (8.4-29%, 3.4-11.6 nmol/L) and significant increases in MeFox (88-320%, 1.82-6.62 nmol/L). The molar loss in 5-methylTHF exceeded the gain in MeFox in these 2 experiments. When we exposed 3 serum pools (tFOL: 16.7-58.3 nmol/L) for 24 h to an elevated temperature of 37°C (Expt. 3), the significant decrease in 5-methylTHF (33% on average) was compensated for by an equimolar gain in MeFox. Repeated freeze/thaw cycles (≤ 3 cycles) of serum [closed (Expt. 4) and open (Expt. 5) vials] showed generally stable folates with small (<1 nmol/L) changes. Long-term (≤ 12 mo) exposure of 3 serum pools (tFOL: 17.5-63.7 nmol/L) to a suboptimal (-20°C) freezing temperature (Expt. 6) showed significant decreases in tFOL (5% on average) already after 3 mo. The molar loss in 5-methylTHF exceeded the gain in MeFox. Folic acid generally showed good stability. CONCLUSIONS To avoid folate losses, unprocessed whole blood should be protected from elevated temperatures and serum should not be refrigerated for >2 d or for a long term stored at -20°C.