National Health and Nutrition Examination Survey

Organophosphorus insecticides, which are active against a broad spectrum of insects, have accounted for a large share of all insecticides used in the United States. Although organophosphorus insecticides are still used for insect control on many food crops, most residential uses have been phased out in the United States as a result of implementation of the Food Quality Protection Act of 1996. Pesticide residues and their metabolites in human tissues and fluids can be indicative of pesticide exposure and the total body burden of these pesticides. About 75% of registered organophosphorus insecticides are metabolized in the body to measurable dialkyl phosphate (DAP) metabolites. Dialkyl phosphates may also occur in the environment as a result of degradation of organophosphorus insecticides, and therefore, the presence in a person’s urine may reflect exposure to the metabolite itself.

Generally, six urinary dialkyl phosphate metabolites of organophosphorus insecticides have been measured: dimethylphosphate (DMP); dimethylthiophosphate (DMTP); dimethyldithiophosphate (DMDTP); diethylphosphate (DEP); diethylthiophosphate (DETP); and diethyldithiophosphate (DEDTP). Each of the six urinary dialkyl phosphate metabolites can be produced from the metabolism of more than one organophosphorus insecticide or may be present following ingestion of the specific DAP metabolite. Therefore, the presence of one or more dialkyl phosphate metabolites without additional information cannot be linked to exposure to a specific organophosphorus insecticide.

Participants aged 6 years and older who met the subsample requirements.

This method was developed 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 mg/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.

Urine specimens are processed, stored, and shipped to the Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention for analysis.
Detailed specimen collection and processing instructions are discussed in the NHANES Laboratory/Medical Technologists Procedures Manual (LPM). Vials are stored under appropriate frozen (–20°C) conditions until they are shipped to National Center for Environmental Health for testing.

Mobile Examination Centers (MECs)

Laboratory team performance is monitored using several techniques. NCHS and contract consultants use a structured quality assurance evaluation during unscheduled visits to evaluate both the quality of the laboratory work and the quality-control procedures. Each laboratory staff person is observed for equipment operation, specimen collection and preparation; testing procedures and constructive feedback are given to each staff. Formal retraining sessions are conducted annually to ensure that required skill levels were maintained.

The NHANES QA/QC protocols meet the 1988 Clinical Laboratory Improvement Act mandates. Detailed QA/QC instructions are discussed in the NHANES LPM.

Analytical Laboratories

NHANES uses several methods to monitor the quality of the analyses performed by the laboratories. In the MEC, these methods include performing blind split samples collected on “dry run” sessions. In addition, contract laboratories randomly perform repeat testing on 2.0% of all specimens.

NCEH developed and distributed a quality control protocol for all the laboratories which outlined the Westgard rules used when running NHANES specimens. Any problems encountered during shipping or receipt of specimens, instrument calibration, reagents, and any special considerations are submitted to NCHS and Westat. Summary statistics for each control pool, QC graphs, are reviewed by NCHS for trends or shifts in the data. The laboratories are required to explain any identified areas of concern.

All QC procedures recommended by the manufacturers were followed. Reported results for all assays meet the Division of Laboratory Sciences’ quality control and quality assurance performance criteria for accuracy and precision (similar to specifications outlined by Westgard (1981).

Subsample weights

Measures of urinary diakyl phosphate metabolites were measured in a one third subsample of persons 6 years and over. Special sample weights are required to analyze these data properly. Specific sample weights for this subsample are included in this data file and should be used when analyzing these data.

Variance estimation

The analysis of NHANES 2003-2004 laboratory data must be conducted with the key survey design and basic demographic variables. The NHANES 2003-2004 Demographic Data File contains demographic and sample design variables. The recommended procedure for variance estimation requires use of stratum and PSU variables (SDMVSTRA and SDMVPSU, respectively) in the demographic data file.

Links to NHANES Data Files

This laboratory data file can be linked to the other NHANES 2003-2004 data files using the unique survey participant identifier SEQN.

Detection Limits

The detection limits were constant for all of the analytes except for diethyldithiophosphate (URXOP6) in the data set. Diethyldithiophosphate (URXOP6) has two detection limits. Two variables are provided for each of these analytes. The variable named URD___LC indicates whether the result was below the limit of detection. There are two values: “0” and “1”. “0” means that the result was at or above the limit of detection. “1” indicates that the result was below the limit of detection.

The other variable named URX___ provides the analytic result for that analyte.

The detection limit divided by the square root of 2 is the value that is provided for results that are below the limit of detection.

The lower limit of detections for the diakyl phosphate metabolites is:

Please refer to the Analytic Guidelines for further details on the use of sample weights and other analytic issues.