National Health and Nutrition Examination Survey

Arsenic is widely distributed in the earth’s crust and is found most often in ground water rather than surface water. People encounter arsenic in many chemical forms that vary greatly in toxicity. The most toxic of the naturally-occurring arsenic compounds are inorganic forms of arsenic and their methylated metabolites. Less toxic are the organic arsenic compounds. Although this method does not reveal the chemical form of arsenic to which a person is exposed, it is sensitive enough to screen urine specimens rapidly from people thought to be exposed to arsenic or to evaluate total environmental or other total non-occupational exposure to arsenic.

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

1. Total arsenc

The method described in this manual assesses arsenic exposure by analyzing urine through the use of inductively coupled-plasma dynamic reaction cell-mass spectrometry (ICP-DRC-MS). Urine is analyzed because urinary excretion is the major pathway for eliminating arsenic from the mammalian body (Vahter ME, 1988). This method achieves rapid and accurate quantification of total urinary arsenic.

Total urine arsenic concentrations are determined by using ICP-DRC-MS. This multielement analytical technique is based on quadrupole ICP-MS technology (Date AR et al., 1989) and includes DRC™ technology (Tanner SD et al. , 1999), which minimizes or eliminates much argon-based polyatomic interference. Coupling radio frequency power into a flowing argon stream seeded with electrons creates the plasma, the heat source, which is ionized gas suspended in a magnetic field. Predominant species in the plasma are positive argon ions and electrons. Diluted urine samples are converted into an aerosol by using a nebulizer inserted within a spray chamber. A portion of the aerosol is transported through the spray chamber and then through the central channel of the plasma, where it is exposed to temperatures of 6000-8000 ºK. This thermal energy atomizes and ionizes the sample. The ions and the argon enter the mass spectrometer through an interface that separates the ICP, which is operating at atmospheric pressure (approximately 760 torr), from the mass spectrometer, which is operating at approximately 10^-5 torr. The mass spectrometer permits detection of ions at each mass-to-charge ratio in rapid sequence, which allows the determination of individual isotopes of an element. Once inside the mass spectrometer, the ions pass through the ion optics, then through DRC™, and finally through the mass-analyzing quadrupole before being detected as they strike the surface of the detector. The ion optics uses an electrical field to focus the ion beam into the DRC™. The DRC™ component is pressurized with an appropriate reaction gas and contains a quadrupole. In the DRC™, elimination or reduction of argon-based polyatomic interferences takes place through the interaction of the reaction gas with the interfering polyatomic species in the incoming ion beam. The quadrupole in the DRC™ allows elimination of unwanted reaction by-products that would otherwise react to form new interferences. Electrical signals, resulting from the detection of the ions, are processed into digital information that is used to indicate the intensity of the ions, and subsequently the concentration of the element. In this method, arsenic (isotope mass 75) and gallium (isotope mass 71) or tellurium (isotope mass 126) is measured in urine by ICP-DRC-MS, using argon/hydrogen (90%/10%, respectively) as a reaction gas (Neubauer K. et al., 1999). Urine samples are diluted 1:9 with 2% (v/v) double-distilled nitric acid containing gallium or tellurium for internal standardization.

2. Speciated arsenics

 (arsenobetaine, arsenocholine, trimethylarsine oxide, monomethylarsonic acid, dimethylarsinic acid, arsenous (III) acid, arsenic (V) acid)

The concentration of speciated arsenics is determined by using high performance liquid chromatography (HPLC) to separate the species coupled to an ICP-DRC-MS to detect the arsenic species. This analytical technique is based on separation by anion-exchange chromatography (IC), followed by detection using quadrupole ICP-MS technology, and includes DRC™ technology (Baranov VI et al., 1999), which minimizes or eliminates many argon-based polyatomic interferences (Tanner S et al., 2000) will require 0.5 mL of urine. Arsenic species column separation is largely achieved due to differences in charge-charge interactions of each negatively-charged arsenic component in the mobile phase, with the positively-charged quaternary ammonium groups bound at the column’s solid-liquid interface. Upon exit from the column, the chromatographic eluent goes through a nebulizer, where it is converted into an aerosol upon entering the spray chamber.

Carried by a stream of argon gas, a portion of the aerosol is transported through the spray chamber and then through the central channel of the plasma, where it is heated to temperatures of 6000-8000° K. This thermal energy atomizes and ionizes the sample. The ions and the argon enter the mass spectrometer through an interface that separates the ICP, which is operating at atmospheric pressure (approximately 760 torr), from the mass spectrometer, which is operating at approximately 10^-5 torr.

The mass spectrometer permits detection of ions at each mass-to-charge ratio in rapid sequence, which allows the determination of individual isotopes of an element. Once inside the mass spectrometer, the ions pass through the ion optics, then through the DRC™, and finally through the mass-analyzing quadrupole before being detected as they strike the surface of the detector. The ion optics uses an electrical field to focus the ion beam into the DRC™.

The DRC™ component is pressurized with an appropriate reaction gas and contains a quadrupole. In the DRC™, elimination or reduction of argon-based polyatomic interferences takes place through the interaction of the reaction gas with the interfering polyatomic species in the incoming ion beam. The quadrupole in the DRC™ allows elimination of unwanted reaction by-products that would otherwise react to form new interferences.

There were no changes (from the previous 2 years of NHANES) to equipment, lab methods or lab site.

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, and testing procedures. Constructive feedback are given to each staff person. 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 contract 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.

NCHS developed and distributed a quality control protocol for all the contract laboratories, which outlined the Westgard rules used when running NHANES specimens. Progress reports, containing any problems encountered during shipping or receipt of specimens, summary statistics for each control pool, QC graphs, instrument calibration, reagents, and any special considerations, are submitted to NCHS and Westat quarterly. The reports are reviewed 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 Science’s quality control and quality assurance performance criteria for accuracy and precision (similar to specifications outlined by Westgard (1981).

Subsample weights

Measures of urinary speciated arsenic were obtained in a one-third subsample of persons 6 years and over. Therefore, special sample weights (WTSA2YR) 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 laboratory data must be conducted with the key survey design and basic demographic variables. The NHANES 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 data files using the unique survey participant identifier SEQN.

Detection Limits

The detection limits were constant for all of the arsenics in the data set.

Lower detection limits for the total and speciated arsenics
Urinary total arsenic 0.74 µg/L
Urinary arsenous acid 1.2 µg/L
Urinary Arsenic acid 1.0 µg/L
Urinary Arsenobetaine 0.4 µg/L
Urinary Arsenocholine 0.6 µg/L
Urinary Dimethylarsonic acid 1.7 µg/L
Urinary Monomethylarsonic acid 0.9 µg/L
Urinary Trimethylarsine Oxide 1.0 µg/L

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

Please refer to the Analytic Guidelines for further details on the use of sample weights and other analytic issues. The Analytic Guidelines are available on the NHANES website.