Data Sources and Data Analysis

Blood, serum, and urine samples from NHANES

Biomonitoring measurements for CDC’s National Report on Human Exposure to Environmental Chemicals and Updated Tables (referred to as the Report and Updated Tables, respectively) are made using samples from participants in a series of surveys conducted by CDC’s National Center for Health Statistics (NCHS). The National Health and Nutrition Examination Survey (NHANES) is designed to collect data on the health and nutritional status of the U.S. population. NHANES collects information about a wide range of health-related behaviors, performs physical examinations, and collects clinical samples for laboratory tests. Beginning in 1999, NHANES became a continuous survey, sampling the U.S. population annually and releasing the data in 2-year cycles. The sampling plan follows a complex, stratified, multistage, probability-cluster design to select a representative sample of the civilian, noninstitutionalized population in the United States based on age, gender, and race/ethnicity.

The NHANES protocol includes a home interview, followed by a standardized physical examination in a mobile examination center. As part of the examination, blood is obtained by venipuncture from participants aged 1 year and older, and urine specimens are collected from participants aged 6 years and older. Starting with the 2015–2016 survey, urine specimens will be collected from children ages 3–5 years. Additional detailed information on the design and conduct of the NHANES survey is available at

NHANES measures environmental chemicals in blood, serum, or urine specimens. Most of the environmental chemicals are measured in randomly selected subsamples within specific age groups. Randomization of subsample selection is built into the NHANES design before sample collection begins. Random subsamples include different participants. Subsampling is needed to ensure an adequate quantity of sample for analysis and the mass spectrometry analytical methods.

  • Data tables in the Report and Updated Tables specify age groups and sample sizes for each exposure measurement.
  • Blood measurements in children younger than 12 years are limited by the amount of blood that can be collected.
  • Blood metals are measured in participants aged 1 year and older.
  • Beginning in 2011, the blood mercury species methyl mercury and ethyl mercury were added.
  • Serum cotinine and hemoglobin adducts of acrylamide and glycidamide are measured in the entire NHANES sample for participants ages 3 years and older.
  • Cotinine data are shown only in nonsmokers in the report.
  • Most measurements of chemicals in serum are made in samples from participant ages 12 years and older.
  • Urine was collected from participants ages 6 years and older until 2015, when it became possible to collect urine from children as young as 3 years.
  • Most urine chemicals are measured in a representative one-third subsample, but a full sample is used for participants ages 3–5 years.

If a particular chemical or an entire chemical group has no detectable results for three survey periods, it is not measured in subsequent survey periods.

  • The sulfonyl urea herbicide group was measured in NHANES 2003–2004, 2005–2006, and 2007–2008 with no detections, even at the 95th percentile, so this group is no longer measured in the ongoing survey.
  • Mono-n-octyl phthalate (MOP), a phthalate urinary metabolite, was not detected above the 95th percentile for more than three survey cycles, so it was no longer reported after 2010.
  • Several banned organophosphate and organochlorine pesticides have become largely undetectable and are no longer measured.
  • Some chemicals are widely detected but have had unchanging levels in the population (e.g., phytoestrogens) and are no longer measured.

By discontinuing measurements of chemicals that are rarely or not detectable, or well-characterized chemicals for which the source of exposure is consistent, CDC’s Environmental Health Laboratory can better allocate and optimize its limited resources. It can use those resources to develop methods to measure new chemicals of concern and continue to measure population exposure to chemicals of concern.

Beginning with NHANES 2005–2006, CDC used a weighted pooled-sample design to measure serum concentrations of dioxins, furans, polychlorinated biphenyls (PCBs), organochlorine pesticides and metabolites, and brominated flame retardants (polybrominated diphenyl ethers and polybrominated biphenyl [PBB] 153). Pooled samples are used when larger sample volumes are needed to improve the sensitivity of the measurements and to reduce the number of samples being analyzed, balancing the cost of the analysis against a low frequency of detectable results.

The following considerations guided the selection of chemicals in the Report and Updated Tables:

  • Scientific data suggested exposure in the U.S. population.
  • Health effects known or suspected to result from some levels of exposure were serious enough to cause concern.
  • Data were needed to assess the effectiveness of public health actions to reduce exposure to a chemical.
  • Biomonitoring analytical methods could provide results with adequate accuracy, precision, sensitivity, specificity, and throughput.
  • Adequate blood or urine samples were available.
  • Incremental analytical costs to perform the biomonitoring analysis for the chemical were reasonable.

The availability of biomonitoring methods with adequate performance and acceptable cost was a major consideration. Details on the prioritization process for scoring nominated chemicals and the resulting scores are available at:

Laboratory Analysis

CDC’s Environmental Health Laboratory (Division of Laboratory Sciences, National Center for Environmental Health) made the blood, serum, and urine exposure measurements in the Report and Updated Tables. The following analytical methods used for measuring the environmental chemicals or their metabolites in blood, serum, and urine:

  • Isotope dilution mass spectrometry
  • Inductively coupled plasma mass spectrometry
  • Graphite furnace atomic absorption spectrometry

Laboratory measurements underwent extensive quality control and quality assurance review:

  • Tolerance limits for operational parameters were maintained.
  • Quality control samples were measured in each analytical run to detect unacceptable performance in accuracy or precision.
  • Traceable calibration materials were verified.

Appendix D lists references for the analytical methods used to measure the different chemicals.

Data Analysis

NHANES is a complex, stratified, multistage, probability-cluster design survey. Because of that, sample weights must be used to adjust for the unequal probability of selection into the survey. Sample weights also are used to adjust for possible bias resulting from nonresponse and are post-stratified to U.S. Census Bureau estimates of the U.S. population. Data were analyzed using Statistical Analysis System (SAS) (SAS Institute Inc., 2002) and the SUDAAN (RTI International, NC) statistical software packages. SUDAAN uses sample weights and calculates variance estimates that account for the complex survey design. The design does not permit straightforward analysis of exposure levels by non-targeted strata such as locality, state, or region; seasons of the year; proximity to sources of exposure; or by use of particular products. NCHS guidelines for analysis of NHANES data are available at

The Report and Updated Tables present descriptive statistics on the blood, serum, or urine levels for each environmental chemical reported. Statistics include unadjusted geometric means and percentiles with confidence intervals. In each table, results are given for the total population and by age group, gender, and race/ethnicity, as defined in NHANES. For these analyses, based on the sample design, race/ethnicity is categorized as Mexican American, non-Hispanic black, and non-Hispanic white. Beginning in 2011–2012, two additional categories were added: all Hispanic and non-Hispanic Asian (referred to as Asian in the tables). Other racial/ethnic groups are sampled, but the proportion of the total U.S. population represented by other racial/ethnic groups is not large enough to produce valid estimates. Other racial/ethnic groups are included in estimates that are based on the entire population sample. Age groups are as described for each chemical in each data table. Gender is coded as male or female.


For chemicals measured in urine, levels are presented per volume of urine and per gram of creatinine. Urinary levels are expressed both ways in the literature and used for different purposes. Levels per gram of creatinine (i.e., creatinine corrected) adjust for urine dilution. For example, a person who drinks more fluids than another person will likely have a higher, but more dilute, urine output. Interpretation of creatinine-corrected results should also recognize that creatinine correction can partially adjust for differences in lean body mass or renal function among persons. Children and young adolescents typically have less muscle mass than adults. Because muscle is an important source of creatinine in urine, children have lower urinary creatinine concentrations than do adults. The lower urine creatinine concentration can result in higher creatinine-corrected results in children in comparison with adults.

For dioxins, furans, PCBs, and organochlorine pesticides, serum levels are presented per gram of total lipid and per whole weight of serum. These compounds are lipophilic and concentrate in the body’s lipid stores, including the lipid in serum. Serum levels reported per gram of total lipid reflect the amount of these compounds stored in body fat. Serum levels per whole weight of serum are also included for easier comparison with studies investigating exposure to these chemicals and reported levels in these units. Other mostly non-lipophilic chemicals measured in serum are expressed per liter of serum (e.g., micrograms per liter). Acrylamide and glycidamide adducts are expressed in picomoles per gram of blood hemoglobin.

Results are reported in standard units, generally conforming to those most commonly used in biomonitoring measurements. Table 2 shows some useful unit conversions.

Table 2. Units of Measurements and Abbreviations

Table 2. Units of Measurements and Abbreviations
Unit Abbreviation Value
liter L
deciliter dL 10-1 liters
milliliter mL 10-3 liters
gram g
milligram mg 10-3 grams
microgram µg 10-6 grams
nanogram ng 10-9 grams
picogram pg 10-12 grams
femtogram fg 10-15 grams

Geometric means

A geometric mean gives a better estimate of central tendency for data that are distributed with a long tail at the upper end of the distribution. This type of distribution is common when measuring environmental chemicals in blood or urine. The geometric mean is influenced less by high values than is the arithmetic mean. In the Report and Updated Tables, geometric means are calculated by taking the log of each concentration and then using SUDAAN software to compute the weighted mean of those log-transformed values. Ninety-five percent confidence intervals around the weighted mean are calculated by adding and subtracting an amount equal to the product of a Student’s t-statistic and the standard error of the weighted mean estimate. The degrees of freedom of the t-statistic was determined by subtracting the number of strata from the number of primary sampling units, according to the data available from the complex survey design. The standard error was computed with SUDAAN’s PROC DESCRIPT procedure and “with replacement” design option (design = WR), with Taylor series linearization used for variance estimation. The weighted geometric mean and its confidence limits were obtained by taking the antilogs of this weighted mean and its upper and lower confidence limits.

The measured value for a pooled sample is comparable to an arithmetic average of measurements in individuals. Consequently, the pooled sample result is expected to be higher than the geometric mean of multiple individual results. In addition, direct calculation of the design effects required for accurate standard error and confidence interval estimation is not possible because samples are pooled across the design cells of the original survey. For these reasons, data tables showing the pooled sample results present only weighted arithmetic means and unadjusted standard errors for each category. The standard errors are unadjusted and therefore, do not reflect the design effects of the survey. In many cases, the standard errors are based on very few pooled sample measurements and cannot be expected to accurately reflect the true imprecision of the weighted arithmetic mean estimates. Therefore, when the unadjusted standard error was more than 30% of the weighted arithmetic mean, this is noted with a double asterisk (**) and footnoted.

Limit of detection

The limit of detection (LOD) is the level at which the measurement has a 95% probability of being greater than zero [Taylor 1987]. In the Report and Updated Tables, the LODs for each chemical and survey period are provided in each data table and collectively in Appendix C. Concentrations less than the LOD are assigned a value equal to the LOD divided by the square root of two for calculation of geometric means. Assigning a value of the LOD divided by two made little difference in geometric mean estimates. If the proportion of results below the LOD was greater than 40%, geometric means were not calculated. For the same chemical, LOD values sometimes change over time as a result of improvements to analytical methods. One possible consequence is that results reported as “< LOD” in the 1999–2000 data might be reported as a concentration value above the LOD in 2001–2002 or 2003–2004 because the analytical method had improved. Thus, for proper interpretation of LODs in the data tables, care must be taken to use the LOD that applies to the survey period. Percentile estimates (see below) that are less than the LOD for the chemical analysis were reported as “< LOD.”

For most chemicals, the LOD is constant for each specimen analyzed. Dioxins, furans, PCBs, organochlorine pesticides, and a few other pesticides have their own LOD. These analyses have an individual LOD for each sample, mostly because the sample volume used for analysis was different. A higher sample volume results in a lower LOD (i.e., a better ability to detect low levels).The maximum LOD values are given in each data table and in Appendix D. The maximum LOD was the highest LOD among all samples analyzed. Typically, the mean LOD is about 40%–50% of the maximum LOD. The same procedure for imputing values below the LOD in calculations of geometric means was used for chemicals with individual LODs for each sample. Concentrations less than the individual LOD were assigned a value equal to the individual LOD divided by the square root of two. When reporting percentiles for chemicals with individual sample LODs, if any sample LOD in the demographic group was above the percentile estimate, then the percentile estimate was not reported.

For chemicals measured in urine, separate tables are presented for the chemical concentration expressed per volume of urine (uncorrected table) and the chemical concentration expressed per gram of creatinine (creatinine corrected table). Geometric mean and percentile calculations were performed separately for each of these concentrations. LOD calculations were performed using the chemical concentration expressed per volume of urine, because this concentration determines the analytical sensitivity. For this reason, LOD results for urine measurements in each data table and in Appendix C are in units of weight per volume of urine. In the creatinine corrected tables, a result for a geometric mean or percentile was reported as < LOD if the corresponding geometric mean or percentile was < LOD in the table using weight per volume of urine. For example, if the 50th percentile for males was < LOD in the table using weight per volume of urine, it would also be < LOD in the creatinine corrected table.

For chemicals measured in serum lipid, separate tables are presented for the chemical concentration expressed per volume of serum (lipid unadjusted table) and the chemical concentration expressed per amount of lipid (lipid adjusted table). Geometric mean and percentile calculations were performed separately for each of these concentrations. LOD calculations were performed using the chemical concentration expressed per amount of lipid, because this concentration determines the analytical sensitivity. For this reason, LOD results for chemicals measured in each data table and in Appendix C are in weight per amount of lipid. In the lipid unadjusted tables, a result for a geometric mean or percentile was reported as < LOD if the corresponding geometric mean or percentile was < LOD in the lipid adjusted table.


Percentiles (50th, 75th, 90th, and 95th) provide additional information about the shape of the distribution. Percentile estimates and 95% confidence interval estimates that are less than the limit of detection are indicated as <LOD in the data tables. In the Third National Report on Human Exposure to Environmental Chemicals, weighted percentile estimates for 1999–2000 and 2001–2002 data were calculated using SAS univariate procedure (PROC UNIVARIATE) and a proportions estimation procedure. A percentile estimate might fall on a value that is repeated multiple times in a particular demographic group defined by age, sex, and race. In non-Hispanic white males ages 12–19 years, for example, five results all have a value of 90.1. Since the Third Report, we have improved the procedure for estimating percentiles to better handle this situation. This improved procedure makes each repeated value unique by adding a negligibly small number to each repeated value. All data from 1999–2004 have been reanalyzed using this new procedure to handle situations where the percentile falls on a repeating value. Therefore, some percentile estimates differ slightly in the current Fourth Report compared with the Third Report. Appendix A gives the details of the new procedure for estimating percentiles.


Taylor JK. 1987. Quality assurance of chemical measurements. Boca Raton, FL: Lewis Publishers.

Page last reviewed: July 27, 2017