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Protocol - Concentrations of Polychlorinated Biphenyls (PCBs) and Persistent Pesticides

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Description:

Blood is collected from the subject in a pre-screened 3 ml red top tube and is not inverted or mixed and kept upright on a tube rack for 20-30 minutes to allow to coagulate. Centrifuge the red-top tubes for 10 minutes, transfer to Wheaton Bottle, and cap. Place serum samples in a -20° C freezer and store until shipment.

Analysis is by high-resolution gas chromatography/isotope-dilution high-resolution mass spectrometry (HRGC/ID-HRMS). This device measures concentrations of polychlorinated biphenyls (PCBs) and persistent pesticides in serum.

Protocol:

Administer the following questions prior to blood collection.

1. Do you have hemophilia or any bleeding disorder?

[ ] Yes

[ ] No

[ ] Don't Know

[ ] Refused

      • If the participant answers "Yes" or "Don't know," or refuses to answer, blood will not be collected.

2. Have you had cancer chemotherapy within the past 4 weeks?

[ ] Yes

[ ] No

[ ] Don't Know

[ ] Refused

      • If the participant answers "Yes" or "Don't know," or refuses to answer, blood will not be collected.

3. Have you had any problems with a blood draw in the past?

[ ] Yes

[ ] No

[ ] Don't Know

[ ] Refused

  • If the participant answers "Yes," go to question 4.
  • If the participant answers "No," "Don't know," or refuses to answer, go to question 5.

4. What problems have you had with a blood draw in the past?

______________________________

  • Record the types of problems that the participant experienced during previous blood draws.
  • If the participant refuses to answer or does not remember specifically what type of problem was experienced in the past, record and go to question 5.

5. When was the last time you had anything to eat or drink other than water?

Date ____________ mm/dd/yyyy

Time ____________ am/pm

6. Have you had sweetener or milk added to a drink, such as coffee or tea, in the last 8 hours?

[ ] Yes

[ ] No

[ ] Don't Know

[ ] Refused

  • Record the participant's response.
  • "Sweetener" includes sugar, honey, and flavored creamers. If the participant consumed an artificial sweetener in coffee, tea, or a diet soda, record "No."

7. Have you had alcohol such as beer, wine, or liquor in the last 8 hours?

[ ] Yes

[ ] No

[ ] Don't Know

[ ] Refused

8. Have you chewed gum, or used breath mints, lozenges, cough drops, or other cough or cold remedies in the last 8 hours?

[ ] Yes

[ ] No

[ ] Don't Know

[ ] Refused

9. Have you used antacid, laxatives, or anti-diarrheal medications in the last 8 hours?

[ ] Yes

[ ] No

[ ] Don't Know

[ ] Refused

10. Have you taken a dietary supplement such as vitamins or minerals in the last 8 hours?

[ ] Yes

[ ] No

[ ] Don't Know

[ ] Refused

11. Has a doctor ever told you that you had diabetes?

[ ] Yes

[ ] No

[ ] Don't Know

[ ] Refused

  • If the participant answers "Yes," go to question 12.
  • If the participant answers "No" and is pregnant probe "This includes gestational diabetes." If the participant still answers "No" after probe, prepare to draw participant's blood.
  • If the participant is not pregnant and answers that she had gestational diabetes while pregnant, indicate that this does not include gestational diabetes and prepare to draw the participant's blood.
  • If the participant answers "No" and is not pregnant, prepare to draw the participant's blood.

12. Have you taken any insulin in the last 8 hours?

[ ] Yes

[ ] No

[ ] Don't Know

[ ] Refused

      • Record the participant's response and prepare to draw the participant's blood.

The entire standard operating procedure that includes the questions, venipuncture supplies, and venipuncture procedure appears here.

Follow a standard venipuncture protocol but red top vacutainer tube must be collected first. Draw the blood with a stainless steel needle and use a pre-screened vacutainer tube.

The optimal amount of blood in red top tube is 1 to 10 ml. For collection, loosen the tourniquet immediately after blood flow is established and release entirely as the last tube fills. Completely fill all the Vacutainer tubes and then withdraw the needle with a slow but firm motion. Red-top tubes should not be inverted or mixed. Label all tubes. Place the red-top tubes upright in a rack and allow them to clot at room temperature for 20-30 minutes. Centrifuge the red-top tubes for 10 minutes at the RPM necessary to attain a force of 1000 x g. Using a transfer pipette, pipette the serum from each participant's red-top tubes into the Wheaton Bottle and cap. Check to make sure that the numbers on the labels are the same. DO NOT ALLOW SERUM TO REMAIN IN CONTACT WITH THE CLOT FOR LONGER THAN 1 HOUR AFTER THE SPECIMEN IS COLLECTED. Mix the serum gently, cap each bottle and place upright in a -20° C freezer and store at the same temperature until shipment. The time between collecting blood and freezing serum should not be more than 1 1/2 hours. Note on the sample log if a sample is turbid or hemolyzed, or if the serum was left in contact with red cells for more than 1 hour or left at room temperature for more than 90 minutes before freezing.

The blood may be aliquoted to cryovials and placed in a -20 C freezer prior to analyses. Specimen stability has been demonstrated for analytes measured by this method for at least 10 years at -30° C or below.

Concentrations of PCBs and persistent pesticides are measured in serum using high performance liquid chromatography coupled to inductively-coupled-plasma dynamic-reaction-cell mass spectrometry (ICP-DRC-MS).

The entire lab procedure is available here.

Protocol Name from Source:

National Children’s Study, Biospecimen Adult Blood Procedures: Standard Operating Procedures

National Health and Nutrition Examination Survey, CDC Laboratory Procedure Manual, PCBs and Persistent Pesticides

Availability:

Publicly available

Personnel and Training Required

The blood draw and processing procedures should be performed by a certified laboratory technician or a phlebotomist.

Equipment Needs

Highly specialized laboratory equipment is necessary to perform accurate analyses. A stainless steel needle should be used for venipuncture. Collection tubes need to be pre-screened for these toxicants prior to venipuncture.

Requirements
Requirement CategoryRequired
Major equipment Yes
Specialized training No
Specialized requirements for biospecimen collection No
Average time of greater than 15 minutes in an unaffected individual No
Mode of Administration

Bioassay

Life Stage:

Adolescent, Adult, Pregnancy

Participants:

Age 12 and older, pregnant women

Specific Instructions:

Follow the venipuncture procedures closely. Collect this blood sample first in a red-top tube.

Standard venipuncture procedures may be used for children aged 1 year and older.

Selection Rationale

The National Children's Study (NCS) was one of the largest pregnancy cohort studies in the United States, and the biospecimen collection methods have been validated in other studies.

The National Health and Nutrition Examination Survey (NHANES) is a major cross-sectional study in the United States, and the methods have been validated on other studies.

Language

English

Standards
StandardNameIDSource
Common Data Elements (CDE) Serum Polychlorinated Biphenyl Persistent Pesticide Concentration in Nanogram Per Gram Lipid Weight 5633750 CDE Browser
Process and Review

The Expert Review Panel has not reviewed this measure yet.

Source

Centers for Disease Control and Prevention (CDC), National Health and Nutrition Examination Survey (NHANES) 2003-2004. (2006). CDC laboratory procedure manual, PCBs and persistent pesticides. Atlanta, GA: Author.

Eunice Kennedy Shriver National Institute of Child Health and Human Development. (2014). National Children’s Study, Biospecimen adult blood procedures: Standard operating procedures. Rockville, MD: Author.

General References

Barr, J. B., Maggio, V. L., Barr, D. B., Turner, W. E., Sjodin, A., Sandau, C. D., Pirkle, J. L., Needham, L. L., & Patterson, D. G., Jr. (2003). New high-resolution mass spectrometric approach for the measurement of polychlorinated biphenyls and organochlorine pesticides in human serum. Journal of Chromatography B, 794, 137-148.

Centers for Disease Control and Prevention. (2013-14). National Health and Nutrition Examination Survey (NHANES), Mobile Examination Center (MEC) Laboratory Procedures Manual.

Eskenazi, B., Chevrier, J., Rosas, L. G., Anderson, H. A., Bornman, M. S., Bouwman, H., Chen, A., Cohn, B. A., de Jager, C., Henshel, D. S., Leipzig, F., Leipzig, J. S., Lorenz, E. C., Snedeker, S. M., & Stapleton, D. (2009). The Pine River Statement: Human health consequences of DDT Use. Environmental Health Perspective, 117, 1359-1367.

Eubig, P. A., Aguiar, A., & Schantz, S. L. (2010). Lead and PCBs as risk factors for attention deficit/hyperactivity disorder. Environmental Health Perspectives, 118(12), 1654-1667.

Sjodin, A., Jones, R. S., Lapeza, C. R., Focant, J. F., McGahee, E. E., & Patterson, D. G., Jr. (2004). Semiautomated high-throughput extraction and cleanup method for the measurement of polybrominated diphenyl ethers, polybrominated biphenyls, and polychlorinated biphenyls in human serum. Analytical Chemistry, 76, 1921-1927.

Turner, W., DiPietro, E., Lapeza, C., Green, V., Gill, J., & Patterson, D.G., Jr. (1997). A fast universal automated cleanup system for the isotope-dilution high resolution mass spectrometric analysis of PCDDs, PCDFs, coplanar PCBs, PCB congeners, and persistent pesticides from the same serum sample. Organohalogen Compounds, 31, 26-31.

Protocol ID:

240401

Variables:
Export Variables
Variable NameVariable IDVariable DescriptionVersiondbGaP Mapping
Research Domain Information
Measure Name:

Concentrations of Polychlorinated Biphenyls (PCBs) and Persistent Pesticides

Release Date:

January 31, 2017

Definition

Collect and analyze a blood sample to determine the concentrations of several polychlorinated biphenyls (PCBs) and persistent pesticides.

Purpose

This biomarker determines the levels of polychlorinated biphenyls (PCBs) and persistent pesticides in the subject’s blood. Prenatal exposure PCBs may lead to poor cognitive function and attention deficit/hyperactivity disorder (ADHD). Prenatal exposures to persistent pesticides (e.g., DDT) may have adverse neurodevelopmental effects on children. PCB and persistent pesticide exposures are harmful to people of all ages, and this biomarker may be used to measure exposures in children, adolescents, and adults.

Keywords

Pregnancy, polychlorinated biphenyls, PCBs, pesticides, National Children’s Study, NCS, National Health and Nutrition Examination Survey, NHANES, Centers for Disease Control and Prevention, CDC