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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.

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.

Protocol:

Administer the following questions prior to blood collection.

1. Do you have hemophilia or any bleeding disorder?

[ ] Yes

[ ] No

[ ] Dont Know

[ ] Refused

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

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

[ ] Yes

[ ] No

[ ] Dont Know

[ ] Refused

      • If the participant answers "Yes" or "Dont 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

[ ] Dont Know

[ ] Refused

  • If the participant answers "Yes," go to question 4.
  • If the participant answers "No," "Dont 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

[ ] Dont Know

[ ] Refused

  • Record the participants 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

[ ] Dont 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

[ ] Dont Know

[ ] Refused

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

[ ] Yes

[ ] No

[ ] Dont Know

[ ] Refused

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

[ ] Yes

[ ] No

[ ] Dont Know

[ ] Refused

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

[ ] Yes

[ ] No

[ ] Dont 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 participants 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 participants blood.
  • If the participant answers "No" and is not pregnant, prepare to draw the participants blood.

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

[ ] Yes

[ ] No

[ ] Dont Know

[ ] Refused

      • Record the participants response and prepare to draw the participants 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 participants 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 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

Selection Rationale

The National Childrens 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
Derived Variables

None

Process and Review

Not applicable.

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 Name Variable IDVariable DescriptiondbGaP Mapping
PX240401_PCB_Pesticide_Alcohol
PX240401070000 Have you had alcohol such as beer, wine, or more
liquor in the last 8 hours? show less
N/A
PX240401_PCB_Pesticide_Antacid
PX240401090000 Have you used antacid, laxatives, or more
anti-diarrheal medications in the last 8 hours? show less
N/A
PX240401_PCB_Pesticide_Blood_Draw
PX240401030000 Have you had any problems with a blood draw more
in the past? show less
N/A
PX240401_PCB_Pesticide_Blood_Draw_Problems
PX240401040000 What problems have you had with a blood draw more
in the past? show less
N/A
PX240401_PCB_Pesticide_Chemotherapy
PX240401020000 Have you had cancer chemotherapy within the more
past 4 weeks? show less
N/A
PX240401_PCB_Pesticide_Concentrations
PX240401130000 Serum Polychlorinated Biphenyl Persistent more
Pesticide Concentration in Nanogram Per Gram Lipid Weight show less
N/A
PX240401_PCB_Pesticide_Diabetes
PX240401110000 Has a doctor ever told you that you had diabetes? N/A
PX240401_PCB_Pesticide_Dietary_Supplement
PX240401100000 Have you taken a dietary supplement such as more
vitamins or minerals in the last 8 hours? show less
N/A
PX240401_PCB_Pesticide_Gum
PX240401080000 Have you chewed gum, or used breath mints, more
lozenges, cough drops, or other cough or cold remedies in the last 8 hours? show less
N/A
PX240401_PCB_Pesticide_Hemophilia
PX240401010000 Do you have hemophilia or any bleeding disorder? N/A
PX240401_PCB_Pesticide_Ingested_Date
PX240401050100 When was the last time you had anything to more
eat or drink other than water? Date show less
N/A
PX240401_PCB_Pesticide_Ingested_Time
PX240401050200 When was the last time you had anything to more
eat or drink other than water? Time show less
N/A
PX240401_PCB_Pesticide_Insulin
PX240401120000 Have you taken any insulin in the last 8 hours? N/A
PX240401_PCB_Pesticide_Sweetener
PX240401060000 Have you had sweetener or milk added to a more
drink, such as coffee or tea, in the last 8 hours? show less
N/A
Pregnancy
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