Abstract
Background: Preanalytical conditions encountered during collection, processing, and storage of biospecimens may influence laboratory results. The National Children’s Study (NCS) is a planned prospective cohort study of 100,000 families to examine the influence of a wide variety of exposures on child health. In developing biospecimen collection, processing, and storage procedures for the NCS, we identified several analytes of different biochemical categories for which it was unclear to what extent deviations from NCS procedures could influence measurement results.
Methods: A pilot study was performed to examine effects of preanalytic sample handling conditions (delays in centrifugation, freezing delays, delays in separation from cells, additive delay, and tube type) on concentrations of eight different analytes. A total of 2825 measurements were made to assess 15 unique combinations of analyte and handling conditions in blood collected from 151 women of childbearing age (≥20 individuals per handling condition).
Results: The majority of analytes were stable under the conditions evaluated. However, levels of plasma interleukin-6 and serum insulin were decreased in response to sample centrifugation delays of up to 5.5 h post-collection (p<0.0001). In addition, delays in freezing centrifuged plasma samples (comparing 24, 48 and 72 h to immediate freezing) resulted in increased levels of adrenocorticotropic hormone (p=0.0014).
Conclusions: Determining stability of proposed analytes in response to preanalytical conditions and handling helps to ensure high-quality specimens for study now and in the future. The results inform development of procedures, plans for measurement of analytes, and interpretation of laboratory results.
We gratefully acknowledge the generosity of our study participants for donating their blood, and the contributions to this project of Bonnie Ferguson, Susan Hassell, Kristen Keating, Ben Laimon, Charlie Lawrence, and Frances Whalen of the Westat NCS Biospecimen Team, the scientists at Quest Diagnostics Nichols Institute, and Elaine Gunter of Specimen Solutions, LLC.
Conflict of interest statement
Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article. Research funding played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
Research funding: This analysis was conducted as part of the National Children’s Study, supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development, and funded, through its appropriation, by the Office of the Director of the National Institutes of Health. Supported by NICHD Contracts HHSN275200503395C, HHSN275201000107U, HHSN275200900010C, and HHSN275201000121U. The manuscript was developed by a Writing Team identified by the National Children’s Study Publications Committee for the purpose of timely sharing of NCS data.
Employment or leadership: None declared.
Honorarium: None declared.
Authors’ declaration: The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the Centers for Disease Control and Prevention, National Children’s Study, National Institutes of Health, or U.S. Department of Health and Human Services.
References
1. Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health. What is the National Children’s Study? The National Children’s Study. Available from: http://www.nationalchildrensstudy.gov. Accessed June 1, 2013.Search in Google Scholar
2. Vaught JB. Blood collection, shipment, processing, and storage. Cancer Epidemiol Biomarkers Prev 2006;15:1582–4.10.1158/1055-9965.EPI-06-0630Search in Google Scholar PubMed
3. Lippi G, Becan-McBride K, Behúlová D, Bowen RA, Church S, Delanghe J, et al. Preanalytical quality improvement: in quality we trust. Clin Chem Lab Med 2013;51:229–41.http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000312497600023&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed
4. Lippi G, Chance JJ, Church S, Dazzi P, Fontana R, Giavarina D, et al. Preanalytical quality improvement: from dream to reality. Clin Chem Lab Med 2011;49:1113–26.http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000292538800004&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed
5. Lippi G, Guidi GC. Risk management in the preanalytical phase of laboratory testing. Clin Chem Lab Med 2007;45:720–7.10.1515/CCLM.2007.167Search in Google Scholar PubMed
6. Holland NT, Pfleger L, Berger E, Ho A, Bastaki M. Molecular epidemiology biomarkers–sample collection and processing considerations. Toxicol Appl Pharmacol 2005;206:261–8.10.1016/j.taap.2004.10.024Search in Google Scholar PubMed
7. Holland NT, Smith MT, Eskenazi B, Bastaki M. Biological sample collection and processing for molecular epidemiological studies. Mutat Res/Rev Mutat Res 2003;543:217–34.10.1016/S1383-5742(02)00090-XSearch in Google Scholar
8. Tworoger SS, Hankinson SE. Collection, processing, and storage of biological samples in epidemiologic studies: sex hormones, carotenoids, inflammatory markers, and proteomics as examples. Cancer Epidemiol Biomarkers Prev 2006;15:1578–81.10.1158/1055-9965.EPI-06-0629Search in Google Scholar PubMed
9. Chaigneau C, Cabioch T, Beaumont K, Betsou F. Serum biobank certification and the establishment of quality controls for biological fluids: examples of serum biomarker stability after temperature variation. Clin Chem Lab Med 2007;45:1390–5.http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000250943800021&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed
10. Zhang DJ, Elswick RK, Miller WG, Bailey JL. Effect of serum-clot contact time on clinical chemistry laboratory results. Clin Chem 1998;44:1325–33.10.1093/clinchem/44.6.1325Search in Google Scholar PubMed
11. Chan AY, Swaminathan R, Cockram CS. Effectiveness of sodium fluoride as a preservative of glucose in blood. Clin Chem 1989;35:315–7.10.1093/clinchem/35.2.315Search in Google Scholar PubMed
12. Narayanan S. The preanalytic phase: an important component of laboratory medicine. Am J Clin Path 2000;113:429–52.10.1309/C0NM-Q7R0-LL2E-B3UYSearch in Google Scholar PubMed
13. Elliott P, Peakman TC. The UK biobank sample handling and storage protocol for the collection, processing and archiving of human blood and urine. Int J Epidemiol 2008;37:234–44.10.1093/ije/dym276http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000254713400004&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed
14. Jackson C, Best N, Elliott P. UK biobank pilot study: stability of haematological and clinical chemistry analytes. Int J Epidemiol 2008;37:i16–22.10.1093/ije/dym280http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000256097500005&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed
15. Centers for Disease Control. Laboratory procedure manual vitamin C. Available from: http://www.cdc.gov/nchs/data/nhanes/nhanes_05_06/vic_d_met.pdf. Accessed January, 2013.Search in Google Scholar
16. Hornung RW, Reed LD. Estimation of average concentration in the presence of nondetectable values. Appl Occup Environ Hyg 1990;5:46–51.10.1080/1047322X.1990.10389587Search in Google Scholar
17. CDC/National Center for Health Statistics. General information for the public files of the 2007–2008 laboratory data. Available from: http://www.cdc.gov/nchs/nhanes/nhanes2007-2008/labdoc_e.htm. Accessed March 15, 2013.Search in Google Scholar
18. Wu A. Tietz: clinical guide to laboratory tests. 4th ed. St. Louis: Saunder Elsevier, 2006.Search in Google Scholar
19. Musselman DL, Miller AH, Porter MR, Manatunga A, Gao F, Penna S, et al. Higher than normal plasma interleukin-6 concentrations in cancer patients with depression: preliminary findings. Am J Psychiatry 2001;158:1252–7.10.1176/appi.ajp.158.8.1252Search in Google Scholar PubMed
20. Flower L, Ahuja RH, Humphries SE, Mohamed-Ali V. Effects of sample handling on the stability of interleukin 6, tumour necrosis factor-α and leptin. Cytokine 2000;12:1712–6.10.1006/cyto.2000.0764Search in Google Scholar PubMed
21. Thavasu PW, Longhurst S, Joel SP, Slevin ML, Balkwill FR. Measuring cytokine levels in blood: importance of anticoagulants, processing, and storage conditions. J Immunol Methods 1992;153:115–24.10.1016/0022-1759(92)90313-ISearch in Google Scholar PubMed
22. Jackman RP, Utter GH, Heitman JW, Hirschkorn DF, Law JP, Gefter N, et al. Effects of blood sample age at time of separation on measured cytokine concentrations in human plasma. Clin Vaccine Immunol 2011;18:318–26.10.1128/CVI.00465-10http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000286653900018&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed
23. Chevenne D, Trivin F, Porquet D. Insulin assays and reference values. Diabetes Metab 1999;25:459–76.Search in Google Scholar PubMed
24. Matthews DR, Hosker JP, Rudenski AS, Naylor BA, Treacher DF, Turner RC. Homeostasis model assessment: insulin resistance and beta-cell function from fasting plasma glucose and insulin concentrations in man. Diabetologia 1985;28:412–9.10.1007/BF00280883Search in Google Scholar PubMed
25. Kempf T, Guba-Quint A, Torgerson J, Magnone MC, Haefliger C, Bobadilla M, et al. Growth differentiation factor 15 predicts future insulin resistance and impaired glucose control in obese nondiabetic individuals: results from the xendos trial. Eur J Endocrinol 2012;167:671–8.10.1530/EJE-12-0466http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000309809400009&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar
26. Kim OY, Paik JK, Lee JY, Lee SH, Lee JH. Follow-ups of metabolic, inflammatory and oxidative stress markers, and brachial-ankle pulse wave velocity in middle-aged subjects without metabolic syndrome. Clin Exp Hypertens 2012;13:382–8.http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000321579000012&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar
27. Telford RD, Cunningham RB, Telford RM, Kerrigan J, Hickman PE, Potter JM, et al. Effects of changes in adiposity and physical activity on preadolescent insulin resistance: the Australian look longitudinal study. PLoS One 2012;7:e47438.10.1371/journal.pone.0047438http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000309809500049&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar
28. Turchiano M, Sweat V, Fierman A, Convit A. Obesity, metabolic syndrome, and insulin resistance in urban high school students of minority race/ethnicity. Arch Pediatr Adolesc Med 2012;166:1030–6.10.1001/archpediatrics.2012.1263http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000310686400006&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar
29. Evans MJ, Livesey JH, Ellis MJ, Yandle TG. Effect of anticoagulants and storage temperatures on stability of plasma and serum hormones. Clin Biochem 2001;34:107–12.10.1016/S0009-9120(01)00196-5Search in Google Scholar PubMed
30. Casati M, Cappellani A, Perlangeli V, Ippolito S, Pittalis S, Romano R, et al. Adrenocorticotropic hormone stability in preanalytical phase depends on temperature and proteolytic enzyme inhibitor. Clin Chem Lab Med 2013;51:E45–7.10.1515/cclm-2012-0431http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000316548000003&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed
31. Ray DW, Gibson S, Crosby SR, Davies D, Davis JR, White A. Elevated levels of adrenocorticotropin (ACTH) precursors in post-adrenalectomy Cushing’s disease and their regulation by glucocorticoids. J Clin Endocrinol Metab 1995;80:2430–6.10.1210/jc.80.8.2430Search in Google Scholar
32. Margolis SA, Ziegler RG, Helzlsouer KJ. Ascorbic and dehydroascorbic acid measurement in human serum and plasma. Am J Clin Nutr 1991;54:1315S–8S.10.1093/ajcn/54.6.1315sSearch in Google Scholar PubMed
33. Key T, Oakes S, Davey G, Moore J, Edmond LM, McLoone UJ, et al. Stability of vitamins A, C, and E, carotenoids, lipids, and testosterone in whole blood stored at 4 degrees c for 6 and 24 hours before separation of serum and plasma. Cancer Epidemiol Biomarkers Prev 1996;5:811–4.Search in Google Scholar
34. Ching SY, Prins AW, Beilby JP. Stability of ascorbic acid in serum and plasma prior to analysis. Ann Clin Biochem 2002;39: 518–20.10.1258/000456302320314566Search in Google Scholar PubMed
35. Karlsen A, Blomhoff R, Gundersen TE. Stability of whole blood and plasma ascorbic acid. Eur J Clin Nutr 2007;61:1233–6.10.1038/sj.ejcn.1602655http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=000249881600013&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=b7bc2757938ac7a7a821505f8243d9f3Search in Google Scholar PubMed
©2013 by Walter de Gruyter Berlin Boston
