Planned variation in preanalytical conditions to evaluate biospecimen stability in the National Children's Study (NCS).

Author(s): Mechanic L, Mendez A, Merrill L, Rogers J, Layton M, Todd D, Varanasi A, O'Brien B, Meyer Iii WA, Zhang M, Schleicher RL, Moye J

Publication: Clin Chem Lab Med, 2013, Vol. 51, Page 2287-94

PubMed ID: 23924524 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of delayed centrifugation of blood, refrigerating blood after centrifugation, and type of plasma separation tube on levels of 9 clinical chemistry analytes in plasma and serum. Non-fasting blood was collected through a butterfly needle into 8.5 mL SST tubes, 5 mL PPT tubes with K2EDTA, 8.5 mL P100 tubes containing K2EDTA and protein stabilizers, 5.0 plain serum tubes, or 5.0 mL K2EDTA tubes. For one experiment, blood specimens in SST (IGF-1, insulin, %CDT, and free T4), PPT (IL-6) and P100 (ACTH) tubes were stored at room temperature and centrifuged 30 min, 1.5 h, 3.5 h, or 5.5 h after collection. 0.3, 0.5 or 1 mL aliquots of serum and plasma were frozen immediately. In a second experiment, blood specimens in SST (IGF-1, insulin, %CDT, and free T4), P100 (ACTH), and PPT (IL-6) tubes were centrifuged after 2 h at room temperature and then the serum was refrigerated in the tube for 0, 24, 48, or 72 before aliquoting and freezing. Still another group of blood specimens in plain serum tubes was centrifuged after 2 h at 2-8 degrees C and stored for 0, 24, 48 or 72 h at [4 degrees C]. Some of these specimens were recentrifuged, and some were not, and then 0.5 mL aliquots were frozen with 2% MPA (vitamin C) or without 2% MPA (estradiol). Blood was collected in P100, PPT and K2EDTA tubes to determine the effects of tube type on ACTH levels.

   Summary of Findings:

   Delayed centrifugation led to decreased levels of IL-6 and insulin (p<0.001) but did not affect levels of ACTH, free T4, %CDT, or IGF-1. When specimens were centrifuged 2 h after collection and then subjected to refrigerated storage in the original tube before freezing, levels of ACTH increased (p=0.0014), but levels of IL-6, insulin, free T4, %CDT and IGF-1 were unaffected. Refrigerated storage in the original tube, after centrifugation but prior to freezing, had no effects on levels of estradiol or vitamin C in serum. Further, tube type had no effect on the levels of ACTH in plasma.

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Serum
   • Bodily Fluid - Plasma

   Preservative Types

   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   Protein	Immunoassay
   Protein	ELISA
   Peptide	Radioimmunoassay
   Peptide	Immunoassay
   Small molecule	HPLC
   Glycoprotein	Clinical chemistry/auto analyzer
   Steroid	ELISA

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Time at room temperature	30 min 1.5 h 2.0 h 3.5 h 5.5 h
   Storage	Storage duration	0 h 24 h 48 h 72 h
   Biospecimen Acquisition	Type of collection container/solution	K2EDTA vacutainer PPT P100
   Biospecimen Aliquots and Components	Centrifugation	Centrifugation delays investigated

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