Influence of centrifugation conditions on the results of 77 routine clinical chemistry analytes using standard vacuum blood collection tubes and the new BD-Barricor tubes.
Author(s): Cadamuro J, Mrazek C, Leichtle AB, Kipman U, Felder TK, Wiedemann H, Oberkofler H, Fiedler GM, Haschke-Becher E
Publication: Biochem Med (Zagreb), 2018, Vol. 28, Page 010704
PubMed ID: 29187797 PubMed Review Paper? No
Purpose of Paper
This paper investigated the effects of collection tube type and centrifugation speed on levels of 60 clinical chemistry analytes in plasma and 21 clinical chemistry analytes in serum.
Conclusion of Paper
Levels of lactate dehydrogenase (LD), bicarbonate, and hemoglobin differed between plasma collected in Gel and Barricor tubes. Free hemoglobin levels in serum were significantly affected by centrifugation protocol, but an effect of centrifugation speed on plasma levels was only significant for specimens collected in gel tubes. The remaining analytes examined were not significantly affected by tube type or centrifugation speed and were below the analytical range for the majority of specimens.
Studies
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Study Purpose
The purpose of this study was to determine the effects of collection tube type and centrifugation speed on levels of 60 clinical chemistry analytes in plasma and 21 clinical chemistry analytes in serum. Blood was drawn using a 21-gauge needle from 41 healthy volunteers into three Vacutainer lithium-heparin gel tubes, three serum gel tubes, and two Vacutainer Barricor lithium heparin tubes with a mechanical separator. Tube order for collection was randomized. Blood for serum was stored at room temperature for 30 min before centrifugation while plasma tubes were centrifuged immediately. Differences in centrifugation protocols were examined in serum and plasma obtained by centrifugation at 2000 x g for 10 min (all tube types), 3000 x g for 7 min (serum and lithium heparin gel tubes only), or 3000 x g for 5 min (all tube types). A total of 60 clinical chemistry analytes were analyzed in the resultant plasma and 21 in serum using a Roche COBAS 8000 device. Values were compared to two different commercial controls.
Summary of Findings:
A significant difference in LD levels in plasma was noted between the protocols (P<0.001), but further analysis revealed the difference to be attributed to higher levels in gel tubes than Barricor tubes (P=<0.001 to 0.003) with no significant effect of centrifugation protocol in either tube type. Similarly, a small but significant difference in bicarbonate levels among specimens was attributable to higher levels in Barricor than gel tubes (P=0.008-0.049) and was not affected by centrifugation speed. Free hemoglobin levels in plasma were higher in gel tubes than Barricor tubes when centrifuged at 3000 x g but significance was not reached when both specimens were centrifuged at 2000 x g for 10 min. Free hemoglobin in plasma was higher when gel tubes were centrifuged at 3000 x g for 7 min rather than 2000 x g for 10 min (P=0.008). Similarly, free hemoglobin levels in serum were significantly higher in specimens centrifuged at 3000 x g for 5 or 7 min than 2000 x g for 10 min (P=0.020 and P<0.001, respectively) and in specimens centrifuged at 3000 x g for 7 min rather than 5 min (P=0.030). Centrifugation protocol and tube type did not significantly affect levels of chloride, potassium, sodium, urea, creatinine, creatinine estimated glomerular filtration rate (GFR), calcium, total protein, glucose, c-reactive protein (CRP), high sensitive c-reactive protein (CRP), uric acid, phosphorus, magnesium, total bilirubin, direct bilirubin, cholesterol, triglycerides, HDL-cholesterol, Apo-ipoprotein B, lipoprotein (a), aspartate transaminase, alanine transaminase, alkaline phosphatase, gamma-glutamyl-transferase(GGT), cholinesterase, glutamate dehydrogenase, amylase, lipase, creatinine kinase, iron, transferrin, soluble transferrin, ferritin, fructosamine, icterus index, lipemia index, creatinine kinase MB immunologic, myoglobin, N-terminal pro brain natriuretic peptide (NT-pro BNP), thyroid stimulating hormone (TSH), free triiodothyronine (T3), albumin, complement C3, complement C4, alpha 1-antitrypsin, haptoglobin, ceruloplasmin, interleukin 6 in plasma nor did to affect levels of icterus index, lipemia index, anti-Mullerian hormone, prealbumin, carcinoembryonic antigen, alpha fetoprotein, cancer antigen (CA) 125, CA 19-9, CA 15-3, S100 protein, neuron specific enolase, CA 72.-4, cystatin C, Cystatin C EGFR, soluble fms-like tyrosine kinase-1, placental growth factor, or sFlt1/PIGF ratio in serum. However, some specimens were excluded from the analysis of C-reactive protein (CRP), lipoprotein a (Lp(a)), N-terminal pro-brain natriuretic peptide (NT-proBNP), glutamate-dehydrogenase (GLDH), anti-streptolysin O (ASL), anti-Müller hormone (AMH), carcinoembryonic antigen (CEA), and alpha-fetoprotein (AFP) as levels were below the analytical range for the assay. As levels of high-sensitive Troponin T (hsTnT), pregnancy-associated plasma protein A (PAPP-A), ß-human chorionic gonadotropin (ßHCG), and rheumatoid factor (RF) were below the limit of detection in more than 50% of specimens and were excluded from the analysis.
Biospecimens
Preservative Types
- None (Fresh)
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Electrolyte/Metal Clinical chemistry/auto analyzer Carbohydrate Clinical chemistry/auto analyzer Small molecule Clinical chemistry/auto analyzer Peptide Clinical chemistry/auto analyzer Lipid Clinical chemistry/auto analyzer Steroid Clinical chemistry/auto analyzer Glycoprotein Clinical chemistry/auto analyzer Protein Clinical chemistry/auto analyzer Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Aliquots and Components Centrifugation Multiple speeds compared
Multiple durations compared
Biospecimen Acquisition Type of collection container/solution Lithium-heparin Gel tube
Lithium-heparin Barricor tube