Effect of temperature and time delay in centrifugation on stability of select biomarkers of nutrition and non-communicable diseases in blood samples

Abstract

Introduction Preanalytical conditions are critical for blood sample integrity and poses challenge in surveys involving biochemical measurements. A cross sectional study was conducted to assess the stability of select biomarkers at conditions that mimic field situations in surveys. Material and methods Blood from 420 volunteers was exposed to 2 – 8 °C, room temperature (RT), 22 – 30 °C and > 30 °C for 30 min, 6 hours, 12 hours and 24 hours prior to centrifugation. After different exposures, whole blood (N = 35) was used to assess stability of haemoglobin, HbA1c and erythrocyte folate; serum (N = 35) for assessing stability of ferritin, C-reactive protein (CRP), vitamins B12, A and D, zinc, soluble transferrin receptor (sTfR), total cholesterol, high density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL), tryglicerides, albumin, total protein and creatinine; and plasma (N = 35) was used for glucose. The mean % deviation of the analytes was compared with the total change limit (TCL), computed from analytical and intra-individual imprecision. Values that were within the TCL were deemed to be stable. Result Creatinine (mean % deviation 14.6, TCL 5.9), haemoglobin (16.4%, TCL 4.4) and folate (33.6%, TCL 22.6) were unstable after 12 hours at 22-30°C, a temperature at which other analytes were stable. Creatinine was unstable even at RT for 12 hours (mean % deviation: 10.4). Albumin, CRP, glucose, cholesterol, LDL, triglycerides, vitamins B12 and A, sTfR and HbA1c were stable at all studied conditions. Conclusion All analytes other than creatinine, folate and haemoglobin can be reliably estimated in blood samples exposed to 22-30°C for 12 hours in community-based studies.

Associations of circulating folate, vitamin B12 and homocysteine concentrations in early pregnancy and cord blood with epigenetic gestational age: the Generation R Study

“…It may be that more extreme variations in folate, vitamin B12 and homocysteine blood concentrations are associated with gestational age acceleration. Blood samples were stored at room temperature for up to 3 h, which may have led to time-dependent increases in homocysteine concentrations due to a continuous production and release of homocysteine from blood cells [ 31 , 32 ]. This may have limited our capacity to detect significant associations between homocysteine and all outcomes.…”

Confounders in Identification and Analysis of Inflammatory Biomarkers in Cardiovascular Diseases

“…One research study has found that the high sensitivity and stability of CRP in multiple freeze-thaw cycles of the assay may make it a particularly useful biomarker of CVDs. A delay up to 6 h in specimen processing and storage temperature did not affect levels of CRP [88]. According to one study, the level of TNF-α decreases 50% when there is a delay in plasma sample processing during the separation of plasma from cells [86].…”

“…Temperature and time between collection and performance of analytical assay are critical for sample integrity and stability [87]. However, most of the research studies have been performed by collecting blood samples within the laboratory, but don't show operational conditions of large-scale field surveys because transportation of samples to the analytical laboratory can be a potential variable [88].…”

“…To avoid pre-analytical variations that occur due to differences in sample handling conditions, ideal pre-analytical conditions should be maintained uniformly. Therefore, the stability of biomarkers should be ensured when fluctuating pre-analytical conditions such as temperature and time taken for centrifugation, in particular, are encountered [88].…”

The CRESS checklist for reporting stability studies: on behalf of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for the Preanalytical Phase (WG-PRE)

“…Whether the blood sample was venous, collected via a vascular catheter or another form of sampling should also be included, as well as how and for how long the tubes were mixed. Authors should also provide information on the length of sample storage before the baseline sample was analysed, how long the contact time with cellular components was, and when the sample was separated prior to storage as this can significantly affect sample stability for some analytes [10, [12][13][14]. Additional information including whether the tubes were kept open or closed during the experiment and whether the sample was exposed to light should be recorded and reported in the manuscript.…”