Bevital AS

Pre-analytical stability

Stability varies and is an important source of preanalytical error. We or others have assessed stability of most biomarkers, at room temperature, in frozen samples, during transportation in cold container, and in various matrices like serum and plasmas. The results are summarized in qualitative terms in the table below.

Stability

Most biomarkers protected from light are stable for years at temperature ≤ -80 °C. In general, biomarkers that are stable under certain non-optimal conditions are often relatively stable under other conditions. Likewise, biomarker instability is also a feature demonstrated under a variety of conditions. But there are notable exceptions to this general rule. For example, many biomarkers are most stable in EDTA plasma, but this is not the case for 5-methyltetrahydrofolate, which degrades faster in EDTA plasma than in serum. Methionine is rather stable, but is occasionally oxidized to methionine sulfoxide in samples frozen at -25 °C. The table below summarizes stability of biomarkers.

Stability curves for platforms B – H

This page provides data on the stability during storage and freezing/thawing of most biomarkers or metabolites included in the repertoire of BEVITAL by April 2019.

Biomarker profile may reflect non-optimal sample handling or storage

The biomarker profile of a serum/plasma sample set gives some indications of how the samples have been handled or stored. Delayed separation of serum/plasma may cause changes in concentrations of metabolites explained by cellular export (homocysteine), uptake (PL) or metabolism (arginine to ornithine conversion). Improper handling or storage of serum/plasma samples may cause degradation, oxidation, metabolism including formation from precursor(s) (choline) of numerous metabolites. Changes in selected metabolites in isolated serum/plasma are indicated in the figure to the right.

High median total homocysteine (>> 15 µmol/L) in combination with adequate circulating folate or cobalamin suggest that whole blood has been left at room temperature for hours before centrifugation to isolate the serum/plasma fraction. Delayed separation also causes an arginine to ornithine conversion giving an Arg/(Arg+Orn) ratio less than 0.5. Low folate and PLP in combination with low-normal total homocysteine suggest vitamin degradation rather than low folate status. Serum/plasma samples that have been exposed to room temperature or repeated freezing-thawing cycles have high (> 20%) 4-alfa-hydroxy-5-methyltetrahydrofolate relative to (micriobiological active) folate, high methionine sulfoxide (> 20%) relative to methionine and/or low 3-hydroxykynurenine and 3-hydroxyanthranilic acid in combination with high anthranilic acid. Sample left at room temperature also have high free choline with median values in the range 15 – 100 µmol/L.

Publications on preanalytical stability of biomarkers

BiomarkerStability (Change)CommentsPubl
Acetateli_2020_a_145_2692.pdf
Acetylcarnitineliu_2008_rcms_22_3434.pdf
anton_2015_po_0121495.pdf
ADMAhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Acetoacetatecuster_1983_ajcp_80_375.pdf
mcneil_2014_cca_433_278.pdf
Alaninekamlage_2014_60_399.pdf
Amyloid AResults of adequate quality available for only equine SAA. Probably stable.hillstrom_2010_avs_52_8.pdf
Anthranilic acidhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
ArginineRelatively stable in isolated serum or plasma, but decreases in (chilled) whole blood.hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
AspargineModerate decay due to conversion to aspartic acid in improperly stored serum/plasma samples, leading to an increase in the Asp/Asn ratio.
Aspartic acidIncrease due to formation from aspargine in improperly stored serum/plasma samples, leading to an increase in the Asp/Asn ratio.
Betainehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Butyrateli_2020_a_145_2692.pdf
CalprotectinCalprotectin (Calp) is several-fold higher in serum than EDTA-plasma, possibly related to rapid release before and during centrifugation of Calp from activated neutrophils and leukocytes containing large amounts of Calp. After removal of blood cells, Calp is rather stable. EDTA-plasma is recommended for epidemiological/clinical studies.
Carboxyethyllysinehull_2013_jcb_53_129.pdf
Carboxymethyllysinehull_2013_jcb_53_129.pdf
Carnitinesowell_2011_jcs_49_463.pdf
anton_2015_po_0121495.pdf
Choline, freePhosphatidylcholine is converted to free choline; the conversion is inhibited by ETDA.hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
holm_2003_cc_49.286.pdf
Choline, total
Citratemalm_2018_bb_14_416.pdf

BiomarkerStability (Change)CommentsPubl
Citrullinedemacker_2009_jc_877_387.pdf
jones_2014_abc_406_4663.pdf
Cobalaminhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Cotininebernert_1997_cc_43_2281.pdf
midttun_2014_jn_144_784.pdf
Creatinecarling_2008_acb_45_575.pdf
Creatininehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
CRP (hsCRP)tanner_2008_acb_45_375.pdf
doumatey_2014_cb_47_315.pdf
Cystathioninehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Cystatin Cprice_2000_cca_297_55.pdf
Cysteine, totalhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Dimethylglycinehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Flavin mononucleotidehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Folate5-Methyltetrahydrofolate degrades fast in EDTA plasma, but is relatively stable in chilled whole blood.hannisdal_2010_jn_140_522.pdf
hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Formateli_2020_a_145_2692.pdf
Glutamic acidMarked increase due formation from glutamine in improperly stored serum/plasma samples, leading to a marked increase in the Glu/Gln ratio.kamlage_2014_60_399.pdf
GlutamineModerate decay due conversion to glutamic acid in improperly stored serum/plasma samples, leading to a marked increase in the Glu/Gln ratio.kamlage_2014_60_399.pdf
Glycinehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
HbA1coddozo_2012_cb_45_464.pdf
liotta_2013_abc_405_429.pdf
Hexanoylcarnitineanton_2015_po_0121495.pdf

BiomarkerStability (Change)CommentsPubl
Histidinedejonge_1996_jc_677_61.pdf
Homoargininemidttun_2014_jn_144_784.pdf
Homocysteine, totalStable in isolated serum or plasma, but is increased in whole blood, because of export of homocysteine from intact cells.Fiskerstrand_93_CC_39_263.pdf
hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
3-Hydroxyanthranilic acidhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
2-Hydroxybutyratemiyazaki_2015_sp_4_494.pdf
3-Hydroxybutyratemiyazaki_2015_sp_4_494.pdf
custer_1983_ajcp_80_375.pdf
α-Hydroxyglutaric acidfernandez-galan_2018_jc_1083_28.pdf
3-Hydroxyisobutyratemiyazaki_2015_sp_4_494.pdf
custer_1983_ajcp_80_375.pdf
3-Hydroxykynureninehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Isobutyrateli_2020_a_145_2692.pdf
IsocitrateRelatively stable, but show a small increase, particularly in whole blood.kamlage_2018_m_8_6pdf
Isovalerateli_2020_a_145_2692.pdf
Isovalerylcarnitineanton_2015_po_0121495.pdf
Kynurenic acidhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Kynureninehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
LactateRelatively stable, but show an increase in whole blood.kamlage_2018_m_8_6pdf
MalateRelatively stable, but show a small increase in whole blood.kamlage_2018_m_8_6pdf
malm_2018_bb_14_416.pdf
MethionineRelatively stable, but oxidized to methionine sulfoxide after storage for years at –25 oC.hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Methionine sulfoxidehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
α-Methylbutyrateli_2020_a_145_2692.pdf
N1-Methylnicotinamideszafarz_2010_jc_878_895.pdf
Methylmalonic acidhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Myristoylcarnitineanton_2015_po_0121495.pdf
BiomarkerStability (Change)CommentsPubl
Neopterin, totalNeopterin plus 7,8-dihydroneopterin are measured. The latter is converted to 7,8-dihydroxanthopterin during non-optimal sample handling.midttun_2014_jn_144_784.pdf
Nicotinamidepfuhl_2005_jpba_36_1045.pdf
szafarz_2010_jc_878_895.pdf
Nicotinic acidpfuhl_2005_jpba_36_1045.pdf
szafarz_2010_jc_878_895.pdf
Octanoylcarnitineanton_2015_po_0121495.pdf
OrnithineRelatively stable in isolated serum or plasma, but increases in whole blood, leading to an increased Orn/Arg ratio.kamlage_2014_60_399.pdf
Palmitoylcarnitineanton_2015_po_0121495.pdf
liu_2008_rcms_22_3434.pdf
Picolinic acidwhiley_2019_ac_91_5207.pdf
Propionateli_2020_a_145_2692.pdf
Propionylcarnitineanton_2015_po_0121495.pdf
PyridoxalIncrease in pyridoxal is not observed in chilled whole blood, possibly because of cellullar uptake.hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Pyridoxal 5′-phosphateConverted to pyridoxal; the conversion is inhibited by EDTA. Relatively stable in chilled whole blood.hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
4-Pyridoxic acidhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
PyruvateRelatively stable in serum, but shows decrease in whole blood.kamlage_2014_60_399.pdf
Quinaldic acidProbably stable.
Quinolinic acidAssumed to be stable.whiley_2019_ac_91_5207.pdf
RiboflavinFMN is converted to riboflavin; the conversion is inhibited by EDTA.hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Sarcosinehustad_2012_cc_58_402.pdf
SDMAhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
BiomarkerStability (Change)CommentsPubl
Serinehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Stearoylcarnitineanton_2015_po_0121495.pdf
Succinatemalm_2018_bb_14_416.pdf
ThiamineProbably stable.liu_2008_c_67_583.pdf
Total cysteinehustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Total homocysteineStable in isolated serum or plasma, but is increased in whole blood, because of export of homocysteine from intact cells.Fiskerstrand_93_CC_39_263.pdf
hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Trimethylamine-N-oxideStable.wang_2014_ab_455_35.pdf
Trimethyllysinemidttun_2014_jn_144_784.pdf
Tryptophanhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Valerateli_2020_a_145_2692.pdf
Vitamin AStable in whole blood and serum.midttun_2014_jn_144_784.pdf
albahrani_2016_cclm_%2054_1609.pdf
Vitamin B95-Methyltetrahydrofolate degrades fast in EDTA plasma, but is relatively stable in chilled whole bloodhannisdal_2010_jn_140_522.pdf
hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
Vitamin B12hustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
25-OH vitamin D3Stable in whole blood and serum.midttun_2014_jn_144_784.pdf
albahrani_2016_cclm_%2054_1609.pdf
Vitamin EStable in whole blood and serum.midttun_2014_jn_144_784.pdf
albahrani_2016_cclm_%2054_1609.pdf
Vitamin KStable.riphagen_2016_cclm_54_1201.pdf
Xanthurenic acidhustad_2012_cc_58_402.pdf
midttun_2014_jn_144_784.pdf
April 22, 2022

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