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Stability of Whole Blood Antioxidants During Overnight Storage at 4°C
Author(s) -
Leonard Scott,
Bobe Gerd,
Traber Maret
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.760.1
Subject(s) - chemistry , whole blood , ascorbic acid , ethylenediaminetetraacetic acid , chromatography , vitamin c , heparin , anticoagulant , blood plasma , antioxidant , biochemistry , food science , medicine , chelation , organic chemistry
To evaluate the reproducibility of antioxidant vitamin assays where sample‐handling logistics might preclude prompt separation of erythrocytes (RBC) from plasma, subjects (n=8, 6M/2F) were recruited and non‐fasting blood collected. We hypothesized that samples could be collected using a single anticoagulant that would not interfere with routine assays of plasma and RBC α‐tocopherol (α‐T), ascorbic acid (AA), and vitamin E metabolites (α‐ and γ‐CEHCs). To test this hypothesis, blood samples were collected into tubes containing ethylenediaminetetraacetic acid (EDTA), Li‐ or Na‐heparin, then either were 1) processed immediately and samples frozen (‐80° C), or 2) stored at 4° C, then at 30 h plasma and RBC isolated, then frozen. Plasma (acidified 1:1 with 5% metaphosphoric acid) AA concentrations were highly unstable; time=0, AA (68±14) in EDTA concentrations were lower than in either heparin (Li‐ 115 ±15 vs Na‐ 111 ± 15 μM, ANOVA P=0.018, Tukey P<0.05); in 30 h stored EDTA samples AA decreased by 75%, but heparin AA samples did not. Neither time prior to processing, nor anticoagulant used had any significant effects upon plasma or RBC α‐Ts, or plasma CEHCs, whether measured by HPLC with electrochemical detection or mass spectrometry. Thus, whole blood collected with heparin refrigerated at 4°C overnight is effectively protected for analysis of plasma AA and α‐T, α‐ and γ‐CEHCs and RBC α‐T. Supported by NIH grant R01 DK081761.