Stability of antioxidant vitamins in whole human blood during overnight storage at 4°C and frozen storage up to 6 months.
Stability of antioxidant vitamins in whole human blood during overnight storage at 4°C and frozen storage up to 6 months.
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人全血中抗氧化维生素在 4°C 过夜储存和冷冻储存长达 6 个月期间的稳定性。
DOI：
10.1024/0300-9831/a000485
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Traber,MaretG
中科院分区：
文献类型：
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作者：
Leonard,ScottW;Bobe,Gerd;Traber,MaretG
为了确定临床试验中采集血液的最佳条件，样品处理物流可能妨碍红细胞从血浆中及时分离，招募了健康受试者（n= 8,6 M/2F），并将非空腹血液样本收集到含有不同抗凝剂（乙二胺四乙酸（EDTA）， li -肝素或na -肝素）的管中。我们假设肝素，而不是EDTA，可以有效地保护血浆生育酚、抗坏血酸和维生素E分解代谢物（α和γ-CEHC）免受氧化损伤。为了验证这一假设，一组试管立即处理，血浆样品在- 80℃下保存，另一组试管在4℃下保存，第二天早上（~ 30小时）处理并分析，或者在6个月后分析样品。血浆抗坏血酸，用HPLC电化学检测（LC-ECD）测定，在使用EDTA作为抗凝剂过夜后，血浆抗坏血酸下降了75%，但当使用肝素时，血浆抗坏血酸没有变化。加工前的时间和抗凝剂对血浆α-或γ-生育酚或α-或γ-CEHC浓度均无显著影响。用LC- ecd或LC/质谱法测定α和γ-生育酚浓度在- 80℃下保存6个月后保持不变。因此，全血在4℃冷藏过夜不会改变血浆α-或γ-生育酚浓度或其分解产物。当EDTA用作抗凝剂时，抗坏血酸在全血中是不稳定的，但当全血与肝素一起收集时，它可以储存过夜并随后进行处理。
To determine optimal conditions for blood collection during clinical trials, where sample handling logistics might preclude prompt separation of erythrocytes from plasma, healthy subjects (n= 8, 6 M/2F) were recruited and non-fasting blood samples were collected into tubes containing different anticoagulants (ethylenediaminetetra-acetic acid (EDTA), Li-heparin or Na-heparin). We hypothesized that heparin, but not EDTA, would effectively protect plasma tocopherols, ascorbic acid, and vitamin E catabolites (α-and γ-CEHC) from oxidative damage. To test this hypothesis, one set of tubes was processed immediately and plasma samples were stored at− 80 C, while the other set was stored at 4 C and processed the following morning (~ 30 hours) and analyzed, or the samples were analyzed after 6 months of storage. Plasma ascorbic acid, as measured using HPLC with electrochemical detection (LC-ECD) decreased by 75% with overnight storage using EDTA as an anticoagulant, but was unchanged when heparin was used. Neither time prior to processing, nor anticoagulant, had any significant effects upon plasma α-or γ-tocopherols or α-or γ-CEHC concentrations. α-and γ-tocopherol concentrations remained unchanged after 6 months of storage at− 80 C, when measured using either LC-ECD or LC/mass spectrometry. Thus, refrigeration of whole blood at 4 C overnight does not change plasma α-or γ-tocopherol concentrations or their catabolites. Ascorbic acid is unstable in whole blood when EDTA is used as an anticoagulant, but when whole blood is collected with heparin, it can be stored overnight and subsequently processed.