Evaluation of an automated spectrophotometric assay for reactive oxygen metabolites in serum
Evaluation of an automated spectrophotometric assay for reactive oxygen metabolites in serum
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DOI：
10.1515/cclm.2002.115
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发表时间：
2002-01-01
影响因子：
6.8
通讯作者：
Vernocchi, A
中科院分区：
文献类型：
--
作者：
Iamele, L;Fiocchi, R;Vernocchi, A
自由基浓度的体内评估受到其不稳定性和极短半衰期的阻碍。Diacron活性氧代谢（DROM）试验是最近引入的评价有机化合物过氧化的方法。由于手动执行检测会导致分析不精密度过大，因此本研究旨在评价该检测的自动化。根据NCCLS提出的指导原则评估运行内和运行间不精密度和干扰。在不同的物理条件下，活性氧代谢产物（ROM）的稳定性进行了评价。对于批内和批间不精密度，变异系数始终低于5%。甘油三酯、血红蛋白和胆红素的最大允许浓度分别为28.2 mmol/l、0.068 mmol/l和171 mmol/l。血清在20 ℃下储存可提供足够的ROM稳定性长达3个月，而在4 ℃下储存则产生不可重现的结果。总之，我们的数据提供了证据，证明DROM测定具有可接受的稳定性和足够的不精密度。该自动化检测方法可作为一种快速、重复性好的氧化应激定量评价方法。由于它很容易进行，该方法是适合常规的临床实验室，并可以提供一个准确的估计体内氧化应激。
The in vivo assessment of free radicals concentration is hampered by their instability and extremely short halflife. The Diacron Reactive Oxygen Metabolites (DROM) test is a recently introduced method to evaluate the peroxidation of organic compounds. Since the manual performance of the test provides excessive analytical imprecision, the aim of this study was to evaluate the automation of this test. Within and betweenrun imprecision and interference were assessed according to the guidelines proposed by the NCCLS. The reactive oxygen metabolites (ROM) stability was evaluated in different physical conditions. For withinrun and betweenrun imprecision the coefficients of variation were consistently lower than 5%. The maximum allowable concentration was 28.2 mmol/l, 0.068mmol/l and 171mmol/l for triglycerides, haemoglobin and bilirubin, respectively. Serum storage at 20degreesC provided adequate ROM stability for up to 3 months, whereas storage at 4degreesC yielded nonreproducible results. In conclusion, our data provide evidence that the DROM assay has both an acceptable stability and an adequate imprecision. The automated assay may be regarded as a fast and reproducible method for the quantitative evaluation of oxidative stress. Since it is easily performed, the method is suitable for routine in clinical laboratories and may provide an accurate estimation of oxidative stress in vivo.