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验证用于测量猪唾液中氧化剂化合物的分析方法:硫代巴比妥酸反应物质(TBARS)、羰基和活性氧物种(ROS)。

Validation of assays for measurement of oxidant compounds in saliva of pigs: Thiobarbituric acid reactive substances (TBARS), carbonyl, and reactive oxygen species (ROS).

机构信息

Interlab-UMU, Regional Campus of International Excellence "Mare Nostrum" University of Murcia, Murcia 30100, Spain.

Deparment of Biochemistry, Faculty of Medicine, Ankara Yildrim Bezayit University, Ankara City Hospital, Ankara 06800, Turkey.

出版信息

Res Vet Sci. 2023 Dec;165:105069. doi: 10.1016/j.rvsc.2023.105069. Epub 2023 Oct 30.

Abstract

Thiobarbituric acid reactive substances (TBARS), carbonyls and reactive oxygen species (ROS) are oxidant compounds that can provide useful information on the oxidative status. Pigs can be affected by oxidative stress in different situations including physiological conditions such as lactation and also in different diseases, and the measurement of these three analytes in saliva could be potentially useful as biomarkers of the redox status in this species. Assays for the measurement of TBARS and carbonyls by spectrophotometry and ROS by luminol-based chemiluminescence in pigs' saliva were analytically validated and were applied in saliva of pigs after an in vitro incubation with different doses of ascorbic acid (AA). All the assays showed a satisfactory analytical precision and accuracy. The 240 h-incubation of saliva samples with 60 mM of AA induced to an increased TBARS and carbonyls production. TBARS, carbonyls and ROS can be estimated in saliva of pigs by the assays validated in this report. In addition, these assays can detect changes in the concentration of these analytes associated to incubation of saliva samples with AA.

摘要

硫代巴比妥酸反应物质(TBARS)、羰基和活性氧(ROS)是氧化剂化合物,可提供有关氧化状态的有用信息。猪在不同情况下,包括泌乳等生理状态,以及在不同疾病中,都可能受到氧化应激的影响,因此唾液中这三种分析物的测量可能是该物种氧化还原状态的潜在生物标志物。通过分光光度法测量唾液中的 TBARS 和羰基,以及通过鲁米诺化学发光法测量 ROS 的分析在猪唾液中得到了验证,并应用于唾液在体外与不同剂量抗坏血酸(AA)孵育后的情况。所有的分析方法都显示出令人满意的分析精密度和准确性。唾液样本在 60mM AA 下孵育 240 小时后,TBARS 和羰基的产生增加。本报告中验证的分析方法可用于检测猪唾液中的 TBARS、羰基和 ROS。此外,这些分析方法可以检测与 AA 孵育相关的唾液样本中这些分析物浓度的变化。

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