Simultaneous analysis of the majority of low-molecular-weight, redox-active compounds from mitochondria
Simultaneous analysis of the majority of low-molecular-weight, redox-active compounds from mitochondria
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DOI：
10.1006/abio.1998.2831
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发表时间：
1998-10-01
影响因子：
2.9
通讯作者：
Matson, WR
中科院分区：
文献类型：
--
作者：
Kristal, BS;Vigneau-Callahan, KE;Matson, WR
氧化应激和线粒体功能障碍之间相互作用的研究因分析限制而变得复杂，特别是需要在相对较小的样本中评估多个参数。我们通过开发一种方法来解决这个问题，该方法使用 HPLC 分离和库仑阵列检测来同时分析线粒体中的大多数低分子量氧化还原活性化合物。所描述的方法也应适用于其他亚细胞器以及完整细胞和组织中氧化还原活性化合物的研究。所述协议能够同时测量抗氧化剂（例如，生育酚、抗坏血酸、硫辛酸、尿酸和谷胱甘肽）、氧化应激标记物（例如，o-酪氨酸、m-酪氨酸、硝基酪氨酸、二酪氨酸、谷胱甘肽二硫化物和8-羟基脱氧鸟苷）以及其他代谢物（例如，嘌呤和吲哚），总共约。可检测 600 种氧化还原活性化合物，大多数检测限类似于柱上 5 pg。对对照大鼠和糖尿病大鼠的肝线粒体进行的研究结果（包括分析参数）旨在证明该方法的实用性，(C) 1998 年学术出版社。
Studies of the interaction between oxidative stress and mitochondrial dysfunction are complicated by analytical limitations, especially the need to assess multiple parameters in relatively small samples. We have addressed this problem by developing a methodology for the simultaneous analysis of the majority of low-molecular-weight, redox-active compounds from mitochondria using HPLC separations followed by coulometric array detection, The method described should also be applicable for the study of redox-active compounds in other subcellular organelles as well as in intact cells and tissues. The protocol described enables simultaneous measurement of antioxidants (e.g., tocopherols, ascorbate, lipoates, uric acid, and glutathione), markers of oxidative stress (e.g., o-tyrosine, m-tyrosine, nitrotyrosine, dityrosine, glutathione disulfide, and 8-hydroxydeoxyguanosine) as well as other metabolites (e.g,, purines and indoles), In all, ca. 600 redox active compounds can be detected, most with a limit of detection of similar to 5 pg on column. Results, including analytical parameters, from a study of liver mitochondria from control and diabetic rats are presented to demonstrate utility of this methodology, (C) 1998 Academic Press.