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The determination of lipoic acid in human plasma using high-performance liquid chromatography method with elelctrochemical detection
Authors: Bencová Soňa | Žáková Pavla
Year: 2012
Type of publication: ostatní - přednáška nebo poster
Page from-to: nestránkováno
Titles:
Language Name Abstract Keywords
eng The determination of lipoic acid in human plasma using high-performance liquid chromatography method with elelctrochemical detection Lipoic acid is a disulfhydryl-containing compound that consists of 8 carbon atoms and 2 sulfur atoms. It contains one chiral center. This is the body´s own substance that is synthesized in the liver in insignificant concentrations, and therefore must be taken from the diet. It is used in clinical medicine and in experimental models as an important antioxidant. It retains its antioxidant properties in both its forms, oxidized and reduced. It was proved that lipoic acid plays an important role in a therapy of many diseases, which are related to enhanced oxidative stress. The aim of this study is to summarize the avalable knowledge of lipoic acid, with a particular focus on diabetes mellitus and its complications, particularly atherosclerosis. The next point of our study is to try to develope simple and effective method for the determination of lipoic acid in human plasma by using HPLC with electrochemical detection on the principle of coulometry. We succeeded in develop of method that is applicable for the determination of oxidized forms of lipoic acid in the concentrations of exogenous supplementation. Lipoic acid supplementation concentration in human plasma is between 50 nmol/l to 50 µmol/l. A calibration curve was linear in the range 500 – 20 000 nmol/l. For this determination we use chromatographic column with reversed-phase LiChroCart® 254-4, Purospher® (RP-C18, 5 µm), and a mobile phase consisting of 65% 20 mM NaH2PO4 and 35 % acetonitrile (pH 2,7). Hexane is used as extraction agent. We are currently working on a method by witch we were able to simultaneous determination of oxidized and reduced forms of lipoic acid. This process is complicated and consists of several sub-steps in difficult sample preparation. Now we are exploring the possibility of extraction with dichlormethane. lipoic acid, high-performance liquid chromatography, electrochemical detection

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