Abstract
This chapter discusses the analysis of water-soluble antioxidants by high-performance liquid chromatography (HPLC) with electrochemical detection (HPLC/EC). Water-soluble antioxidants—such as ascorbic acid and glutathione—receive substantial attention because of their potential role in maintaining optimal health. The endogenous level of one or more antioxidants may reflect the health of a particular tissue. Compared to colorimetric or spectrophotometric methods, HPLC/EC provides a relatively fast, quantitative method that is reliable, highly sensitive, specific, and allows no interference from nonrelated substances within the sample and is currently the method of choice. The sensitivity and selectivity of HPLC/EC are appropriate for evaluating small tissue samples. Vitreous or aqueous humor samples are obtained by a needle connected to a syringe. The size of the needle depends on the animal species and whether vitreous humor or aqueous humor is being obtained. The chapter concludes with a discussion of the hydrodynamic voltammetry plot.
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