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Fluorescamine use in high-performance liquid chromatographic determination of aminocaproic acid in serum
Abstract:
A sensitive and specific high-performance liquid chromatographic determination of aminocaproic acid in serum is described. omega-Aminocaprylic acid is used as an internal standard. To 10 microliter of serum, 10 microliter of the internal standard solution and 50 microliter of ethanol are added. After centrifugation, a portion of the supernate is evaporated. The residue is dissolved in 750 microliter of 50 mM dibasic sodium phosphate, and then 250 microliter of fluorescamine in acetonitrile (35 mg/100 ml) is added. The reaction mixture is chromatographed using a column of octadecylsilane bonded to silica and 44% acetonitrile in 0.5 mM phosphoric acid as the eluent. Quantitation is achieved by monitoring either the absorbance of the effluent at 405 nm or the fluorescence of the compounds with a fluorometer equipped with a flowcell. The method is reproducible, simple, and fast and has a precision of 4.4%.