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Reversed-phase liquid chromatography was applied to separate the selenium species with 0.01M ammonium acetate buffer (pH=4) eluent containing 0.5% (v/v) methanol and 0.1moll-1 of DDAB. The operating conditions for chromatographic separation (gradient elution, run time) and hydride generation (concentration of HCl and NaBH4 solution, argon and hydrogen flow rates) were carefully optimized. The efficiency of hydride generation for all of the four species was measured. Detection limits (DLs) obtained for SeCys, SeMet, SeEt and Se(IV) are 18, 70, 96, and 16μgl-1, respectively. The analytical performance of the method was controlled by measuring the selenium content of spiked selenium food supplements. 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