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Electroreduction and adsorption behavior of pyruvic acid were studied in the presence of cetyltrimethylammonium bromide by using cyclic voltammetry, differential pulse adsorptive stripping voltammetry and square-wave adsorptive stripping voltammetry at HMDE. The reduction peak current increases in the presence of CTAB. These fully validated sensitive and reproducible adsorptive stripping voltammetric techniques were applied for the trace determination of the pyruvic acid in biological samples. Pyruvic acid shows a single irreversible reduction peak at −1.35 V in ammonia buffer of pH 8.2 ± 0.01. Different experimental conditions were examined by using differential pulse adsorptive stripping voltammetry and square wave adsorptive stripping voltammetry. These electroanalytical procedures enabled to determine pyruvic acid in the concentration range 0.004–0.036 mM for both DPAdSV and SWAdSV. The detection and quantification limits were found to be 6.12 × 10−6 and 2.0 × 10−5 mM for DPAdSV and 1.12 × 10−7 and 4.4 × 10−6 mM for SWAdSV, respectively.