Electromembrane extraction coupled with liquid chromatography–tandem mass spectrometry (LC–MS/MS) was developed for determination of ten volatile organic compound metabolites in dried urine spot samples. The dried urine spot approach is a convenient and economical sampling method, wherein urine is spotted onto a filter paper and dried. This method requires only a small amount of sample, but the analysis sometimes suffers from low sensitivity, which can lead to analytical problems in the detection of minor components in samples. The newly developed dried urine spot analysis using electromembrane extraction exhibited improved sensitivity and extraction, and enrichment of the sample was rapidly achieved in one step by applying an electric field. Aliquots of urine were spotted onto Bond Elut DMS cards and dried at room temperature. After drying, the punched out dried urine spot was eluted with water. Volatile organic compound metabolites were extracted from the sample through a supported liquid membrane into an alkaline acceptor solution inside the lumen of a hollow fiber with the help of an electric potential. The optimum extraction conditions were determined by using design of experiments (fractional factorial design and response surface methodology). Satisfactory sensitivity was achieved and the limits of quantification (LOQ) obtained were lower than the regulatory threshold limits. The method was validated by assessing the linearity, precision, accuracy, recovery, reproducibility, stability, and matrix effects. The results were acceptable, and the developed method was successfully applied to biological exposure monitoring of volatile organic compound metabolites in fifty human urine samples.