It is important for separation and determination of plant growth regulators in the research of plant physiology. A study of the separation of five plant growth regulators, such as abscisic acid (ABA), gibberellic acid (GA), indole-butyric acid (IBA), indole acetic acid (IAA) and naphthaleneacetic (NAA) by micellar electrokinetic chromatography (MEKC) is presented. The effects of pH, SDS concentration, applied voltage and organic additives on migration behavior were investigated. Results showed that satisfactory separation was achieved on optimization of electrophoretic condition: 80 mmol/L SDS, 10 mmol/L borate-phosphate, 5% isopropanol, 18 kV applied voltage, pH 8.0. The SDS concentration and organic additives played an important role in separating plant growth regulators, and the applied voltage and pH gave less effect. The detection limit of ABA, GA, IBA, IAA and NAA were 5.0 x 10(-3), 3.0 x 10(-3), 5.8 x 10(-4), 1.5 x 10(-4) and 1.4 x 10(-4) g/L respectively. Increasing of the SDS concentration or organic additives would change separation factor (alpha) and result an improvement of the resolution of difficult separation pairs ABA-GA and IAA-NAA.