As a result of pharmacological investigations of the Ephedra herb, it has been shown that most of the clinical efficacy is exerted by its alkaloidal components, which include (-)-ephedrine (E), (+)-pseudoephedrine (PE), (-)-methylephedrine (ME), (+)-methylpseudoephedrine (MPE), (-)-norephedrine (NE) and (+)-norpseudoephedrine (NPE). Several methods have been reported for the simultaneous determination of some of these six alkaloids: i.e. salting out chromatography, thin-layer chromatography, gas-liquid chromatography and high-performance liquid chromatography. However, the separation of all these alkaloids by these methods is unknown. Furthermore, in the case of the analysis of the ephedrine alkaloids in the crude drug, Ephedra herb, the known methods require tedious pretreatments of Ephedra extracts before analysis, leading to possible sources of error. Isotachophoresis (ITP) has already been successfully applied to the analysis of ionic components contaminated with other components, and which require simple pretreatments of samples before analysis because no absorbents are employed in the separation tubes. We have thus examined the utility of ITP for the simultaneous determination of all the six ephedrine alkaloids. This paper describes the development of a suitable method for the analysis of the ephedrine alkaloids and its application to analysis of the crude drug, Ephedra herb.