Ephedra (Ma huang) has been used as traditional medicinal herbs and recently found to be effective in weight-loss and used for boosters of energy/performance. The pharmacological effects of Ephedra appear to be attributed to Ephedra alkaloids, mainly ephedrine (EP) and pseudo- ephedrine (PSE). EP is a diastereomer of PSE, known as source for illegal production of methamphetamine (Meth). Also, Ephedra- containing dietary supplements have caused many serious health problems such as heart attack, stroke and sudden death by misuse or abuse. Therefore, analysis of Ephedra herba is an important issue in the fields of Forensics, food and drug safety. The rapid and easy thin-layer chromatographic (TLC) co-spot (in situ) method for the identification of Ephedra (Ma huang) was performed. The liquid-liquid extraction products of Ephedra were identified, compared with standard EP and PSE. The Ephedra extract was reacted with each of these three derivatizing reagents (FMOC-Cl, Dansyl-Cl (DAN-Cl) or Dabsyl-Cl (DAB-Cl)) in situ (co-spot) to generate derivatives in TLC having different identification properties at room temperature and 70 ℃. Derivatives of EP and PSE that are major active ingredients of Ephedra were well separated from each other using the TLC method. This separation allowed easy identification of such derivatives compared to the non-derivatized EP and PSE, which comprises a diastereomeric pair. Then, the detection range of the amount of Ephedra powder was determined and compared, based on the type of reagents used and the reaction temperature for the current derivatization method. When the reagent FMOC-Cl and DNS-Cl were used to identify EP in Ephedra extract (reaction spots were exposed to air at room temperature or covered with slide glass and heated at 70 ℃), less than 1 mg of Ephedra powder was needed, but higher needed Ephedra powder at least 2.5 mg when DAB-Cl was used even when the raction spots were covered and heated at 70 ℃. To demonstrate the validity of the current TLC method to be used for identifying Ephedra at the scene and be useful as a preliminary analysis at the forensic laboratory, further studies are required such as the various changes of the conditions to improve detection limit(LOD).