the application of hair analysis for the detection of drugs in the human body has become an important method, because it can provide evidence of drug use over a much longer period than the conventional testing in blood and urine. Aim:The present work was designed to detect amitriptyline HCl and clozapine in hair of albino rats. Hair samples were examined during drug administration and after stopping. Method: Fifty adult albino rats of both sex were divided into three groups. Each of the first two groups included fifteen rats which were given the therapeutic doses of amitriptyline HCl at a concentration of 3.4 mg/day and clozapine at a concentration of 4.2 mg/day .The third group included twenty rats, which was subdivided into two subgroups, each included ten rats, subgroup (A) which was given distilled water (positive control), and subgroup (B) which was the blank group that lived in the same environmental conditions without being treated with any medications. The drugs were given orally by gavage for three consecutive months. Hair samples were taken on the second day of drug administration then on the fifth day and then weekly till the end of the three months to detect the onset of appearance and the rate of increase of these drugs, and after stopping the drugs, we continued to take hair samples every week to detect the time of drug disappearance. Combination of thin-layer chromatography and color reactions have been used. Samples were confirmed by using high performance liquid chromatography. Results: The results revealed that, amitriptyline HCl and clozapine were detected in the hair in the second day after drug administration. After stopping drug administration ,hair samples taken revealed that, clozapine, can be detected in the hair up to 28 days. Amitriptyline HCl disappeared after clozapine, as it can be detected in the hair up to 35 days after stopping drug administration. Conclusion: This study can concluded that psychotropic drugs can be detected in hair. Also hair analysis can be used to differentiate between drug consumers, chronic poisoning and drug abusing.