Amygdalin is a glycoside which commonly found in the seeds of stone fruits. This active ingredient has a great importance in drug industry. Amygdalin is commonly obtained from the seeds of three different Prunus species (almond, cherry laurel and apricot) by producing in vitro callus cultures. In this study, analyses of amygdalin was performed with HPLC from the explants, calluses and kernels of these three species. According to the results obtained, the maximum amygdalin was noted from the whole kernel of the apricot fruits as 5888.45 mg per 100 gwhereas the maximum amygdalin content of cotyledons was 213.47 mg per 100 g which was measured from the almond explants. Amygdalin content of the Prunus calluses were then noted as 1376.13 mg, 140.74 mg and 74.08 mg in 100 g at cherry laurel, apricot and almond, respectively. Present results suggested that the callus cultures of Prunus species can be used in order to obtain amygdalin.