Determination of Hyoscyamine and Scopolamine in Four Hyoscyamus Species from Iran

The purpose of the present study was to determine the tropane alkaloid content of four Hyoscyamus species from Iran, i.e. H. pusillus L., H. niger L., H. reticulatus L. and H. kurdicus Bornm. Determination of alkaloids was performed by the HPLC method. Samples were extracted with chloroform-methanol-25% ammonium hydroxide 15:5:1 (V/V/V). HPLC separation was performed on Eurospher C18 reversed phase column. An isocratic mixture of triethylammonium phosphate buffer (30 Mm, pH 6.2) and acetonitrile (75:25), was used as the eluent. Hyoscyamine and scopolamine were determined by the external standard method at 210 nm. All the four mentioned Hyoscyamus species contained hyoscyamine and scopolamine, but in different amounts. Scopolamine was the predominant tropane alkaloid in H. pusillus, H. niger and H. kurdicus, while, H. reticulatus contained a higher amount of hyoscyamine.