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A SIMPLE ISOCRATIC HPLC METHOD FOR THE SIMULTANEOUS DETERMINATION OF ANTIPSYCHOTIC INDOLE ALKALOIDS IN RAUWOLFIA TETRAPHYLLA

A simple isocratic HPLC method has been developed for the simultaneous quantitation of three antipsychotic indole alkaloid (IA), α-yohimbine (1), isoreserpiline (2), and 10-methoxy tetrahydroalstonine (3) in Rauwolfia tetraphylla leaf. Samples were analyzed by reverse-phase chromatography on a Waters spherisorb ODS2 column using isocratic elution with acetonitrile containing 0.1% TEA and water containing 0.1% TFA (35:65, v/v) at a flow rate of 1 mL/min, a column temperature of 30°C, and UV detection at 210 nm. The method was validated and applied for quantification of individual alkaloids in various leaf extracts of R. tetraphylla. The method allowed simultaneous determination of alkaloids, 1, 2, and 3 in the plant. The limits of detection and quantification were 1.30, 1.16, 1.01, and 4.35, 3.88, and 3.39 µg/mL (20 µL injection) for compounds 1, 2, and 3, respectively. The method is simple, accurate, and precise which may be recommended for the routine quality control analysis of R. tetraphylla leaf extract containing these three IA as antipsychotic principles in the herb.