ONLINE HPLC-DAD/UV-MS DETERMINATION OF MAJOR FLAVONOIDS RUTIN AND ISOQUERCITRIN IN TWO MORUS SPECIES.

Mulberry is one such plant that has tremendous therapeutic applications but not much exploited for its medicinal value. The taxonomy of Mulberry (Morus) is complex and disputed. There are no established methods to differentiate among existing species. The present paper demonstrates the work focused on the quality aspects of medicinal plant research for standardizing Mulberry Species. A reversed-phase high performance liquid chromatographic method with simultaneous diode array detection and mass spectral analysis (HPLC-DAD/UV-MS) was developed to detect and quantify two flavonoids, rutin and isoquercitrin as two major flavonoids from two Morus species, Morus alba Linn. and Morus australis Poir. collected from Maharashtra, India. The chromatographic separation was performed on a Cosmosil C8 column (150 x 4.6 mm, 5 μm) with an isocratic mobile phase, comprising 0.1 % formic acid in distilled water, acetonitrile and methanol (75:15:10 v/v/v) at a flow rate of 1.0 mL/min. The wavelength selected for simultaneous quantitation of both analytes was 259 nm. The method developed with careful validation was successfully used for simultaneous quantification of two major flavonoids rutin and isoquercitrin from hydroalcoholic extracts of two Morus species. The hydroalcoholic extraction of the finely powdered dried leaves of both the plants was carried out using ultrasonic extraction technique. The further confirmation of rutin and isoquercitrin in both Morus leaves was done by developed online mass spectral analysis. An Online HPLC-DAD/UV-MS method thus established could be considered as a possible quality control tool for standardizing Morus species and even in routine quality testing of various formulations having rutin and isoquercitrin in them