Determination of Seven Compounds in Tang Maikang Granule by High-Performance Liquid Chromatography

Abstract

An accurate and sensitive high-performance liquid chromatography method is developed and applied to the determination of seven compounds in a kind of traditional Chinese medicinal preparation of Tang Maikang Granule. The method is performed on Hypersil C(18) column (250- x 4.6-mm i.d., 5 microm), and different mobile phases and detectors are selected according to the various compounds. For astragaloside IV, an evaporative light scattering detector (ELSD) is used with a gradient of methanol-water at an eluent gas rate of 2.0 mL/min, under a drift tube temperature of 80 degrees C. Formononetin and calycosin are also eluted by a gradient of methanol-water, but a photodiode array (PDA) detector is used at a wavelength of 254 nm for formononetin and calycosin. A PDA detector at a wavelength of 230 nm is used for paeoniflorin, with methanol-water (30:70, v/v) as the mobile phase. For danshensu and protocate chualdehyde, an eluent of methanol-0.5% acetic acid (12:88, v/v) is used, with PDA detection at 280 nm. For berberine, methanol and water containing 0.1% sodium dodecanesulphonate (SDS) and 0.1% phosphorous acid (70:30, v/v) is employed as the mobile phase, also using a PDA detector, but the detection wavelength is 265 nm. The intra- and interrun precision (relative standard deviation) of this method is less than 5% for seven analytes.