Reversed phase HPLC method for separation and identification of steroidal glycoalkaloids (SGAs) from Form (solanum) extract

An accurate and fast high-performance liquid chromatography (HPLC) method were adapted to separate and determine the steroidal glycoalkaloids (SGAs) (solasonine, solanine, chaconine and solamargine), and the steroidal glycoalkaloid aglycones (SGAAs) solasodine. The present work challenges to indicate the suitable mobile phases combined with using fast and efficient liquid chromatographic column (short length with using high surface area 3 μm particle size) to develop a simple gradient elution HPLC method which were efficient to separate both SGAs or SGAAs of interest in solanum extract. However, to separate solamargine and chaconine successfully, an effected approach using a mobile phase modifier such as acetonitrile was still required. by exploring the polarity of and pH of mobile phase buffers, tritely ammonium dihydrogen phosphate (TEAP) was found as the most efficient eluent, and therefore was chosen. The optimization separation conditions were applied to successfully obtained baseline separation in less than 7.0 minutes for all contents of alkaloids. The best separation occurred of all extract constituents at column temperature of 50 °C. The effects of low pH 3.14 were tried to be the best to give sharp and efficient separation. The best separation parameters such a as limit of detection (LOD) which is 0.5 μg/ml, limit of quantitation (LOQ) were 0.7 μg/ml, and linear dynamic range are obtained, by using fast liquid chromatographic column C-18 (50 x 4.6 mm I.D) 3μm particle size.