Hassan Askary, Syed2018-08-062018-08-062017https://escholar.umt.edu.pk/handle/123456789/3101Supervised by: Dr. Sohail NadeemSanguinaria canadensis, common name bloodroot have eight isoquinoline alkaloids with six main alkaloids names as quaternary ammonium benzophenanthridine are sanguinarine, chelerythrine, sanguilutin, chelilutin, sanguherin, and quetririn in them sanguinarine alkaloid concentration is highest that is approximatly 50 %. Sanguinarin alkaloid was separated by precipitation method and sanguinarine nitrate was synthesized by repeated acid base reaction. In which concentrated amonia and dilute nitric acid was used. Conformation test was performed on HPLC and UV-vis. A shortest time period method was introduced on HPLC. Sanguinarine was separated on Zorbax Eclipse Plus C18, 4.6 x 100mm, 3.5 micron column, by using gradient elution of ACN (A) in 1% acetic acid in water (B). For quantification, Chromatograms were acquired at 254nm. 10µg/ml injection was passed repeatedly three times. Results showed peak area at 1.43±0.02 retention time. Nitrate confirmation was confirmed by salt analysis while in UV-vis spectrophotometer standard and sample was run at 300 nm from which concentration of nitrate was also determined.enchelerythrine, sanguilutin, chelilutinQuaternary ammonium benzophenanthridineMSThesis