Integrated Quality by Design Approach for Quantification and Standardization of Beta-Sitosterol and Lupeol in a Cissus quadrangularis Linn. Plant Extracts and its Marketed Formulation by Reverse Phase High Performance Liquid Chromatography

The objective of the study was to develop and validate a simple, robust accurate method for detection simultaneous estimation lupeol beta-sitosterol as dual bioactive markers in ethanol n-hexane extracts Cissus quadrangularis Linn. its marketed formulation using reverse phase highperformance liquid chromatography standardization by integrated quality design approach. A high-performance with photodiode array developed two major phytosterols compounds, betasitosterol lupeol, experiments generated Box-Behnken three level 3 factorial optimize pH mobile phase, percentage buffer organic modifier better separation between beta-sitosterol. optimized chromatographic performed on Merck Hibar LiChrospher® C18 (250 mm×4.6 mm, 5 μm) column stationary isocratic elution program mixture A: Methanol acetonitrile ratio 35:15 v/v B: 20 mM ammonium acetate 40 % (pH 4.5) flow rate 1.0 ml/ min at 210 nm. experimental results predicted experiment were similar suggested responses all fall acceptance criteria desirability 0.961. validated per International Conference Harmonisation guidelines Q2(R1). showed good regression (R2>0.9995) within test ranges percent recovery found be 1.77 w/w 3.007 formulation. extract 0.926 3.63 %, 0.065 0.19 respectively, extract. highly specific without interference impurities. Thus, could extended marker-based routine control their