title: Systematic Development and Validation of a Thin-Layer Densitometric Bioanalytical Method for Estimation of Mangiferin Employing Analytical Quality by Design (AQbD) Approach.
abstract: The present work aims at the systematic development of a simple, rapid and highly sensitive densitometry-based thin-layer chromatographic method for the quantification of mangiferin in bioanalytical samples. Initially, the quality target method profile was defined and critical analytical attributes (CAAs) earmarked, namely, retardation factor (Rf), peak height, capacity factor, theoretical plates and separation number. Face-centered cubic design was selected for optimization of volume loaded and plate dimensions as the critical method parameters selected from screening studies employing D-optimal and Plackett-Burman design studies, followed by evaluating their effect on the CAAs. The mobile phase containing a mixture of ethyl acetate : acetic acid : formic acid : water in a 7 : 1 : 1 : 1 (v/v/v/v) ratio was finally selected as the optimized solvent for apt chromatographic separation of mangiferin at 262 nm withRf 0.68 ± 0.02 and all other parameters within the acceptance limits. Method validation studies revealed high linearity in the concentration range of 50-800 ng/band for mangiferin. The developed method showed high accuracy, precision, ruggedness, robustness, specificity, sensitivity, selectivity and recovery. In a nutshell, the bioanalytical method for analysis of mangiferin in plasma revealed the presence of well-resolved peaks and high recovery of mangiferin.
authors.affiliation: University Institute of Pharmaceutical Sciences, UGC Centre of Advanced Studies, Punjab University, Chandigarh 160 014, India.
authors.forename: Rajneet Kaur
authors.initials: RK
authors.lastname: Khurana
authors.affiliation: Division of Radiation Biology and Toxicology, School of Life Sciences, Manipal University, Manipal, Karnataka 576 104, India.
authors.forename: Satish
authors.initials: S
authors.lastname: Rao
authors.affiliation: University Institute of Pharmaceutical Sciences, UGC Centre of Advanced Studies, Punjab University, Chandigarh 160 014, India.
authors.forename: Sarwar
authors.initials: S
authors.lastname: Beg
authors.affiliation: University Institute of Pharmaceutical Sciences, UGC Centre of Advanced Studies, Punjab University, Chandigarh 160 014, India.
authors.forename: O P
authors.initials: OP
authors.lastname: Katare
authors.affiliation: University Institute of Pharmaceutical Sciences, UGC Centre of Advanced Studies, Punjab University, Chandigarh 160 014, India UGC-Centre of Excellence in Applications of Nanomaterials, Nanoparticles and Nanocomposites (Biomedical Sciences), Punjab University, Chandigarh 160 014, India bsbhoop@yahoo.com bsbhoop@pu.ac.in.
authors.forename: Bhupinder
authors.initials: B
authors.lastname: Singh
mesh_terms: D003720:Densitometry / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D015203:Reproducibility of Results; D044004:Xanthones / Q000032:analysis*
chemical_list: D044004:Xanthones; C013592:mangiferin