Estimation of rutin‐loaded chitosan sodium alginate nanoparticles in rat plasma using a chemometrics‐assisted bioanalytical high‐performance liquid chromatography method

Abstract

AbstractRutin, a flavonoid glycoside is mostly made up of phenolic compounds and is involved in a variety of biological processes. Rutin‐loaded chitosan‐alginate nanoparticles were prepared for enhancing the site specificity and bioavailability. The main focus of the current research work was to develop and validate a simple, accurate, and robust bioanalytical reverse‐phase high‐performance liquid chromatography method using the Design of Experiments principles for the estimation of rutin‐loaded chitosan‐alginate nanoparticles in plasma. A mixture of methanol and phosphate buffer in the ratio of 45:55 v/v was used as the mobile phase. The pH of the mobile phase is adjusted to 3.2 with 80% orthophosphoric acid; the flow rate was optimized to 1.0 ml/min and the ultraviolet absorption wavelength at 254 nm. Critical analytical attributes were screened with Taguchi orthogonal array model design. To develop and analyze a “Design Space,” Box‐Behnken three‐level, three‐factorial design in the design of experiment was employed. Analysis of variance, contour plots, and three‐dimensional surface response plots were used to statistically assess this design, and the model was shown to be statistically significant. The devised approach was validated according to the recommendations of the International Council for Harmonization (Q2 R1). The linearity of the developed method showed excellent in the range of 100 to 900 ng/ml with good regression (R2 > 0.998), the limit of quantitation (30 ng/ml), and the limit of detection (10 ng/ml). The validation findings of the parameters were examined and found to be within acceptable limits. The study findings stated that the Quality by Design‐driven bioanalytical approach is applicable for the in vivo and in vitro quantification of rutin derived from bulk and rutin‐loaded chitosan‐alginate nanoparticles formation.