A simple, precise, accurate, rapid, and sensitive reverse phase high performance liquid chromatography (RP-HPLC) method with UV detection has been developed and validated for quantification of naringin (NAR) in novel pharmaceutical formulation. NAR is a polyphenolic flavonoid present in most of the citrus plants having variety of pharmacological activities. Method optimization was carried out by considering the various parameters such as effect of pH and column. The analyte was separated by employing a C18 ( ？mm, 5？μm) column at ambient temperature in isocratic conditions using phosphate buffer pH 3.5: acetonitrile (75？:？25%？v/v) as mobile phase pumped at a flow rate of 1.0？mL/min. UV detection was carried out at 282 nm. The developed method was validated according to ICH guidelines Q2(R1). The method was found to be precise and accurate on statistical evaluation with a linearity range of 0.1 to 20.0？μg/mL for NAR. The intra- and interday precision studies showed good reproducibility with coefficients of variation (CV) less than 1.0%. The mean recovery of NAR was found to be 99.33 ± 0.16%. The proposed method was found to be highly accurate, sensitive, and robust. The proposed liquid chromatographic method was successfully employed for the routine analysis of said compound in developed novel nanopharmaceuticals. The presence of excipients did not show any interference on the determination of NAR, indicating method specificity. 1. Introduction The alternative system of medicine is gaining importance recently. The aim of using plant isolates such as flavonoids, terpenoids, alkaloids, resins, and proanthocyanidins is increasing nowadays. Flavonoids, commonly used in human diet, are a group of naturally occurring polyphenolic compounds that are abundant in many vascular plants. These bioactive compounds have gained interest recently because of their broad pharmacological activities including antioxidant, blood lipid and cholesterol lowering, anti-inflammatory, anticarcinogenic, antiulcer, and antimicrobial, superoxide scavenging actions. Recent attention towards these substances has been stimulated by their potential health benefits [1–7]. Pure herbal constituents are used these days as components in targeted drug delivery system for achieving various pharmaceutical benefits. These flavonoids have prominent bioactivity but the major problems associated with them are limited solubility and permeability accounting for their poor bioavailability [8]. To improve the bioavailability of these components, the use of novel pharmaceutical technology is important.
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