RP-HPLC Method Development and Validation for Simultaneous Estimation of Rutin and Quercetin in Morus alba L. leaf extract

Abstract

Mulberry, which pertains to the Moraceae family and the genus Morus,has been used in medicine for ages because of its chemical composition and pharmacological action. The plant is recognized in traditional Chinese medicine for its antiphlogistic, diuretic, expectorant, and antidiabetic characteristics. The mulberry leaf is high in flavonoids, which have a variety of biological functions, including antioxidant potential. Quercetin, isoquercetin, rutin, isoquercitrin, quercitrin, luteolin, chlorogenic acid, and other flavonoids can be found in their leaves. The major constituents of total flavones in mulberry leaf extract are rutin and quercetin. The goal of the RP-HPLC approach was to design and validate a method for identifying these elements. HPLC (Shimadzu Technologies LC series) with UV-visible detector at 259nm and C18 column (250mm 4.6mm, 5µm particle size) was used to achieve chromatographic separation (Phenomenex Luna). Solvents (acetonitrile and 0.1 percent v/v solution of glacial acetic acid) were used in the isocratic elution mode. The run time was 10 minutes. The column temperature was 37 degrees Celsius with sample insertion volume 20µl and the flow velocity was 1ml/min. For quercetin and rutin, the devised method produced strong linearity, high accuracy, high precision, high repeatability, high robustness, and high ruggedness. For quercetin and rutin, the LOD and LOQs values were determined to be within acceptable levels.