Method development, optimization and validation of RP-UFLC method for bioactive flavonoids from Cassia auriculata

Abstract

Cassia auriculata (CA) Linn (vernacular name: Avartaki,) is a traditional ayurvedic medicine used in the treatment of Diabetes mellitus. The present study was aimed for extraction, isolation, method development, optimization and validation of RP-UFLC method for bioactive flavonoids such as quercetin-3-O-rutinoside & quercetin from Cassia auriculata. Reverse phase Ultra Fast Liquid Chromatography method was developed with help of UV detector for isolated compounds. Method validation was performed as per international conferenece of harmonization guidelines for system suitability, linearity, accurcay, precision, specificity, robustness, limit of detection and limit of quantification of quercetin and quercetin-3-O-rutinoside. Intra-day and inter-day precisions for quercetin 3-O-rutinoside and quercetin was found precise as ≤ 2.0% &< 4.0% in intraday exercise & ≤2.0% &< 3.0% in inter day exercise. The concentration found in Cassia auriculata are 0.05 μg/mL of quercetin and 0.16 μg/mL of quercetin-3-O-rutinoside. The proposed method for simultaneous estimation of quercetin 3-O-rutinoside and quercetin seems to be accurate, precise, reproducible and repeatable.