Rapid and sensitive High-Performance Thin-Layer Chromatographic (HPTLC) method for identification and quantification of luteolin by densitometry in Kasamarda (Cassia occidentalis L.)

Abstract

Kasamarda (C. occidentalis L.) is a traditional herb recently recognized as a potential nutraceutical in bone health. The current botanical nutraceutical regulations require consistent standardization for biological applications. The present study reported the standardization of bioactive flavonoid luteolin from Cassia occidentalis L. using validated high-performance thin-layer chromatographic (HPTLC) densitometric (DS) method. The mobile phase composition of toluene, ethyl acetate, and formic acid was optimized to separate and identify luteolin using silica gel 60F₂₅₄ aluminum plates. The densitometric (DS) scanning was performed at 353 nm. This HPTLC-DS method was further validated as per ICH guidelines. The linearity was 200–700 ng/band with a correlation coefficient value of 0.994. The LOD and LOQ were found to be 54.06 ng/band and 163.84 ng/band, respectively. The recovery (88.38% and 100.72%) and precision (RSD,<5%) indicated method performance is robust and accurate for the routine analysis. Further, this bioactive flavonoid presence was confirmed and quantified by UV-spectrumin the sample matrix using this validated HPTLC-DS method. This HPTLC-DS method was robust, precise and accurate for quality control of active constituents present in C. occidentalis L.