HPLC-PDA Method Development and Validation of Lagenaria siceraria (Molina) Standl. Fruit Extracts Supported by Multianalytical Techniques

The fruit of Lagenaria siceraria belonging to Cucurbitaceae family commonly recognized as bottle gourd is rich in bioactive compounds exhibited an extensive range of therapeutic and pharmacological properties that include antioxidant, lipid lowering, cardioprotective, diuretic, central nervous system stimulant and immunosuppressive, etc. Due to the broad spectrum of bioactivities, researchers have been consistently working on identification, qualitative and quantitative analyses of a new series of bioactive compounds. Therefore, the current investigation was performed to develop and subsequently validation of a rapid, simple, reliable, sensitive and multianalytical methods to standardize constituents of L. siceraria fruits. The simultaneous identification and quantitation of phenolic and flavonoid glycosides, including (E)-4 hydroxymethyl phenyl-6-O-caffeoyl-B-D-glucopyranoside (LS1) and kaempferol 3-O-rutinoside (LS2) were done by ultra-high-performance liquid chromatography (UHPLC) method for the L. siceraria fruits extract. A high-performance thin-layer chromatography-based (HPTLC) approach for phenolic and flavonoid glycosides has been developed as a tool for quality assessment for chemical fingerprinting and quick screening. Additionally, mass spectrometry was used to characterize compounds, LS1 and LS2 in sample matrices. The aforementioned compounds LS1 and LS2 were successfully identified using the qualitative HPTLC-based approach. The results of the method's performance and data validation showed that the UHPLC method was robust (RSD < 5.0%), linear (r2 > 0.99), precise (RSD < 5.0%) and accurate (recoveries 80-110%). It could also be used for quantification. The standardization, quality evaluation, quantification, authenticity and foundation for future research in the field would be benefited from this multianalytical method for identifying the compounds in L. siceraria fruit extract.