Comparison between HPLC and HPTLC densitometry for the determination of spinosin from Ziziphus jujuba Mill. fruit extracts

Abstract

Two validated, rapid and precise methods based on HPTLC and RP-HPLC with UV detection for quantitative determination of spinosin in edible extracts of Ziziphus jujuba fruit, is described. The HPTLC chromatograms were developed using a mobile phase of ethyl acetate –dichloromethane – methanol – water (18:10:15:5, v/v/v/v) on a pre-coated plate of silica gel 60 F254 and quantified by densitometry absorbance/remission mode at 334 nm in the range of 10–120 ng/mL. A gradient elution scheme with methanol-water at a flow rate of 1 mL/min and C18-Hypersil (250 × 4.6 mm, 5 µm) analytical column in the range of 32–400 ng/mL were used at 334 nm in the HPLC method. The limit of detection and limit of quantification were 11.59 and 35.12 ng/mL for HPTLC and 33.78 and 102.36 ng/mL for HPLC method, respectively. The relative standard deviation ranged between 0.94–1.27 (intra-day) and 0.39–1.69 (inter-day) for the HPTLC method and 1.02–1.56 (intra-day) and 0.89–1.71 (inter-day) for HPLC method. The results indicated that the developed methods were accurate and precise. The spinosin contents from different extracts were determined and compared by HPLC and HPTLC densitometry. The quantitative results of both analytical methods did not show any statistically significant differences between them.