Determination of rosmarinic acid and rutin in Hymenocrater bituminosus by using TLC

A simple, rapid, precise, accurate and repeatable thin-layer chromatographic (TLC) method has been established for the determination of rosmarinic acid and rutin in whole plant powder of Hymenocrater bituminosus Fisch. and C.A. Mey. Rosmarinic acid and rutin have been reported to have strong antioxidant properties and also anti-diabetic, anti-thrombotic, anti-inflammatory and anti-carcinogenic activity [1,2]. The aqueous methanolic extract of aerial parts of plant powder was prepared using Sonication Extraction Method (SEM). The concentration of rosmarinic acid and rutin in the whole plant powder were found to be 1.37 and 0.5% (w/w) respectively. Separation was performed on TLC aluminum sheets silica gel 60 F254 plates with ethyl acetate-methanol-distilled water-formic acid (7.2:0.8:1.3:0.7) for rutin and acetone-toluene-formic acid (4:5:1) for rosmarinic acid as mobile phase. Sample solutions for TLC analyses were applied by means of a CAMAG Linomat 5 automated spray-on band applicator. The determination was carried out using the densitometry absorbance mode at 366nm using a CAMAG TLC Scanner 3. The linear range for rutin and rosmarinc acid were 50–450ng with correlation coefficient (r-value) of 0.991 and 150–600ng with correlation coefficient 0.991 respectively. The variability of the method was expressed as intra-day and inter-day precision

References: [1] Petersen, M., Simmonds M.S.J. (2003) Phytochemistry 62:121–125.

[2] Lacopini, P. et al. (2008)J. Food Comp. Anal. 21:589–598.