Abstract:

A reversed-phase high performance liquid chromatographic (RP-HPLC) method for the determination of 9 polyphenols in plant was developed. The separation conditions of the chromatography were optimized with a Sunfire-C18 (0.5 μm, 4.6 mm×150 mm) separation column. The relationship of acetic acid concentration in mobile phase and the retention time, the effect of mobile phase composition and the ratio of acetic acid solution and methanol on retention time were studied separately, and a gradient elution was established. Quantitative results showed that the limits of detection were 13.26-59.29 mg/kg for the 9 analytes in plant. The linear range for the quantification of the 9 analytes was 3.0-100.0 mg/L and the correlation coefficients (r2) were between 0.9979-0.9999. Tobacco sample was extracted by ultrasonication using 80% methanol, and determined using HPLC. The spiked recoveries from tobacco samples were 96.8%-108%, and their relative standard deviations were less than 3.8% (n=3). Chlorogenic acid and rutin in real tobacco samples were also determined.

Key words: ultrasonic extraction, polyphenol, tobacco , high performance liquid chromatography (HPLC)