Determination of polyphenols in tobacco samples by ultra-fast HPLC

In order to achieve high-throughput and accurate determination of polyphenols in tobacco, a sample extraction vial with a solid-phase extraction(SPE)function was designed and used as a pre-treatment device to establish an ultra-fast high performance liquid chromatography (HPLC) method for determining eight polyphenols, including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, scopoletin, rutin, kaempferol-3-O-rutinoside and quercetin. The results showed that:1)The sample extraction vial integrated the functions of sample extraction, SPE purification, filtration and transfer, which significantly improved the preparation speed. 2)The prepared sample was separated on a C₁₈ column(50 mm×4.6 mm, 1.8 μm)by using 0.1% formic acid aqueous solution and acetonitrile as the mobile phase. The eight analytes were separated smoothly with good linearity in the range of 0.05-350 μg/mL and the correlation coefficients larger than 0.999. The limits of detection(LODs)were 0.35-1.03 μg/g. The mean recoveries ranged from 91.1% to 101.5%, and the intra-day and inter-day precision was less than 5%. 3)The contents of the eight polyphenols in five tobacco samples were between 0.43 and 15.22 mg/g, which were consistent with the results obtained using the tobacco industry standard method. This new method is simple, rapid, sensitive, reproducible, and suitable for the high-throughput detection of polyphenols in tobacco samples.