Simultaneous determination of eight polyphenols in tobacco and tobacco products by quantitative analysis of multi-components via single marker

In order to achieve rapid determination of polyphenols in tobacco and tobacco products for quality assessment, a quantitative multi-component analysis by single marker (QAMS) was proposed to simultaneously determine 8 polyphenols including neochlorogenic acid, scopolamine, chlorogenic acid, cryptochlorogenic acid, caffeic acid, scopoletin, rutin and kaempferol 3-O-rutinoside. The analysis was performed using high-performance liquid chromatography with diode array detection (HPLC-DAD) for a segmented detection program. Separation of the 8 polyphenols was achieved on a C₁₈ column using a mobile phase consisting of methanol, water and acetic acid under a gradient elution mode. The flow rate was set at 1.0 mL/min, and the column temperature was maintained at 30 ℃. Using chlorogenic acid as the internal reference, the relative correction factors for the other 7 polyphenols were calculated. The mass fractions of the 8 polyphenols in tobacco and tobacco products were determined using both the external standard method and the QAMS method (calibrated via multi-point, slope, and single-point methods). The results showed that: 1) Under the optimized conditions, the 8 polyphenols exhibited good linear relationships within their concentration ranges (R² > 0.999 9), and the limits of detection (LODs) and quantification (LOQs) were 0.01‒0.15 and 0.02-0.48 mg/L. The spiked recoveries ranged from 92.00% to 102.38% with relative standard deviations (RSDs) of 1.56%-4.10%. 2) Using chlorogenic acid as the internal reference, the relative correction factors calculated by the slope correction method demonstrated good durability and reproducibility (RSDs < 3.00%). Furthermore, the slope correction method achieved the highest consistency with the external standard method, yielding an average relative deviation no more than 3.21%. 3) The established method is simple, accurate and suitable for the batch determination and quality evaluation of the 8 polyphenols in tobacco and tobacco products.