反相高效液相色谱法分离测定烟草中的多酚类化合物

Analysis of Polyphenols in Tobacco Using Reversed-Phase High Performance Liquid Chromatography

对植物中9种多酚类化合物的色谱分离条件进行了优化,分别探讨了流动相组成、流动相中醋酸浓度、醋酸溶液与甲醇的比例对保留时间的影响,确定了梯度分离条件,并对9种天然多酚类化合物进行了定量分析。该方法的检测限为13.26-59.29m g/kg（S/N=3）。在3.0-100.0m g/L范围内呈良好的线性关系,相关系数r^2为0.9979-0.9999。9种待测化合物的加标回收率为96.8%-108%,相对标准偏差（RSD）小于3.8%（n=3）。用80%甲醇超声提取烟草样品,并通过优化的色谱条件对其进行分析,测定了实际烟草样品中芸香苷和绿原酸的含量。结果表明,该方法具有一定的实用价值。

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 （0.5μm, 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 （ r^2 ） were between 0. 997 9 - 0. 999 9. Tobacco sample was extracted by ultrasoni- cation 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.