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一测多评法同时测定烟草及烟草制品中8种多酚类化合物

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

  • 摘要: 为实现烟草及烟草制品中多酚类化合物的快速测定和质量评价,建立了一测多评法同时测定新绿原酸、东莨菪苷、绿原酸、隐绿原酸、咖啡酸、莨菪亭、芸香苷、山奈酚-3-O-芸香糖苷等8种多酚类化合物。采用高效液相色谱-二极管阵列(HPLC-DAD)分段检测技术,以甲醇、水和乙酸为流动相,采用梯度洗脱方式,流速1.0 mL/min,柱温30 ℃,在C18色谱柱上分离8种多酚类化合物。以绿原酸为内参物,建立其他7种多酚类化合物的相对校正因子,分别采用外标法和一测多评法(多点校正法、斜率校正法和单点校正法)测定烟草及烟草制品中8种成分的质量分数。结果表明:①在优化条件下,8种多酚类化合物在各自线性范围内线性关系良好,相关系数R2>0.999 9,检出限和定量限分别为0.01~0.15和0.02~0.48 mg/L,加标回收率为92.00%~102.38%,相对标准偏差为1.56%~4.10%。②以绿原酸为内参物,斜率校正法计算相对校正因子具有良好的耐用性和重现性(RSD < 3.00%)。斜率校正法所获结果与外标法的一致性最佳,平均相对偏差均不大于3.21%。③该方法操作简便、准确,适用于烟草及烟草制品中8种多酚类化合物的批量测定和质量评价。

     

    Abstract: 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.

     

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