College of Enology, Northwest A&F University, Yangling 712100, Shaanxi, China.
Molecules. 2013 Nov 18;18(11):14241-57. doi: 10.3390/molecules181114241.
The paper described a novel chromatographic method for the simultaneous determination of phenolic compounds such as gallic, protocatechuic, vanillic, caffeic, syringic, p-coumaric and salicylic acid, (+)-catechin, (‒)-epicatechin, rutin, morin, quercetin, coumarin and trans-resveratrol at their maximum absorbance wavelengths (MAW) employing reverse-phase high performance liquid chromatography combined with DAD and UV detection via detection wavelength switching. The method was based on MAW acquisition by DAD and quantification by UV. The separation process was performed on a Shim-Pack VP-ODS C18 column (250 mm × 4.6 mm, 5 μm) held at 30 °C, utilizing 3.0% acetic acid and acetonitrile as mobile phase at a flow rate of 0.8 mL/min in the gradient elution mode. The method was fully validated in terms of linearity (r2 > 0.9990, 10‒350 mg/L), precision (both intra-day and inter-day RSD < 4.22%), accuracy (97.31%‒104.66%), specificity, robustness (0.59% < RSD < 2.86%), limit of detection and quantification. The switching method significantly improved the sensitivities of most phenolics studied in comparison with the standard constant wavelength detection (280 nm). The proposed method has been successfully applied to the determination of 14 phenolic compounds in 89 varieties of one-year-old Chinese grape one-year-canes. Grape canes contain many phenolics, especially trans-resveratrol, (‒)-epicatechin, and (+)-catechin.
本文描述了一种新颖的色谱方法,用于同时测定酚类化合物,如没食子酸、原儿茶酸、香草酸、咖啡酸、丁香酸、对香豆酸和水杨酸、(+)-儿茶素、(‒)-表儿茶素、芦丁、杨梅素、槲皮素、香豆素和反式白藜芦醇,在其最大吸收波长(MAW)下,采用反相高效液相色谱法结合 DAD 和紫外检测,通过检测波长切换。该方法基于 DAD 获取 MAW 和 UV 定量。分离过程在 Shim-Pack VP-ODS C18 柱(250mm×4.6mm,5μm)上进行,在 30℃下,以 3.0%乙酸和乙腈为流动相,以 0.8mL/min 的流速在梯度洗脱模式下进行。该方法在以下方面进行了全面验证:线性(r2>0.9990,10‒350mg/L)、精密度(日内和日间 RSD<4.22%)、准确度(97.31%‒104.66%)、特异性、稳健性(0.59%<RSD<2.86%)、检测限和定量限。与标准固定波长检测(280nm)相比,切换方法显著提高了大多数研究的酚类化合物的灵敏度。该方法已成功应用于 89 个一年生中国葡萄一年生茎中 14 种酚类化合物的测定。葡萄藤含有许多酚类化合物,特别是反式白藜芦醇、(‒)-表儿茶素和(+)-儿茶素。