Università Campus Bio-Medico, Rome, Italy.
J Sep Sci. 2011 Jan;34(2):150-9. doi: 10.1002/jssc.201000665.
Nano-LC and conventional HPLC techniques were applied for the analysis of anthocyanins present in commercial fruit juices using a capillary column of 100 μm id and a 2.1 mm id narrow-bore C(18) column. Analytes were detected by UV-Vis at 518 nm and ESI-ion trap MS with HPLC and nano-LC, respectively. Commercial blueberry juice (14 anthocyanins detected) was used to optimize chromatographic separation of analytes and other analysis parameters. Qualitative identification of anthocyanins was performed by comparing the recorded mass spectral data with those of published papers. The use of the same mobile phase composition in both techniques revealed that the miniaturized method exhibited shorter analysis time and higher sensitivity than narrow-bore chromatography. Good intra-day and day-to-day precision of retention time was obtained in both methods with values of RSD less than 3.4 and 0.8% for nano-LC and HPLC, respectively. Quantitative analysis was performed by external standard curve calibration of cyanidin-3-O-glucoside standard. Calibration curves were linear in the concentration ranges studied, 0.1-50 and 6-50 μg/mL for HPLC-UV/Vis and nano-LC-MS, respectively. LOD and LOQ values were good for both methods. In addition to commercial blueberry juice, qualitative and quantitative analysis of other juices (e.g. raspberry, sweet cherry and pomegranate) was performed. The optimized nano-LC-MS method allowed an easy and selective identification and quantification of anthocyanins in commercial fruit juices; it offered good results, shorter analysis time and reduced mobile phase volume with respect to narrow-bore HPLC.
采用纳升液相色谱和常规高效液相色谱技术,使用 100μm id 和 2.1mm id 窄径 C(18)柱的毛细管柱分析商业果汁中的花色苷。采用 HPLC 和纳升液相色谱法,分别在 518nm 处用 UV-Vis 和 ESI 离子阱 MS 检测分析物。使用商业蓝莓汁(检测到 14 种花色苷)优化分析物的色谱分离和其他分析参数。通过比较记录的质谱数据与已发表文献的数据,对花色苷进行定性鉴定。在这两种技术中使用相同的流动相组成,表明微型化方法比窄径色谱法具有更短的分析时间和更高的灵敏度。两种方法均获得了良好的日内和日间精密度,保留时间的 RSD 值均小于 3.4%和 0.8%,分别为纳升液相色谱和高效液相色谱。通过对氰花翠苷-3-O-葡萄糖苷标准品的外标曲线校准进行定量分析。校准曲线在研究浓度范围内呈线性,HPLC-UV/Vis 为 0.1-50μg/mL,纳升液相色谱-MS 为 6-50μg/mL。两种方法的 LOD 和 LOQ 值均良好。除了商业蓝莓汁外,还对其他果汁(如覆盆子、甜樱桃和石榴)进行了定性和定量分析。优化的纳升液相色谱-MS 方法能够轻松、选择性地鉴定和定量商业果汁中的花色苷;与窄径高效液相色谱相比,该方法具有较短的分析时间和减少的流动相体积,结果良好。