Department of Food Science, Fu Jen University, Taipei 242, Taiwan.
J Pharm Biomed Anal. 2010 Feb 5;51(3):549-56. doi: 10.1016/j.jpba.2009.09.006. Epub 2009 Sep 11.
A high-performance liquid chromatography-diode array detection-mass spectrometry method with electrospray ionization mode (HPLC-DAD-ESI-MS) was developed for simultaneous determination of phenolic acids and flavonoids in fruits of Lycium barbarum Linnaeus, a widely used traditional Chinese herb possessing vital biological activity. Both phenolic acids and flavonoids were extracted with 50% ethanol and purified using a polymeric solid phase extraction cartridge followed by HPLC-DAD-ESI-MS analysis. By employing a Vydac C18 column, a total of 52 phenolic acids and flavonoids were separated within 70min using a gradient mobile phase of 0.5% (v/v) formic acid in water and acetonitrile-water (94:6, v/v) with flow rate at 1mL/min, column temperature at 30 degrees C and detection wavelength at 280nm. Of 52 compounds, 15 phenolic acids and flavonoids were positively identified based on both absorption and mass spectra, with the remaining 37 tentatively identified by comparison of absorption spectra with reported values in the literature. Internal standards 3-hydroxybenzoic acid and hesperidin were used for quantitation of phenolic acids and flavonoids, respectively. Among the 15 positively identified compounds, quercetin-rhamno-di-hexoside was present in largest mass fraction (438.6microg/g), followed by quercetin-3-O-rutinoside (281.3microg/g), dicaffeoylquinic acid isomers (250.1microg/g), chlorogenic acid (237.0microg/g), quercetin-di-(rhamnohexoside) (117.5microg/g), quercetin-di-(rhamno)-hexoside (116.8mug/g), kaempferol-3-O-rutinoside (97.7microg/g), isorhamnetin-3-O-rutinoside (72.1microg/g), p-coumaric acid (64.0microg/g), caffeic acid (23.7microg/g) and vanillic acid (22.8microg/g).
建立了高效液相色谱-二极管阵列检测-电喷雾电离质谱法（HPLC-DAD-ESI-MS），用于同时测定广泛应用于具有重要生物活性的传统中药枸杞中的酚酸和黄酮类化合物。采用 50%乙醇提取，聚合物固相萃取小柱纯化，采用 Vydac C18 柱，在 30°C 柱温下，以 0.5%（v/v）甲酸水和乙腈-水（94:6，v/v）为流动相，梯度洗脱，流速为 1mL/min，70min 内分离出 52 种酚酸和黄酮类化合物。在 52 种化合物中，根据吸收光谱和质谱，鉴定了 15 种酚酸和黄酮类化合物，其余 37 种化合物通过与文献报道的吸收光谱比较进行了初步鉴定。采用 3-羟基苯甲酸和橙皮苷作为酚酸和黄酮类化合物的内标进行定量。在 15 种被确证的化合物中，槲皮素-鼠李二糖苷的质量分数最高（438.6μg/g），其次是槲皮素-3-O-芸香糖苷（281.3μg/g）、二咖啡酰奎宁酸异构体（250.1μg/g）、绿原酸（237.0μg/g）、槲皮素-二（鼠李糖苷）（117.5μg/g）、槲皮素-二（鼠李糖苷）（116.8μg/g）、山奈酚-3-O-芸香糖苷（97.7μg/g）、异鼠李素-3-O-芸香糖苷（72.1μg/g）、对香豆酸（64.0μg/g）、咖啡酸（23.7μg/g）和香草酸（22.8μg/g）。
