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    高效液相色谱法同时测定不同来源柑橘幼果中10种类黄酮化合物和香豆素类化合物

    Simultaneous Determination of 10 Flavonoids and Coumarins in Green Aurantii Fructus from Different Origins by HPLC

    • 摘要: 建立了高效液相色谱法(HPLC)同时测定柑橘幼果中10种类黄酮化合物和香豆素类化合物含量的方法。用50%(体积分数)甲醇溶液超声提取柑橘样品中的10种化合物,离心过滤后,滤液供高效液相色谱分析。采用Extend XDB C18色谱柱(250 mm×4.6 mm,5 μm)为固定相,以不同体积比的乙腈和0.1%(体积分数)甲酸溶液的混合溶液为流动相进行梯度洗脱,在检测波长330 nm处测定。结果显示:芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、橘皮内酯水合物、木犀草素、川陈皮素、橘皮内酯、橘皮素、葡萄内酯的质量浓度均在一定的线性范围内与其对应的色谱峰面积呈线性关系,检出限为0.019~0.105 mg·g-1。方法用于S3号样品的分析,24 h内,样品溶液中10种化合物峰面积的相对标准偏差(n=7)不大于2.0%;平行测定6次,峰面积的相对标准偏差小于2.2%;方法的稳定性和重复性良好。以样品为基质进行3个浓度水平的加标回收试验,回收率为94.6%~104%。13批样品的分析结果显示,橙皮苷、芸香柚皮苷、川陈皮素、橘皮素均有检出,且这些化合物在不同来源样品中含量差别较大。以柑橘中的特征物质川陈皮素和橘皮素的测定结果为变量进行链接聚类分析,得到的结果可判断各样品的种间亲疏关系。

       

      Abstract: A method was established for simultaneous determination of 10 flavonoids and coumarins in the green aurantii fructus from different orignis by HPLC. 10 compounds in the sample were untrasonically extracted with 50% (volume fraction) methanol solution. After centrifugation and filtration, the filtrate was analyzed by HPLC. Extend XDB C18 (250 mm×4.6 mm, 5 μm) column was adopted as stationary phase, the mixed solution composed of acetonitrile and 0.1% (volume fraction) formic acid solution with various volume ratios was used as mobile phase in gradient elution, and the target compounds were determined at the detected wavelength of 330 nm. As shown by results, linearity relationships between the mass concentration of 10 compounds (narirutin, naringin, hesperidin, neohesperidin, meranzin hydrate, luteolin, meranzin, nobiletin, hesperetin and aurapten) and respective peak area were kept in the definite ranges, with detection limits in the range of 0.019-0.105 mg·g-1. The proposed method was applied to the analysis of sample No. S3, RSDs (n=7) of peak areas for 10 compounds in sample solutions were less than 2% in 24 h, and RSDs of peak areas for 6 parallel determination were less than 2.2%. This showed that the method had good stability and repeatability. The spiked recovery test was made at the 3 concentration levels using sample as matrix, giving recoveries in the range of 94.6%-104%. Narirutin, hesperidin, nobiletin and hesperetin were detected in 13 samples, the content of these compounds in sample from different origins varied greatly. The link cluster analysis was made by using the determined results of the characteristic substances of citrus clementine and tangeretin as variables, and results obtained could judge the relationship between the species.

       

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