广东省化橘红中黄酮类物质的HPLC测定

Determination of Flavanones in Tomentose Pummelo from Guangdong Province by High-Performance Liquid Chromatography(HPLC)

采用高效液相／紫外-可见波长检测器（HPLC／VWD）法，测定了化橘红中黄酮类物质的含量。7种黄酮类标准品分别是芦丁、金丝桃苷、柚皮苷、橙皮苷、柚皮素、川皮苷和甜橙黄酮。分离条件：分离柱为Nucleosil C18反相柱，5μm，250mm×4．6min，流动相A为20％的甲醇，流动相B为30％的乙腈，流动相C为乙腈，三元梯度洗脱，流速1．O～1．2mL／min，柱温35℃，分析波长285-350nm，梯度波长。测定结果表明：标准品浓度范围为0．098-25．000μg／mL，回归方程相关系数为0．999-1．000；标准品连续5次进样和在48h内多次进样，保留时间和峰面积的相对标准偏差均≤4．518％；供试的10个化橘红样品中均含有柚皮苷，其中4个样品中含有少量的柚皮素；化橘红中黄酮类物质主要成分是柚皮苷。

A high-performance liquid chromatography（HPLC） method for separating and analyzing seven standard flavanones was developed and used to measure flavanones in tomentose pummelo（Citrus grandis var. tanentosa Hort. ） collected from ten areas in Huazhou County,Guangdong Province. Seven standard flavanones were chosen as follows： rutin, hyperoside, naringin, hesperidin, naringenin, nobiletin and sinensetin. Samples were injected onto reversed-phase Nueleosil C18 column（5 μm,250 mm× 4. 6 mm i. d. ）. Mobile phase A and B were 20% methanol and 30% acetonitrile respectively. Both of them were adjusted to pH 3.2＋0. 05 with 50% phosphoric acid. Acetonitrile was set as mobile C. Triple gradient elution proceeded at 1.0-1.2 mL/min. Operation conditions were set as follows, column temperature 35 ℃, detection at 285-350 nm. The results showed that. Linear calibration of seven standard samples were within 0. 098-25. 000μg/mL with correlation（r） 0. 999-1.000; relative standard deviation（RSD） of retained time and peak area were both below 4. 518% after the compound of seven standard flavanones was injected five times in consecution or repeatedly analyzed in 48 h; and naringin was detected in all of ten tomentose pummelo samples with maximum 17. 6% dried mass,whiel little naringenin was determined in four samples. Naringin was the main flavanone present in tomentose pummelo.