大鼠血浆中芦丁和槲皮素的RP-HPLC同时分析方法

RP - HPLC Method for Simultaneously Determination of Rutin and Qucercetin in Rat Plasma

建立测定大鼠血浆中芦丁和槲皮素含量的反相高效液相色谱(RP—HPLC)方法．用甲醇-乙酸(9:1, v／v)去血浆样品中蛋白质处理,以香兰素作内标,采用Century SIL C18BDS(250 mm×4．6 mm,5μm)色谱柱, 以甲醇-水-磷酸(45:53:0．02)为流动相,流速为0．7 ml／min;紫外检测波长为370nm,柱温为室温,进样量 10 μl．芦丁的定量限为0．068 μg／ml,线性范围为0．068～8．6 μg／ml;槲皮素的定量限为0．0502μg／ml,线性范围为0．0502～13μg／ml．芦丁和槲皮素的平均回收率分别为95．3％,RSD=4．1和90．3％,RSD=2．53(n =6),芦丁的日内及日间精密度的相对标准差分别小于13．2％和9．9％,而槲皮素的日内及日间精密度的相对标准差分别为小于13．9％和11．3％．该方法简单、快速、灵敏度高、重复性良好,可作为芦丁和槲皮素体内样品的分析方法．

To develop a reversed- phase high performance liquid chromatography （RPHPLC） method to simultaneously determine rutin and quacitn in rat plasma. Plasma samples taken from rats were pretreated by protein precipitation with methyl - aceticAcid （9： 1（v/v））. Separation of the main effective constituents rutin and quercetin were accomplished on a reversed - phase Century SIL C18BDS（250mm × 4.6 mm, 5μm） column and mobile phase of methyl - water - phosphoric acid （47：53：0.02（v/v）） with ultraviolet detection at 370 nm. Vanallic aldehyde was used as the internal standard （IS）. The linear range of the calibration curves were 0. 068 - 17.20μg/ml for rutin and 0. 0502-13. 00μg/ml for quercetin. The limit of quantification was 0. 068μg/ml for rutin and 0. 0502μg/ml for quercetin, respectively. The intra- and interday precisions （R. S. D） were ≤ 13.2 % , 49.9 % for rutin and ≤13.9 % , ≤11.3 % for quercetin , respectively. Mean recovery was determined to be 95.3% for rutin and 90.3% for quercetin. The method is simple, rapid, sensitive and has a good productivity, which could be used as the analysis method of rutin and quercetin in body samples.