反相高效液相色谱法分离测定5种黄酮类化合物

Separation and Determination of Five Flavone Compounds by RP-HPLC

建立了用反相高效液相色谱法测定槲皮素、芦丁、桑色素、木犀草素和染料木素等黄酮类化合物的方法．色谱条件是：ZORBAX ECLIPSE XPB—C8 5μm（4．6mm×150mm）色谱柱，V甲醇：Ｖ水（含0．2％磷酸）＝50：50的溶液为流动相，流速为0．8mL／min。检测波长260nm．结果表明槲皮素、芦丁、桑色素、木犀草素和染料木素能较好的分离，分别在0．04～20，0．03～15，0．02～20，0．03～15，0．02～16μg／mL的浓度范围内与峰面积成良好线性关系,检测限分别为12．6，10．0，15．2，13．3，6．1ng／mL，方法可用于槐米中黄酮类物质的测定．

A reversed-phase high performance liquid chromatography （RP-HPLC） method for the separation and dertermination of the five flavonoids （rutin, morin, quercetin, luteolin and genistein） was developed. The column was 4.6 mm× 150 mm i. d. , packed with Zorbax Eclipse XPB-C8（5 mm）. The mobile phase was a mixture of MeOH-H2O（ 50 ： 50, V/V）. The flow rate was 0.8 mL/min. UV detection was performed at 260 nm. External standard was used and the calibration curves showed good linearity over the range of 0. 04-20 mg/mL（R=0. 999 1）,0.03-15 mg/mL（R=0. 998 6）,0.02-20 mg/mL（R=0. 999 0）, 0.03-15 mg/mL（R=0. 998 7）and 0.02-16 mg/mL（R=0. 9989）for quercetin, rutin, morin, luteolin and genistein, respectively. Finally, this method was applied for separating and determining quercetin, rutin and genistein in Japanese pagodatree （Sophora japonica ）.