HPLC法测定杠板归中槲皮素-3-O-β-D-葡萄糖醛酸及槲皮素的含量

Quantitative Determination of Quercetin-3-O-β-D-glucuronide and Quercetin in Polygonum Perfoliatum L. by HPLC

建立了高效液相色谱法同时测定杠板归中槲皮素-3-O-β-D-葡萄糖醛酸和槲皮素含量的方法。采用Lichrosher 5-C18色谱柱,以甲醇-0.05%磷酸水溶液为流动相,梯度洗脱,流速为1 mL/min,紫外检测波长为258 nm,柱温为25℃,2种化合物在30 min内均得到较好分离。槲皮素-3-O-β-D-葡萄糖醛酸和槲皮素的线性范围分别为0.037 6～3.920 0 mg和0.004 2～0.440 0 mg,检出限和定量下限分别为2.509、0.282 ng和6.272、0.702 ng,方法的平均回收率分别为99%和98%。运用该方法对不同产地杠板归药材中的2种黄酮类化合物进行测定,并得出二者含量的比例。方法快速简便,灵敏准确,重现性好,可作为杠板归药材的质量控制方法。

An high performance liquid chromatographic method for the quantitative determination of quercetin-3-O-β-D-glucuronide and quercetin in Polygonum perfoliaturn L. was developed. The separation was performed on a Lichrosher 5 -C18 column（250 mm× 4. 6 mm, 5 μm） together with a C18 guard column by gradient elution using methanol -0. 05% phosphoric acid as mobile phase at a flow rate of 1 mL/min. The detection wavelength was set at 258 nm and the colume temperature was 25 ℃. Two compounds were successfully separated in 30 min. The calibration curves were linear in the ranges of 0. 037 6 - 3.920 0 mg for quercetin-3-O-β-D-glucuronide and 0. 004 2 - 0. 440 0 mg for quercetin. The limits of detection （LOD） and limits of quantitation （ LOQ ） were 2. 509, 0. 282 ng and 6. 272, 0. 702 ng, respectively. The mean recoveries of two analytes were 99% and 98% , respectively. The established method was applied in the determination of contents of both two compounds in 15 samples obtained from different regions, and the results showed that the proportion of both compounds in different samples were great different. The proposed method is simple, rapid and accurate, and can be used for the quality control of Polygonum perfoliatum L.