HPLC法测定白芍和赤芍中1,2,3,4,6-五-O-倍酰-D-葡萄糖含量

Determination of 1,2,3,4,6-penta-O-galloyl-D-glucose in Radix Paeoniae Alba and Radix Paeoniae Rubra with HPLC

目的用高效液相色谱(HPLC)法测定白芍和赤芍中1,2,3,4,6-五-O-倍酰-D-葡萄糖(PGG)的含量。方法用50%乙醇水溶液,通过超声提取法从白芍和赤芍中提取PGG。采用Diamonsil C18色谱柱,流动相为乙腈-2.5%醋酸水溶液(18∶82,V/V),pH为2.85,紫外检测波长为280nm,流速为1mL/min。结果PGG在5μg/mL^150μg/mL范围内呈良好的线性关系(r=0.9999,n=5),平均回收率为100.9%,RSD为3.57%(n=9)。在白芍和赤芍中PGG的含量分别为2.48mg/g和2.24mg/g。结论本实验所建立的测定PGG含量的HPLC法能够有效消除白芍和赤芍中其他组分的干扰,操作简单快速,精密度高、重现性好、回收率高,适用于中药提取物中PGG含量的测定。

Objective In recent reports, it has been indicated that 1, 2, 3, 4, 6-penta-O-galloyl-D-glucose（PGG） in Chinese herbal medicines is a natural active component which exhibited distinct anticancer, antidiabete and anti-obesity activities. A rapid, sensitive and reliable method using high-performance liquid chromatography is to be developed for determination of PGG in Radix Paeoniae Alba and Radix Paeoniae Rubra. Methods PGG was extracted with 50% ethanol from Radix Paeoniae Alba and Radix Paeoniae Rubra. For the chromatographic separation and analysis of PGG, a reversed phase Diamonsil C18 column（4.6 mm ×250 mm, 5 μm） was used at 25 ℃ with an isocratic mobile phase consisting of aeetonitrile and 2.5% acetic acid solution（ 18： 82, V/V, pH = 2.85 ）. Flow rate was set at 1.0 mL/min. An ultraviolet detector was used and wavelength of detection was at 280 nm. Volume was 20 μL. Results The calibration curve for PGG was linear over the range of 5 μg/mL - 150 μg/mL（ r = 0. 999 9, n = 5 ）. PGG was stable within 48 hours and showed good precision. The average recovery was 100.9% （ RSD = 3.57% , n = 9）. The contents of PGG in Radix Paeoniae Alba and Radix Paeoniae Rubra were 2.48 mg/g and 2.24 rag/g, respectively. Conclusion In the present work, a high-performance liquid chromatography method is developed for quantitative analysis of PGG in Radix Paeoniae Alba and Radix Paeoniae Rubra. The method is rapid, sensitive, reliable, and precise. It is applicable to determine PGG in Chinese herbal medicines.