Chen Juan, Shi Yanping
Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, China.
Zhongguo Zhong Yao Za Zhi. 2009 Feb;34(4):423-7.
To develop a high performance liquid chromatographic method for the simultaneous determination of quercetin and kaempferol in Polygonum aviculare.
The optimized method was achieved for the separation and detection of quercetin and kaempferol using Kromasil C18 column (4.6 mm x 250 mm, 5 microm) as the stationary phase, methanol-0.1% aqueous phosphoric acid solution (65:35) as the mobile phase at a flow rate of 1.0 mL x min(-1), 366 nm as the detection wavelength. The main factors, extraction solvent extraction time, hydrolysis tine, temperature and hydrochloric acid concentration, which influenced the sample extraction and hydrolysis procedure, were intensively explored.
Methanol and 65% methanol were chosen as the extracting solvent for the free compounds and for the total compounds, respectively. The optimized hydrolysis procedure was that the sample was hydrolyzed 1.0 h with 4.0 mol x L(-1) hydrochloric acid at 80 degrees C. Quercetin and kaempferol showed good relationship at the range of 0.58-362 mg x L(-1) and 0.51-320 mg x L(-1), respectively. Both of the correlation coefficients of the calibration curves were 0.999 9. The detection limits of quercetin and kaempferol were 0.03 mg x L(-1) and 0.02 mg x L(-1) at a signal-to-noise ratio of 3, respectively. The average recoveries of quercetin and kaempferol were 99.7% and 99.0% with relative standard derivation (RSD) of 1.7% and 3.9%.
The results showed that the method is simple, accurate and repeatable and it is suitable for quality control of Polygonum aviculare.
建立同时测定扁蓄中槲皮素和山柰酚含量的高效液相色谱法。
采用Kromasil C18柱（4.6 mm×250 mm，5μm）为固定相，甲醇-0.1%磷酸水溶液（65:35）为流动相，流速为1.0 mL·min-1，检测波长为366 nm，对槲皮素和山柰酚进行分离检测，优化方法。深入探讨了影响样品提取和水解过程的主要因素，包括提取溶剂、提取时间、水解时间、温度和盐酸浓度。
分别选择甲醇和65%甲醇作为游离化合物和总化合物的提取溶剂。优化的水解方法为：样品在80℃下用4.0 mol·L-1盐酸水解1.0 h。槲皮素和山柰酚在0.58 - 362 mg·L-1和0.51 - 320 mg·L-1范围内线性关系良好，校准曲线的相关系数均为0.999 9。槲皮素和山柰酚在信噪比为3时的检测限分别为0.03 mg·L-1和0.02 mg·L-1。槲皮素和山柰酚的平均回收率分别为99.7%和99.0%，相对标准偏差（RSD）分别为1.7%和3.9%。
结果表明该方法简便、准确、可重复，适用于扁蓄的质量控制。
