高效液相色谱法评价贵州金钱草代谢产物槲皮素和山奈素的多样性

目的： 建立金钱草中槲皮素和山奈素的HPLC测定方法，用以测定贵州省不同来源地的21份金钱草中槲皮素和山奈素的含量，并对其多样性进行分析。方法： 采用Alltima C₁₈柱（5 μm，250 mm×4.6 mm），以甲醇-0.4%磷酸（50∶50，V/V）为流动相，柱温：25℃，流速：1.0 mL·min^-1，检测波长为360 nm；用SPSS 19.0软件对不同来源地金钱草中槲皮素和山奈素进行聚类分析。结果： 槲皮素和山奈素分别在0.028 6~0.196 5 mg （r=0.999 7）和0.047 7~0.381 4 mg （r=0.999 9）的定量范围内，色谱峰面积与进样量呈良好的线性关系，相关系数均大于0.999。槲皮素和山奈素在精密度实验、重复性实验和稳定性试验中的RSD分别为0.24%和0.58%、0.95%和1.32%、1.02%和1.08%，RSD均小于1.40％，加样平均回收率为99.42%和98.36%。不同来源地的21份金钱草中槲皮素和山奈素含量差异较大。结论： 用HPLC法测定金钱草中槲皮素和山奈素含量简单、精确、重复性高，贵州金钱草中槲皮素和山奈素含量差异较大，具有丰富的多样性及一定的地域性。

Evaluation of the diversity of quercetin and kaempferol in Lysimachia christinae Hance in Guizhou province by HPLC

OBJECTIVE To establish a method for the determination of quercetin and kaempferol in Lysimachia christinae Hance by HPLC, and the method was used to determine the contents of quercetin and kaempferol in 21 samples from different sources in Guizhou Province were determined, and its diversity was analyzed. METHODS Alltima C₁₈ column(5 micron, 250 mm×4.6 mm) was used with methanol-0.4% phosphoric acid(50:50, V/V) as mobile phase, column temperature:25℃, flow rate:1.0 mL·min^-1, detection wavelength 360 nm. SPSS 19.0 software was used for cluster analysis of quercetin and kaempferol in Lysimachia christinae Hance from different sources. RESULTS Within the quantitative ranges of 0.028 6-0.196 5 mg(r=0.999 7) and 0.047 7-0.381 4 mg(r=0.999 9), respectively, the peak area of quercetin and kaempferol showed a good linear relationship with the injection volume. The correlation coefficients were greater than 0.999. RSD of quercetin and kaempferol in precision, repeatability and stability tests were 0.24% and 0.58%, 0.95% and 1.32%, 1.02% and 1.08%, respectively. The average recovery was 99.42% and 98.36%. The content of quercetin and kaempferol between 21 samples from different sources varied greatly. CONCLUSION The HPLC method for the determination of quercetin and kaempferol in Lysimachia christinae Hance is simple, accurate and highly reproducible. The content of quercetin and kaempferol in Lysimachia christinae Hance from different sources in Guizhou varied greatly with rich diversity and certain regional characteristics.