摘要
目的：建立松叶中槲皮素、山柰酚的HPLC含量测定方法,为松叶药材质量标准的建立提供参考。同时对不同产地、不同种属松叶进行比较研究。方法：采用高效液相色谱法,使用Agilent ZORBAX SB-C18色谱柱（4.6 mm×250 mm,5μm）,以0.5%磷酸（A）-乙腈（B）为流动相,梯度洗脱（0～9 min,25%B→30%B;9～19 min,30%B;19～20 min,30%B→25%B;20～40 min,25%B）,流速0.8 mL·min^-1,检测波长380 nm,柱温30℃。通过优化参数（脱色、脱色溶剂、甲醇预提、盐酸水溶液浓度、溶剂用量、提取时间）确定槲皮素、山柰酚含量测定预处理的最佳条件,并测定不同产地马尾松及同一产地不同种属松叶中槲皮素、山柰酚的含量。结果：槲皮素、山柰酚的含量测定预处理方法为：石油醚（30～60℃）脱色,甲醇-盐酸水溶液（5.0→10）（4∶1）50 mL,回流提取110 min;松叶中槲皮素进样量在0.13～2.24μg范围内线性关系良好（r=0.9997）,平均回收率为98.6%（RSD=1.6%）;山柰酚进样量在0.20～1.04μg范围内线性关系良好（r=0.9999）,平均回收率为100.5%（RSD=2.4%）;两者方法精密度及重复性均良好。在所测的不同产地马尾松中槲皮素、山柰酚总含量为0.715%～0.371%,其中广西产马尾松最高,江苏产马尾松含量最低;在吉林产不同种属松叶中槲皮素、山柰酚的含量为0.846%～0.515%,以红松最高,樟子松最低。结论：本方法经过方法学考察证明可用于松叶药材中槲皮素、山柰酚的含量测定;不同产地马尾松和不同种属松叶中槲皮素、山柰酚含量有一定差异。
Objective： To establish an HPLC method for determining quercetin and kaempferol contained in pine needles,so as to provide preliminary experimental data for quality control standard and to compare the content of Pinus massoniana Lamb. from different areas and different species of pine needles from Jilin. Methods： HPLC and the Agilent ZORBAX SB- C18column（ 4. 6 mm × 250 mm,5 μm） were adopted,with 0. 5% H3PO4（ A）- acetonitrile（ B） as the mobile phase with gradient elution（ 0- 9 min,25% B→30% B; 9- 19 min,30% B; 19- 20 min,30% B→25% B; 20- 40 min,25% B）. The flow rate was 0. 8 mL·min^-1,the column temperature was set at 30 ℃,and the UV detection wavelength was 380 nm. The optimum condition was decided for determining quercetin and kaempferol in pine needles by optimizing the parameters of preparation（ decolorization,decolorization solvent,methanol pre-extraction,hydrochloric acid concentration,solvent dosage,extraction time） and the content of Pinus massoniana Lamb. from different areas and different species of from Jilin was determined. Results： The optimum preparation condition for samples was petroleum ether（ 30- 60 ℃） decolorization,methanol- hydrochloric acid（ 5. 0→10）（ 4∶ 1） 50 mL,reflux extraction for 110 min. Quercetin showed a good linear relationship within the range of 0. 13- 2. 24 μg（ r =0. 9997）,with the average recovery of 98. 6%（ RSD = 1. 6%）. Kaempferol showed a good linear relationship within the range of 0. 20- 1. 04 μg（ r = 0. 9999）,with the average recovery of 100. 5%（ RSD = 2. 4%）. The precision and repeatability were both good. The content range of quercetin and kaempferol was 0. 715%- 0. 371% in Pinus massoniana Lamb. from different areas,and the highest and the lowest content appeared in those from Guangxi and Jiangsu respectively. The content range of quercetin and kaempferol was 0. 846%- 0. 515% in different species of pine needles from Jilin,and species with the highest and the lowest content was Pinus koraiensis Sieb. et Zucc. and Pinus sylvestris var. mongolica Litv. respectively. Conclusion： The methodological validation indicated that the method can be used to determine the content of quercetin,kaempferol in pine needle. The content of quercetin,kaempferol has certain difference among Pinus massoniana Lamb. from different areas and different species.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2014年第11期1969-1974,共6页
Chinese Journal of Pharmaceutical Analysis
基金
国家十二五科技重大项目(2014ZX09301-307-13)
浙江省中医药科技计划(2012ZA030)
浙江省重点实验室(No.2012E10002)
浙江中医药大学校级创新团队
关键词
松叶
槲皮素
山柰酚
含量比较
不同产地样品
不同种属样品
pine needle
quercetin
kaempferol
content comparison
samples from different areas
samples of different species
