摘要
目的:建立同时测定三叶青块根中芦丁、异槲皮苷、山柰酚-3-O-芸香糖苷、紫云英苷、槲皮素和山柰酚含量的方法。方法:采用高效液相色谱(HPLC)法。色谱柱为Alliance SilGreen C18,流动相为0.2%磷酸水溶液-乙腈溶液(梯度洗脱),流速为1.0mL/min,柱温为35℃,检测波长为360 nm,进样量为15μL。结果:芦丁、异槲皮苷、山柰酚-3-O-芸香糖苷、紫云英苷、槲皮素和山柰酚的检测质量浓度线性范围分别为21.77~217.77、12.37~123.75、13.23~132.31、4.63~46.30、5.75~57.50、3.36~33.66μg/mL(r均=0.999 9);检测限分别为0.217 8、0.123 8、0.066 2、0.046 3、0.191 7、0.112 3μg/mL;定量限分别为0.435 6、0.247 5、0.165 4、0.154 3、0.575 0、0.421 2μg/mL;精密度(n=6)、稳定性(24 h,n=7)、重复性(n=6)试验的RSD均≤3.20%;平均加样回收率分别为96.23%、86.88%、97.51%、97.67%、97.50%、87.46%,RSD分别为1.85%、1.90%、1.84%、1.87%、1.25%、2.01%(n=9)。结论:该方法快速、简便,可用于同时测定三叶青块根中芦丁、异槲皮苷、山柰酚-3-O-芸香糖苷、紫云英苷、槲皮素和山柰酚的含量。
OBJECTIVE:To establish a method for simultaneous determination of rutin,isoquercitrin,kaempferol-3-Orutinoside,daunorubicin,quercetin and kaempferol in the roots of Tetrastigma hemsleyanum. METHODS:HPLC method was adopted. The determination was performed on Alliance SilGreen C18 column with mobile phase consisted of 0.2% phosphoric acid solution-acetonitrile solution(gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was 360 nm and the column temperature was at 35 ℃. The sample size was 15 μL. RESULTS:The linear range of rutin,isoquercitrin,kaempferol-3-O-rutinoside,daunorubicin,quercetin and kaempferol were 21.77-217.77,12.37-123.75,13.23-132.31,4.63-46.30,5.75-57.50,3.36-33.66 μg/mL(all r=0.999 9),respectively. limit of detection of them were 0.217 8,0.123 8,0.066 2,0.046 3,0.191 7,0.112 3 μg/mL,respectively. limit of quantitation of them were 0.435 6,0.247 5,0.165 4,0.154 3,0.575 0,0.421 2 μg/mL,respectively. RSDs of precision(n=6),stability(24 h,n=7) and reproducibility tests(n=6) were lower than 3.20%. The average recoveries of them were 96.23%,86.88%,97.51%,97.67%,97.50%,87.46%,RSDs were 1.85%,1.90%,1.84%,1.87%,1.25%,2.01%(n=9),respectively. CONCLUSIONS:The method is fast and simple,and could be applied for simultaneous determination of rutin,isoquercitrin,kaempferol-3-O-rutinoside,daunorubicin,quercetin and kaempferol in the roots of T. hemsleyanum.
引文
[1]南京中医药大学.中药大辞典:下册[M].上海:上海科学技术出版社,1979:3007-3008.
[2]蔡韦炜,陈丹,范世明,等.中药三叶青化学成分及药理作用研究进展[J].天津药学,2014,26(1):38-41.
[3]浦锦宝,梁卫青,郑军献,等. HPLC法测定三叶青中槲皮素、山柰酚的含量[J].中国中医药科技,2011,18(2):134-135.
[4]郑军献,胡轶娟,梁卫青,等.紫外可见分光光度法测定三叶青中总黄酮的含量[J].中国中医药科技,2009,16(5):386-387.
[5]徐硕,金鹏飞,徐文峰,等.民间中药三叶青的研究进展[J].中南药学,2016,14(12):1336-1341.
[6]许文,傅志勤,林婧,等. UPLC-MS/MS法同时测定三叶青中10种黄酮类成分[J].药学学报,2014,49(12):1711-1717.
[7]许文,傅志勤,林婧,等. HPLC-Q-TOF-MS和UPLC-QqQMS的三叶青主要成分定性与定量研究[J].中国中药杂志,2014,39(22):4365-4372.
[8]刘洋,钱丽华. HPLC法测定三叶青中槲皮素和山柰酚含量研究[J].安徽农学通报,2015,21(17):26-28,35.
[9]何丽针,夏骏,姜文娟,等.高效液相色谱内标法测定三叶青块根中槲皮素和β-谷甾醇的含量[J].中兽医学杂志,2017(5):3-5.
[10]丁丽,章璐幸,邱彦. HPLC-ELSD法同时测定三叶青中胡萝卜苷和β-谷甾醇的含量[J].安徽医药,2015,19(11):2083-2084.
[11]林婧,纪明妹,黄泽豪,等.三叶青的化学成分及其体外抗肿瘤活性研究[J].中国药学杂志,2015,50(8):658-663.