HPLC法测定菊苣中芦丁、槲皮素和山柰酚含量
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摘要
建立HPLC法同时测定维药菊苣茎、根、种子总黄酮提取液中芦丁、槲皮素和山柰酚含量的方法。用安捷伦HC-C_(18)柱(250 mm×4.6 mm, 5μm)色谱柱,流动相为0.1%乙腈(A)-磷酸(B)(50︰50, V/V),流速0.8 mL/min,检测波长365 nm,柱温30℃。结果表明,芦丁、槲皮素和山柰酚分别在2.0～20μg/mL (r~2=0.999 9), 0.5～40μg/m L (r~2=0.999 8)和0.5～60μg/m L (r~2=0.999 1)浓度范围内呈现良好线性关系,加标回收率(n=3)分别为97.7%(RSD=1.89%), 97.3%(RSD=1.01%)和102.6%(RSD=0.33%)。菊苣茎中芦丁含量为116.3×10~3μg/kg,槲皮素为122.4×10~3μg/kg,山柰酚为32.7×10~3μg/kg;菊苣根中存在芦丁,含量为82.0×10~3μg/kg;菊苣子中存在芦丁和槲皮素,芦丁含量为85.0×10~3μg/kg,槲皮素为45.3×10~3μg/kg。该方法简便、快速、有效、灵敏、准确,具有良好回收率,可作为该植物的定量分析方法,试验结果可为以后菊苣资源的开发利用提供依据。
The content of rutin, quercetin and kaempferol in extract of total flavonoids of stem, root and seed of Cichorium intybus L. was determined by HPLC. HPLC with HC-C_18 column(250 mm×4.6 mm, 5 μm), and the mobile phase was0.1% CH_3 CN, H_3 PO_4(50︰50, V/V) for gradient elution, flow rate 0.8 mL/min, detection wavelength 365 nm, and column temperature 30 ℃. The result showed that the linear responses were ranged from 2.0-20 μg/m L for rutin(r~2=0.999 9), 0.5-40 μg/m L for quercetin(r~2=0.999 8), and 0.5-60 μg/m L for kaempferol(r~2=0.999 1), respectively. The average recoveries of rutin, quercetin and kaempferol were 97.7%(RSD=1.89%), 97.3%(RSD=1.01%) and 102.6%(RSD=0.33%). The content of rutin in stems was 116.3×10~3 μg/kg; The quercetin was 122.4×10~3 μg/kg; The kaempferol was 32.7×10~3 μg/kg; The content of rutin in roots was 82.0×10~3 μg/kg; The content of rutin in seeds was 85.0×10~3 μg/kg; The quercetin was 45.3×10~3μg/kg. The method was simple, rapid, effective, sensitive and accurate, with good recovery rate, which could be used as a quantitative analysis method for the plant. The experimental results might provide important theoretical basis for the future development and utilization of cichorium resources.
The content of rutin, quercetin and kaempferol in extract of total flavonoids of stem, root and seed of Cichorium intybus L. was determined by HPLC. HPLC with HC-C_18 column(250 mm×4.6 mm, 5 μm), and the mobile phase was0.1% CH_3 CN, H_3 PO_4(50︰50, V/V) for gradient elution, flow rate 0.8 mL/min, detection wavelength 365 nm, and column temperature 30 ℃. The result showed that the linear responses were ranged from 2.0-20 μg/m L for rutin(r~2=0.999 9), 0.5-40 μg/m L for quercetin(r~2=0.999 8), and 0.5-60 μg/m L for kaempferol(r~2=0.999 1), respectively. The average recoveries of rutin, quercetin and kaempferol were 97.7%(RSD=1.89%), 97.3%(RSD=1.01%) and 102.6%(RSD=0.33%). The content of rutin in stems was 116.3×10~3 μg/kg; The quercetin was 122.4×10~3 μg/kg; The kaempferol was 32.7×10~3 μg/kg; The content of rutin in roots was 82.0×10~3 μg/kg; The content of rutin in seeds was 85.0×10~3 μg/kg; The quercetin was 45.3×10~3μg/kg. The method was simple, rapid, effective, sensitive and accurate, with good recovery rate, which could be used as a quantitative analysis method for the plant. The experimental results might provide important theoretical basis for the future development and utilization of cichorium resources.
引文
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[8]张雪,褚文静,黄喜茹,等.反相高效液相色谱二极管阵列检测法同时测定丹参饮片中5种水溶性成分[J].理化检验(化学分册), 2010(11):1323-1325.
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[10]陈晨,张兴旺,罗智敏,等. RP-HPLC法测定不同地区金露梅中槲皮素和山柰酚[J].分析试验室, 2009(S2):54-56.
[11]木合塔尔·吐尔洪,热萨莱提·伊敏,木尼热·阿不都克里木,等.液相色谱法同时测定维药蜀葵花中芦丁、槲皮素和山柰酚[J].色谱, 2015(12):1269-1273.
[12]买买提·吐尔逊,古丽巴哈尔·达吾提,陈娟,等.HPLC-PAD法同时测定维药祖卡木颗粒中甘草苷、柚皮苷、槲皮素和山柰酚的含量[J].应用化工, 2015(7):1354-1357, 1362.
[13]边清泉. HPLC法测定凉山杜鹃花中芦丁、槲皮素的含量[J].西南科技大学学报, 2004, 19(4):83-85.
[14]朱轶姮,赵鸿雁,罗贤标,等.反相高效液相色谱法测定槐米中槲皮素的含量[J].南京医(科大学学报), 2004,24(5):551-552.
[15]毛琼玲,艾来提·苏里坦,夏热帕提·吐孙,等.新疆菊苣籽中总黄酮提取工艺研究[J].生物技术, 2009(3):78-80.
[2]陈乃富.蕨菜黄酮类化合物的提取及其抗氧化作[J].食品与发酵工程, 2003(11):63-66.
[3]饶璐璐.菊苣[J].蔬菜, 1997(3):2.
[4]买买提·艾买提,木合布力·阿布力孜,孟磊.新疆昆仑雪菊水溶性总黄酮的含量测定[J].海峡药学, 2010(10):56-58.
[5] GILANI A H, JANBAZ K H. Evaluation of the liver protective potential of Cichorium intybus seed extract on acetaminophen and CCl4-induced damage[J]. Phyto-medicine,1994, 1(3):193-197.
[6] KAMBOJ V P. A review of Indian medicinal plants with interceptive activity[J]. Indian J Med Res, 1988, 87:336-355.
[7]夏虹,彭茂民.高效液相色谱法同时测定槐花中芦丁、槲皮素和山柰酚的含量[J].应用化工, 2014(10):1919-1921.
[8]张雪,褚文静,黄喜茹,等.反相高效液相色谱二极管阵列检测法同时测定丹参饮片中5种水溶性成分[J].理化检验(化学分册), 2010(11):1323-1325.
[9]张丽贤,周光明,姜兰芳,等.非离子表面活性剂萃取-高效液相色谱法测定枇杷叶中的绿原酸、槲皮素和山柰酚[J].食品科学, 2012(6):150-153.
[10]陈晨,张兴旺,罗智敏,等. RP-HPLC法测定不同地区金露梅中槲皮素和山柰酚[J].分析试验室, 2009(S2):54-56.
[11]木合塔尔·吐尔洪,热萨莱提·伊敏,木尼热·阿不都克里木,等.液相色谱法同时测定维药蜀葵花中芦丁、槲皮素和山柰酚[J].色谱, 2015(12):1269-1273.
[12]买买提·吐尔逊,古丽巴哈尔·达吾提,陈娟,等.HPLC-PAD法同时测定维药祖卡木颗粒中甘草苷、柚皮苷、槲皮素和山柰酚的含量[J].应用化工, 2015(7):1354-1357, 1362.
[13]边清泉. HPLC法测定凉山杜鹃花中芦丁、槲皮素的含量[J].西南科技大学学报, 2004, 19(4):83-85.
[14]朱轶姮,赵鸿雁,罗贤标,等.反相高效液相色谱法测定槐米中槲皮素的含量[J].南京医(科大学学报), 2004,24(5):551-552.
[15]毛琼玲,艾来提·苏里坦,夏热帕提·吐孙,等.新疆菊苣籽中总黄酮提取工艺研究[J].生物技术, 2009(3):78-80.