HPLC法同时测定白花蛇舌草中4种多酚物质
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摘要
建立一种同时测定白花蛇舌草中4种天然抗氧化活性成分(槲皮素、山奈酚、咖啡酸、芦丁等)的高效液相色谱法(HPLC)分析方法.采用AgilentXDB-C_(18)(75 mm×2.1 mm,2.7μm)色谱柱,流动相A为甲醇、流动相B为磷酸/水(0.5%,V/V);紫外检测器检测波长260 nm;流速0.3 mL/min;柱温35℃.结果表明,槲皮素、山奈酚、咖啡酸和芦丁4种活性成分的线性范围分别为0.9～20、4.5～100、2.7～60、3.6～80 mg/L,回收率分别为99.4%(RSD=1.84)、98.5%(RSD=2.15)、98.7%(RSD=1.90)、99.3%(RSD=2.00),且4种抗氧化活性成分均得到很好的分离.该方法快速、准确,重现性好.
This research aims to establish a HPLC method for simultaneous determination of four polyphones(quercetin, kaempferol, caffeic acid and rutin) in the extract of Hedyotis diffusa Willd. XDB-C_(18)(75 mm×2.1 mm, 2.7 μm) column was used, and the chromatography was carried out with a linear gradient program. The mobile phase was A: methanol and B: 0.5% phosphoric acid at the flow rate of 0.3 mL·min~(-1), the detection wave-length was 260 nm, and the column temperature was 35 ℃. The results showed that the linear ranges were 0.9 — 20 mg/L for quercetin, 4.5 — 100 mg/L for kaempferol, 2.7 — 60 mg/L for caffeic acid and 3.6 — 80 mg/L for rutin. The average recoveries(n=5) were 99.40% with a relative standard deviation(RSD) of 1.84% for quercetin, 98.5% with RSD of 2.15% for kaempferol, 98.7% with RSD of 1.90% for caffeic acid and 99.3% with RSD of 2.00% for rutin. And four polyphenols were separated successfully from each other. This method is rapid, accurate and reproducible for simultaneous determination of four polyphenols in the extract of Hedyotis diffusa Willd. It is the first time to establish a quantitative method for simultaneous determination of four polyphenols in the extract of Hedyotis diffusa Willd.
This research aims to establish a HPLC method for simultaneous determination of four polyphones(quercetin, kaempferol, caffeic acid and rutin) in the extract of Hedyotis diffusa Willd. XDB-C_(18)(75 mm×2.1 mm, 2.7 μm) column was used, and the chromatography was carried out with a linear gradient program. The mobile phase was A: methanol and B: 0.5% phosphoric acid at the flow rate of 0.3 mL·min~(-1), the detection wave-length was 260 nm, and the column temperature was 35 ℃. The results showed that the linear ranges were 0.9 — 20 mg/L for quercetin, 4.5 — 100 mg/L for kaempferol, 2.7 — 60 mg/L for caffeic acid and 3.6 — 80 mg/L for rutin. The average recoveries(n=5) were 99.40% with a relative standard deviation(RSD) of 1.84% for quercetin, 98.5% with RSD of 2.15% for kaempferol, 98.7% with RSD of 1.90% for caffeic acid and 99.3% with RSD of 2.00% for rutin. And four polyphenols were separated successfully from each other. This method is rapid, accurate and reproducible for simultaneous determination of four polyphenols in the extract of Hedyotis diffusa Willd. It is the first time to establish a quantitative method for simultaneous determination of four polyphenols in the extract of Hedyotis diffusa Willd.
引文
[1] 方允中,郑荣梁. 基生物学的理论与应用[M]. 北京:科学出版社,2002:1-58.
[2] 贝玉祥,郭英,范逸平,等. 诃子多酚清除活性氧自由基及体外抗氧化作用研究[J]. 云南民族大学学报(自然科学版),2009,18(1):51-54.
[3] LIU X, ARDO S,BUNNING M, et al. Total phenolic content and DPPH radical scavenging activity of lettuce (Lactuca sativa L.) grown in Colorado [J]. LWT-Food. Science and Technology, 2007, 40 (3):552-557.
[4] 戴云,邱晓燕,何耀莹,等. 齐墩果酸和熊果酸对DPPH 自由基的清除作用[J]. 云南民族大学学报(自然科学版),2012,21(6):395-398.
[5] COTELLE N, BERNIER J L, CATTEAU J P, et al. Antioxidant properties of hydroxy-flavones[J]. Free Radical Biology and Medicine, 1996, 20(1):35-43.
[6] BHARANI A, GANGULY A, BHARGAVA K D. Salutary effect of Terminalia arjuna in patients with severe refractory heart failure [J]. International Journal of Cardiology, 1995, 49 (3):191-199.
[7] PFUNDSTEIN B, El DESOUKY S K,HULL W E,et al. Polyphenolic compounds in the fruits of Egyptian medicinal plants (Terminalia bellerica, Terminalia chebula and Terminalia horrida):characterization, quantitation and determination of antioxidant capacities[J].Phytochemistry. 2010, 71(10):1132-1148.
[8] OKUDA T. Antioxidant in herbs:polyphends[J]. Antioxidant Food suplements in Human Health, 1999, 39(4):393-395.
[9] 中华人民共和国国家药典委员会. 中国药典(一部)[S]. 北京:化学工业出版社,2005:385-387.
[10] 张海娟,陈业高. 白花蛇舌草黄酮成分的研究[J]. 中药材,2005,28(5):385-387.
[11] 周军平,潘任满. 白花蛇舌草抗癌成分研究[J]. 数量医药杂志,1996,9(4):362-364.
[12] 周建波,龙斯华. 白花蛇舌草素的微量元素分析[J]. 中国中药杂志,1990,15(12):36-39.
[13] 刘志刚,罗佳波. 不同产地白花蛇舌草挥发性成分的初步研究[J]. 中药新药及临床药理,2005,16(2):132-134.
[14] 袁志诚,李巧玉.白花蛇舌草在裸鼠胶质瘤间质化疗的试验研究[J]. 江苏医药杂志,2003,29(11):808-810.
[15] 邵振中,贾晓斌,施峰,等. HPLC同时测定白花蛇舌草中咖啡酸、槲皮素和山奈酚[J]. 中成药,2009,31(12):1894-1897.
[16] 陈莉琳,钱雪良,谢卫锋,等. 辣椒风湿凝胶质量标准研究[J]. 中药,2008,30(4):529-532.
[2] 贝玉祥,郭英,范逸平,等. 诃子多酚清除活性氧自由基及体外抗氧化作用研究[J]. 云南民族大学学报(自然科学版),2009,18(1):51-54.
[3] LIU X, ARDO S,BUNNING M, et al. Total phenolic content and DPPH radical scavenging activity of lettuce (Lactuca sativa L.) grown in Colorado [J]. LWT-Food. Science and Technology, 2007, 40 (3):552-557.
[4] 戴云,邱晓燕,何耀莹,等. 齐墩果酸和熊果酸对DPPH 自由基的清除作用[J]. 云南民族大学学报(自然科学版),2012,21(6):395-398.
[5] COTELLE N, BERNIER J L, CATTEAU J P, et al. Antioxidant properties of hydroxy-flavones[J]. Free Radical Biology and Medicine, 1996, 20(1):35-43.
[6] BHARANI A, GANGULY A, BHARGAVA K D. Salutary effect of Terminalia arjuna in patients with severe refractory heart failure [J]. International Journal of Cardiology, 1995, 49 (3):191-199.
[7] PFUNDSTEIN B, El DESOUKY S K,HULL W E,et al. Polyphenolic compounds in the fruits of Egyptian medicinal plants (Terminalia bellerica, Terminalia chebula and Terminalia horrida):characterization, quantitation and determination of antioxidant capacities[J].Phytochemistry. 2010, 71(10):1132-1148.
[8] OKUDA T. Antioxidant in herbs:polyphends[J]. Antioxidant Food suplements in Human Health, 1999, 39(4):393-395.
[9] 中华人民共和国国家药典委员会. 中国药典(一部)[S]. 北京:化学工业出版社,2005:385-387.
[10] 张海娟,陈业高. 白花蛇舌草黄酮成分的研究[J]. 中药材,2005,28(5):385-387.
[11] 周军平,潘任满. 白花蛇舌草抗癌成分研究[J]. 数量医药杂志,1996,9(4):362-364.
[12] 周建波,龙斯华. 白花蛇舌草素的微量元素分析[J]. 中国中药杂志,1990,15(12):36-39.
[13] 刘志刚,罗佳波. 不同产地白花蛇舌草挥发性成分的初步研究[J]. 中药新药及临床药理,2005,16(2):132-134.
[14] 袁志诚,李巧玉.白花蛇舌草在裸鼠胶质瘤间质化疗的试验研究[J]. 江苏医药杂志,2003,29(11):808-810.
[15] 邵振中,贾晓斌,施峰,等. HPLC同时测定白花蛇舌草中咖啡酸、槲皮素和山奈酚[J]. 中成药,2009,31(12):1894-1897.
[16] 陈莉琳,钱雪良,谢卫锋,等. 辣椒风湿凝胶质量标准研究[J]. 中药,2008,30(4):529-532.