HPLC法同时测定仙鹤草及委陵菜提取物中4种黄酮类成分的含量
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摘要
目的:建立同时测定仙鹤草及近缘植物委陵菜提取物中山柰酚-3-O-α-L-吡喃鼠李糖苷、汉黄芩素、木犀草素和芹菜素的HPLC法,并比较两种提取物中各成分的含量差异。方法:采用HPLC法,Hypersil ODS C_(18)(250×4.6 mm,5μm),0.2%醋酸水(A)-乙腈(B)进行梯度洗脱:0～12 min,15%B～43%B;12～18 min,43%B～58%B;18～30 min,58%B～80%B;30～35 min,80%B～90%B;35～42 min,90%B～15%B;检测波长:265 nm,流速:1 ml·min~(-1);柱温:30℃。结果:山柰酚-3-O-α-L-吡喃鼠李糖苷、汉黄芩素、木犀草素和芹菜素分别在2.385～95.400μg·ml~(-1)、6.092～243.700μg·ml~(-1)、4.187～167.500μg·ml~(-1)和4.635～185.400μg·ml-1范围内线性关系良好(r=0.999 6、0.999 8、0.999 7、0.999 6)。仙鹤草中上述4种成分的平均回收率分别为99.47%(RSD=1.27%)、99.18%(RSD=1.26%)、99.38%(RSD=1.37%)和98.99%(RSD=1.66%),委陵菜中木犀草素和芹菜素的平均回收率分别为98.45%(RSD=1.31%)、98.83%(RSD=1.20%)。仙鹤草提取物中山柰酚-3-O-α-L-吡喃鼠李糖苷、汉黄芩素、木犀草素和芹菜素的含量分别为2.172 0 mg·g~(-1)、6.363 0 mg·g~(-1)、4.215 0 mg·g~(-1)和4.499 0 mg·g~(-1),而委陵菜提取物中仅能检测到木犀草素和芹菜素,含量分别为5.248 0 mg·g~(-1)和5.757 0 mg·g~(-1),此二者含量均显著高于仙鹤草中的含量。结论:该方法准确、可靠,重复性好,可用于仙鹤草及委陵菜提取物的质量评价与控制标准。
Objective: To establish an HPLC method for simultaneous determination of the content of kaempferol-3-O-α-L-rhamnoside,wogonin,luteolin and apigeni in extract of Agrimonia pilosa and closely-related plant Potentilla chinensis,and to compare the content differences of the two extracts. Methods: The HPLC analysis was carried out on Hypersil ODS C_(18) column( 4. 6 mm × 250 mm,5 μm,) with 0. 2% acetic acid( A)-acetonitrile( B) gradient elution: 0-12 min,15% B-43% B; 12-18 min,43% B-58% B;18-30 min,58% B-80% B; 30-35 min,80% B-90% B; 35-42 min,90% B-15% B at a flow rate of 1 ml·min~(-1). The detection wavelength was 265 nm and the column temperature was 30 ℃. Results: Good linear relationship were found in the the range of 2. 385-95. 400 μg·ml~(-1) for kaempferol-3-O-α-L-rhamnoside,6. 092-243. 700 μg·ml~(-1) for wogonin,4. 187-167. 500 μg·ml~(-1) for luteolin and 4. 635-185. 400 μg·ml~(-1) for apigeni( r = 0. 999 6,0. 999 8,0. 999 7,0. 999 6). The average recoveries of the four components in extract of Agrimonia pilosa were 99. 47%,99. 18%,99. 38% and 98. 99%,respectively. And the RSD values were 1. 27%,1. 26%,1. 37% and 1. 66%. The average recoveries of luteolin and apigeni in extract of Potentilla chinensis were 98. 45% and 98. 83%,and the RSD values were 1. 31% and 1. 20%. The study found that the content of kaempferol-3-O-α-L-rhamnoside,wogonin,luteolin and apigeni in extract of Agrimonia pilosa were 2. 172 0,6. 363 0,4. 215 0,4. 499 0 mg·g~(-1),but only can be detected luteolin and apigenin in extracts of Potentilla chinensis. Their contents were 5. 248 0 mg·g~(-1) and 5. 757 0 mg·g~(-1),and they were significantly higher than the content in extracts of Agrimonia pilosa. Conclusion: The method is accurate,reliable,and can be used for the quality control and evaluation of extract of Agrimonia pilosa and Potentilla chinensis.
Objective: To establish an HPLC method for simultaneous determination of the content of kaempferol-3-O-α-L-rhamnoside,wogonin,luteolin and apigeni in extract of Agrimonia pilosa and closely-related plant Potentilla chinensis,and to compare the content differences of the two extracts. Methods: The HPLC analysis was carried out on Hypersil ODS C_(18) column( 4. 6 mm × 250 mm,5 μm,) with 0. 2% acetic acid( A)-acetonitrile( B) gradient elution: 0-12 min,15% B-43% B; 12-18 min,43% B-58% B;18-30 min,58% B-80% B; 30-35 min,80% B-90% B; 35-42 min,90% B-15% B at a flow rate of 1 ml·min~(-1). The detection wavelength was 265 nm and the column temperature was 30 ℃. Results: Good linear relationship were found in the the range of 2. 385-95. 400 μg·ml~(-1) for kaempferol-3-O-α-L-rhamnoside,6. 092-243. 700 μg·ml~(-1) for wogonin,4. 187-167. 500 μg·ml~(-1) for luteolin and 4. 635-185. 400 μg·ml~(-1) for apigeni( r = 0. 999 6,0. 999 8,0. 999 7,0. 999 6). The average recoveries of the four components in extract of Agrimonia pilosa were 99. 47%,99. 18%,99. 38% and 98. 99%,respectively. And the RSD values were 1. 27%,1. 26%,1. 37% and 1. 66%. The average recoveries of luteolin and apigeni in extract of Potentilla chinensis were 98. 45% and 98. 83%,and the RSD values were 1. 31% and 1. 20%. The study found that the content of kaempferol-3-O-α-L-rhamnoside,wogonin,luteolin and apigeni in extract of Agrimonia pilosa were 2. 172 0,6. 363 0,4. 215 0,4. 499 0 mg·g~(-1),but only can be detected luteolin and apigenin in extracts of Potentilla chinensis. Their contents were 5. 248 0 mg·g~(-1) and 5. 757 0 mg·g~(-1),and they were significantly higher than the content in extracts of Agrimonia pilosa. Conclusion: The method is accurate,reliable,and can be used for the quality control and evaluation of extract of Agrimonia pilosa and Potentilla chinensis.
引文
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10 乔卫,赵川,卢滨,等.委陵菜黄酮对正常小鼠及四氧嘧啶所致糖尿病小鼠血糖与血脂的影响[J].中草药,2010,41(4):612-614
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12 孙晓阳.委陵菜抗菌活性及其质量评价[D].黑龙江佳木斯:佳木斯大学硕士学位论文,2016
13 殷海霞,邬秋萍,平欲晖,等.HPLC法同时测定黄精中5-羟基麦芽酚、5-羟甲基糠醛和黄精碱A的含量[J].中国药师,2018,21(9):1683-1686
2 刘位杰,梁敬钰,孙建博,等.仙鹤草化学成分及药理作用研究进展[J].海峡药学,2016,28(2):1-7
3 李利英,邓瑞雪,刘普,等.委陵菜属植物的化学成分及药理作用研究进展[J].中国现代中药,2008,10(9):3-6
4 张婷婷,刘文洪,周琢艳,等.基于线粒体凋亡通路探讨仙鹤草促Hep G2凋亡的机制研究[J].中草药,2018,41(9):1933-1937
5 仙鹤草水提液对胰腺癌细胞BXPC-3和PANC-1增殖的抑制作用研究[J].中国现代应用药学,2018,35(8):1208-1211
6 秦灵灵,王磊,李娟娥,等.仙鹤草对2型糖尿病大鼠胰岛素抵抗及炎性因子的干预作用[J].环球中医药,2013,6(5):333-336
7 朱慧渊,别玉龙,王江,等.仙鹤草不同方式提取产物抗脑缺血再灌注损伤保护作用的实验研究(英文)[J].中国医学科学杂志,2017,32(4):239-247
8 康林之,袁敏,邵峰,等.仙鹤草不同提取物抗疲劳抗氧化及免疫调节作用研究[J].时珍国医国药,2016,27(5):1079-1081
9 徐威,王耀斌,周彦宇,等.仙鹤草收敛止血功能及体外抑菌实验[J].天然产物研究与开发,2016,28(7):1020-1023
10 乔卫,赵川,卢滨,等.委陵菜黄酮对正常小鼠及四氧嘧啶所致糖尿病小鼠血糖与血脂的影响[J].中草药,2010,41(4):612-614
11周媛,李荣,姜子涛.食用委陵菜黄酮的抗氧化性及清除自由基能力研究[J].食品工业科技,2012,33(17):102-105
12 孙晓阳.委陵菜抗菌活性及其质量评价[D].黑龙江佳木斯:佳木斯大学硕士学位论文,2016
13 殷海霞,邬秋萍,平欲晖,等.HPLC法同时测定黄精中5-羟基麦芽酚、5-羟甲基糠醛和黄精碱A的含量[J].中国药师,2018,21(9):1683-1686