地骨皮中咖啡酰丁二胺、地骨皮乙素和绿原酸含量测定方法研究
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摘要
目的建立地骨皮中咖啡酰丁二胺、地骨皮乙素与绿原酸的含量测定方法。方法采用单因素考察方法建立供试品溶液制备方法;采用高效液相色谱(HPLC)法测定咖啡酰丁二胺、地骨皮乙素与绿原酸的含量,色谱条件为Waters Xbridge C18(4.6 mm×250 mm,5μm)色谱柱,甲醇-0.05%三氟乙酸溶液梯度洗脱,流速1.0 m L/min,检测波长为290 nm,柱温30℃。结果地骨皮药材采用100倍量的70%甲醇回流提取60 min,制备供试品溶液;咖啡酰丁二胺在0.013 75~0.220 0 g/L范围内线性关系良好,r=0.999 5,平均加样回收率为99.25%,RSD=1.81%(n=6);地骨皮乙素在0.057 875~0.926 0 g/L范围内线性关系良好,r=0.999 8,平均加样回收率为101.01%,RSD=2.81%(n=6);绿原酸在0.009 25~0.148 0 g/L范围内线性关系良好,r=0.999 5,平均加样回收率为99.51%,RSD=2.32%(n=6)。结论所建立的含量测定方法简便准确,重现性好,为地骨皮药材质量控制和评价奠定基础。
Objective To establish a method for quantitative determination of N-caffeoylputrescine,kukoamine B and chlorogenic acid in Digupi(Chinese Wolfberry Root-bark,Cortex Lycii). Methods The preparation method for solution of test sample was established by using single-factor investigation.HPLC procedure was performed for determining the content of N-caffeoylputrescine,kukoamine B and chlorogenic acid on chromatographic column of Waters Xbridge C18(4. 6 mm × 250 mm,5 μm),mobile phase was methanol-0. 05% trifluoroacetic acid in gradient elution. The flow velocity was 1. 0 m L/min and the detection wavelength was 290 nm at 30℃. Results The solution of test sample was prepared after Digupi was extracted with 100 times amount of 70% methanol by using reflux extraction for 60 min.N-caffeoylputrescine showed a good linear relationship at a range from 0. 013 75 mg/m L to 0. 220 0 g/L(r = 0. 999 5),and average recovery was 99. 25%(RSD = 1. 81%,n = 6). Kukoamine B showed agood linear relationship at a range from 0. 057 875 g/L to 0. 926 0 mg/m L(r = 0. 999 8),and average recovery was 101. 01%(RSD = 2. 81%,n = 6). Chlorogenic acid showed a good linear relationship at a range from 0. 009 25 g/L to 0. 148 0 g/L(r = 0. 999 5),and average recovery was 99. 51%(RSD =2. 32%,n = 6). Conclusion The established quantitative determination method is easy and accurate with a good repeatability,which can lay a foundation for the quality control and reviewing of Digupi.
Objective To establish a method for quantitative determination of N-caffeoylputrescine,kukoamine B and chlorogenic acid in Digupi(Chinese Wolfberry Root-bark,Cortex Lycii). Methods The preparation method for solution of test sample was established by using single-factor investigation.HPLC procedure was performed for determining the content of N-caffeoylputrescine,kukoamine B and chlorogenic acid on chromatographic column of Waters Xbridge C18(4. 6 mm × 250 mm,5 μm),mobile phase was methanol-0. 05% trifluoroacetic acid in gradient elution. The flow velocity was 1. 0 m L/min and the detection wavelength was 290 nm at 30℃. Results The solution of test sample was prepared after Digupi was extracted with 100 times amount of 70% methanol by using reflux extraction for 60 min.N-caffeoylputrescine showed a good linear relationship at a range from 0. 013 75 mg/m L to 0. 220 0 g/L(r = 0. 999 5),and average recovery was 99. 25%(RSD = 1. 81%,n = 6). Kukoamine B showed agood linear relationship at a range from 0. 057 875 g/L to 0. 926 0 mg/m L(r = 0. 999 8),and average recovery was 101. 01%(RSD = 2. 81%,n = 6). Chlorogenic acid showed a good linear relationship at a range from 0. 009 25 g/L to 0. 148 0 g/L(r = 0. 999 5),and average recovery was 99. 51%(RSD =2. 32%,n = 6). Conclusion The established quantitative determination method is easy and accurate with a good repeatability,which can lay a foundation for the quality control and reviewing of Digupi.
引文
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[13]魏明,杨晓梅,刘佳红,等.绿原酸的药理作用研究进展[J].陕西中医,2016,37(4):511-512.Wei M,Yang XM,Liu JH,et al.Progresses in the pharmacology of chlorogenic acid[J].Shaanxi Journal of Traditional Chinese Medicine,2016,37(4):511-512.
[2]郑海生,牛阳,王荣,等.高血压病和糖尿病治疗中地骨皮的作用[J].宁夏医科大学学报,2010,32(6):651-653.Zheng HS,Niu Y,Wang R,et al.The effect of Lycii Cortex in treatment of hypertensive disease and diabetes[J].Journal of Ningxia Medical University,2010,32(6):651-653.
[3]李春英,姜海霞,张晶,等.地骨皮中有效成分的提取及其降血糖调血脂作用研究[J].中国林副特产,2014(4):27-29.Li CY,Jiang HX,Zhang J,et al.Extraction of active ingredients from Cortex Lycii and their regulative effects on blood sugar and fat[J].Forest By-Product and Speciality in China,2014(4):27-29.
[4]李惠,王卫杰,何栋.地骨皮蒽醌对高脂血症大鼠胆汁酸代谢机理研究[J].饲料博览,2016(11):40-43.Li H,Wang WJ,He D.Study on the bile acid metabolism mechanism of anthraquinone Cortex Lycii in hyperlipidemia rats[J].Feed Review,2016(11):40-43.
[5]谢练武,李顺祥,谢宇霞,等.活性跟踪法研究地骨皮中抑制NF-κB的化学成分[J].中国中药杂志,2014,39(4):689-694.Xie LW,Li SX,Xie YX,et al.Bioassay-guided fraction of constituents targeting mediators of inflammation from Lycii Cortex as inhibitors of NF-κB[J].China Journal of Chinese Materia Medica,2014,39(4):689-694.
[6]刘洪超,郭庆梅,周凤琴.地骨皮的功效考证与现代药理学研究比较[J].山东中医药大学学报,2009,33(6):533-534.Liu HC,Guo QM,Zhou FQ.Comparison between efficacy research and modern pharmacology of Lycii Cortex[J].Journal of Shandong University of Traditional Chinese Medicine,2009,33(6):533-534.
[7]孟令杰,刘百联,张英,等.地骨皮化学成分研究[J].中草药,2014,45(15):2139-2142.Meng LJ,Liu BL,Zhang Y,et al.Chemical constituents in root barks of Lycium chinense[J].Chinese Traditional and Herbal Drugs,2014,45(15):2139-2142.
[8]李行诺,楚楚,毛慧琼,等.地骨皮化学成分研究[J].中国现代应用药学,2012,29(9):805-808.Li XN,Chu C,Mao HQ,et al.Chemical constituents from Lycii Cortex[J].Chinese Journal of Modern Applied Pharmacy,2012,29(9):805-808.
[9]Yang YN,An YW,Zhan ZL,et al.Nine new compounds from the root bark of Lycium chinense and theirα-glucosidase inhibitory activity[J].RSC Adv,2017,7(2):805-812.
[10]Lee DG,Park Y,Kim MR,et al.Anti-fungal effects of phenolic amides isolated from the root bark of Lycium chinense[J].Biotechnol Lett,2004,26(14):1125-1130.
[11]Telang N,Li G,Sepkovic D,et al.Comparative efficacy of extracts from Lycium Barbarum bark and fruit on estrogen receptor positive human mammary carcinoma MCF-7cells[J].Nutr Cancer,2014,66(2):278-284.
[12]赵文红,邓泽元,范亚苇,等.阿魏酸体外抗氧化作用研究[J].食品科学,2010,31(1):219-223.Zhao WH,Deng ZY,Fan YW,et al.Antioxidant properties of ferulic acid in vitro[J].Food Science,2010,31(1):219-223.
[13]魏明,杨晓梅,刘佳红,等.绿原酸的药理作用研究进展[J].陕西中医,2016,37(4):511-512.Wei M,Yang XM,Liu JH,et al.Progresses in the pharmacology of chlorogenic acid[J].Shaanxi Journal of Traditional Chinese Medicine,2016,37(4):511-512.