药材问荆异槲皮苷提取工艺的研究
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摘要
采用薄层色谱法对三批次问荆中的异槲皮苷进行定性鉴别。采用高效液相色谱法对异槲皮苷标准品进行标准曲线的绘制,得出异槲皮苷在10.7～86.2μg/mL范围内具有良好的线性关系。通过实验得出加样回收率为99.50%,精密度相对标准偏差(RSD)为1.65%,稳定性相对标准偏差(RSD)为1.60%,重现性相对标准偏差(RSD)为1.62%。使用碱溶酸沉对问荆异槲皮苷的提取条件进行研究,为了研究提取问荆异槲皮苷的最优条件,以异槲皮苷的收率为指标,在此基础上,进行单因素实验和正交实验设计,以问荆与质量分数75%甲醇质量比、温度、超声时间、溶液pH为影响因素。得出使用碱溶酸沉对问荆的异槲皮苷在碱性条件下提取的最优条件为问荆与质量分数75%甲醇质量比为1∶50,浸提温度为90℃,超声时间为60 min,pH=11。
Qualitative research of isoquercitrin in three batches of equisetum was performed by thin layer chromatography. The standard curve of isoquercitrin standard was plotted via high performance liquid chromatography, and it was found that isoquercitrin has a good linear relationship in the range of 10.7～86.2 μg/mL. The experiment results showed that the sample recovery was 99.50%, the precision standard deviation(RSD) was 1.65%, the stability RSD was 1.60% and the reproducible RSD was 1.62%. The extraction conditions of quercetin were investigated by using alkali-soluble acid precipitation. In order to study the optimal conditions for extracting isoquercitrin, the yield of isoquercitrin was used as an indicator for factor experiment and orthogonal experiment design using mass ratio of equistum to 75% methanol, temperature, ultrasonic time and solution pH as the influencing factors. It is concluded that the optimum conditions for the extraction of isoquercitrin are as follows: mass ratio of equistum to 75% methanol of 1∶50, leaching temperature of 90 ℃, extraction time of 60 min and pH of 11.
Qualitative research of isoquercitrin in three batches of equisetum was performed by thin layer chromatography. The standard curve of isoquercitrin standard was plotted via high performance liquid chromatography, and it was found that isoquercitrin has a good linear relationship in the range of 10.7～86.2 μg/mL. The experiment results showed that the sample recovery was 99.50%, the precision standard deviation(RSD) was 1.65%, the stability RSD was 1.60% and the reproducible RSD was 1.62%. The extraction conditions of quercetin were investigated by using alkali-soluble acid precipitation. In order to study the optimal conditions for extracting isoquercitrin, the yield of isoquercitrin was used as an indicator for factor experiment and orthogonal experiment design using mass ratio of equistum to 75% methanol, temperature, ultrasonic time and solution pH as the influencing factors. It is concluded that the optimum conditions for the extraction of isoquercitrin are as follows: mass ratio of equistum to 75% methanol of 1∶50, leaching temperature of 90 ℃, extraction time of 60 min and pH of 11.
引文
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[12] Yang R R,Suo Y R,Qiao Y H,et al.Simultaneous Determination of Formononetin,Medicarpin and Other 4 Flavonoidsin Spatholobi Caulisby HPLC [J].Chin J Pharm Anal,2017,37(12):2 139-2 144 (in Chinese).(杨冉冉,索亚然,乔艺涵,等.高效液相色谱法同时测定鸡血藤中芒柄花素和美迪紫檀素等6个黄酮类成分[J].药物分析杂志,2017,37(12):2 139-2 144.)
[13] Weng J Y,Chen J,Liu K L,et al.Research Progress of Natural Flavonoids in Prevention and Treatment of Diabetic Nephropathy [J].J Pharm Res,2018,37(10):593-596 (in Chinese).(翁竞玉,陈俊,刘坤玲,等.天然黄酮类化合物防治糖尿病肾病的研究进展[J].药学研究,2018,37(10):593-596.)
[14] Zhu W Z,Ma L,Li G R.Prevention and Molecular Mechanisms of Flavonoids Against Cancer [J].Chin Bull Life Sci,2012,24(5):444-449 (in Chinese).(朱文振,马龙,李国荣.黄酮类化合物的抗癌作用及作用机制[J].生命科学,2012,24(5):444-449.)
[15] He J K,Yu Y,Chen X J,et al.Research Progress on Drug Metabolism of Flavanoids [J].China J Chin Mater Med,2010,35(21):2 789-2 794 (in Chinese).(何佳珂,于洋,陈西敬等.黄酮类化合物的药物代谢研究进展[J].中国中药杂志,2010,35(21):2 789-2 794.)
[16] Tao A E,Zhang X C,Du Z F,et al.Research Progress on Flavonoids in Plants of Polygonatum Mill.and Their Pharmacological Activities [J].Chin Tradit Herb Drugs,2018,49(9):2 163-2 171 (in Chinese).(陶爱恩,张晓灿,杜泽飞,等.黄精属植物中黄酮类化合物及其药理活性研究进展[J].中草药,2018,49(9):2 163-2 171.)
[17] Chinese Pharmacopoeia Commission.Pharmacopoeia of the People’s Republic of China:Volume 1 [M].Beijing:China Medical Science Press,2015:248 (in Chinese).(国家药典委员会.中华人民共和国药典:一部[M].北京:中国医药科技出版社,2015:248.)
[2] Syrchina A I,Voronkov M G,Tyukavkina N A.Naringenin,Dihydrokaempferol,and Dihydroquercetin from Equisetum arvense [J].Chem Nat Compd,1975,11(3):439-439.
[3] Wang S,Yuan J Q,Zhou X L,et al.The Pre-test on Chemical Composition of Equisetum diffusum D.Don [J].Guangxi J Tradit Chin Med,2009,32(6):55-56 (in Chinese).(王硕,袁经权,周小雷,等.散生问荆的化学成分预试验[J].广西中医药,2009,32(6):55-56.)
[4] Cui Y B,Ma X J,Liang S,et al.Study on the Qualitative and Quantitative Analysis Methods of Isoquercitrin in Equisetum arvens L.[J].Chin J Drug Eval,2016,33(6):485-488 (in Chinese).(崔业波,马晓静,梁帅等.问荆药材中异槲皮苷的定性和定量分析方法[J].中国药物评价,2016,33(6):485-488.)
[5] Zhang Y L,Ablaev A G,Wang Y F.Distribution of Equisetum pratense Ehrhart (Equisetaceae) [J].Chin Bull Bot,2008,25(3):307-314 (in Chinese).(张玉玲,Albert G.Ablaev,王宇飞.草问荆(木贼科)的地理分布 [J],植物学通报,2008,25(3):307-314.)
[6] Ji Y B,Gao S Y.Effect of Total Alkaloids of Equisetum pretense on Contents of Monoamine Neurotransmitters in Rat Brain [J].Chin J Tissue Eng Res,2005,9(45):185-187 (in Chinese).(季宇彬,高世勇.草问荆总生物碱对大鼠脑内单胺类神经递质含量的影响[J].中国临床康复,2005,9(45):185-187.)
[7] Wang Q,Du G Y.Topical Application of Natural Substances with Anti-inflammatory Activity (Report 7):Equisetum Active Ingredients that Block Platelet Aggregation [J].Int J Tradit Chin Med,1995,17(3):34-35 (in Chinese).(王谦,杜贵友.局部适用之具有消炎活性的天然物质研究(第7报):问荆阻碍血小板聚集的活性成分[J].国际中医中药杂志,1995,17(3):34-35.)
[8] Hu P,Luo G A,Zhao Z,et al.Study on the Evaluating Method and the Affecting Factors of Measurement Uncertainty for the Determination of Marker Compounds in Chinese Materia Medica [J].Chin J Pharm Anal,2005,25(4):455-460 (in Chinese).(胡坪,罗国安,赵中振,等.中药材定量分析中测量不确定度的评定方法及其影响因素的研究[J].药物分析杂志,2005,25(4):455-460.)
[9] Yuan C.Studies on Spectrum Characteristic,Antioxidative Property and Pharmacokinetics of Isoquercitrin and Its Metal Complexes [D].Taiyuan:Shanxi University,2010 (in Chinese).(袁超.异槲皮苷及其金属配合物的光谱特征、抗氧化特性及药代动力学研究[D].太原:山西大学,2010.)
[10] Wu P.Molecular Mechanism of Isoquercitrin Inhibiting Bladder Cancer Cells Growth [D].Guangzhou:South China University of Technology,2017.(吴萍.异槲皮苷抑制膀胱癌细胞增殖的分子机制研究 [D]广州:华南理工大学,2017.)
[11] Xu Y J,Lu Z S,Luo J.Study on Effect of Soy Isoflavones in Reducing Lesion of Rat Non-alcoholic Fatty Liver Disease [J].J Mod Med Health,2019(2):188-191 (in Chinese).(许永杰,卢志顺,罗洁.大豆异黄酮减轻大鼠非酒精性脂肪肝病变的作用研究[J].现代医药卫生,2019(2):188-191.)
[12] Yang R R,Suo Y R,Qiao Y H,et al.Simultaneous Determination of Formononetin,Medicarpin and Other 4 Flavonoidsin Spatholobi Caulisby HPLC [J].Chin J Pharm Anal,2017,37(12):2 139-2 144 (in Chinese).(杨冉冉,索亚然,乔艺涵,等.高效液相色谱法同时测定鸡血藤中芒柄花素和美迪紫檀素等6个黄酮类成分[J].药物分析杂志,2017,37(12):2 139-2 144.)
[13] Weng J Y,Chen J,Liu K L,et al.Research Progress of Natural Flavonoids in Prevention and Treatment of Diabetic Nephropathy [J].J Pharm Res,2018,37(10):593-596 (in Chinese).(翁竞玉,陈俊,刘坤玲,等.天然黄酮类化合物防治糖尿病肾病的研究进展[J].药学研究,2018,37(10):593-596.)
[14] Zhu W Z,Ma L,Li G R.Prevention and Molecular Mechanisms of Flavonoids Against Cancer [J].Chin Bull Life Sci,2012,24(5):444-449 (in Chinese).(朱文振,马龙,李国荣.黄酮类化合物的抗癌作用及作用机制[J].生命科学,2012,24(5):444-449.)
[15] He J K,Yu Y,Chen X J,et al.Research Progress on Drug Metabolism of Flavanoids [J].China J Chin Mater Med,2010,35(21):2 789-2 794 (in Chinese).(何佳珂,于洋,陈西敬等.黄酮类化合物的药物代谢研究进展[J].中国中药杂志,2010,35(21):2 789-2 794.)
[16] Tao A E,Zhang X C,Du Z F,et al.Research Progress on Flavonoids in Plants of Polygonatum Mill.and Their Pharmacological Activities [J].Chin Tradit Herb Drugs,2018,49(9):2 163-2 171 (in Chinese).(陶爱恩,张晓灿,杜泽飞,等.黄精属植物中黄酮类化合物及其药理活性研究进展[J].中草药,2018,49(9):2 163-2 171.)
[17] Chinese Pharmacopoeia Commission.Pharmacopoeia of the People’s Republic of China:Volume 1 [M].Beijing:China Medical Science Press,2015:248 (in Chinese).(国家药典委员会.中华人民共和国药典:一部[M].北京:中国医药科技出版社,2015:248.)