加味四妙颗粒的药效成分含量测定
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摘要
目的:建立HPLC-DAD波长切换法同时测定加味四妙颗粒中延胡索乙素、盐酸小檗碱、盐酸黄柏碱、木兰花碱、杯苋甾酮、甘草苷、甘草次酸共7个成分的含量。方法:采用Insert Sustain C_(18)色谱柱(250 mm×4.6 mm,5μm),以乙腈(A)-0.05 mol·L~(-1)磷酸二氢钾(B)为流动相,梯度洗脱,DAD检测器,检测波长268 nm(0～22.5 min,检测木兰花碱,67～71 min,检测盐酸小檗碱)、280 nm(22.5～30 min,检测延胡索乙素)、237 nm(30～45 min,检测甘草苷)、247 nm(45～55 min,检测杯苋甾酮,71～85 min,检测甘草次酸)、284 nm(55～67 min,检测盐酸黄柏碱)。结果:各待测组分分离度良好;延胡索乙素、盐酸小檗碱、盐酸黄柏碱、木兰花碱、杯苋甾酮、甘草苷、甘草次酸7个成分的进样质量浓度分别在2.060～51.49μg·mL~(-1)(r=1.000)、3.111～77.77μg·mL~(-1)(r=1.000)、1.813～45.31μg·mL~(-1)(r=1.000)、1.960～49.00μg·mL~(-1)(r=0.999 9)、2.019～50.48μg·mL~(-1)(r=1.000)、2.468～61.70μg·mL~(-1)(r=0.999 9)、2.408～60.20μg·mL~(-1)(r=0.999 9)范围内与各自峰面积呈良好线性关系;平均加样回收率(n=6)分别为100.6%(RSD=1.6%)、100.3%(RSD=0.4%)、101.0%(RSD=1.3%)、100.7%(RSD=0.8%)、100.1%(RSD=1.9%)、101.6%(RSD=1.3%)、99.8%(RSD=1.7%)。结论:本试验建立的含量测定方法符合方法学验证要求,简便可靠,重现性好,可用于加味四妙颗粒的7个指标性成分的同时测定。
Objective:To establish a method for simultaneous determination of 7 active components(tetrahydropalmatine,berberine hydrochloride,phellodendrine chloride,magnoflorine,cyasterone,liquiritin and glycyrrhetinic acid)in Jiawei Simiao particles by HPLC-DAD method with wavelength switching. Methods:The separation was performed on a Insert Sustain C_(18)(250 mm×4.6 mm,5 μm)column. The mobile phase was composed of acetonitrile(A)-0.05 mol·L~(-1) potassium dihydrogen phosphate(B)with gradient elution. The detection wavelength was set at 268 nm(0-22.5 min,detection of magnoflorine,67-71 min,detection of berberine hydrochloride),280 nm(22.5-30 min,detection of tetrahydropalmatine)and 237 nm(30-45 min,detection of liquiritin,247 nm(45-55 min,detection of cyasterone,71-85 min,detection of glycyrrhetinic acid)and 284 nm(55-67 min,detection of phellodendrine chloride). Results:The ingredients could be effectively detected and separated. For tetrahydropalmatine,berberine hydrochloride,phellodendrine chloride,magnoflorine,cyasterone,liquiritin and glycyrrhetinic acid,the linear ranges were 2.060-51.49 μg·mL~(-1)(r=1.000),3.111-77.77 μg·mL~(-1)(r=1.000),1.813-45.31 μg·mL~(-1)(r=1.000),1.960-49.00 μg·mL~(-1)(r=0.999 9),2.019-50.48 μg·mL~(-1)(r=1.000),2.468-61.70 μg·mL~(-1)(r=0.999 9)and 2.408-60.20 μg·mL~(-1)(r=0.999 9),respectively. The average recovery rate(n=6)were 100.6%(RSD=1.6%),100.3%(RSD=0.4%),101.0%(RSD=1.3%),100.7%(RSD=0.8%),100.1%(RSD=1.9%),101.6%(RSD=1.3%)and 99.8%(RSD=1.7%),respectively. Conclusion:The assay method established in this experiment met the requirements of methodological verification. The method was simple,reliable and reproducible. It could be used for simultaneous determination of 7 active components in Jiawei Simiao particles.
Objective:To establish a method for simultaneous determination of 7 active components(tetrahydropalmatine,berberine hydrochloride,phellodendrine chloride,magnoflorine,cyasterone,liquiritin and glycyrrhetinic acid)in Jiawei Simiao particles by HPLC-DAD method with wavelength switching. Methods:The separation was performed on a Insert Sustain C_(18)(250 mm×4.6 mm,5 μm)column. The mobile phase was composed of acetonitrile(A)-0.05 mol·L~(-1) potassium dihydrogen phosphate(B)with gradient elution. The detection wavelength was set at 268 nm(0-22.5 min,detection of magnoflorine,67-71 min,detection of berberine hydrochloride),280 nm(22.5-30 min,detection of tetrahydropalmatine)and 237 nm(30-45 min,detection of liquiritin,247 nm(45-55 min,detection of cyasterone,71-85 min,detection of glycyrrhetinic acid)and 284 nm(55-67 min,detection of phellodendrine chloride). Results:The ingredients could be effectively detected and separated. For tetrahydropalmatine,berberine hydrochloride,phellodendrine chloride,magnoflorine,cyasterone,liquiritin and glycyrrhetinic acid,the linear ranges were 2.060-51.49 μg·mL~(-1)(r=1.000),3.111-77.77 μg·mL~(-1)(r=1.000),1.813-45.31 μg·mL~(-1)(r=1.000),1.960-49.00 μg·mL~(-1)(r=0.999 9),2.019-50.48 μg·mL~(-1)(r=1.000),2.468-61.70 μg·mL~(-1)(r=0.999 9)and 2.408-60.20 μg·mL~(-1)(r=0.999 9),respectively. The average recovery rate(n=6)were 100.6%(RSD=1.6%),100.3%(RSD=0.4%),101.0%(RSD=1.3%),100.7%(RSD=0.8%),100.1%(RSD=1.9%),101.6%(RSD=1.3%)and 99.8%(RSD=1.7%),respectively. Conclusion:The assay method established in this experiment met the requirements of methodological verification. The method was simple,reliable and reproducible. It could be used for simultaneous determination of 7 active components in Jiawei Simiao particles.
引文
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[10]朱彩云,夏道宗,金晓敏,等.HPLC-DAD波长切换法建立加味三妙丸的指纹图谱[J].药物分析杂志,2016,36(8):1377ZHU CY,XIA DZ,JIN XM,et al.Wavelength switching method by HPLC-DAD is used to establish the fingerprint of the added flavor trimaru pill[J].Chin J Pharm Anal,2016,36(8):1377
[11]孙磊.川牛膝HPLC指纹图谱构建与药材质量评价研究[D].四川:四川农业大学,2005SUN L.A Study on the Establishment of HPLC Fingerprint and the Quality Evaluation of Medicinal Materials[D].Sichuan:Sichuan Agricultural University,2015
[12]于霞,吴娟,姜雪,等.HPLC法同时测定消乳散中甘草苷、橙皮苷和甘草酸铵的含量[J].沈阳药科大学学报,2017,34(5):369YU X,WU J,JIANG X,et al.HPLC method was used to determine the content of glycyrrhizin,hesperidin and ammonium glycyrrhizin[J].J Shenyang Pharm Univ,2017,34(5):369
[13]杨铭,李嘉旗,焦丽静,等.基于复杂网络结合生存分析的中医药治疗肺癌的核心有效处方的发现研究[J].中国中药杂志,2015,40(22):4482YANG M,LI JQ,JIAO LJ,et al.A study on the core effective prescription of TCM treatment for lung cancer based on complex network and survival analysis[J].J Chin Mater Med,2015,40(22):4482
[14]周旭,郭倩,白璐.数据挖掘技术在中医处方经验研究中的应用[J].电子产品世界,2016,2-3:37ZHOU X,GUO Q,BAI L.The application of data mining technique in traditional Chinese medicine prescription experience[J].Electronic Eng Prod World,2016,2-3:37
[15]石玲,邓琳雯,魏绍斌.湿热瘀结型盆腔炎中医综合治疗方案的标准化研究[J].时珍国医国药,2015,26(12):2961SHI L,DENG LW,WEI SB.Study on the standardization of TCM comprehensive treatment of dampness and heat stasis[J].Lishizhen Med Mater Med Res,2015,26(12):2961
[16]宋英,黄永亮,盛蓉,等.医疗机构中药制剂系列研发的规划与实践[J].中国实验方剂学杂志,2016,22(7):19SONG Y,HUANG YL,SHENG R,et al.Planning and practice of the research and development of TCM preparations for medical institutions[J].J Exp Chin Med,2016,22(7):19
[17]刘莉,袁强华,宋英.HPLC波长切换测定小儿鼻敏口服液中9个成分的含量[J].药物分析杂志,2016,36(11):1981LIU L,YUAN QH,SONG Y.Wavelength switch by HPLC was used to determine the content of 9 components in the oral liquid[J].Chin J Pharm Anal,2016,36(11):1981
[2]徐正莉.陈伟隆临床运用大血藤验案[J].四川中医,2012,30(10):117XU ZL.The clinical application of Sargentodoxa cuneate case by Chenweilong[J].Sichuan Tradit Chin Med,2012,30(10):117
[3]邱鹏程.元胡止痛片治疗溃疡型胃痛的物效机制[D].陕西:第四军医大学博士论文,2013QIU PC.The effect mechanism of Yuanhu pain-relieving tablet in treating ulcerative stomachache[D].Shaanxi:Doctoral Thesis of The Fourth Military Medical University,2013
[4]邱鹏程,张笑恺,王四旺.元胡止痛方的研究进展[J].时珍国医国药,2015,26(12):2993QIU PC,ZHANG XK,WANG SW.Progress in the study of Yuanhu pain prescription[J].Lishizhen Med Mater Med Res,2015,26(12):2993
[5]杨清,杨明.四妙散加减治疗妇科疾病举隅[J].中国医学创新,2014,11(7):113YANG Q,YANG M.Simiaosan treat gynecological diseases for example[J].Chin Med Innov,2014,11(7):113
[6]文建飞.王维英应用四妙散的临床经验[J].陕西中医学院学报,2013,36(2):33WEN JF.The clinical experience of the application of simiaosan by Wang Wei-ying[J].J Shaanxi Coll Tradit Chin Med,2013,36(2):33
[7]李秋月.延胡索生物碱谱-效关系和相互作用研究[D].北京:北京协和医学院,2014LI QY.Study on the Relationship and Interaction of the alkaloid spectrum of Corydalis Rhizoma[D].Beijing:Doctoral Dissertation of Peking Union Medical College,2014
[8]张振巍,石磊.RP-HPLC法测定元胡饮片中延胡索乙素的含量[J].实用医药杂志,2012,29(2):131ZHANG ZW,SHI L.The content of tetrahydroxanthine in Yuanhu herbal was determined by RP-HPLC method[J].2012,29(2):131
[9]杨静,马斐,李海霞,等.元胡止痛片中延胡索乙素和欧前胡素的含量测定[J].中国药师,2014,17(9):1512YANG J,MA F,LI HX,et al.Determination of the content of tetrahydroxanthine and eproxil in the pain-relieving tablets of Yuanhu[J].China Pharm,2014,17(9):1512
[10]朱彩云,夏道宗,金晓敏,等.HPLC-DAD波长切换法建立加味三妙丸的指纹图谱[J].药物分析杂志,2016,36(8):1377ZHU CY,XIA DZ,JIN XM,et al.Wavelength switching method by HPLC-DAD is used to establish the fingerprint of the added flavor trimaru pill[J].Chin J Pharm Anal,2016,36(8):1377
[11]孙磊.川牛膝HPLC指纹图谱构建与药材质量评价研究[D].四川:四川农业大学,2005SUN L.A Study on the Establishment of HPLC Fingerprint and the Quality Evaluation of Medicinal Materials[D].Sichuan:Sichuan Agricultural University,2015
[12]于霞,吴娟,姜雪,等.HPLC法同时测定消乳散中甘草苷、橙皮苷和甘草酸铵的含量[J].沈阳药科大学学报,2017,34(5):369YU X,WU J,JIANG X,et al.HPLC method was used to determine the content of glycyrrhizin,hesperidin and ammonium glycyrrhizin[J].J Shenyang Pharm Univ,2017,34(5):369
[13]杨铭,李嘉旗,焦丽静,等.基于复杂网络结合生存分析的中医药治疗肺癌的核心有效处方的发现研究[J].中国中药杂志,2015,40(22):4482YANG M,LI JQ,JIAO LJ,et al.A study on the core effective prescription of TCM treatment for lung cancer based on complex network and survival analysis[J].J Chin Mater Med,2015,40(22):4482
[14]周旭,郭倩,白璐.数据挖掘技术在中医处方经验研究中的应用[J].电子产品世界,2016,2-3:37ZHOU X,GUO Q,BAI L.The application of data mining technique in traditional Chinese medicine prescription experience[J].Electronic Eng Prod World,2016,2-3:37
[15]石玲,邓琳雯,魏绍斌.湿热瘀结型盆腔炎中医综合治疗方案的标准化研究[J].时珍国医国药,2015,26(12):2961SHI L,DENG LW,WEI SB.Study on the standardization of TCM comprehensive treatment of dampness and heat stasis[J].Lishizhen Med Mater Med Res,2015,26(12):2961
[16]宋英,黄永亮,盛蓉,等.医疗机构中药制剂系列研发的规划与实践[J].中国实验方剂学杂志,2016,22(7):19SONG Y,HUANG YL,SHENG R,et al.Planning and practice of the research and development of TCM preparations for medical institutions[J].J Exp Chin Med,2016,22(7):19
[17]刘莉,袁强华,宋英.HPLC波长切换测定小儿鼻敏口服液中9个成分的含量[J].药物分析杂志,2016,36(11):1981LIU L,YUAN QH,SONG Y.Wavelength switch by HPLC was used to determine the content of 9 components in the oral liquid[J].Chin J Pharm Anal,2016,36(11):1981