消栓肠溶胶囊的HPLC指纹图谱及5个活性成分的测定
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摘要
目的:建立消栓肠溶胶囊HPLC指纹图谱并测定苦杏仁苷、芍药苷、阿魏酸、毛蕊异黄酮苷和藁本内酯的含量,为消栓肠溶胶囊的质量控制提供参考。方法:采用Diamonsil C_(18)(2)色谱柱(4.6 mm×250 mm,5μm),以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱(0～5 min,8%A;5～12 min,8%A→15%A;12～20 min,15%A→37%A;20～28 min,37%A;20～45 min,37%A→50%A;45～65 min,50%A→65%A),流速1.0 mL·min~(-1),检测波长220 nm(0～30 min)和310 nm(30～65 min),柱温35℃。结果:在特征图谱研究中,共确定消栓肠溶胶囊HPLC指纹图谱21个共有峰,通过与混合对照品比较,指认其中5个指标成分:苦杏仁苷(2号峰)、芍药苷(10号峰)、阿魏酸(14号峰)、毛蕊异黄酮苷(17号峰)和藁本内酯(20号峰),利用相似度软件对12批样品指纹图谱进行分析,各批样品相似度均在0.99以上。在建立的色谱条件下,5个成分分离度良好,方法精密度和重复性RSD均<2.0%,供试品溶液在24 h内稳定,各成分具有良好的线性关系和较宽线性范围;12批消栓肠溶胶囊中苦杏仁苷、芍药苷、阿魏酸、毛蕊异黄酮苷和藁本内酯质量分数分别为0.27～0.39、2.54～2.66、2.71～2.79、0.44～0.56和1.47～1.59 mg·g~(-1)。结论:所建立的消栓肠溶胶囊HPLC指纹图谱和定量测定分析方法快速、准确,灵敏度高,专属性强,可以有效地评价该制剂的质量。
Objective:To establish the HPLC fingerprint and determine the contents of amygdalin,paeoniflorin,ferulic acid,calycosin and ligustilide in Xiaoshuan enteric-coated caplsules,so as to provide the reference for quality control of Xiaoshuan enteric-coated capsules.Methods:The analysis was carried out on an analytical Diamonsil C_(18)(2) column(4.6 mm×250 mm,5 μm)with gradient elution of acetonitrile(A)-0.1% phosphoric acid solution(B)(0-5 min,8%A;5-12 min,8%A → 15%A;12-20 min,15%A → 37%A;20-28 min,37%A; 20-45 min,37%A → 50%A;45-65 min,50%A → 65%A).The detection wavelengths were 220 nm(0-30 min)and 310 nm(30-65 min)and the flow rate was 1.0 mL·min~(-1).The column temperature was 35 ℃.Results:The common mode of the fingerprint was set up with 21 common peaks,and 5 of them were identified by comparison with the reference. They were amygdalin(peak 2),paeoniflorin(peak 10),ferulic acid(peak 14),calycosin(peak17) and ligustilide(peak 20).The similar degrees of 12 batches of samples were over 0.99.The 5 compounds were well separated under the determined chromatographic conditions. The RSDs of precision and repeatability experiments were all less than 2.0%. And the sample solution was stable during 24 h. All the compounds had good linearity and wide linear range.The content of amygdalin,paeoniflorin,ferulic acid,calycosin and ligustilide in 12 batches of Xiaoshuan enteric-coated caplsules were 0.27-0.39 mg·g~(-1),2.54-2.66 mg·g~(-1),2.71-2.79 mg·g~(-1),0.44-0.56 mg·g~(-1) and 1.47-1.59 mg·g~(-1),respectively.Conclusion:The established method is fast,accurate,sensitive and specific,and can be used for the quality control of Xiaoshuan enteric-coated caplsules.
Objective:To establish the HPLC fingerprint and determine the contents of amygdalin,paeoniflorin,ferulic acid,calycosin and ligustilide in Xiaoshuan enteric-coated caplsules,so as to provide the reference for quality control of Xiaoshuan enteric-coated capsules.Methods:The analysis was carried out on an analytical Diamonsil C_(18)(2) column(4.6 mm×250 mm,5 μm)with gradient elution of acetonitrile(A)-0.1% phosphoric acid solution(B)(0-5 min,8%A;5-12 min,8%A → 15%A;12-20 min,15%A → 37%A;20-28 min,37%A; 20-45 min,37%A → 50%A;45-65 min,50%A → 65%A).The detection wavelengths were 220 nm(0-30 min)and 310 nm(30-65 min)and the flow rate was 1.0 mL·min~(-1).The column temperature was 35 ℃.Results:The common mode of the fingerprint was set up with 21 common peaks,and 5 of them were identified by comparison with the reference. They were amygdalin(peak 2),paeoniflorin(peak 10),ferulic acid(peak 14),calycosin(peak17) and ligustilide(peak 20).The similar degrees of 12 batches of samples were over 0.99.The 5 compounds were well separated under the determined chromatographic conditions. The RSDs of precision and repeatability experiments were all less than 2.0%. And the sample solution was stable during 24 h. All the compounds had good linearity and wide linear range.The content of amygdalin,paeoniflorin,ferulic acid,calycosin and ligustilide in 12 batches of Xiaoshuan enteric-coated caplsules were 0.27-0.39 mg·g~(-1),2.54-2.66 mg·g~(-1),2.71-2.79 mg·g~(-1),0.44-0.56 mg·g~(-1) and 1.47-1.59 mg·g~(-1),respectively.Conclusion:The established method is fast,accurate,sensitive and specific,and can be used for the quality control of Xiaoshuan enteric-coated caplsules.
引文
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[8]刘杰,陈琳,范彩荣,等.基于HPLC-DAD-Q-TOF-MS/MS的白芍和赤芍主要成分定性定量研究[J].中国中药杂志,2015,40(9):1762LIU J,CHEN L,FAN CR,et al.Qualitative and quantitative analysis of major constituents of Paeoniae Radix Alba and Paeoniae Radix Rubra by HPLC-DAD-Q-TOF-MS/MS[J].China J Chin Mater Med,2015,40(9):1762
[9]张翠英,章洪,戚琼华.川芎的有效成分及药理研究进展[J].辽宁中医杂志,2014,41(10):2264ZHANG CY,ZHANG H,QI QH.Advance on active ingredients and pharmacological action of Chuanxiong Rhizoma[J].Liaoning JTradit Chin Med,2014,41(10):2264
[10]田璐,闫海霞,傅欣彤,等.一测多评法同时测定川芎、当归饮片中多种化学成分的含量[J].药物分析杂志,2014,34(5):848TIAN L,YAN HX,FU XT,et al.Simultaneous determination of multiple components in decoction pieces of Chuanxiong Rhizoma and Angelicae Sinensis Radix by quantitative analysis of multicomponents by single maker[J].Chin J Pharm Anal,2014,34(5):848
[11]许筱凰,李婷,王一涛,等.桃仁的研究进展[J].中草药,2015,46(17):2649XU XH,LI T,WANG YT,et al.Research progress in Persicae Semen[J].Chin Tradit Herb Drugs,2015,46(17):2649
[12]李强,杜思邈,张忠亮,等.中药指纹图谱技术进展及未来发展方向展望[J].中草药,2013,44(22):3095LI Q,DU SM,ZHANG ZL,et al.Progress in fingerprint technology on Chinese materia medica and prospect of its future development[J].Chin Tradit Herb Drugs,2013,44(22):3095
[13]孙国祥,闫波,侯志飞,等.中药色谱指纹图谱评价方法研究进展[J].中南药学,2015,13(7):673SUN GX,YAN B,HOU ZF,et al.Advances in evaluating chromatographic fingerprints of traditional Chinese medicine[J].Central South Pharm,2015,13(7):673
[14]刘桂花,邢建国,何承辉,等.反相高效液相色谱法同时测定芪香胶囊中红景天苷、芍药苷及毛蕊异黄酮葡萄糖苷的含量[J].中国医院药学杂志,2017,37(15):1440LIU GH,XIN JG,HE CH,et al.Simultaneous determination of salidroside,paeoniflorin and calycosin-7-O-β-D-glucoside in Qixiang capsules by HPLC[J].Chin Hosp Pharm J,2017,37(15):1440
[15]刘国红,赵路,吴纯伟,等.HPLC法测定桂枝茯苓丸有效部位中苦杏仁苷、芍药苷的含量[J].广东药学院学报,2015,31(1):50LIU GH,ZHAO L,WU CW,et al.Determination of amygdali and paeoniflorin of the active fraction in Guizhi Fuling pills by HPLC[J].J Guangdong Pharm Univ,2015,31(1):50
[2]徐东勋,伍雪英,何佳,等.消栓肠溶胶囊对脑梗死急性期神经功能及PAF、ET、NO的影响[J].中药药理与临床,2016,32(3):188XU DX,WU XY,HE J,et al.Effects of Xiaoshuan enteric-coated caplsules on neurological function and PAF,ET,NO with acute cerebral infarction[J].Pharmacol Clin Chin Mater Med,2016,32(3):188
[3]孙政华,邵晶,郭玫.黄芪化学成分及药理作用研究进展[J].中医临床研究,2015,7(25):22SUN ZH,SHAO J,GUO M.A review on chemical components and pharmacological effects of Huangqi[J].Clin J Chin Med,2015,7(25):22
[4]梁瑾,刘小花,任远,等.HPLC-DAD-ELSD法同时测定黄芪中5个成分的含量[J].药物分析杂志,2013,33(2):210LAING J,LIU XH,REN Y,et al.Simultaneous determination of five compounds in Radix Astragali by high-performance liquid chromatography coupled with diode arry and evaporative light scattering detectors[J].Chin J Pharm Anal,2013,33(2):210
[5]温学逊,牛研,王书芳.当归不同药用部位活性成分含量的HPLC分析方法及特征图谱研究[J].药物分析杂志,2014,34(2):317WEN XS,NIU Y,WANG SF.HPLC analysis and characteristic chromatograms of active components in different medicinal parts of Angelicae sinensis radix[J].Chin J Pharm Anal,2014,34(2):317
[6]尹辉.当归化学成分及药理活性研究进展[J].重庆科技学院学报(自然科学版),2015,17(1):100YIN H.The latest research progress on chemical composition and pharmacological action of angelica[J].J Chongqing Univ Sci Technol(Nat Sci Ed),2015,17(1):100
[7]陆小华,马骁,王建,等.赤芍的化学成分和药理作用研究进展[J].中草药,2015,46(4):595LU XH,MA X,WANG J,et al.Research progress on chemical constituents of Paeoniae Rubra Radix and their pharmacological effects[J].Chin Tradit Herb Drugs,2015,46(4):595
[8]刘杰,陈琳,范彩荣,等.基于HPLC-DAD-Q-TOF-MS/MS的白芍和赤芍主要成分定性定量研究[J].中国中药杂志,2015,40(9):1762LIU J,CHEN L,FAN CR,et al.Qualitative and quantitative analysis of major constituents of Paeoniae Radix Alba and Paeoniae Radix Rubra by HPLC-DAD-Q-TOF-MS/MS[J].China J Chin Mater Med,2015,40(9):1762
[9]张翠英,章洪,戚琼华.川芎的有效成分及药理研究进展[J].辽宁中医杂志,2014,41(10):2264ZHANG CY,ZHANG H,QI QH.Advance on active ingredients and pharmacological action of Chuanxiong Rhizoma[J].Liaoning JTradit Chin Med,2014,41(10):2264
[10]田璐,闫海霞,傅欣彤,等.一测多评法同时测定川芎、当归饮片中多种化学成分的含量[J].药物分析杂志,2014,34(5):848TIAN L,YAN HX,FU XT,et al.Simultaneous determination of multiple components in decoction pieces of Chuanxiong Rhizoma and Angelicae Sinensis Radix by quantitative analysis of multicomponents by single maker[J].Chin J Pharm Anal,2014,34(5):848
[11]许筱凰,李婷,王一涛,等.桃仁的研究进展[J].中草药,2015,46(17):2649XU XH,LI T,WANG YT,et al.Research progress in Persicae Semen[J].Chin Tradit Herb Drugs,2015,46(17):2649
[12]李强,杜思邈,张忠亮,等.中药指纹图谱技术进展及未来发展方向展望[J].中草药,2013,44(22):3095LI Q,DU SM,ZHANG ZL,et al.Progress in fingerprint technology on Chinese materia medica and prospect of its future development[J].Chin Tradit Herb Drugs,2013,44(22):3095
[13]孙国祥,闫波,侯志飞,等.中药色谱指纹图谱评价方法研究进展[J].中南药学,2015,13(7):673SUN GX,YAN B,HOU ZF,et al.Advances in evaluating chromatographic fingerprints of traditional Chinese medicine[J].Central South Pharm,2015,13(7):673
[14]刘桂花,邢建国,何承辉,等.反相高效液相色谱法同时测定芪香胶囊中红景天苷、芍药苷及毛蕊异黄酮葡萄糖苷的含量[J].中国医院药学杂志,2017,37(15):1440LIU GH,XIN JG,HE CH,et al.Simultaneous determination of salidroside,paeoniflorin and calycosin-7-O-β-D-glucoside in Qixiang capsules by HPLC[J].Chin Hosp Pharm J,2017,37(15):1440
[15]刘国红,赵路,吴纯伟,等.HPLC法测定桂枝茯苓丸有效部位中苦杏仁苷、芍药苷的含量[J].广东药学院学报,2015,31(1):50LIU GH,ZHAO L,WU CW,et al.Determination of amygdali and paeoniflorin of the active fraction in Guizhi Fuling pills by HPLC[J].J Guangdong Pharm Univ,2015,31(1):50