固相萃取-定量核磁共振波谱法测定板蓝根饮片中的表告依春
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摘要
建立了固相萃取(SPE)-定量核磁共振波谱(qNMR)技术测定板蓝根饮片中有效成分表告依春含量的方法。样品用水超声提取两次,采用SPE对提取液进行富集浓缩,用q NMR测定表告依春的含量。考察了样品预处理和qNMR实验条件对测定结果的影响,选择氘代二甲基亚砜为溶剂,用基准试剂邻苯二甲酸氢钾标定的2,3,5-三碘苯甲酸为内标,选择脉冲宽度P_1=14.1μs,延迟时间d_1=5 s,扫描次数N_S=256为qNMR定量测定表告依春的最佳实验条件。表告依春的定量峰为δ5.365~5.399(H-7b,d,1H)。结果表明,日内测量精密度(RSD)为0.5%,日间精密度为0.8%,表告依春与三碘苯甲酸峰面积比与质量比的零截距标准曲线线性相关系数为0.9991,且斜率与理论值相符。根据响应值标准偏差和标准曲线斜率法确定此法测定表告依春的检测限(LOD)为0.05 mg/g;定量限(LOQ,S/N≥150)为0.19 mg/g。包括样品提取过程的表告依春的回收率为97.4%~101.7%。采用本方法测定板蓝根饮片中的表告依春的含量为<0.19~1.26 mg/g。研究结果表明,采用SPE进行富集,扩大了q NMR的应用范围,可用于低含量复杂样品的定量分析。
The method of quantitative nuclear magnetic resonance spectroscopy( qNMR) for determination of epigoitrin in Radix Isatidis was established based on solid phase extraction( SPE). The twice ultrasonic extraction method using pure water was used for fully extracting epigoitrin in sample,and then the extraction was enriched and concentrated by poly-Sery MCX SPE cartridge. The effect of sample pretreatment and qNMR experimental conditions was investigated. The qNMR experiment conditions were selected using DMSO as solvent,calibrated 2,3,5-triiodobenzoate as internal standard,and P_1( pulse width) = 14.1 μs,d_1( pulse delay time) = 5 s,N_S( number of scan) = 256. The ~1H-NMR peaks of δ 5. 365-5. 399( H-7b,d,1H) of epigoitrin were chosen as the quantitative peaks. Method validation was performed including precision( intraday precision RSD was 0. 5%,and the inter-day precision was 0. 8%),linearity( correlation coefficient r>0.9991),LOD( 0.05 mg/g,standard curve method) and LOQ( 0.19 mg/g,S/N≥150). The recoveries of the SPE-qNMR were 97.4%-101.7%. The result showed that the method was stable,accurate and reliable.With this method the epigoitrin in a real Radix Isatidis was determined to be < 0. 19-1. 26 mg/g. SPE combining with qNMR could extend the application field of qNMR,especially in the detection of low-content component in complex samples.
The method of quantitative nuclear magnetic resonance spectroscopy( qNMR) for determination of epigoitrin in Radix Isatidis was established based on solid phase extraction( SPE). The twice ultrasonic extraction method using pure water was used for fully extracting epigoitrin in sample,and then the extraction was enriched and concentrated by poly-Sery MCX SPE cartridge. The effect of sample pretreatment and qNMR experimental conditions was investigated. The qNMR experiment conditions were selected using DMSO as solvent,calibrated 2,3,5-triiodobenzoate as internal standard,and P_1( pulse width) = 14.1 μs,d_1( pulse delay time) = 5 s,N_S( number of scan) = 256. The ~1H-NMR peaks of δ 5. 365-5. 399( H-7b,d,1H) of epigoitrin were chosen as the quantitative peaks. Method validation was performed including precision( intraday precision RSD was 0. 5%,and the inter-day precision was 0. 8%),linearity( correlation coefficient r>0.9991),LOD( 0.05 mg/g,standard curve method) and LOQ( 0.19 mg/g,S/N≥150). The recoveries of the SPE-qNMR were 97.4%-101.7%. The result showed that the method was stable,accurate and reliable.With this method the epigoitrin in a real Radix Isatidis was determined to be < 0. 19-1. 26 mg/g. SPE combining with qNMR could extend the application field of qNMR,especially in the detection of low-content component in complex samples.
引文
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20 WANG Rui,YANG Hai-Ying,YANG Qi-Wei,HUANG Shan-Jun,WANG Zheng-Tao.Chinese Traditional and Herbal Drugs,2010,41(3):478-480王瑞,杨海英,杨琪伟,黄山君,王峥涛.中草药,2010,41(3):478-480
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24 YU Shan,GUO Qiang-Sheng,WANG Hui-Lin,GAO Jian-Ping,XU Xu.Chinese Journal of Analytical Chemistry,2015,43(1):69-74禹珊,郭强胜,王会琳,高建平,许旭.分析化学,2015,43(1):69-74
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27 Pieri V,Stuppner H.Planta Med.,2011,77(15):1756-1758
28 Li C Y,Xu H X,Han Q B,Wu T S.J.Chromatogr.A,2009,1216(11):2124-2129
29 Moura S,Ultramari M D A,Paula D M L D,Yonamine M,Pinto E.Toxicon,2009,53(5):578-583
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31 Wagner L,Kall C,Fridjonsson E O,May E F,Stanwix P L,Graham B F,Carroll M R J,Johns M L.Meas.Sci.Technol.,2016,27(10):105501
32 Godejohann M,Heintz L,Daolio C,Berset J D,Muff D.Environ.Sci.Technol.,2009,43(18):7055-7061
33 MA Li,SUN Qin,LI You,XIAO Xiao-He.Chinese Journal of Pharmaceutical Analysis,2010,30(9):1642-1645马莉,孙琴,李友,肖小河.药物分析杂志,2010,30(9):1642-1645
34 WANG Xiao-Liang,CHEN Ming-Hua,WANG Fang,BU Peng-Bin,LIN Sheng,ZHU Cheng-Gen,LI Yu-Huan,JIANG Jian-Dong,SHI Jian-Gong.China Journal of Chinese Materia Medica,2013,38(8):1172-1182王晓良,陈明华,王芳,卜鹏滨,林生,朱承根,李玉环,蒋建东,石建功.中国中药杂志,2013,38(8):1172-1182
35 Gardrat C,Latxague L,Picard J P.J.Heterocyclic Chem.,1990,27(3):811-812
36 Fan G,Zhang M Y,Zhou X D,Lai X R,Yue Q H,Tang C,Luo W Z,Zhang Y.Analytica Chimica Acta,2012,747:76-83
37 The British Pharmacopoeia Commission.British Pharmacopoeia(2013 Ed).London:The Stationary Office,2013:Appendix IIC Nuclear Magnetic Resonance Spectrometry
2 Pauli G F,G9decke T,Jaki B U,Lankin D C.J.Nat.Prod.,2012,75(4):834-851
3 Simmler C,Napolitano J G,Mc Alpine J B,Chen S N,Pauli G F.Curr.Opin.Biotechnol.,2014,25:51-59
4 LIU Ying,HU Chang-Qin.Chinese Journal of Pharmaceutical Analysis,2001,21(6):447-452刘英,胡昌勤.药物分析杂志,2001,21(6):447-452
5 Mahajan S,Singh I P.Magn.Reson.Chem.,2013,51(2):76-81
6 Liu X,Kolpak M X,Wu J,Leo G C.Anal.Chem.,2012,84(15):6914-6918
7 Liu N Q,Choi Y H,Verpoorte R,Kooy F.Phytochem.Anal.,2010.21(5):451-456
8 Yang M,Wang J,Kong L.J.Pharm.Biomed.Anal.,2012.70:87-93
9 Li Z Y,Welbeck E,Wang R F,Liu Q,Yang Y B,Chou G X,Bi K S,Wang Z T.Phytochem.Anal.,2015,26(1):8-14
10 Staneva J,Denkova P,Todorova M,Evstatieva L.Pharm.Biomed.Anal.,2011,54(1):94-99
11 Chauthe S K,Sharmal R J,Aqil F,Gupta R C,Singh I P.Phytochem.Anal.,2012,23(6):689-696
12 Barding Jr G A,Salditos R,Larive C K.Anal.Bioanal.Chem.,2012,404(4):1165-1179
13 Salem A A,Abdou I M,Saleh H A.J.AOAC Int.,2012,95(6):1644-1651
14 Moazzami A A,Andersson R E,Kamal-Eldin A.J.Nutr.,2007,137(4):940-944
15 National Pharmacopoeia Committee.The Pharmacopoeia of the People's Republic of China(Part IV,2015 Ed),Beijing:China Medical Science Press,2015:52-54,376国家药典委员会编.中华人民共和国药典(四部,2015年版).北京:中国医药科技出版社,2015:52-54,376
16 National Pharmacopoeia Committee.The Pharmacopoeia of the People's Republic of China(Part II,2010 Ed),Beijing:China Medical Science Press,2010:Appendix Page 81-83国家药典委员会.中华人民共和国药典(二部,2010年版).北京:中国医药科技出版社,2010:附录81-83
17 National Pharmacopoeia Committee.The Pharmacopoeia of the People's Republic of China(Part I,2015 Ed),Beijing:China Medical Science Press,2015:205-206国家药典委员会.中华人民共和国药典(一部,2015年版).北京:中国医药科技出版社,2015:205-206
18 HUANG He-Fei,LIU Hong-Bing,WANG Yan-Yan,ZHOU Gang,ZHANG Hua,HUANG Zhi-Xiong.Journal of Hunan Normal University(Medical Science),2014,11(2):46-48黄鹤飞,刘红兵,王燕燕,周刚,张烨,黄志雄.湖南师范大学学报(医学版),2014,11(2):46-48
19 XU Li-Hua,HUANG Fang,CHEN Ting,WU Jie.Chinese Journal of Natural Medicines,2005,3(6):359-360徐丽华,黄芳,陈婷,吴洁.中国天然药物,2005,3(6):359-360
20 WANG Rui,YANG Hai-Ying,YANG Qi-Wei,HUANG Shan-Jun,WANG Zheng-Tao.Chinese Traditional and Herbal Drugs,2010,41(3):478-480王瑞,杨海英,杨琪伟,黄山君,王峥涛.中草药,2010,41(3):478-480
21 ZHANG Ye,ZHANG Lei,WANG Yu.Chinese Journal of Pharmaceutical Analysis,2008,28(11):1848-1850张叶,张蕾,王玉.药物分析杂志,2008,28(11):1848-1850
22 AN Yi-Qiang,JIA Xiao-Bin,YUAN Hai-Jian,SUN E,XU Zhen-Zhen.China Journal of Chinese Materia Medica,2008,33(18):2074-2076安益强,贾晓斌,袁海建,孙娥,许珍珍.中国中药杂志,2008,33(18):2074-2076
23 NIE Li-Xing,WANG Gang-Li,DAI Zhong,LIN Rui-Chao.Chinese Journal of Chromatography,2010,28(10):1001-1004聂黎行,王钢力,戴忠,林瑞超.色谱,2010,28(10):1001-1004
24 YU Shan,GUO Qiang-Sheng,WANG Hui-Lin,GAO Jian-Ping,XU Xu.Chinese Journal of Analytical Chemistry,2015,43(1):69-74禹珊,郭强胜,王会琳,高建平,许旭.分析化学,2015,43(1):69-74
25 Andrade-Eiroa A,Canle M,Leroy-Cancellieri V,CerdàV.TRAC-Trends Anal.Chem.,2016,80:655-667
26 HE Jiu-Ming,DAI Dong-Mei,LIU Ying,SUN Rui-Xiang,ABLIZ Zeper.Chinese J.Anal.Chem.,2009,37(suppl):D185贺玖明,代冬梅,刘影,孙瑞祥,再帕尔·阿不力孜.分析化学,2009,37(增刊):D185
27 Pieri V,Stuppner H.Planta Med.,2011,77(15):1756-1758
28 Li C Y,Xu H X,Han Q B,Wu T S.J.Chromatogr.A,2009,1216(11):2124-2129
29 Moura S,Ultramari M D A,Paula D M L D,Yonamine M,Pinto E.Toxicon,2009,53(5):578-583
30 van der Hooft J J J,de Vos R C H,Mihaleva V,Bino R J,Ridder L,de Roo N,Jacobs D M,van Duynhoven J P M,Vervoort J.Anal.Chem.,2012,84(16):7263-7271
31 Wagner L,Kall C,Fridjonsson E O,May E F,Stanwix P L,Graham B F,Carroll M R J,Johns M L.Meas.Sci.Technol.,2016,27(10):105501
32 Godejohann M,Heintz L,Daolio C,Berset J D,Muff D.Environ.Sci.Technol.,2009,43(18):7055-7061
33 MA Li,SUN Qin,LI You,XIAO Xiao-He.Chinese Journal of Pharmaceutical Analysis,2010,30(9):1642-1645马莉,孙琴,李友,肖小河.药物分析杂志,2010,30(9):1642-1645
34 WANG Xiao-Liang,CHEN Ming-Hua,WANG Fang,BU Peng-Bin,LIN Sheng,ZHU Cheng-Gen,LI Yu-Huan,JIANG Jian-Dong,SHI Jian-Gong.China Journal of Chinese Materia Medica,2013,38(8):1172-1182王晓良,陈明华,王芳,卜鹏滨,林生,朱承根,李玉环,蒋建东,石建功.中国中药杂志,2013,38(8):1172-1182
35 Gardrat C,Latxague L,Picard J P.J.Heterocyclic Chem.,1990,27(3):811-812
36 Fan G,Zhang M Y,Zhou X D,Lai X R,Yue Q H,Tang C,Luo W Z,Zhang Y.Analytica Chimica Acta,2012,747:76-83
37 The British Pharmacopoeia Commission.British Pharmacopoeia(2013 Ed).London:The Stationary Office,2013:Appendix IIC Nuclear Magnetic Resonance Spectrometry