~(31)P核磁共振内标法测定甘油磷酸胆碱的含量
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摘要
目的:建立核磁共振磷谱定量法(~(31)Pq NMR)测定三类新药甘油磷酸胆碱(glycerophosphocholine,GPC)中GPC的绝对含量。方法:选择磷酸三苯酯(triphenyl phosphate,TPP)为内标,以其谱峰(δ-17.0)为内标峰,待测物定量峰化学位移在δ0.56处,溶剂为MEOD-d_4。结果:通过考察NMR实验条件的影响,选择采样次数NS为32次,弛豫延迟时间D_1为20.0 s,脉冲宽度P_1为9.8μs。(样品/内标)质量比对NMR峰面积比的零截距标准曲线斜率与理论值非常接近,线性回归系数r=0.996 7,重复性实验测得RSD为0.27%;核磁共振法测得GPC99药物中GPC的含量为99.67%,在无基准物质作对照的情况下,比HPLC-MS外标法(94.57%)更准确。结论:采用~(31)P q NMR方可测定甘油磷酸胆碱的含量,且此方法简单易行、结果准确。
Objective:To determine the content of Glycerophosphocholine(GPC)in its tablet,a quantitative~(31)P nuclear magnetic resonance(~(31)P q NMR)approach was presented.Methods:Triphenyl phosphate(TPP)was chosen as an internal standard and its ~(31)P NMR peak atδ-17.0 as the internal standard peak.Quantitative NMR peak of GPC was selected atδ0.56.Deuterated methanol was used as solvent.Results:The optional conditions were selected as follows:number of scan(NS):32,relaxation delay time(D_1):20 s,90~o pulse width(P_1):9.8μs.A good linear relationship between weight ration and peak area ratio of the internal standard TPP to GPC with correlative coefficient of 0.996 7;Results:The content of GPC determined by~(31)P q NMR was 99.67%,which was more accurate than the results of HPLC-MS(94.57%).Conclusions:This method is specific,accurate,simple,and it is suitable for the quantitative determination of the reference substance content.This method can be used for the determination of the content of GPC.
Objective:To determine the content of Glycerophosphocholine(GPC)in its tablet,a quantitative~(31)P nuclear magnetic resonance(~(31)P q NMR)approach was presented.Methods:Triphenyl phosphate(TPP)was chosen as an internal standard and its ~(31)P NMR peak atδ-17.0 as the internal standard peak.Quantitative NMR peak of GPC was selected atδ0.56.Deuterated methanol was used as solvent.Results:The optional conditions were selected as follows:number of scan(NS):32,relaxation delay time(D_1):20 s,90~o pulse width(P_1):9.8μs.A good linear relationship between weight ration and peak area ratio of the internal standard TPP to GPC with correlative coefficient of 0.996 7;Results:The content of GPC determined by~(31)P q NMR was 99.67%,which was more accurate than the results of HPLC-MS(94.57%).Conclusions:This method is specific,accurate,simple,and it is suitable for the quantitative determination of the reference substance content.This method can be used for the determination of the content of GPC.
引文
[1]WALTER A,KORTH U,HILGERT M,et al.Glycerophosphocholine is elevated in cerebrospinal fluid of Alzheimer patients[J].Neurobiol Aging,2004,25(10):1299
[2]WURTMAN R.Biomarkers in the diagnosis and management of Alzheimer’s disease[J].Metabolism,2015,64:47
[3]赵艳艳,张洪斌,毛慧芳,等.甘油磷酸胆碱的鉴别与含量测定[J].安徽医药,2012,16(5):601ZHAO YY,ZHANG HB,MAO HF,et al.Identification and determination of a new drug glycerophosphocho Hne[J].Anhui Med Pharm J,2012,16(5):601
[4]PAULI GF,G?DECKE T,JAKI BU,et al.Quantitative 1H NMR:development and potential of an analytical method-an update[J].J Nat Products,2012,75(4):834
[5]王强,王茂田,张志权.核磁共振法定量测定替米考星含量[J].分析测试学报,2003,22(6):101WANG Q,WANG MT,ZHANG ZQ.Quantitative determination of tilmicosin by 1H NMR[J].J Instrum Anal,2003,22(6):101
[6]张琪,朱红波,吴先富,等.邻苯二甲酸酯类标准物质核磁共振定量方法的建立[J].药物分析杂志,2015,35(7):1287ZHANG Q,ZHU HB,WU XF,et al.Establishment of a quantitative1H-NM R method for phthalateesters standard material analysis[J].Chin J Pharm Anal,2015,35(7):1287
[7]刘阳,周颖,张才煜,等.核磁共振定量法测定脒基吡唑含量[J].药物分析杂志,2014,34(6):1087LIU Y,ZHOU Y,ZHANG CY,et al.Determination of the quantitative content of 1H-pyrazole-1-carboxamidine bynuclear magnetic resonance(q NMR)[J].Chin J Pharm Anal,2014,34(6):1087
[8]陈春丽,田兰,马晓丽,等.核磁共振内标法测定维生素C片中维生素C的含量[J].中国药房,2010,21(32):3063CHEN CL,TIAN L,MA XL,et al.Quantitative determination of vitamin C in vitamin C tablets by proton nuclear magnetic resonancewith internal standard method[J].China Pharm,2010,21(32):3063
[9]郭强胜,石高旗,宋巍,等.NMR定量测定片剂中的对乙酰氨基酚[J].分析测试学报,2012,31(1):117GUO QS,SHI GQ,SONG W,et al.Determination of paracetamol in tablets by NMR[J].J Instrum Anal,2012,31(1):117
[10]中国药典2015年版.四部[S].2015:52Ch P 2015.IV[S].2015:52
[11]黄挺,张伟,全灿,等.定量核磁共振法研究进展[J].化学试剂,2012,34(4):327HUANG T,ZHANG W,QUAN C,et al.Review on quantitative nuclearmagnetic resonance[J].Chem Reag,2012,34(4):327
[12]张芬芬,蒋孟虹,沈文斌,等.定量核磁共振(QNMR)技术及其在药学领域的应用进展[J].南京师范大学学报(工程技术版),2014,14(2):8ZHANG FF,JIANG MH,SHEN WB,et al.Progress in quantitative nuclear magnetic resonance technology in pharmaceutical applications[J].Nanjing Norm Univ(Eng Technol Ed),2014,14(2):8
[13]DEENTS A,HIBBERT DB,HOOK JM,WELLS RJ.Quantitative nuclear magnetic resonance spectrometry II.Purity of phosphorusbased agrochemicals glyphosate(N-(phosphonomethyl)-glycine)and profenofos(O-(4-bromo-2-chlorophenyl)O-ethyl S-propyl phosphorothioate)measured by 1H and 31P q NMR spectrometry[J].Anal Chim Acta,2002,474:125
[14]WEBER M,HELLRIEGEL C,RUECK A,et al.Method development in quantitative NMR towardsmetrologically traceable organic certified reference materialsused as 31P q NMR standards[J].Anal Bioanal Chem,2015,407:3115
[15]MOURA S,CARVALHO FG,de OLIVEIRA CDR,et al.q NMR:An applicable method for the determination of dimethyltryptaminein ayahuasca,a psychoactive plant preparation[J].Phytochem Let,2010,3:79
[2]WURTMAN R.Biomarkers in the diagnosis and management of Alzheimer’s disease[J].Metabolism,2015,64:47
[3]赵艳艳,张洪斌,毛慧芳,等.甘油磷酸胆碱的鉴别与含量测定[J].安徽医药,2012,16(5):601ZHAO YY,ZHANG HB,MAO HF,et al.Identification and determination of a new drug glycerophosphocho Hne[J].Anhui Med Pharm J,2012,16(5):601
[4]PAULI GF,G?DECKE T,JAKI BU,et al.Quantitative 1H NMR:development and potential of an analytical method-an update[J].J Nat Products,2012,75(4):834
[5]王强,王茂田,张志权.核磁共振法定量测定替米考星含量[J].分析测试学报,2003,22(6):101WANG Q,WANG MT,ZHANG ZQ.Quantitative determination of tilmicosin by 1H NMR[J].J Instrum Anal,2003,22(6):101
[6]张琪,朱红波,吴先富,等.邻苯二甲酸酯类标准物质核磁共振定量方法的建立[J].药物分析杂志,2015,35(7):1287ZHANG Q,ZHU HB,WU XF,et al.Establishment of a quantitative1H-NM R method for phthalateesters standard material analysis[J].Chin J Pharm Anal,2015,35(7):1287
[7]刘阳,周颖,张才煜,等.核磁共振定量法测定脒基吡唑含量[J].药物分析杂志,2014,34(6):1087LIU Y,ZHOU Y,ZHANG CY,et al.Determination of the quantitative content of 1H-pyrazole-1-carboxamidine bynuclear magnetic resonance(q NMR)[J].Chin J Pharm Anal,2014,34(6):1087
[8]陈春丽,田兰,马晓丽,等.核磁共振内标法测定维生素C片中维生素C的含量[J].中国药房,2010,21(32):3063CHEN CL,TIAN L,MA XL,et al.Quantitative determination of vitamin C in vitamin C tablets by proton nuclear magnetic resonancewith internal standard method[J].China Pharm,2010,21(32):3063
[9]郭强胜,石高旗,宋巍,等.NMR定量测定片剂中的对乙酰氨基酚[J].分析测试学报,2012,31(1):117GUO QS,SHI GQ,SONG W,et al.Determination of paracetamol in tablets by NMR[J].J Instrum Anal,2012,31(1):117
[10]中国药典2015年版.四部[S].2015:52Ch P 2015.IV[S].2015:52
[11]黄挺,张伟,全灿,等.定量核磁共振法研究进展[J].化学试剂,2012,34(4):327HUANG T,ZHANG W,QUAN C,et al.Review on quantitative nuclearmagnetic resonance[J].Chem Reag,2012,34(4):327
[12]张芬芬,蒋孟虹,沈文斌,等.定量核磁共振(QNMR)技术及其在药学领域的应用进展[J].南京师范大学学报(工程技术版),2014,14(2):8ZHANG FF,JIANG MH,SHEN WB,et al.Progress in quantitative nuclear magnetic resonance technology in pharmaceutical applications[J].Nanjing Norm Univ(Eng Technol Ed),2014,14(2):8
[13]DEENTS A,HIBBERT DB,HOOK JM,WELLS RJ.Quantitative nuclear magnetic resonance spectrometry II.Purity of phosphorusbased agrochemicals glyphosate(N-(phosphonomethyl)-glycine)and profenofos(O-(4-bromo-2-chlorophenyl)O-ethyl S-propyl phosphorothioate)measured by 1H and 31P q NMR spectrometry[J].Anal Chim Acta,2002,474:125
[14]WEBER M,HELLRIEGEL C,RUECK A,et al.Method development in quantitative NMR towardsmetrologically traceable organic certified reference materialsused as 31P q NMR standards[J].Anal Bioanal Chem,2015,407:3115
[15]MOURA S,CARVALHO FG,de OLIVEIRA CDR,et al.q NMR:An applicable method for the determination of dimethyltryptaminein ayahuasca,a psychoactive plant preparation[J].Phytochem Let,2010,3:79