定量核磁共振法测定4种磷酸二酯酶-5抑制剂的含量

目的：建立定量核磁共振法（qNMR）测定那莫西地那非、豪莫西地那非、硫代艾地那非和伪伐地那非4种磷酸二酯酶-5（PDE-5）抑制剂的含量。方法：采用Bruker Ascend 500 MHz核磁共振仪采集一维定量氢谱，90°脉冲，延迟时间为20 s，采样次数为64次，测定温度25℃，以对苯二甲酸二甲酯为内标，氘代氯仿（CDCl₃）为溶剂，对4种PDE-5抑制剂进行定量研究。结果：样品与内标物定量峰分离良好，4种成分在各自线性范围内线性关系良好（R²>0.999 2），进样精密度和重复性好。那莫西地那非、豪莫西地那非、硫代艾地那非和伪伐地那非的含量（n=6）分别99.2%（RSD=0.27%）、99.4%（RSD=0.28%）、99.1%（RSD=0.18%）和99.3%（RSD=0.24%），与质量平衡法测定结果比较，两者基本一致。结论：本文建立的qNMR法测定4种PDE-5抑制剂的含量准确可靠，简便快速，为该类成分的含量测定和对照品的标定提供了新的测定方法。  

Objective:To establish a quantitative nuclear magnetic resonance(qNMR)method for the determination of norneosildenafil,homosildenafil,thioaildenafil,and pseudovardenafil. Methods:Proton nuclear magnetic resonance spectroscopy(¹H NMR)spectra were obtained in CDCl₃ with the internal standard of dimethyl terephthalate using a Bruker Ascend 500 spectrometer,the parameters were setup as follows:the pulse sequence was 90o,the relaxation delay time was 20 s,the number for scanning was 64,and the experiment temperature was controlled at 25. Results:The linearity of four components were linear in the concentration ranges(R²>0.999 2),and the precision,stability,and repeatability were good. The contents (n=6) of norneosildenafil,homosildenafil,thioaildenafil,and pseudovardenafil by qNMR were 99.2%(RSD=0.27%),99.4%(RSD=0.28%),99.1%(RSD=0.18%),and 99.3%(RSD=0.24%),respectively,which were almost consistent with those by mass balance method. Conclusion:The qNMR method is accurate,reliable,simple,and rapid for the quantitative determination of the above four phosphodiesterase 5(PDE-5)inhibitors. The study provides a new way for the content determination and calibration of reference materials. 

[1] 张立元,卢一平.5型磷酸二酯酶抑制剂在ED治疗及其他领域应用的研究现状和前景[J].中华医学杂志,2008,88(12):861 ZHANG LY,LU YP.Research status and prospects of phosphodiesterase 5 inhibitors in ED therapy and other fields[J].Natl Med J China,2008,88(12):861
[2] 潘涛,赵庆杰,吕小伟,等.磷酸二酯酶5抑制剂的研究进展[J].药学实践杂志,2002,20(6):365 PAN T,ZHAO QJ,Lü XW,et al.Research progress of phosphodiesterase 5 inhibitors[J].J Pharm Pract,2002,20(6):365
[3] 张石革,梁建华,马国辉.磷酸二酯酶5抑制剂的进展与评价[J].中国医院用药评价与分析,2004,4(3):133 ZHANG SG,LIANG JH,MA GH.Evaluation and research progress in inhibitor of phosphodiesterase type 5[J].Eval Anal Drug Use Hosp China,2004,4(3):133
[4] 陈昌亮,黄爽.磷酸二酯酶及其抑制剂的研究进展[J].中国药理学通报,2014,30(2):283 CHEN CL,HUANG S.Research progress of phosphodiesterases and phosphodiesterase inhibitors[J].Chin Pharmacol Bull,2014,30(2):283
[5] 陈吉汉,侯彩云,路勇,等.液相色谱-串联质谱法同时测定保健食品中四种违禁药物含量[J].食品安全质量检测学报,2014,5(7):2027 CHEN JH,HOU CY,LU Y,et al.Simultaneous determination of 4 kinds of illicit drugs in health-care foodstuffs by ultra performance liquid chromatography-mass spectrometry[J].J Food Safe Qual,2014,5(7):2027
[6] 郭常川,石峰,巩丽萍,等.Q-Orbitrap高分辨质谱用于中成药和保健食品中非法添加PDE-5抑制剂的快速筛查、确证和定量[C]//2013年中国药学大会暨第十三届中国药师周,南宁:2013 GUO CC,SHI F,GONG LP,et al.Application of a Q-Orbitrap high resolution mass spectrometry to the rapid screening,confirmation and quantitation of the illegal adulterated phosphodiesterase-5 inhibitors in herbal medicines and dietary supplements[C]//2013 Chinese Pharmaceutical Conference and the 13th Chinese Pharmacist Week Conference,Nanninig:2013
[7] SALEM AA,MOSSA HA,BARSOUM BN.Application of nuclear magnetic resonance spectroscopy for quantitative analysis of miconazole,metronidazole and sulfamethoxazole in pharmaceutical and urine samples[J].J Pharm Biomed Anal,2006,41(2):654
[8] MAGGIO RM,CALVO NL,VIGNADUZZO SE,et al.Pharmaceutical impurities and degradation products:Uses and applications of NMR techniques[J].J Pharm Biomed Anal,2014,101:102
[9] PAULI GF,JAKI BU,LANKIN DC.Quantitative ¹H NMR:development and potential of a method for natural products analysis[J].J Nat Prod,2005,68(1):133
[10] BHARTI SK,ROY R.Quantitative ¹H NMR spectroscopy[J].Trends Anal Chem,2012,355
[11] 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):8 ZHANG FF,JIANG MH,SHEN WB,et al.Progress in quantitative nuclear magnetic resonance technology in pharmaceutical applications[J].J Nanjing Norm Univ(Eng Technol Ed),2014,14(2):8
[12] WU Y,HE Y,HE WY,et al.Application of quantitative ¹H NMR for the calibration of protoberberine alkaloid reference standards[J].J Pharm Biomed Anal,2014,90:92
[13] LI ZY,WELBECK E,WANG RF,et al.A universal quantitative ¹H nuclear magnetic resonance(qNMR)method for assessing the purity of dammarane-type ginsenosides[J].Phytochem Anal,2015,26(1):8
[14] SHIN MH,HONG MK,KIM WS,et al.Identification of a new analogue of sildenafil added illegally to a functional food marketed for penile erectile dysfunction[J].Food Addit Contam,2003,20(9):793
[15] 张才煜,张娜,何兰.核磁共振内标法测定波生坦的含量[J].药学学报,2014,49(2):249 ZHANG CY,ZHANG N,HE L.Quantitative determination of bosentan by proton nuclear magnetic resonance with internal standard method[J].Acta Pharm Sin,2014,49(2):249
[16] SHEN S,YAO J,SHI Y.Quantitative proton nuclear magnetic resonance for the structural and quantitative analysis of atropine sulfate[J].J Pharm Biomed Anal,2014,89:118