质谱讲义4-5(解析-软离子化)

1、第五节、软离子化质谱解析一、一般规律二、ESI质谱多电荷离子峰去卷积法求分子量三、分子和离子间相互作用四、应用示例一、一般规律 准分子离子：MH+, MNa+, MK+, MLi+, MAg+ 碎片离子： (MH+-H2O), (MH+-CO), (MH+-CO2),SCIOOOOONOOOHOMeOHR2R1HR3OOOOONOOHR2R1OOMeHR3OOOOONOOHR2R1HR3H+HH+ONR2+ONOHR2+(f)N+NOH+(b)(c)(d)(e)(a)H+ESIH+ON+HONOOHO MeOOOOOC2H5OOOHHH+HONOOOOOC2H5OOHHH+HONOOOOOC2

2、H5OOHOHH+ONOOOOC2H5OOHHOOOC2H5OOHOHONO(a)(b)(c)H2O(d)(e)H+HONOOOOOC2H5OOHOHO MeO(d)-(f)H+RRRRRRR2222222H2N CH C NH CH C NH CH C OHR1R2R3OOOa1a2b1b2c1c2x1x2y1y2z1z2by二、ESI质谱多电荷离子峰去卷积法求分子量 P267三、分子和离子间相互作用F生物大分子非共价复合物 酶-底物，酶-抑制剂，药物筛选F超分子化学 人工酶，人工受体，自组装F主客体分子 手性识别，包络物分 子 识 别 的 内 在 机 制分 子 识 别 的 内 在 机 制-

3、相互作用离子-相互作用范德华力作用氢键特定结构和功能的分子体系NMR x-ray 热量计 电位计 圆二色散 色谱SPR仪质谱的优势（质谱的优势（5S5S）C 软电离技术（软电离技术（softsoft）C 快速（快速（SpeedSpeed）C 灵敏（灵敏（sensitivitysensitivity）C 样品量少（样品量少（small amount of samplesmall amount of sample）C 化学计量比信息（化学计量比信息（stoichiometrystoichiometry）质谱提供的分子识别信息v 分子和分子之间以和数量比作用v 分子识别的选择性v 分子识别中非共价作

4、用力的大小v 直接反映分子识别的本质1、以金属离子辅助气相手性识别研究.意义生命的起源生物功能的实质手性药物手性识别的机制手性识别的机制 具有手性的参考分子（主体分子）对要分析的目标手性分子（客体分子）的识别作用。 形成了以弱键相连的复合物，是一个非对映异构体 物理性质有差异，自由能有差异。高灵敏、快速的手性质谱分析方法高灵敏、快速的手性质谱分析方法 一级质谱的差异（同位素标记的应用） RPI=（IAL.R/Iinternal）/ （IAD.R/Iinternal） 二级质谱的差异 R=(Ifragment/IAL.R)/ (Ifragment/IAD.R) 反应平衡测定热平衡函数 分子离子反

5、应 二级质谱裂解分支途径比较 R=(Ifrgment1/ Ifrgment2) AL.R/ (Ifrgment1/ Ifrgment2) AD.R 1.1. L-色胺酸(A)对二苯甲酰酒石酸二正丁酯(T)的手性识别 Figure 1 ESI-FTMS/MS spectra of TAH+ containing (1) L-T or (2) D-THao-Jie Lu,Yin-Long Guo* Journal of the American Society for Mass Spectrometry 2003;14(6):571-580Figure 2 ESI-FTMS/MS spectra

6、of TACu-H22+ containing (1) L-T or (2) D-T Hao-Jie Lu,Yin-Long Guo* Journal of the American Society for Mass Spectrometry 2003;14(6):571-580Figure 3 ESI-FTMS/MS spectra of TAZn-H22+ containing (1) L-T or (2) D-T Hao-Jie Lu,Yin-Long Guo* Journal of the American Society for Mass Spectrometry 2003;14(6

7、):571-5801.2、锌离子辅助L-色胺酸对龙脑的手性识别 Hao-Jie Lu,Yin-Long Guo* Anal Chim Acta 2003;482(1):1-7Figure ESI-FTMS/MS spectra of solutions of 50:50 water:methanol containing zinc() salt, L-tryptophan and (A)(+)-borneol or (B)(-)-borneolHao-Jie Lu,Yin-Long Guo* Anal Chim Acta 2003;482(1):1-7Figure ESI-FTMS/MS sp

8、ectra of solutions of 50:50 water:methanol containing zinc() salt, L-tryptophan and (A)(+)-borneol or (B)(-)-borneolHao-Jie Lu,Yin-Long Guo* Anal Chim Acta 2003;482(1):1-7Figure ESI-FTMS/MS spectra of solutions of 50:50 water:methanol containing zinc() salt, L-tryptophan and (A)(+)-borneol or (B)(-)

9、-borneolHao-Jie Lu,Yin-Long Guo* Anal Chim Acta 2003;482(1):1-71.3.锌离子辅助萘普生对bicyclo3,3,0 octane-2,6-dinol的手性识别 分析目标手性药物 bicyclo3,3,0 octane-2,6-dinol （O）使用参考手性分子 （S）萘普生 Yu Chongtian,Yin-Long Guo* Journal of the American Society for Mass Spectrometry 2004;(5):795-802 COOHH3COHOOHHHHHHOOHfigure ESI FT

10、MS tandem mass spectra of N2O2Zn(II)-H+ from solutions of 20:80(v:v) water:methonal containing zinc(II) salts, S-naproxen and (+)-Octane(upper spectrum) or (-)-Octane(lower spectrum). Yu Chongtian,Yin-Long Guo* Journal of the American Society for Mass Spectrometry 2004;(5):795-802 =2.40 the value “R

11、” of intensity ratio between different diastereomeric complex ions obtained in tetrameric ion or trimeric ion: from left to right are NO2Zn(II)-H+ to N2Ozn(II)-H+ in N2O2Zn(II)-H+, NOZn(II)-H+ to N2OZn(II)-H+ in N2OZn(II)-H+, NOZn(II)-H+ to O2Zn(II)-H+ in N2OZn(II)-H+ and NOZn(II)-H+ to NO2Zn(II)-H+

12、 in NO2Zn(II)-H+. ( white column represent these complex including (+)-O, and black column represent these complexes including (-)-O). N: S-Naproxen, O: 2,6-dihydroxy-biscyclio3.3.0 octane4 . 8 57 . 7 32 . 0 20 . 2 71 . 6 57 . 2 51 . 3 50 . 4 401234567895 7 7 / 6 6 54 3 5 / 6 6 54 3 5 / 5 2 34 3 5 /

13、 5 7 7value of R+-=H-OZn(II)- )(N)-(S)H-Zn(II)O)- )N(-(S)H-OZn(II)-(-)N)-(S)H-Zn(II)O)-N(-)-(S)2222/IIIIRRRchiral2、环糊精非共价复合物的质谱研究2.1、在气相中研究超分子开关超分子颜色变化现象： pH10的水溶液时，酚酞显红色 加入 -CD 显无色，加入适量甲醇后显红色， 再滴加过量甲醇，红色渐渐消失； 加入-CD 颜色不变； 加入-CD 颜色也不变意 义 提出了新的机理 该颜色变化的超分子开关有望用于微量或痕量有机污染物的监测 Hao-Jie Lu,Yin-Long Guo* A

14、nal Chim Acta 2003; 495(1) :1-10 Figure The ESI-TOF mass spectrum of the phenolphthalein with A、-CD（无复合峰） ；B、 -CD ；C、 -CDHao-Jie Lu,Yin-Long Guo* Anal Chim Acta 2003; 495(1) :1-10 Figure The ESI-TOF mass spectra of the 1 : 1 phenolphthalein-CD adduct at nozzle pottential 150 V(A) and 200 V(B)Hao-Jie

15、 Lu,Yin-Long Guo* Anal Chim Acta 2003; 495(1) :1-10 Figure The ESI-TOF mass spectra of the 1 : 1 phenolphthalein-CD adduct at nozzle pottential 150 V(A) and 200 V(B)Hao-Jie Lu,Yin-Long Guo* Anal Chim Acta 2003; 495(1) :1-10 Figure The ESI-FTMSn spectra of the parent ion phenolphthalein in the soluti

16、on (A)without methanol and with methanol (B) colored; (C) color decreased; (D) colorlessHao-Jie Lu,Yin-Long Guo* Anal Chim Acta 2003; 495(1) :1-10 Hao-Jie Lu,Yin-Long Guo* Anal Chim Acta 2003; 495(1) :1-10 苯酚式和醌式碎裂方式Figure 6 ESI-FTMSn spectrum of the parent ion of 1 : 1 phenolphthalein-CD adduct. (A

17、) isolation sweep attenuation and activation attenuation were 25 dB and 31 dB respectively; (B) only isolated the parent ion with isolation sweep attenuation as 25 dB; (C) isolation sweep attenuation and activation attenuation were 25 dB and 30 dB respectively2.22.2、CDs和双氯芬酸钠的包合行为研究 (1) Effect of Ca

18、vity Size on the Non-covalent Complex(2) Effect of Material Ratio on the Stoichiometric Ratio(3) Effect of the Nozzle Potential on the Stability of the Complexes(4) Effect of Solution pH Value on the Complex Structure of CD-DS Ping He, Haojie Lu, Yinlong Guo Analytical Letters 2003;36(2):491-508Figu

19、re 1. ESI-TOF mass spectrum of No.3 (in Table 1) in ethanol solution (-CD) Ping He, Haojie Lu, Yinlong Guo Analytical Letters 2003;36(2):491-508 Ping He, Haojie Lu, Yinlong Guo Analytical Letters 2003;36(2):491-508Figure 4. Effects of different nozzle potential on the complexes in the No.1 (-CD) Pin

20、g He, Haojie Lu, Yinlong Guo Analytical Letters 2003;36(2):491-5082.32.3、电喷雾飞行时间质谱研究、电喷雾飞行时间质谱研究CDsCDs对伪麻黄碱的手性识别对伪麻黄碱的手性识别手性拆分剂： -cyclodextrin, -cyclodextrin, -cyclodextrin 研究结果： nozzle potential对手性识别有独特的变化特征 三种CD对右旋体的识别能力明显强于左旋体 陆豪杰、余陆豪杰、余翀翀天、郭寅龙天、郭寅龙* * 化学学报化学学报 2002;60(5):882-52002;60(5):882-5 Fi

21、gure Mass spectrum of -cyclodextrin and pseudoephedrine (a) -CD-(d)-pseudoephedrine；（b）-CD-(dl)-pseudoephedrine陆豪杰、余陆豪杰、余翀翀天、郭寅龙天、郭寅龙* * 化学学报化学学报 2002;60(5):882-52002;60(5):882-5 Figure Mass spectrum of -cyclodextrin and pseudoephedrine (c) -CD-(d)-pseudoephedrine；（d）-CD-(dl)-pseudoephedrine陆豪杰、余陆豪杰

22、、余翀翀天、郭寅龙天、郭寅龙* * 化学学报化学学报 2002;60(5):882-52002;60(5):882-5 Figure Mass spectrum of -cyclodextrin and pseudoephedrine (e) -CD-(d)-pseudoephedrine；（f）-CD-(dl)-pseudoephedrine；陆豪杰、余陆豪杰、余翀翀天、郭寅龙天、郭寅龙* * 化学学报化学学报 2002;60(5):882-52002;60(5):882-5 3、蛋白质复合物研究 (1) 溶菌酶与环糊精复合物的初步研究 (2) 细胞色素 b5与heme的复合 (3) 环糊精

23、对细胞色素 b5及其突变体稳定性的研究鸡蛋清溶菌酶-环糊精1:1和1：5浓度比形成复合物的ESI-TOFMS图谱余翀天、郭寅龙*等 化学学报 2001;59(4):615 3.13.2、细胞色素tb5及其突变体中多肽链对辅基heme的特征识别生物功能生物体内的氧化还原反应电子传递链的成员将高铁血红蛋白还原成正常血红蛋白余翀天、郭寅龙*等 化学学报 2001;59(6):956-960YU Chong-Tian, GUO Yin-Long*, et al Chinese J of Chemistry 2002;20(12):1540-1545 Tb5及其突变体待测蛋白脱辅基蛋白分子量(Dalto

24、n)全蛋白分子量(Dalton)野生型Tb5946110077F35Y947710093V61E949110107V61H949910115V61K949010106V61Y952510141E44-48-56-D60A92449860Fig.1 Stereoscopic Cdrawing of cytochrome Tb5 F35Y superimposed with that of cytochrome Tb5. F35Y is shown as a thin line and cytochrome Tb5 as a thick line.Fig.2 Positive ion ESI-MS

25、 of wild type cytochrome Tb5 and its mutant F35Y in pure water. 余翀天、郭寅龙*等 化学学报 2001;59(6):956-960YU Chong-Tian, GUO Yin-Long*, et al Chinese J of Chemistry 2002;20(12):1540-1545 Fig. 3 Stereoscopic C drawing of cytochrome Tb5 V61H superimposed with that of cytochrome Tb5. V61H is shown as a thin lin

26、e and cytochrome Tb5 as a thick line.余翀天、郭寅龙*等 化学学报 2001;59(6):956-960YU Chong-Tian, GUO Yin-Long*, et al Chinese J of Chemistry 2002;20(12):1540-1545 Figure 4: positive ion ESI-MS of Four sites mutant at nozzle potential 70 volt. The labeling scheme for all spectra is as follows: a = apo-protein an

27、d h = holo-protein.余翀天、郭寅龙*等 化学学报 2001;59(6):956-960YU Chong-Tian, GUO Yin-Long*, et al Chinese J of Chemistry 2002;20(12):1540-1545 电喷雾质谱得出气相稳定性结果 F35y突变体弱于野生性 Wild typeV61H V61KV61YV61E E44,48,56,D60Awild type YU Chong-Tian, GUO Yin-Long*, et al Chinese J of Chemistry 2002;20(12):1540-1545 环糊精对野生型细胞色素b5稳定性的影响环糊精对细胞色素b5四点突变体稳定性的影响4、分子离子反应和无