質(zhì)譜講義4-5(解析-軟離子化)

1、第五節(jié)、軟離子化質(zhì)譜解析一、一般規(guī)律二、ESI質(zhì)譜多電荷離子峰去卷積法求分子量三、分子和離子間相互作用四、應(yīng)用示例一、一般規(guī)律 準(zhǔn)分子離子：MH+, MNa+, MK+, MLi+, MAg+ 碎片離子： (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質(zhì)譜多電荷離子峰去卷積法求分子量 P267三、分子和離子間相互作用F生物大分子非共價(jià)復(fù)合物 酶-底物，酶-抑制劑，藥物篩選F超分子化學(xué) 人工酶，人工受體，自組裝F主客體分子 手性識(shí)別，包絡(luò)物分 子 識(shí) 別 的 內(nèi) 在 機(jī) 制分 子 識(shí) 別 的 內(nèi) 在 機(jī) 制-

3、相互作用離子-相互作用范德華力作用氫鍵特定結(jié)構(gòu)和功能的分子體系NMR x-ray 熱量計(jì) 電位計(jì) 圓二色散 色譜SPR儀質(zhì)譜的優(yōu)勢(shì)（質(zhì)譜的優(yōu)勢(shì)（5S5S）C 軟電離技術(shù)（軟電離技術(shù)（softsoft）C 快速（快速（SpeedSpeed）C 靈敏（靈敏（sensitivitysensitivity）C 樣品量少（樣品量少（small amount of samplesmall amount of sample）C 化學(xué)計(jì)量比信息（化學(xué)計(jì)量比信息（stoichiometrystoichiometry）質(zhì)譜提供的分子識(shí)別信息v 分子和分子之間以和數(shù)量比作用v 分子識(shí)別的選擇性v 分子識(shí)別中非共價(jià)作

4、用力的大小v 直接反映分子識(shí)別的本質(zhì)1、以金屬離子輔助氣相手性識(shí)別研究.意義生命的起源生物功能的實(shí)質(zhì)手性藥物手性識(shí)別的機(jī)制手性識(shí)別的機(jī)制 具有手性的參考分子（主體分子）對(duì)要分析的目標(biāo)手性分子（客體分子）的識(shí)別作用。 形成了以弱鍵相連的復(fù)合物，是一個(gè)非對(duì)映異構(gòu)體 物理性質(zhì)有差異，自由能有差異。高靈敏、快速的手性質(zhì)譜分析方法高靈敏、快速的手性質(zhì)譜分析方法 一級(jí)質(zhì)譜的差異（同位素標(biāo)記的應(yīng)用） RPI=（IAL.R/Iinternal）/ （IAD.R/Iinternal） 二級(jí)質(zhì)譜的差異 R=(Ifragment/IAL.R)/ (Ifragment/IAD.R) 反應(yīng)平衡測(cè)定熱平衡函數(shù) 分子離子反

5、應(yīng) 二級(jí)質(zhì)譜裂解分支途徑比較 R=(Ifrgment1/ Ifrgment2) AL.R/ (Ifrgment1/ Ifrgment2) AD.R 1.1. L-色胺酸(A)對(duì)二苯甲酰酒石酸二正丁酯(T)的手性識(shí)別 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、鋅離子輔助L-色胺酸對(duì)龍腦的手性識(shí)別 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.鋅離子輔助萘普生對(duì)bicyclo3,3,0 octane-2,6-dinol的手性識(shí)別 分析目標(biāo)手性藥物 bicyclo3,3,0 octane-2,6-dinol （O）使用參考手性分子 （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、環(huán)糊精非共價(jià)復(fù)合物的質(zhì)譜研究2.1、在氣相中研究超分子開關(guān)超分子顏色變化現(xiàn)象： pH10的水溶液時(shí)，酚酞顯紅色 加入 -CD 顯無色，加入適量甲醇后顯紅色， 再滴加過量甲醇，紅色漸漸消失； 加入-CD 顏色不變； 加入-CD 顏色也不變意 義 提出了新的機(jī)理 該顏色變化的超分子開關(guān)有望用于微量或痕量有機(jī)污染物的監(jiān)測(cè) 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（無復(fù)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和雙氯芬酸鈉的包合行為研究 (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、電噴霧飛行時(shí)間質(zhì)譜研究、電噴霧飛行時(shí)間質(zhì)譜研究CDsCDs對(duì)偽麻黃堿的手性識(shí)別對(duì)偽麻黃堿的手性識(shí)別手性拆分劑： -cyclodextrin, -cyclodextrin, -cyclodextrin 研究結(jié)果： nozzle potential對(duì)手性識(shí)別有獨(dú)特的變化特征 三種CD對(duì)右旋體的識(shí)別能力明顯強(qiáng)于左旋體 陸豪杰、余陸豪杰、余翀翀?zhí)?、郭寅龍?zhí)臁⒐? * 化學(xué)學(xué)報(bào)化學(xué)學(xué)報(bào) 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陸豪杰、余陸豪杰、余翀翀?zhí)?、郭寅龍?zhí)?、郭寅? * 化學(xué)學(xué)報(bào)化學(xué)學(xué)報(bào) 2002;60(5):882-52002;60(5):882-5 Figure Mass spectrum of -cyclodextrin and pseudoephedrine (c) -CD-(d)-pseudoephedrine；（d）-CD-(dl)-pseudoephedrine陸豪杰、余陸豪杰

22、、余翀翀?zhí)?、郭寅龍?zhí)臁⒐? * 化學(xué)學(xué)報(bào)化學(xué)學(xué)報(bào) 2002;60(5):882-52002;60(5):882-5 Figure Mass spectrum of -cyclodextrin and pseudoephedrine (e) -CD-(d)-pseudoephedrine；（f）-CD-(dl)-pseudoephedrine；陸豪杰、余陸豪杰、余翀翀?zhí)?、郭寅龍?zhí)臁⒐? * 化學(xué)學(xué)報(bào)化學(xué)學(xué)報(bào) 2002;60(5):882-52002;60(5):882-5 3、蛋白質(zhì)復(fù)合物研究 (1) 溶菌酶與環(huán)糊精復(fù)合物的初步研究 (2) 細(xì)胞色素 b5與heme的復(fù)合 (3) 環(huán)糊精

23、對(duì)細(xì)胞色素 b5及其突變體穩(wěn)定性的研究雞蛋清溶菌酶-環(huán)糊精1:1和1：5濃度比形成復(fù)合物的ESI-TOFMS圖譜余翀?zhí)?、郭寅?等 化學(xué)學(xué)報(bào) 2001;59(4):615 3.13.2、細(xì)胞色素tb5及其突變體中多肽鏈對(duì)輔基heme的特征識(shí)別生物功能生物體內(nèi)的氧化還原反應(yīng)電子傳遞鏈的成員將高鐵血紅蛋白還原成正常血紅蛋白余翀?zhí)臁⒐?等 化學(xué)學(xué)報(bào) 2001;59(6):956-960YU Chong-Tian, GUO Yin-Long*, et al Chinese J of Chemistry 2002;20(12):1540-1545 Tb5及其突變體待測(cè)蛋白脫輔基蛋白分子量(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?zhí)?、郭寅?等 化學(xué)學(xué)報(bào) 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?zhí)?、郭寅?等 化學(xué)學(xué)報(bào) 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?zhí)?、郭寅?等 化學(xué)學(xué)報(bào) 2001;59(6):956-960YU Chong-Tian, GUO Yin-Long*, et al Chinese J of Chemistry 2002;20(12):1540-1545 電噴霧質(zhì)譜得出氣相穩(wěn)定性結(jié)果 F35y突變體弱于野生性 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 環(huán)糊精對(duì)野生型細(xì)胞色素b5穩(wěn)定性的影響環(huán)糊精對(duì)細(xì)胞色素b5四點(diǎn)突變體穩(wěn)定性的影響4、分子離子反應(yīng)和無