本文選題：常壓質(zhì)譜技術(shù) + 解吸附電噴霧質(zhì)譜�。� 參考：《中國(guó)食品藥品檢定研究院》2014年碩士論文

【摘要】：近年來(lái)，隨著藥物-血漿蛋白親和力以及游離血藥濃度的研究領(lǐng)域的發(fā)展，研究者對(duì)其檢測(cè)手段不斷提出更高的要求，發(fā)展新的高通量、便捷的檢測(cè)方法意義重大。常壓質(zhì)譜技術(shù)能直接、實(shí)時(shí)、快速、原位的提供分子的結(jié)構(gòu)信息，將它應(yīng)用于藥物-血漿蛋白親和力以及游離血藥濃度的檢測(cè)中，能為其研究提供一條新的途徑。本論文著重研究了基于解吸附電噴霧離子源和文丘里簡(jiǎn)易常壓超聲噴霧離子源檢測(cè)藥物-血漿蛋白親和力以及游離血藥濃度的新方法。主要包括以下工作： 一、利用解吸附電噴霧質(zhì)譜（Desorption Electrospray Ionization MassSpectrometry, DESI-MS）解決了目前藥物-血漿蛋白研究存在的三方面的問題。第一，在負(fù)電區(qū)域抑制解離機(jī)理的基礎(chǔ)上，有負(fù)電位點(diǎn)的蛋白不會(huì)釋放特異性結(jié)合的配體，本身可以作為參比蛋白，省去了為每一種藥物尋找參比蛋白的過(guò)程。第二，提取離子流圖檢測(cè)的是藥物分子的準(zhǔn)分子離子峰，而不是藥物-蛋白復(fù)合物，這樣，氣相庫(kù)倫爆炸誘導(dǎo)解離可以起到一個(gè)積極的作用，而不是影響檢測(cè)的作用。第三，我們自行搭建的全自動(dòng)移動(dòng)平臺(tái)可以解決分析速度慢的問題。我們選擇僅有一個(gè)特異性結(jié)合位點(diǎn)的α1-酸性糖蛋白作為模式蛋白，先前文獻(xiàn)已經(jīng)報(bào)導(dǎo)了DMOA可以作為α1-酸性糖蛋白的置換劑。我們的工作分為三部分，第一，用（R）、（S）型氨氯地平作為模式配體，證明了負(fù)電區(qū)域抑制解離機(jī)理，它們與α1-酸性糖蛋白的立體選擇性特異性結(jié)合已經(jīng)被報(bào)導(dǎo)。第二，我們用5種α-腎上腺素受體阻滯劑和10種β-腎上腺素受體阻滯劑證明了我們方法的高通量屬性。我們?cè)?.3分鐘內(nèi)獲得了15*3個(gè)樣本點(diǎn)的全部數(shù)據(jù)信息。第三，幾乎不與α1-酸性糖蛋白發(fā)生作用的阿替洛爾和莫索尼定被選為內(nèi)標(biāo)物，以校正噴入離子傳輸管體積的差別，與先前的實(shí)驗(yàn)結(jié)果相一致，證明我們的方法至少是半定量的方法。 二、利用文丘里-簡(jiǎn)易常壓超聲噴霧質(zhì)譜（Venturi EasyAmbient Sonic-SprayIonization Mass Spectrometry, V-EASI-MS）建立了一種實(shí)時(shí)在線監(jiān)測(cè)藥物-負(fù)電位點(diǎn)蛋白結(jié)合的方法，體系由兩部分組成：商業(yè)化的線性離子阱（LTQ）質(zhì)量分析器和自行搭建的文丘里-簡(jiǎn)易常壓超聲噴霧離子源。我們的實(shí)驗(yàn)結(jié)果進(jìn)一步證明了我們首先提出并驗(yàn)證的負(fù)電區(qū)域抑制解離機(jī)理（Anionic region inhibiteddissociation mechanism）。機(jī)理描述了負(fù)電區(qū)域在特異性結(jié)合配體解離中的作用。為了區(qū)分特異性以及非特異性的相互作用，，我們首先在沒有特異性結(jié)合藥物解離的條件下評(píng)估了非特異性的相互作用。所以我們優(yōu)化了解吸附電噴霧直接分析的質(zhì)譜條件，避免特異性結(jié)合藥物解離。我們主要采用了兩個(gè)策略來(lái)防止特異性結(jié)合藥物從復(fù)合物上解離：第一，我們選擇負(fù)電位點(diǎn)蛋白——α1-酸性糖蛋白，作為模式蛋白。使用負(fù)電位點(diǎn)蛋白獲得質(zhì)譜信號(hào)強(qiáng)度與親和力的相關(guān)性比使用正電位點(diǎn)蛋白更好。第二，使用的文丘里-簡(jiǎn)易常壓超聲噴霧離子源是一種最軟電離源，有利于保護(hù)對(duì)照試驗(yàn)中的特異性相互作用。
[Abstract]:In recent years, with the development of the research field of drug plasma protein affinity and free blood concentration, researchers have constantly put forward higher requirements for their detection methods, and the development of new high throughput and convenient detection methods is of great significance. The atmospheric mass spectrometry technology can provide the structural information of molecules in real time, fast speed and in situ, and apply it to the system. A new approach for the detection of drug plasma protein affinity and free blood concentration can be provided for its research. This paper focuses on the study of new methods for detecting the affinity of plasma protein and the concentration of free blood drugs based on desorption ion source and Venturi simple atmospheric pressure ultrasonic spray ion source. Make:
First, Desorption Electrospray Ionization MassSpectrometry (DESI-MS) is used to solve the three problems in the current drug plasma protein study. First, on the basis of the mechanism of the negative region inhibition, the protein with negative potential will not release the specific binding ligand itself, which itself can be used as a reference. Specific protein, it saves the process of searching for the reference protein for every drug. Second, the extraction ion flow graph is the excimer ion peak of the drug molecule, not the drug protein complex, so that the gas phase Kulun explosion induced dissociation can play a positive role, not the effect of detection. Third, we build it by ourselves. The full automatic mobile platform can solve the problem of slow analysis. We choose only one specific binding site of alpha 1- acid glycoprotein as a pattern protein. Previous literature has reported that DMOA can be used as a replacement agent for alpha 1- acid glycoprotein. Our work is divided into three parts. First, (R), (S) type amlodipine as a pattern match. The body, which proved the mechanism of the negative region inhibition of dissociation, and their stereoselective binding with alpha 1- acid glycoprotein has been reported. Second, we demonstrated our method with 5 alpha adrenergic receptor blockers and 10 beta adrenergic receptor blockers. We obtained 15*3 sample points within 2.3 minutes. All data information. Third, atenolol and mol SONY, which almost do not interact with alpha 1- acid glycoproteins, were selected as the internal standard to correct the difference in the volume of the injected ion transfer tube. It was consistent with the previous experimental results, proving that our method is at least a semi quantitative method.
Two, a method of real-time online monitoring of drug negative potential protein binding is established by using Venturi simple atmospheric pressure Sonic-SprayIonization Mass Spectrometry (V-EASI-MS). The system consists of two parts: commercial linear off Zi Jing (LTQ) quality analyzer and self built Venturi - Our experimental results further demonstrate the mechanism of Anionic region inhibiteddissociation mechanism which we first proposed and verified. The mechanism describes the role of the negative region in the dissociation of specific binding ligands in order to distinguish specific and nonspecific phases. Interaction, we first evaluated the nonspecific interaction without specific combination of drug dissociation. Therefore, we optimized the mass spectrum conditions of the direct analysis of the adsorption electrospray to avoid the specific binding of drugs. We mainly adopted two strategies to prevent the dissociation of the specific binding drugs from the complex: First, we choose the negative potential point protein, alpha 1- acid glycoprotein, as a pattern protein. The use of negative potential protein to obtain the correlation between the intensity and affinity of the mass spectrum is better than the positive potential point protein. Second, the Venturi simple atmospheric pressure ultrasonic spray ion source used is the most soft ionization source, which is beneficial to the protection of the control trial. Specific interaction.
【學(xué)位授予單位】：中國(guó)食品藥品檢定研究院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：O657.63;R927

【共引文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 李霽;基于生物樣品在線富集技術(shù)的臨床毛細(xì)管電泳方法學(xué)研究[D];第二軍醫(yī)大學(xué);2013年

本文編號(hào)：2031162