本文選題：限進(jìn)性填料 + 高效液相色譜　； 參考：《蘭州大學(xué)》2014年碩士論文

【摘要】：生物樣品的前處理方法對(duì)體內(nèi)藥物分析來(lái)說(shuō)是至關(guān)重要的步驟,傳統(tǒng)的生物樣品前處理方法有蛋白沉淀法,離線(xiàn)固相萃取法(SPE),液液萃取法(LLE)等,這些方法不僅需要大量的有機(jī)試劑,還會(huì)因?yàn)樵陔x線(xiàn)狀態(tài)下操作,導(dǎo)致生物樣品中的待測(cè)物被稀釋,損失和污染,使得分析結(jié)果不準(zhǔn)確。此外,生物樣品因?yàn)榛|(zhì)復(fù)雜,含有大量的蛋白質(zhì)等生物大分子,所以對(duì)固相萃取柱污染嚴(yán)重,使分析費(fèi)用加大,不常用于色譜分析中。限進(jìn)性填料(RAM)是一種新型的樣品前處理材料,它具有兩個(gè)不同的表面,外表面兼具分子排阻功能和親水性,內(nèi)表面鍵合有不同的功能基團(tuán),因此,生物樣品中只有小分子物質(zhì)可以進(jìn)入內(nèi)表面并與內(nèi)表面上的功能基團(tuán)相互作用,進(jìn)而保留在限進(jìn)性填料柱上,而蛋白質(zhì)等大分子物質(zhì)無(wú)法進(jìn)入RAM的內(nèi)表面,又因?yàn)橥獗砻娴挠H水性,這些大分子物質(zhì)也不會(huì)吸附沉積在外表面上,而是被流動(dòng)相洗脫除去,因此,RAM通過(guò)柱切換技術(shù)與高效液相色譜(HPLC)或高效液相色譜-質(zhì)譜(HPLC-MS)聯(lián)用,可實(shí)現(xiàn)生物樣品的在線(xiàn)直接進(jìn)樣,相比于蛋白沉淀,SPE等方法,RAM能夠避免待測(cè)物的損失和污染,也能避免生物大分子物質(zhì)對(duì)萃取柱的損害。此外,RAM還能作為預(yù)富集柱富集生物樣品中的小分子物質(zhì)。 生物樣品中待測(cè)物的濃度通常較低,這是體內(nèi)藥物分析面臨的又一問(wèn)題,通常是利用HPLC-MS的高靈敏度來(lái)解決的,但是HPLC-MS存在價(jià)格昂貴,分析條件嚴(yán)格的問(wèn)題,所以,性能穩(wěn)定,易于操作的HPLC依然在體內(nèi)藥物分析中占有重要地位,但其靈敏度無(wú)法完全滿(mǎn)足體內(nèi)藥物分析的要求,因此,本文建立了RAM-HPLC法,通過(guò)大體積直接進(jìn)樣(LVI)的方式,考察了RAM柱對(duì)血漿、尿液、腦脊液中小分子藥物的富集能力,并在線(xiàn)富集檢測(cè)這些生物體液中的藥物濃度,有效提高了HPLC的檢測(cè)靈敏度,為生物樣品的測(cè)定提供簡(jiǎn)單、快速、準(zhǔn)確的在線(xiàn)分析方法。此外,我們還探討了RAM對(duì)藥物的保留機(jī)理,為更加有效利用RAM柱及合成新型RAM提供一定的實(shí)驗(yàn)依據(jù)。
[Abstract]:The pretreatment of biological samples is an important step for drug analysis in vivo. The traditional pretreatment methods of biological samples include protein precipitation, off-line solid phase extraction (SPE), liquid-liquid extraction (Lle) and so on. These methods not only require a large number of organic reagents, but also lead to dilution, loss and contamination of the substances to be tested in biological samples due to off-line operation, which makes the analytical results inaccurate. In addition, because of the complex matrix and the large amount of protein and other biological macromolecules, biological samples contaminate the solid phase extraction column seriously and increase the cost of analysis, so it is not often used in chromatographic analysis. Ram is a new kind of sample pretreatment material. It has two different surfaces. The outer surface has both molecular resistance and hydrophilicity, and the internal surface bonding has different functional groups. In biological samples, only small molecules can enter the inner surface and interact with the functional groups on the inner surface, and then remain on the confined packed column, while the protein and other macromolecular substances cannot enter the inner surface of RAM. Because of the hydrophilicity of the outer surface, these macromolecular substances are not adsorbed and deposited on the outer surface, but are eluted by mobile phase, so RAM is used in combination with HPLC or HPLC-MS by means of column switching technique. Compared with other methods such as protein precipitation and SPE, RAM can avoid the loss and pollution of the material to be tested, and the damage to the extraction column caused by biomolecules. In addition, Ram can also be used as a preconcentration column for the enrichment of small molecules in biological samples. The concentration of the substances to be tested in biological samples is usually low, which is another problem in drug analysis in vivo, which is usually solved by using the high sensitivity of HPLC-MS. However, HPLC-MS has the problems of high price and strict analytical conditions, so its performance is stable. The easy-to-operate HPLC still plays an important role in drug analysis in vivo, but its sensitivity can not fully meet the requirements of drug analysis in vivo. Therefore, the method of RAM-HPLC is established in this paper. The enrichment ability of RAM column on small molecular drugs in plasma, urine and cerebrospinal fluid was investigated. The concentration of these drugs in biological body fluid was detected online. The detection sensitivity of HPLC was improved effectively, which provided a simple and rapid method for the determination of biological samples. Accurate online analysis method. In addition, we also discussed the retention mechanism of RAM on drugs, which provided some experimental basis for the more efficient use of RAM column and the synthesis of new RAM.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R917

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