利用溶膠-凝膠法製作毛細管柱及其在毛細管電層析之應用研究

Abstract

利用溶膠-凝膠法製作毛細管柱及其在毛細管電層析之應用研究 研究生：劉育錚 指導教授：謝有容 國立交通大學應用化學研究所 摘要 本研究中設計製作了兩種毛細管電層析用分離管柱：第一種為利用光聚合溶膠-凝膠法所製作的塊材毛細管柱，應用於塑化劑、防腐劑的毛細管電層析；第二種則為利用溶膠-凝膠法製作的聚合物/環糊精混成物(polymer/cyclodextrin hybrid)塗層管柱，應用於數種胺基酸的毛細管電層析。 所謂整體式管柱(monolithic column)，或稱為塊材管柱，又稱為連續床式管柱(continuous-bed column)，為近幾年來在毛細管電層析領域中被相當廣泛研究的一種新型管柱。本研究首先利用光聚合溶膠-凝膠法(photopolymerization)製造出具有多孔性聚合物塊材之毛細管柱(sol-gel monolithic capillary)，應用於毛細管電層析(capillary electrochromatography, CEC)分離管柱。其製作方式為將適量的含雙鍵矽氧烷單體和鹽酸水溶液均勻攪拌混合使之進行溶膠-凝膠反應，再取一定量的溶液和特定比例甲苯混合後，加入光起始劑，注入毛細管中，置於紫外線光源下進行光聚合反應，反應後在毛細管內部生成整體式塊材結構，最後再用乙醇做為沖洗溶劑，沖洗並置換出毛細管內部含甲苯部分的空間，形成多孔性的溶膠-凝膠塊材毛細管柱。 經由掃描式電子顯微鏡(SEM)對毛細管內聚合物表面型態進行分析，可觀察出此溶膠-凝膠塊材確實為具有多孔性網狀結構，且與毛細管壁互相連結。再使用幫浦加壓來沖堤此毛細管柱時，發現管內塊材結構具有高滲透性、穩固性和耐高壓等特性。 在應用方面，本研究中使用溶膠-凝膠多孔性塊材毛細管柱進行鄰-苯二甲酸酯塑化劑和防腐劑的分析，同時探討等梯度沖提毛細管電層析 ( Isocratic CEC)與階段梯度沖提毛細管電層析 (Step-gradient CEC)兩種方法，發現利用階段梯度沖提CEC的方式可明顯縮短分析時間，改善分析物拖尾的情形，也提高了分離的解析度與理論板數。六種鄰苯二甲酸酯塑化劑可在7分鐘快速進行分離，理論板數可高達201,000 (N/m)，滯留時間的RSD值小於0.68%，顯示再現性良好。用於分析七種防腐劑時可在15分鐘快速分離，理論板數可高達106,000 (N/m)，滯留時間的RSD值小於0.91%。 此外，實驗中另研製出表面具有聚合物/環糊精混成物塗層的毛細管柱，應用於酪胺酸(tyrosine)、苯丙胺酸(phenylalanine)、色胺酸(tryptophan)等胺基酸的毛細管電層析分離，皆可在3分鐘內被快速沖提出，理論板數可達78,000 (N/m)，連續測試下，滯留時間的RSD值小於0.84%。 本研究結果顯示：自行研製的塊材毛細管柱，對於塑化劑,防腐劑均有相當高的分析鑑別力，若將此管柱應用於分析生化樣品、環境污染物之上，應該是極有潛力的分析檢測利器。

Preparation of Sol-Gel derived Stationary Phases for Capillary Electrochromatography Student：Yu-Cheng Liu Advisor： You-Zung Hsieh Department of Applied Chemistry National Chiao Tung University Abstract In this study, two kind of sol-gel based capillary were prepared for application in capillary electrochromatography. The first column is a photopolymerized sol-gel monolithic capillary for separation of six phthalate ester plasticizers and seven preservatives; the second is a polymer/cyclodextrin hybrid coated column for separation of three amino acids. The sol-gel monolithic capillary was prepared by photopolymerization as the following procedures: The monomer stock solution was prepared by adding methacryloxypropyltrimethoxysilane monomer to appropriate amount of porogenic solvent, an acid catalyst solution was added and stirred. The photoinitiator was then mixed with the corresponding amount of monomer solution. A UV-transparent capillary was filled with reaction mixture by a disposable syringe. Polymerization was carried out by irradiation of ultraviolet radiation. A porous monolithic sol-gel column was obtained by an one-step, in situ, process. Using this column, a variety of low-molecular weight neutral compounds, including six phthalate ester plasticizers, and seven preservatives are separated from their mixtures. Various different operational parameters, such as buffer composition, field strength, and column temperature, mobile phase elution methods were varied to assess their influence on column performance. Separation efficiencies of up to 201,000 plates/m were achieved on a 31 cm ×75 μm i.d. column using phthalate ester plasticizers as test solutes. Whenever the step-gradient approach was performed in an automated mode, the retention time RSD for repeated injections was below 0.68 %. The separation of three amino acids was tested by a polymer/cyclodextrin hybrid coated columns. Performed of Open-tubular capillary electrochromatography, in which the γ-CD was immobilized onto the capillary wall during thermal polymerization. Separation efficiencies of up to 78,000 plates/m were achieved on a 31 cm ×50 μm i.d. column using tryptophan、tyrosine and phenylalanine as test solutes. Small variations in the elution times were observed for tryptophan (0.21% RSD), tyrosine (0.23 % RSD), phenylalanine (0.75 % RSD) (n=4).