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  • 學位論文

毛細管電層析之研究 與牛奶中色素之分析

Study on Capillary Electrochromatography and Determining Colorants in Milk Beverages by Capillary Electrophoresis

指導教授 : 黃悉雅
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摘要


毛細管電泳(Capillary Electrophoresis,CE)是將毛細管充滿緩衝液經由高電壓系統在毛細管兩端提供一直流電壓,在此外加電場作用下,使分析物在毛細管中因不同的遷移速率而進行分離的方法。在分析領域中,毛細管電泳成為重要的分離技術,提供快速、高效率的分離。本研究是應用毛細管電泳技術來分析牛奶飲品中所添加的食用色素,但由於牛奶飲品中含有大量蛋白質不易去除,進行毛細管電泳分析時,容易產生蛋白質吸附管壁的現象,造成毛細管阻塞而無法進行偵測,故當樣品進入電泳儀之前,需先使用固態萃取管(solid phase extraction,SPE)對樣品做前處理,如此能有效取得色素,並降低牛奶中蛋白質在分離時的干擾。 現今市面上牛奶飲品提供多樣口味,來增加消費者的購買動機,而為了配合各種口味的銷售,勢必在飲品外觀上做美化,所以常在飲品中添加食用色素,而食用色素的提煉需經一連串的步驟,故色素的取得難以達到很高純度,其中的不純物有致癌的可能性。食用色素的用量在各個國家有不同的規定,其添加量的多寡對消費者的健康有很大的影響,故對市售添加食用色素的牛奶飲品分析其色素含量是相當重要的。 本研究將針對常用的8種食用色素進行分離,並探討pH值和環糊精(β-Cyclodextrin,β-CD)用量對分離的影響,找出最佳分離條件及固相萃取條件應用在分析牛奶飲品上。 同時探討毛細管電層析(Capillary Electrochromatography, CEC)方法: CEC是在毛細管中填入能幫助分離作用的材料,使毛細管內部存有固定相(stationary phase),在進行分離時,可提供和高效液相層析(High- Performance Liquid Chromatography, HPLC)相同的分配效應(partitioning effect),再結合毛細管區帶電泳分析(Capillary Zone Electrophoresis,CZE)的優勢,所展現具有更多選擇性及更高效率的分離技術。 本實驗是使用乙醇當填料時溶劑(packing solvent),並將正十八烷二氧化矽 (octadecyl silica, C18)粉末製備成泥漿狀溶液(slurry solution) 放置在填料管柱中,再施加壓力將粉末填入毛細管中,此毛細管作為層析管柱,配合毛細管電泳分析技術,對芳香族碳氫化合物(aromatic hydrocarbon)進行電層析分離。

並列摘要


Capillary electrophoresis (CE) possesses many advantages, including high separation efficiency, excellent resolution, short analytical time, low consumption of sample, and easy automation. Hence capillary electrophoresis (CE) has became a highly attractive separation technique. In this method, analytes carried with different charges (neutral, negative, or positive) can migrate with a buffer solution under an electric field, its separation principle is based on different migration velocities of analytes themselves. Milk products with various fruit-flavored are high demand in Asia. In order to enhance attraction and appetence, then food colorants usually are added into these milk products. However, these food colorants were harmful for human health according previous reports, such that these colorants and their metabolic products in the body could even cause cancer, deformation etc.. Because food colorants are potentially toxic, their determination in milk beverages are necessary for the assurance of food safety. In the paper, capillary electrophoretic method would be applied to analyze eight food colorants in milk beverages. In order to reduce the matrix interference resulted from milk constituents, a sample pretreatment method for milk beverages containing food colorants would be developed. As a result, a solid-phase extraction (SPE) based on polyamide column was established for milk sample pretreatment. In this study, eight different food colorants commonly used as food additives were determined. These food colorants were separated successfully within 9 min using a borax-NaOH (pH=10.0) modified with 7 mM β-Cyclodextrin (β-CD). The combination of the simple SPE pretreatment and the optimized separation conditions of capillary electrophoresis, the content of these colorant additives in commercial milk beverages was able to determine successfully without milk’s matrix interference. In the meanwhile studying on capillary electrochromatography (CEC), CEC is a developing technique that combines the advantages of high-performance liquid chromatography (HPLC) with those of capillary electrophoresis. CEC utilizes electroosmosis to transport the mobile phase through the capillary column, rather than hydraulic flow in HPLC. Duing to electroosmosis with plug-like flow profile , CEC has higher separation efficiency over than HPLC technique. However, most CEC columns were obtained by home-made, commercial source is rarely. A slurry pressure packing technique is usually used in manufacturing CEC column. Nevertheless, skill and experience are required to complete all the steps involved in the fabrication of packed columns for CEC even though many papers have been reported how to make a good CEC column. Under the study, a 75-μm i.d. capillary columns were packed with a 3-μm octadecyl silica (C18) stationary phase by slurry pressure packing method. A successful procedure in fabricating CEC column was established in our laboratory. Finally, the CEC columns were employed to separate several neutral analytes (aromatic hydrocarbon).

參考文獻


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Yang, Y. K. (2007). 利用溶膠-凝膠法製備毛細管電層析管柱之研究 [master's thesis, Chung Yuan Christian University]. Airiti Library. https://doi.org/10.6840%2fcycu200700138
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