Studies on the Use of Immobilized Enzymes for Analysis of Glucose, Fructose and Sucrose in Foods

• Main Authors: WANG,SHI-YUN, 王詩韻
• Other Authors: Chang, Chen-Tien
• Format: Others
• Language: zh-TW
• Published: 1992
• Online Access: http://ndltd.ncl.edu.tw/handle/76920739551487073869

碩士 === 靜宜大學 === 食品營養學系 === 80 === Two enzyme system"glucose oxidase(GOD) and horseradish peroxidase(HRPO)" , and"hexokinase(HK) and glucose-6-phosphate dehydrogenase(G6P DH) " were immobilized on Sepharose CL-6B and packed in a small column respectively for determination of glucose. Both columns were incorporated into a high-performance liquid chromatography(HPLC) system. The limit for detection of glucose by the column packed with Sepharose-GOD-HRPO is 6 nmole. Linear range is 6-30 nmole. The limit for detection of glucose by the column packed with Sepharose-HK-G6P DH is 0.5 nmole. Linear range is 0.5-5.0 nmole. HK,G6P DH and phosphoglucose isomerase(PGI) were immobilized on Sepharose CL-6B and packed in a small column for determination of fructose. The column was incorporated into the HPLC system as above. The limit and the range for detection of fructose by the column are the same as the column packed with Sepharose-HK-G6P DH for determination of glucose. Invertase(IT) and mutarotase(MT) were immobilized on Sepharose CL-6B and packed in a small column. The column was connected with the column packed with Sepharose-GOD-HRPO and incorporated into HPLC system as above for determination of sucrose. The limit for detection of sucrose by above two column systems is 28 nmole. Linear range is 28-140 nmole. The above immobilized reactors were used for determination of glucose, fructose and sucrose in some vegetables, fruits and commercial foods. The results correlated statisfactorily with those obtained by the commercial kit method. The immobilized enzyme reactors showed good operational stability for one to two month, although the sensitivity was slightly decreased.