Study of the Separation of Protein from the Edible Bird's Nest Based on Capiliary Electrophoresis
|School||Tianjin University of Science and Technology|
|Course||Nutrition and Food Hygiene|
|Keywords||the proteins from edible bird’s nest capillary zone electrophoresis two-dimensional capillary electrophoresis sepration|
A new capillary electrophoresis (CE) method has been developed for separation of the protein extractive from edible bird’s nest. An effective way was provided to identify the edible bird’s nest by confirming the electropherogram of the protein extractive from the bird’s nest.The protein was extractived from four brands of edible bird’s nest samples (Tong Ren Tang, Bai Yun Shen Rong, Yan Hang Jia and Yan Zhi Wu) according to60℃water bath. The protein extraction rate was6.14%,5.54%,5.90%and5.48%respectively. The concentration of protein was767.16μg/mL,692.54μg/mL,737.31μg/mL and685.07μg/mL respectively, which was determined by using the Bradford. The gel electrophoresis experiments showed that the molecular weight of proteins extracted between35-116kDa. By UV absorption spectrum of proteins from four brands of edible bird’s nest, we also choosed215nm detection wavelength for CE. Effect of several parameters, such as inner diameter of capillary, buffer style, buffer pH, buffer concentration, injection concentration, injection time and separation voltage on separation of proteins were systematically investigated. The protein extractive from four brands of bird’s nest were successfully separated in50mmol/L phosphate buffer (pH2) at25℃with7.5s (at0.5psi) as injection time,2000μg/mL as injection concentration,75μm as inner diameter of capillary and UV detection at215nm. The electropherogram of the protein from Tong Ren Tang bird’s nest was two characteristic peaks of the protein extractive from the bird’s nest were confirmed by the electropherogram of the protein from Tong Ren Tang bird’s nest as a criterion. The other three brands of edible bird’s nest were identified by contrasting retention time of the characteristic peaks and the ratio of peak area.An on-column, etched fused-silica porous junction interface was designed and fabricated, and an integrated two-dimensional separation system involving coupling of capillary isoelectric focusing (CIEF) with capillary zone electrophoresis (CZE) was constructed by the interface. The two-dimensional capillary electrophoresis was applied preliminary to separate the protein extractive from edible bird’s nest. The results indicated that the two-dimensional capillary (CIEF-CZE) was found to increase the sepration power on the protein extractive from edible bird’s nest over single dimensional mode (CZE).