Differences Of Whey Protein From South China Buffalo And Other Breeds Milk And The Etablishment Of The Plot System

In this paper, particle size analysis was carried out on whey protein from Buffalo Milk within Guangdong Province, to study the effect of different treatment and storage time that can impose on the particle distribution of buffalo milk whey protein. Malvern Nano-particle size analyzer was adopted, and had shown that the buffalo milk without dialysis treatment,85.6% of whey protein particle size distribution between 58-100nm, whereas 87.5% distribution between 58-105nm after the dialysis treatment. Thus a conclusion can be drawn that buffalo milk whey protein can maintain its stability under the two treatments and stay unaffected. The data collected from the sample after ten days storage showed that 85.8% particle size of buffalo milk whey protein distributed between 91-165nm, which indicate that the increase in storage time can result in protein particles aggregation, together with the destabilization of the system. Sample milk A had its 91.9% of whey protein particle size distributed between 68-120nm. Sample milk B had its 82.2% whey protein particle size distributed between 68-120nm. By comparing to the distribution range of buffalo milk whey protein, all these three samples have a minor difference on its particle size and its distribution had showed no significant difference. By the analysis of AFM, particle size distribution of whey protein, buffalo milk had shown an aggregation level of less than sample A and B, and UHT milk whey protein had a highest degree of aggregation above all.A new method for buffalo milk whey protein detection under alkaline condition was established by using borax alkaline buffer system for capillary zone electrophoresis. The buffer concentration, pH, additives, type of operating voltage were controlled to establish high performance capillary electrophoresis separation system. Sample was detected under the condition of 1.2% borax,0.2% SDS, pH8.95 buffer,25KV operating voltage,25℃column temperature,214nm detection wavelength. Under these conditions, the four main proteins from whey protein were separated well with good linearity and repeatability, its migration time and peak area RSD were less than 1.5% and 0.5% respectively, and its addition standard recovery is 83-94%. This helped establish the standard map for buffalo milk whey protein analysis by capillary electrophoresis. Holstein milk whey proteins had shown significant differences on migration time and protein content with buffalo whey protein on capillary electrophoresis. UHT milk also share this property which indicate that there was a marked impact on protein structure and charge-mass ratio and morphology by ultra-high temperature treatment.A quantitative analysis on buffalo whey protein was carried out byRP-HPLC. Jupiter C18 column, acetonitrile-TFA as the mobile phase was adopted for whey protein separation, and was detected at room temperature,0.8ml/min mobile phase flow rate,214nm wavelength. Under these conditions, the major proteins in whey protein are well separated and the linear correlation was greater than 0.996 with good reproducibility. Also the retention time and peak area were less than 0.45% RSD and 2.7% respectively, and the addition standard recovery is 99.5-101%. This helped establish the standard map for buffalo milk whey protein analysis by RP-HPLC. Holstein milk had shown significant differences with buffalo milk in content, retention time and the variants. UHT milk also shares these differences with buffalo milk.Two-dimensional gel electrophoresis used on buffalo milk whey protein and A, B fresh milk whey protein sample analysis had shown that buffalo milk had 38 different points with sample A, of which points 163,162,159,137,107,104,98,97,94,81 are unique in its existence in buffalo milk, and the rest of points just indicate same type but various in its content. However, sample B had shown 52 different points with buffalo milk, of which the points 76,81,88,91,94,97,98,100,104, 107,110,128,162,163 are unique in its existence in buffalo milk, and the rest of points just indicate same type but various in its content. UHT milk have a greater difference with buffalo milk. According to the software analysis,27 proteins that were absent in UHT milk were detected in buffalo milk 75,76,80,81,86,87,88,91,92,94,96,98,99,100,101,102,103,107,110,113, 121,122,137,152,159,162,163.14 other point just express the content variance.