天然橡胶手套中水抽提蛋白含量测定影响因素的研究

Study on the factors affecting the determination of water-extractable protein content in natural rubber gloves

当前对天然胶乳手套(以下简称手套)中蛋白含量测定方法主要引用国家标准GB/T 21870—2008,该方法未明确提取过程(本研究主要针对方法A-剪碎手套抽题方法)条件,测定影响因素众多。本文通过单因素实验研究抽提过程中各个因素,如手套剪碎程度、抽提液的用量、提取温度、提取时间以及震荡频率等因素的影响,得到最优提取条件。用缓冲液抽提水溶性蛋白质,经通过沉淀、浓缩,再溶解沉淀的蛋白质,以标准蛋白质做参照,用改进Lowry方法进行比色,定量测定蛋白质含量。结果显示,在手套剪碎大小为(3~10)mm×(3~10)mm,液料比为20∶1,提取次数为2次,温度为35℃,振荡速度为120 r/min时,标准曲线回归方程为y=0.01138x+0.08734,r2=0.99965,在5.40μg/mL~54.03μg/mL范围内,有良好的线性关系,平均重复性RSD为0.96%,平均加样回收率为101.81%。表明该试验方法结果更准确可靠,能够为手套蛋白提取工艺的明确提供实验依据。

At present,the main reference standard for the determination of protein content in natural latex gloves(hereinafter referred to as gloves)is GB/T 21870-2008,the method does not specify the conditions of the extraction process(this study is mainly aimed at the method A-shearing glove extraction method),and there are many influencing factors.In this paper,we consider the influence of various factors,such as the degree of glove cutting,the amount of extraction fluid,extraction temperature,extraction time,oscillation frequency,etc,in the process of single-factor experiments,so as to obtain the optimal extraction conditions.The water-soluble protein is extracted with buffer,and then the precipitated,concentrated,and then dissolved the precipitated protein.The standard protein is used as a reference,and the improved Lowry method was used for color comparison to quantitatively determine the protein content.Results When the size of the glove is(3-10)mm×(3-10)mm,the liquid ratio is 20∶1,the extraction times is 2 times,the temperature is 35℃,and the oscillation speed is 120rpm,the standard curve regression equation is y=0.01138x+0.08734,r2=0.99965,indicating that there is a good linear relationship in the range of(5.40-54.03)ug/mL,with an average repeatability RSD of 0.96%and an average sampling recovery rate of 101.81%.The conditions of this test method are clearer than those of the original method,and the results are more accurate and reliable,it provides experimental basis for clear the glove protein extraction process.