摘要
为快速准确地测定烟草中的蛋白质含量,建立并优化了BCA-TCA法,即提取烟叶蛋白质的最佳体系条件为pH值7.0的0.05 mol/L PBS缓冲液、液料比40 m L/g,使用TCA沉淀蛋白质,利用差减法去除烟叶中还原性物质对BCA方法准确性的影响,从而准确测定样品的蛋白质含量。结果表明:该方法测得的烟叶样品蛋白质含量变异系数小于5%,重复性较好;样品的加标回收率为96.39%,加标回收变异系数为0.75%<2%,准确度高;BCA-TCA法和克达尔法测定值的t检验结果为t=1.554 5<t_(0.05),8=2.306,说明2种方法在95%的置信区间内无显著差异,但建立的BCATCA法将耗时缩短至2.5~3.5 h,且操作步骤简捷、所用设备简单。可见,建立的BCA-TCA法能够快速准确地测定烟草中的蛋白质含量。
For the rapid and accurate determination of the protein content in tobacco,an accurate and efficient BCA-TCA method was established and optimized. The optimum conditions for extracting tobacco protein were 0. 05 mol/L PBS buffer at pH 7. 0,and liquid-solid ratio was 40 m L/g. Differential subtraction was used to accurately determine the sample protein content,through removing the impact of interfering substances by using TCA to precipitate proteins. The results showed that the coefficient of variation of protein content was less than 5% and the reproducibility was good. The standard recovery rate of the samples were 96. 39%,the coefficient of variation was 0. 75% < 2% and the accuracy was high. The t test results of the BCA-TCA method and the Kedar method were t = 1. 554 5 < t_(0.05),8= 2. 306,indicating that there was no significant difference between the two methods in the 95% confidence interval,but the detection time of BCA-TCA method would be shortened to 2. 5—3. 5 h,and the steps were simple,the equipment used was simple. The established BCA-TCA method can quickly and accurately determine the protein content in tobacco.
出处
《河南农业科学》
CSCD
北大核心
2017年第9期156-160,共5页
Journal of Henan Agricultural Sciences
基金
川渝中烟工业有限责任公司科技项目(2014Q116)
