【目的】豆粕是动物饲料的主要原料,但其含多种抗营养因子(anti-nutritional factors,ANF),阻碍营养成分的消化、吸收和利用,从而影响动物的生长发育和健康。研究表明豆粕经微生物发酵可有效地降低抗营养因子含量。但由于发酵工艺、发酵菌种、豆粕本身的因素,不同生产厂家的豆粕及发酵豆粕中各抗营养因子含量差别较大,现有研究中也少有关于二者中抗营养因子水平的研究报道。为此,抽取了市售的65批次豆粕和54批次发酵豆粕,对6抗营养因子:大豆球蛋白、β-伴大豆球蛋白、胰蛋白酶抑制因子、棉籽糖、水苏糖、脲酶进行分析测定,以了解饲料行业使用的豆粕及发酵豆粕中的抗营养因子含量。【方法】用ELISA法(enzyme-linked immuno sorbent assay)对样品中的大豆球蛋白、β-伴大豆球蛋白、胰蛋白酶抑制因子含量进行测定,其分析方法和操作要求均与所购ELISA试剂盒的说明相一致,主要过程为:样品前处理、加样、洗板、加酶标试剂、显色、终止。棉籽糖和水苏糖的检测采用高效液相色谱(high performance liquid chromatography,HPLC)检测微波提取的棉籽糖和水苏糖。脲酶分析参照国标方法:加入尿素缓冲液后恒温水浴,一定时间后加入盐酸溶液停止反应后冷却,清洗试管内容物,以氢氧化钠标准溶液滴定至p H4.7后根据体积计算得出脲酶活性。【结果】调查分析后发现:豆粕和发酵豆粕中的大豆球蛋白平均含量分别为129.3、54.7 mg·g^-1,发酵后大豆球蛋白平均含量降低了57.7%,根据百分位数法对数据进行统计分析,得出豆粕和发酵豆粕中的大豆球蛋白正常值范围分别为58.9—P(90)(177.3 mg·g^-1)、ND—P(90)(109.4 mg·g^-1)。豆粕中的β-伴大豆球蛋白平均含量为102.2 mg·g^-1,而发酵豆粕中的β-伴大豆球蛋白为37.6 mg·g^-1,相比豆粕降低了63.2%,使用相同的数据统计方法?
[Objective] Soybean meal is the main raw material of feed. However, various anti-nutritional factors (ANF) in soybean meal hinder the digestion, absorption and utilization for nutrients, which would have negative effects on animal growth and health. Studies have shown that soybean meal fermented by microorganism could decrease the undesirable effect of ANF. While, fermentation strains, soybean species, and manufacturers may have an effect in fermentation process. Moreover, seldom reports referred to the levels of ANF in soybean meal and fermented soybean meal. Based on this, 65 batches of available soybean meal and 54 batches of fermented soybean meal were collected in this study, followed by analysis of 6 kinds of ANF, including glycinin, β-conglycinin, trypsin inhibitors, raffinose, stachyose and urease, in order to investigate the actual levels of ANF in soybean meal and fermented soybean meal. [Method] In this study, glycinin, β-conglycinin and trypsin inhibitors were analyzed by Enzyme-link Immunosorbent Assay (ELISA) method. Experimental procedures were conducted according to instructions which were sample pretreatment, plus, washing, adding enzymatic reagent, color reaction, and termination. The analysis method of raffinose and stachyose was HPLC after extracted by microwave. Analysis of urease refers to GB/T 8622-2006, and titration was utilized foranalysis of urease activity. [Result] The average concentration of glycinin in soybean meal (129.3 mg.g^-1) is 57.7% higher than fermented soybean meal (54.7 mg.g^-1). The percentile method was used for analysis. The normal values in soybean meal and fermented soybean were 58.9-177.3mg.g^-1 and ND (Not Detected)-109.4 mg.g^-1, respectively. Moreover, the fermented soybean meal (37.6 mg.g^-1) is 63.2% lower than the concentration of β-conglycinin in soybean meal (102.2 mg.g^-1). The normal values in soybean meal and fermented soybean were 42.8-147.2 mg.g^-1 and ND-61.8 mg.g^-1, respectively. The average concentration of trypsin inh