Chinese Journal of Animal Nutrition >

2025 , Vol. 37 >Issue 9: 6348 - 6357

DOI: https://doi.org/10.12418/CJAN2025.515

RESEARCH PAPER

Effects of Steam Explosion Treatment for Red Clover on Extraction Efficiency of Isoflavone

  • XIONG Zhanbo , 1 ,
  • LI Kexin 1 ,
  • WANG Dian 2 ,
  • ZHENG Nan 1 ,
  • WANG Jiaqi 1 ,
  • ZHAO Shengguo , 1, *
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  • 1 State Key Laboratory of Animal Nutrition and Feeding, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
  • 2 National Center of Technology Innovation for Dairy, Hohhot 010000, China
* professor, E-mail:

Received date: 2024-12-31

  Online published: 2025-09-12

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Abstract

This study aimed to investigate the effects of steam explosion treatment for red clover on extraction efficiency of isoflavone. A three-factor, three-level response surface methodology was employed, with the factors affecting the extraction efficiency of isoflavone of explosion pressure, treatment time and moisture content, and the red clover isoflavone yield as the response variable, and the optimal steam explosion treatment conditions were determined using heat reflux extraction technology. The results showed as follows: 1) the optimal steam explosion treatment conditions for isoflavone extraction from red clover were explosion pressure of 0.6 MPa, treatment time of 150 s, and moisture content of 40%. Under these conditions, the red clover isoflavone yield was 93.71%, the red clover isoflavone yield was increased by 62.55% compared with no steam explosion treatment. 2) Scanning electron microscopy analysis showed that the significant structural disruption of red clover after steam explosion treatment, resulting in a loose and disordered structure, which exposed internal pores and increased surface area, thereby enhancing isoflavone extraction. In conclusion, the steam explosion treatment can enhance the extraction efficiency of red clover isoflavone, and can providing reference for the industrial production of red clover isoflavone as feed additives.

Key words： red clover; isoflavone; steam explosion; response surface methodology; scanning electron microscopy

Cite this article

XIONG Zhanbo , LI Kexin , WANG Dian , ZHENG Nan , WANG Jiaqi , ZHAO Shengguo . Effects of Steam Explosion Treatment for Red Clover on Extraction Efficiency of Isoflavone[J]. Chinese Journal of Animal Nutrition, 2025 , 37(9) : 6348 -6357 . DOI: 10.12418/CJAN2025.515

红三叶草(Trifolium pratense L.)是一种重要的反刍动物饲草,因为其具有粗蛋白质含量高(17.74%~21.88%,干物质基础)、可消化纤维丰富(中性洗涤纤维37.51%~44.50%,酸性洗涤纤维25.80%~35.22%,干物质基础)的特点而被广泛应用[1-5]。红三叶草含有大量活性成分异黄酮,因此具有类雌激素活性[6]。作为异黄酮来源,红三叶草产品也被用于人类膳食补充剂,以减少女性更年期症状[7]。红三叶草中主要的异黄酮是刺芒柄花素、鹰嘴豆芽素A、大豆苷元和染料木素,其中刺芒柄花素和鹰嘴豆芽素A占异黄酮的90%以上[8]。在反刍动物应用中,红三叶草异黄酮已经被列入《饲料添加剂品种目录(2024)》。多项研究表明,红三叶草异黄酮不仅可以提高奶畜的产奶量和乳品质[9-10],对奶牛的激素调节和免疫能力有益[11],并且对奶牛的瘤胃发酵和瘤胃健康有积极作用[12-15]。
蒸汽爆破作为一种环保、经济且高效的预处理技术,已广泛应用于植物材料的预处理[16-17]。蒸汽爆破是将原料置于高温高压的环境中,当瞬间解除高压时,原料空隙中的过热蒸汽迅速汽化,体积急剧膨胀而使细胞破裂,以此改变坚硬材料的内部结构,使目标成分更容易溶解,从而提高生物活性和综合利用效率[18-21]。目前蒸汽爆破对异黄酮类化合物的影响研究发现,蒸汽爆破提取银杏异黄酮,提取率可提高2.1倍[22];粉葛经蒸汽爆破处理后总异黄酮提取量可增加1.3倍[23];豆渣采用适宜的蒸汽爆破强度处理可显著提升异黄酮提取率[24]。然而,蒸汽爆破处理对红三叶草中异黄酮的影响尚不清楚。Box-Behnken设计(BBD)是一种广泛应用于多领域的响应面设计方法[25],通过较少的试验次数来估计响应面模型[26]。本研究的目的是应用Box-Behnken设计与响应面法模型相结合,以确定红三叶草异黄酮提取的蒸汽爆破处理最优条件,探究蒸汽爆破处理对红三叶草异黄酮提取效率的影响,为相关提取工艺的研究提供理论基础。

1 材料与方法

1.1 材料与试剂

大豆苷元标准品(含量≥98%,西格玛奥德里奇贸易有限公司)、染料木素标准品(含量≥98%,西格玛奥德里奇贸易有限公司)、刺芒柄花素标准品(含量≥98%,西格玛奥德里奇贸易有限公司)、鹰嘴豆芽素A标准品(含量≥98%,西格玛奥德里奇贸易有限公司)、甲醇(色谱纯,国药集团化学试剂北京有限公司)、乙腈(色谱纯,国药集团化学试剂北京有限公司)、无水乙醇(分析纯,西格玛奥德里奇贸易有限公司)、三氟乙酸(分析纯,西格玛奥德里奇贸易有限公司)、双蒸水。

1.2 仪器与设备

蒸汽爆破工艺试验台(QBS-80,鹤壁正道生物能源有限公司)、恒温光照培养箱(MGC-1000BP-2L,上海一恒科学仪器有限公司)、样品粉碎机(BF-04,北京莱万生物技术有限公司)、电热恒温烘干箱(DHG-9030AL,北京陆希科技有限公司)、集热式恒温加热磁力搅拌器(DF-101S,北京鑫益成科技有限公司)、数显型旋转蒸发仪(RV 10 digital V,广州艾卡仪器设备有限公司)、恒温水浴锅(JULABO PURA22,北京优莱博技术有限公司)、扫描电子显微镜(Regulus 8100,日本日立公司)、高效液相色谱仪(Waters 2695,上海沃特世科技有限公司)。

1.3 红三叶草原料预处理

采集盛花期的全株红三叶草,于65 ℃烘干至恒重,粉碎至40~60目后室温保存。

1.4 红三叶草异黄酮提取流程

从工业生产节约能耗、降低成本、操作简便角度出发,且保证红三叶草异黄酮得率情况下,设计热回流提取条件:料液比1∶6,提取温度50 ℃,提取时间0.5 h。准确称取20.0 g红三叶草粉末于圆底烧瓶,每份样品进行3次重复,按照热回流提取条件,加入80%乙醇进行热回流提取。提取完成后真空抽滤取滤液弃滤渣,提取液通过旋蒸仪减压真空浓缩,回收溶剂,将提取物真空干燥后即得红三叶草浸膏,加入10 mL甲醇复溶,取1 mL过0.22 μm尼龙滤膜过滤后待高效液相色谱仪测定红三叶草异黄酮含量。

1.5 红三叶草异黄酮得率的计算

按下式计算红三叶草异黄酮得率:
红三叶草异黄酮得率(%)=[(Wt×Nt)/(Wy×Ny)]×100。
式中:Wt表示提取物重量(g);Nt表示提取物异黄酮含量(mg/g);Wy表示原料重量(g);Ny表示原料异黄酮含量(mg/g)。

1.6 蒸汽爆破处理Box-Behnken设计

将平衡水分后的红三叶草原料置于蒸汽爆破台,密封。蒸汽爆破处理条件以爆破压力、维压时间和水分含量为自变量,红三叶草异黄酮得率为响应值,采用Box-Behnken设计3因素3水平的响应面试验,蒸汽爆破Box-Behnken设计因素及水平见表1,每份样品重复测定3次,蒸汽爆破处理后65 ℃烘干至恒重。
表1 蒸汽爆破处理Box-Behnken设计因素及水平

Table 1 Factors and levels of Box-Behnken design for steam explosion treatment

水平
Levels		 因素Factors
爆破压力
Steam pressure/MPa (A)		 维压时间
Treatment time/s (B)		 水分含量
Moisture content/% (C)
-1 0.6 50 10
0 1.2 125 30
1 1.8 200 50

1.7 扫描电镜分析

取适量红三叶草原料和最佳条件蒸汽爆破处理后红三叶草试样粉碎到40目,分别将样品进行镀膜处理。采用扫描电子显微镜Regulus 8100,在加速电压为3 kV的条件下观察样品纤维表面形态[27]。

1.8 红三叶草异黄酮测定

1.8.1 色谱条件

色谱柱:XBridge C18色谱柱(250 mm×4.6 mm,5 μm);流动相A:25%乙腈水溶液(含0.05%三氟乙酸);流动相B:100%乙腈(含0.05%三氟乙酸);柱温:40 ℃;检测波长:270 mm;流速:1 mL/min;进样量:10 μL;流动相梯度洗脱程序见表2。
表2 流动相梯度洗脱程序

Table 2 Gradient elution procedure of mobile phases

时间
Time/min		 流动相A
Mobile phase
A/%		 流动相B
Mobile phase
B/%
0 90 10
4 90 10
8 87 13
14 80 20
16 73 27
18 55 45
22 50 50
23 40 60
24.5 26 73
26 0 100
29 90 10

1.8.2 标准品溶液的制备

准确称取大豆苷元、染料木素、刺芒柄花素和鹰嘴豆芽素A标准品20.00 mg(误差≤0.05 mg)于100 mL棕色容量瓶中,加入甲醇超声10 min溶解并定容,配制浓度为200 μg/mL的储备液,于-20 ℃以下保存。分别准确量取1 mL上述标准品储备液于10 mL离心管,混匀后准确移取适量混合标准品溶液稀释,配制浓度分别为50、25、12.5、6.25、3.125、1.562 5 μg/mL的系列标准工作溶液。

1.8.3 供试样品溶液的制备

分别准确称取供试样品10 mg于50 mL棕色容量瓶中,加入甲醇超声10 min溶解并定容,配制成200 μg/mL的供试样品溶液。

1.8.4 高效液相色谱检测

分别量取1 mL上述系列混合标准工作溶液、供试样品溶液,通过0.2 μm滤膜过滤,按照从低浓度到高浓度的顺序依次进样,进样量为10 μL,每个样品连续进样2次,取平均值。经高效液相色谱对供试品进行分析,通过与标准品比对,面积归一化法确认供试样品高效液相色谱纯度,以标准品的浓度为横坐标(X)、峰面积为纵坐标(Y)绘制标准曲线,得到标准品线性回归方程,大豆苷元标准曲线方程:Y=63 394X-4 363.8(R2=1.000 0),染料木素标准曲线方程:Y=64 799X-1 962.6(R2=0.999 9),刺芒柄花素标准曲线方程:Y=127 495X-12 903(R2=0.999 4),鹰嘴豆芽素A标准曲线方程:Y=67 320X-1 547.1(R2=0.999 9),并以此计算供试样品的异黄酮含量。

1.9 数据统计与响应面分析

试验数据在Excel 2021中进行初步统计,应用Design-Expert 13.0进行Box-Behnken试验设计和分析并作图。

2 结果

2.1 蒸汽爆破处理响应面模型和方差分析

以爆破压力、维压时间和水分含量为考察因素,红三叶草异黄酮得率为响应值,利用Design-Expert 13.0进行Box-Behnken设计,优化蒸汽爆破处理红三叶草试验设计及结果。如表3所示,当爆破压力为0.6 MPa,维压时间为125 s,水分含量为50%时,红三叶草异黄酮得率达到最大,为93.82%。
表3 蒸汽爆破处理的响应面试验设计及结果

Table 3 Response surface test design and result for steam explosion treatment

序号
No.		 爆破压力
Steam pressure/MPa(A)		 维压时间
Treatment time/s(B)		 水分含量
Moisture content/%(C)		 红三叶草异黄酮得率
Red clover isoflavone yield/%(R)
1 0.6 125 10 85.74
2 1.2 200 50 79.35
3 1.2 125 30 82.87
4 0.6 125 50 93.82
5 1.2 125 30 81.53
6 0.6 50 30 87.73
7 1.2 125 30 80.44
8 1.2 125 30 80.44
9 1.2 50 50 80.88
10 1.2 125 30 80.99
11 1.8 125 50 78.25
12 1.2 50 10 68.52
13 1.8 125 10 71.57
14 1.2 200 10 68.42
15 1.8 50 30 72.55
16 0.6 200 30 65.60
17 1.8 200 30 57.79
利用Design-Expert 13.0对数据进行方差分析(ANOVA),如表4所示,该模型显著(P<0.05),失拟项显著(P<0.05),说明该模型的拟合程度较好。从显著性方面分析,爆破压力、维压时间和水分含量均对红三叶草异黄酮得率有显著影响(P<0.05),爆破压力、维压时间和水分含量之间的交互作用对红三叶草异黄酮无显著影响(P>0.05),维压时间与红三叶草异黄酮得率存在显著二次关系(P<0.05)。根据各因素的P值可得到各因素对红三叶草异黄酮提取率的影响大小顺序为:爆破压力>维压时间>水分含量。
表4 蒸汽爆破处理的响应面试验方差分析

Table 4 Response surface test ANOVA for steam explosion treatment

方差来源
Variance sources		 平方和
Sum of squares		 自由度
Variance		 均方
Mean square		 F值
F-value		 P值
P-value		 显著性
Significant
模型Model 1 107.35 9 123.04 4.94 0.023 4 *
爆破压力Steam pressure (A) 347.56 1 347.56 13.96 0.007 3 *
维压时间Treatment time (B) 185.47 1 185.47 7.45 0.029 4 *
水分含量Moisture content (C) 180.98 1 180.98 7.27 0.030 8 *
AB 13.58 1 13.58 0.55 0.484 2
AC 0.49 1 0.49 0.02 0.892 4
BC 0.51 1 0.51 0.02 0.890 1
A2 5.49 1 5.49 0.22 0.652 9
B2 355.95 1 355.95 14.30 0.006 9 *
C2 20.99 1 20.99 0.84 0.389 0
残差Residual 174.28 7 24.90
失拟项Lack of fit 170.20 3 56.73 55.59 0.001 0 *
误差项Pure error 4.08 4 1.02
总和Sum 1 281.64 16

*表示差异显著(P<0.05)。

* indicated significant difference (P<0.05).

2.2 蒸汽爆破处理响应面交互作用分析

响应值与各因素交互作用构成等高线和响应面,等高线形状反映两因素之间的相互交互作用的强弱,圆形表示交互作用不明显,向椭圆形形变越强则交互作用越明显[28]。由图1反映爆破压力和维压时间的交互作用对红三叶草异黄酮得率的影响,由图2反映维压时间和水分含量的交互作用对红三叶草异黄酮得率的影响,由图3反映爆破压力和水分含量的交互作用对红三叶草异黄酮得率的影响,结合方差分析可知,维压时间和爆破压力的交互作用对红三叶草异黄酮得率的影响最大,其次为维压时间和水分含量的交互作用,爆破压力和水分含量的交互作用对红三叶草异黄酮得率的影响最小。
图1 爆破压力和维压时间的交互作用对红三叶草异黄酮得率影响的响应面图与等高线图

Fig.1 Response surface plot and contour plot of interaction between steam pressure and treatment time on red clover isoflavone yield

图2 维压时间和水分含量的交互作用对红三叶草异黄酮得率影响的响应面图与等高线图

Fig.2 Response surface plot and contour plot of interaction between treatment time and moisture content on red clover isoflavone yield

图3 爆破压力和水分含量的交互作用对红三叶草异黄酮得率影响的响应面图与等高线图

Fig.3 Response surface plot and contour plot of interaction between steam pressure and moisture content on red clover isoflavone yield

2.3 蒸汽爆破处理验证试验

按照响应面软件优化给出的数据,可知预测的最优条件为爆破压力0.6 MPa,维压时间150 s,水分含量40%。在此操作条件下重复3次试验测得红三叶草异黄酮得率为94.39%,与响应面数据模型预测值95.74%的误差值仅为1.35%,证明该模型可较好预测红三叶草异黄酮得率。

2.4 蒸汽爆破处理对红三叶草异黄酮得率的影响

如表5所示,红三叶草在未经蒸汽爆破处理进行热回流提取异黄酮,重复3次试验测得红三叶草异黄酮得率为57.65%;最优条件下蒸汽爆破处理后,测得红三叶草异黄酮得率为94.39%,与未经蒸汽爆破处理相比,红三叶草异黄酮得率提高了62.55%。
表5 蒸汽爆破处理对红三叶草异黄酮得率的影响

Table 5 Effects of steam explosion treatment on red clover isoflavone yield %

项目Items 红三叶草异黄酮得率Red clover isoflavone yield
未经蒸汽爆破处理No steam explosion treatment 57.65
蒸汽爆破处理Steam explosion treatment 94.39

2.5 扫描电镜分析

如图4所示,蒸汽爆破处理后红三叶草表面超微结构发生了明显变化。图4-A为50倍放大效果下未经蒸汽爆破处理的红三叶草纤维结构,结果显示结构完整,纤维排列有序;图4-B为50倍放大效果下蒸汽爆破处理的红三叶草纤维结构,结果显示结构破坏严重,松散无序,纤维束分离,表层破裂形成多孔结构,类似“蜂窝”状,暴露出内部孔隙;图4-C为300倍放大效果下未经蒸汽爆破处理的红三叶草纤维结构,结果显示纤维表面光滑平整;图4-D为300倍放大效果下蒸汽爆破处理的红三叶草纤维结构,结果显示表面褶皱且不规则,出现螺旋状卷曲,纤维束断裂暴露出孔状结构。
图4 扫描电镜下观察到红三叶草的表面超微结构

A:未经蒸汽爆破处理(50×);B:蒸汽爆破处理(50×);C:未经蒸汽爆破处理(300×);D:蒸汽爆破处理(300×)。

Fig.4 Surface ultrastructure of red clover observed under scanning electron microscopy

A: no steam explosion treatment (50×); B: steam explosion treatment (50×); C: no steam explosion treatment (300×); D: steam explosion treatment (300×).

3 讨论

本试验以传统热回流提取为基础,以新型物理化学前处理方式蒸汽爆破处理对红三叶草异黄酮的提取进行优化,结果表明,新鲜红三叶草经65 ℃风干后,以150 s维压时间,0.6 MPa爆破压力,40%含水量的蒸汽爆破处理后,红三叶草异黄酮得率可达94.39%,大幅度高于未经蒸汽爆破处理同等条件提取的红三叶草异黄酮得率57.65%,提高了36.74%。蒸汽爆破作为一种常见的植物提取前处理技术,主要分为蒸煮和降压爆破2个阶段[29]。蒸煮是在高温和高压作用下对原料内部产生机械分离作用[30-31],然后降压爆破会瞬间释放压力导致膨胀的气体使结构形变,释放生物质[32-33]。蒸汽爆破处理促进红三叶草异黄酮析出的原理可能包括以下几个方面:1)细胞壁结构破坏。蒸汽爆破处理通过高温高压的蒸汽破坏植物细胞壁的致密结构,使得细胞壁变得多孔和粗糙,从而增加了比表面积和溶剂的可达性,有利于黄酮类化合物的释放。这一点,在本研究中也可以通过扫描电镜分析结果得到印证。Fan等[34]通过对黄芪进行蒸汽爆破处理,发现其乙醇可溶性和水可溶性黄酮类化合物的提取量分别达到6.34和1.48 mg/g,分别是未经蒸汽爆破处理黄芪的5.22倍和2.88倍。蒸汽爆破处理破坏了黄芪细胞壁的紧密结构,形成了多尺度的孔隙网络,改善了质量传递路径。Fu等[35]研究发现,低温空气蒸汽爆破处理会对杜仲叶片中的纤维网络和细胞壁进行破坏,从而导致叶片中的绿原酸提取效率显著提升,得率和提取率分别提高了62.0%和423.0%。2)氢键破坏和结构重组。蒸汽爆破处理通过高温高压和饱和蒸汽加剧了分子内氢键的破坏,释放出更多的羟基,增加了分子链的活动性,有利于形成有序的分子结构,同时也使得分子链更容易重新排列,有助于打破原有的分子间相互作用,使得黄酮类化合物更容易从植物材料中释放出来[36]。Song等[37]研究了蒸汽爆破处理对松针中黄酮类化合物提取效率的影响,发现经过1.5 MPa蒸汽爆破处理60 s后,松针中黄酮类化合物的含量达到50.8 mg/g,是对照组的2.54倍。蒸汽爆破处理后,植物细胞壁中的纤维素和半纤维素部分降解,木质素发生一定程度的水解,分子键断裂,颗粒变小,使得黄酮从植物材料中提取效率更高。3)去糖基化。蒸汽爆破处理可以促进黄酮糖苷去糖基化成苷元,苷元的生物活性通常优于相应的糖苷,水溶性通常会增加,这有助于它们在水性提取介质中的溶解和释放[38]。Chen等[39]通过试验发现,在2.0 MPa和90 s的蒸汽爆破处理条件下进行大豆种皮预处理,通过破坏植物细胞壁中的糖苷键、酯键和β-O-4醚键,促进了酚类化合物的释放与转化;同时,处理后的蛋白质分子结构发生改变,表面疏水性增加,其总黄酮含量得到显著提高。综上所述,蒸汽爆破处理对红三叶草异黄酮提取效率有显著影响,但具体机制尚不清楚,还需要进一步探究机理。

4 结论

① 响应面法对蒸汽爆破处理红三叶草异黄酮提取条件进行优化可行,最优条件为爆破压力0.6 MPa,维压时间150 s,水分含量40%。在此条件下,红三叶草异黄酮得率为93.71%,相比于未经蒸汽爆破的红三叶草,异黄酮得率提高了62.55%。
② 通过对红三叶草的扫描电镜分析可知,蒸汽爆破处理后红三叶草结构破坏严重,松散无序,暴露出内部孔隙,表面积增大,有利于异黄酮的析出。

References

Publishing order | Descend order by publishing year | Descend order by cited within
[1]
FREY L A, VLEUGELS T, RUTTINK T, et al. Phenotypic variation and quantitative trait loci for resistance to southern anthracnose and clover rot in red clover[J]. Theoretical and Applied Genetics, 2022, 135(12):4337-4349.

[2]
WOODFIELD D R, CLARK D A. Do forage legumes have a role in modern dairy farming systems[J]. Irish Journal of Agricultural and Food Research, 2009, 48(2):137-147.

[3]
PHELAN P, MOLONEY A P, MCGEOUGH E J, et al. Forage legumes for grazing and conserving in ruminant production systems[J]. Critical Reviews in Plant Sciences, 2015, 34(1/2/3):281-326.

[4]
EEVA M. Red clover isoflavonoids in feed,plasma and milk of ruminants[D]. Helsinki:University of Helsinki, 2009.

[5]
HOMOLKA P, KOUKOLOVÁ V, PODSEDNÍČ EK M, et al. Nutritive value of red clover and lucerne forages for ruminants estimated by in vitro and in vivo digestibility methods[J]. Czech Journal of Animal Science,2012, 57(10): 454-468.

[6]
KUIPER G G, LEMMEN J G, CARLSSON B, et al. Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor beta[J]. Endocrinology, 1998, 139(10):4252-4263.

DOI PMID

[7]
ANDRES S, HANSEN U, NIEMANN B, et al. Determination of the isoflavone composition and estrogenic activity of commercial dietary supplements based on soy or red clover[J]. Food & Function, 2015, 6(6):2017-2025.

[8]
PREEDY V R. Isoflavones:chemistry,analysis,function and effects[M]. Cambridge: The Royal Society of Chemistry, 2013.

[9]
XU Q B, LI Y J, DU W J, et al. Effect of dietary biochanin A on lactation performance, antioxidant capacity,rumen fermentation and rumen microbiome of dairy goat[J]. Frontiers in Microbiology, 2023,14:1101849.

[10]
XIONG Z B, LI Y J, ZHANG X Y, et al. Effects of biochanin A on lactational performance,nitrogen metabolism,and blood metabolites in dairy cows[J]. Animal Nutrition, 2024,18:441-449.

[11]
ZHANG S Q, ZHANG X Y, XIONG Z B, et al. Effect of red clover isoflavones on hormone,immune,inflammatory,and plasma biochemistry in lactating dairy cows[J]. Animal Nutrition, 2024,16:306-312.

DOI PMID

[12]
HARLOW B E, FLYTHE M D, AIKEN G E. Biochanin a improves fibre fermentation by cellulolytic bacteria[J]. Journal of Applied Microbiology, 2018, 124(1):58-66.

[13]
HARLOW B E, FLYTHE M D, AIKEN G E. Effect of biochanin A on corn grain (Zea mays) fermentation by bovine rumen amylolytic bacteria[J]. Journal of Applied Microbiology, 2017, 122(4):870-880.

DOI PMID

[14]
HARLOW B E, FLYTHE M D, KLOTZ J L, et al. Effect of biochanin A on the rumen microbial community of Holstein steers consuming a high fiber diet and subjected to a subacute acidosis challenge[J]. PLoS One, 2021, 16(7):e0253754.

[15]
LIU S J, ZHANG Z Y, HAILEMARIAM S, et al. Biochanin a inhibits ruminal nitrogen-metabolizing bacteria and alleviates the decomposition of amino acids and urea in vitro[J]. Animals, 2020, 10(3):368.

[16]
ALVIRA P, TOMÁS-PEJÓ E, BALLESTEROS M, et al. Pretreatment technologies for an efficient bioethanol production process based on enzymatic hydrolysis:a review[J]. Bioresource Technology, 2010, 101(13):4851-4861.

[17]
CHEN H Z, LIU L Y. Unpolluted fractionation of wheat straw by steam explosion and ethanol extraction[J]. Bioresource Technology, 2007, 98(3):666-676.

DOI PMID

[18]
KUROSUMI A, SASAKI C, KUMADA K, et al. Novel extraction method of antioxidant compounds from Sasa palmate (Bean) Nakai using steam explosion[J]. Process Biochemistry, 2007, 42(10):1449-1453.

[19]
ERREIRA I C F R, BAPTISTA P, VILAS-BOAS M, et al. Free-radical scavenging capacity and reducing power of wild edible mushrooms from northeast Portugal:individual cap and stipe activity[J]. Food Chemistry, 2007,100:1511-1516.

[20]
YU Z D, ZHANG B L, YU F Q, et al. A real explosion: the requirement of steam explosion pretreatment[J]. Bioresource Technology, 2012,121:335-341.

[21]
ZHAO Z M, WANG L, CHEN H Z. Physical structure changes of solid medium by steam explosion sterilization[J]. Bioresource Technology, 2016,203:204-210.

[22]
张兵兵, 曾国明, 傅亚, 等. 蒸汽爆破提取银杏叶黄酮类化合物的工艺研究[J]. 纤维素科学与技术, 2012, 20(1):39-44,71.

ZHANG B B, ZENG G M, FU Y, et al. Optimization of steam explosion assisted extractionof flavonoids from Ginkgo biloba leaves by response surface methodology[J]. Cellulose Science and Technology, 2012, 20(1):39-44,71. (in Chinese)

[23]
张棋, 易军鹏, 李欣, 等. 蒸汽爆破预处理对粉葛总黄酮及抗氧化性的影响[J]. 食品科学, 2016, 37(9):40-44.

DOI

ZHANG Q, YI J P, LI X, et al. Effect of steam explosion on total isoflavone content and antioxidant capacity of Puerariae thomsonii Radix[J]. Food Science, 2016, 37(9):40-44. (in Chinese)

DOI

[24]
靳羽慧, 刘长忠, 徐响, 等. 蒸汽爆破对豆渣中大豆异黄酮的影响研究[J]. 中国粮油学报, 2017, 32(10):16-20,27.

JIN Y H, LIU C Z, XU X, et al. Effect of steam explosion treatment on soybean isoflavones of okara[J]. Journal of the Chinese Cereals and Oils Association, 2017, 32(10):16-20,27. (in Chinese)

[25]
熊孟雪, 杨敏, 陈前林. 基于Box-Behnken响应面法的钠离子吸附剂的制备工艺条件优化[J]. 硅酸盐通报, 2022, 41(7):2360-2367.

XIONG M X, YANG M, CHEN Q L. Optimization of preparation process conditions for sodium ion adsorbents based on Box-Behnken response surface methodology[J]. Bulletin of the Chinese Ceramic Societ, 2022, 41(7):2360-2367. (in Chinese)

[26]
徐欣娜, 王婧怡, 吕欣成, 等. 响应面法优化超声-微波协同提取黄连素的工艺研究[J]. 轻工科技, 2023, 39(5):42-47.

XU X N, WANG J Y, LV X C, et al. Process optimization of berberine extraction using ultrasound-microwave synergy based on response surface methodology[J]. Light Industry Science and Technology, 2023, 39(5):42-47. (in Chinese)

[27]
任天宝, 徐桂转, 马孝琴, 等. 蒸汽爆破对玉米秸秆理化特性的影响[J]. 高压物理学报, 2012, 26(2):227-234.

REN T B, XU G Z, MA X Q, et al. Influence of steam explosion on physical-chemical characteristic of corn stalk[J]. Chinese Journal of High Pressure Physics, 2012, 26(2):227-234. (in Chinese)

[28]
田明. 基于Box-Behnken响应面设计结合网络药理学的雪菊复合饮料的研究[D]. 硕士学位论文. 乌鲁木齐: 新疆农业大学, 2022.

TIAN M. Research on a compound beverage technology of Coreopsis tinctoria based on Box-Behnken response surface design combined with network pharmacology[D]. Master’s Thesis. Urumqi: Xinjiang Agricultural University, 2022. (in Chinese)

[29]
ZHANG Y P, YANG R J, ZHAO W, et al. Physicochemical and emulsifying properties of protein extracted from soybean meal assisted by steam flash-explosion[J]. Innovative Food Science & Emerging Technologies, 2014,23:131-137.

[30]
AGBOR V B, CICEK N, SPARLING R, et al. Biomass pretreatment:fundamentals toward application[J]. Biotechnology Advances, 2011, 29(6):675-685.

[31]
SUI W J, ZHOU M J, XU Y, et al. Hydrothermal deglycosylation and deconstruction effect of steam explosion:application to high-valued glycyrrhizic acid derivatives from liquorice[J]. Food Chemistry, 2020,307:125558.

[32]
王田林. 蒸汽爆破技术对甘薯渣膳食纤维改性研究[D]. 硕士学位论文. 新乡: 河南科技学院, 2017.

WANG T L. The study on modification of dietary fiber from sweet potato residue by steam explosion[D]. Master’s Thesis. Xinxiang: Henan Institute of Science and Technology, 2017. (in Chinese)

[33]
SUI W J, XIE X, LIU R, et al. Effect of wheat bran modification by steam explosion on structural characteristics and rheological properties of wheat flour dough[J]. Food Hydrocolloids, 2018,84:571-580.

[34]
FAN X Y, SU Y L, WU Y, et al. Comprehensive understanding of impacts of steam explosion on facilitated extraction and transformation of flavonoids from Astragali Radix[J]. Food Chemistry, 2025, 463(Pt 3):141410.

[35]
FU X G, CHEN H Z. Air-steam explosion enhancing the extraction efficiency of chlorogenic acid from leaves of Eucommia ulmoides Oliver[J]. Separation and Purification Technology, 2015,146:317-325.

[36]
WAN F C, FENG C F, LUO K Y, et al. Effect of steam explosion on phenolics and antioxidant activity in plants:a review[J]. Trends in Food Science & Technology, 2022,124:13-24.

[37]
SONG H D, YANG R J, ZHAO W, et al. Innovative assistant extraction of flavonoids from pine (Larix olgensis Henry) needles by high-density steam flash-explosion[J]. Journal of Agricultural and Food Chemistry, 2014, 62(17):3806-3812.

[38]
肖咏梅, 李明, 毛璞, 等. 黄酮类化合物生物改性及活性的研究进展[J]. 河南工业大学学报(自然科学版), 2019, 40(2):123-131,139.

XIAO Y M, LI M, MAO P, et al. Research progress on the biological modification and activity of flavonoids[J]. Journal of Henan University of Technology (Natural Science Edition), 2019, 40(2):123-131,139. (in Chinese)

[39]
CHEN Y S, SHAN S R, CAO D M, et al. Steam flash explosion pretreatment enhances soybean seed coat phenolic profiles and antioxidant activity[J]. Food Chemistry, 2020,319:126552.

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