中国农业科技导报 ›› 2023, Vol. 25 ›› Issue (5): 96-105.DOI: 10.13304/j.nykjdb.2021.0855
收稿日期:
2021-10-06
接受日期:
2022-04-12
出版日期:
2023-05-20
发布日期:
2023-07-13
通讯作者:
刘晓燕
作者简介:
徐菁 E-mail:1715175329@qq.cm;
基金资助:
Jing XU1(), Xiufen WANG1, Xiaoyan LIU1,2(
)
Received:
2021-10-06
Accepted:
2022-04-12
Online:
2023-05-20
Published:
2023-07-13
Contact:
Xiaoyan LIU
摘要:
为了探讨姜酚的大孔树脂纯化方法,选择4种不同极性树脂(NKA-9、AB-8、D4020 和D101)作静态吸附-解吸动力学研究。结果表明,AB-8为最佳树脂,以AB-8树脂为吸附剂,对其最佳纯化工艺进行探讨,并比较纯化前后姜酚对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-trinitrophenylhydrazine, DPPH)自由基、2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(2,2-diazo-bis (3-ethyl-benzothiazole-6-sulfonic acid) diammonium,ABTS)自由基等的清除能力和对胰脂肪酶抑制活性,采用高效液相色谱法测定纯化前粗提物和纯化后纯化物中6-姜酚、8-姜酚、10-姜酚含量。结果表明,AB-8树脂动态纯化姜酚最佳条件为:上样质量浓度1.6 mg·mL-1,pH=4,上样流速1.5 mL·min-1,以80%乙醇溶液流速1.5 mL·min-1洗脱,经纯化后姜酚纯度为纯化前的3.3倍,高效液相色谱分析结果表明3种姜酚类单体物质含量相比纯化前提高了3倍以上,纯化后的姜酚对猪胰脂肪酶的抑制率和自由基清除能力明显高于纯化前,说明AB-8树脂对姜酚有较好的分离纯化作用,能够实现成分的富集,对提高姜酚的利用附加值具有重要意义。
中图分类号:
徐菁, 王秀芬, 刘晓燕. 姜酚纯化前后对胰脂肪酶抑制活性、抗氧化活性比较研究[J]. 中国农业科技导报, 2023, 25(5): 96-105.
Jing XU, Xiufen WANG, Xiaoyan LIU. Comparative Study on Pancreatic Lipase Inhibitory Activity and Antioxidant Activity of Gingerol Before and After Purification[J]. Journal of Agricultural Science and Technology, 2023, 25(5): 96-105.
时间Time/min | 流动相Mobile phase | |
---|---|---|
A(0.1%磷酸溶液) A(0.1% phosphoric acid)/% | B(乙腈) B(Acetonitrile)/% | |
0.00 | 14 | 86 |
7.00 | 12 | 88 |
17.50 | 10 | 90 |
表1 高效液相色谱法梯度洗脱程序
Table 1 HPLC gradient elution schedule
时间Time/min | 流动相Mobile phase | |
---|---|---|
A(0.1%磷酸溶液) A(0.1% phosphoric acid)/% | B(乙腈) B(Acetonitrile)/% | |
0.00 | 14 | 86 |
7.00 | 12 | 88 |
17.50 | 10 | 90 |
组号 Group number | 样品 Sample | 胰脂肪酶 Pancreatic lipase | 对硝基苯磷酸二钠六水合物 PNPP | Tris-盐酸缓冲液 Tris-HCl |
---|---|---|---|---|
A1 | 30 | 100 | 100 | 70 |
A0 | 30 | 0 | 100 | 170 |
B1 | 0 | 100 | 100 | 100 |
B0 | 0 | 0 | 100 | 200 |
表 2 反应体系各溶液加入量 (μL)
Table 2 Amount of each solution in the reaction system
组号 Group number | 样品 Sample | 胰脂肪酶 Pancreatic lipase | 对硝基苯磷酸二钠六水合物 PNPP | Tris-盐酸缓冲液 Tris-HCl |
---|---|---|---|---|
A1 | 30 | 100 | 100 | 70 |
A0 | 30 | 0 | 100 | 170 |
B1 | 0 | 100 | 100 | 100 |
B0 | 0 | 0 | 100 | 200 |
树脂型号 Resin type | 方程类型 Equation type | 拟合方程 Fitting equation | R2 | 动力学参数 Kinetic parameter |
---|---|---|---|---|
AB-8 | 一级动力学 First-order dynamic | In(Qe-Qt)=0.640-0.852t | 0.876 9 | Qe=4.13 mg·g-1, K2=1.17 |
二级动力学 Second-order dynamic | t/Qt=0.050+0.242t | 0.999 7 | ||
颗粒内扩散 diffusion particle | Qt=0.302t1/2 +3.209 | 0.684 7 | ||
D101 | 一级动力学 First-order dynamic | In(Qe-Qt)=-0.572-0.807t | 0.866 2 | |
二级动力学 Second-order dynamic | t/Qt=0.101+0.244t | 0.999 0 | Qe=4.09 mg·g-1, K2=0.59 | |
颗粒内扩散 diffusion particle | Qt =0.487t1/2 +2.572 | 0.714 3 | ||
D4020 | 一级动力学 First-order dynamic | In(Qe-Qt)=1.284-1.019t | 0.862 6 | Qe=4.03 mg·g-1, K2=0.42 |
二级动力学 Second-order dynamic | t/Qt=0.248+0.146t | 0.998 2 | ||
颗粒内扩散 diffusion particle | Qt=0.592t1/2 +2.147 | 0.729 8 | ||
NKA-9 | 一级动力学 First-order dynamic | In(Qe-Qt)=0.768-0.952t | 0.868 2 | Qe=3.25 mg·g-1, K2=0.71 |
二级动力学 Second-order dynamic | t/Qt=0.133+0.307t | 0.999 3 | ||
颗粒内扩散 diffusion particle | Qt=0.388t1/2 +2.325 | 0.759 0 |
表3 AB-8、D101、D4020和NKA-9大孔树脂多酚吸附动力学方程和参数
Table 3 Adsorption kinetic equations and parameters of polyphenols on AB-8, D101, D4020 and NKA-9 macroporous resins
树脂型号 Resin type | 方程类型 Equation type | 拟合方程 Fitting equation | R2 | 动力学参数 Kinetic parameter |
---|---|---|---|---|
AB-8 | 一级动力学 First-order dynamic | In(Qe-Qt)=0.640-0.852t | 0.876 9 | Qe=4.13 mg·g-1, K2=1.17 |
二级动力学 Second-order dynamic | t/Qt=0.050+0.242t | 0.999 7 | ||
颗粒内扩散 diffusion particle | Qt=0.302t1/2 +3.209 | 0.684 7 | ||
D101 | 一级动力学 First-order dynamic | In(Qe-Qt)=-0.572-0.807t | 0.866 2 | |
二级动力学 Second-order dynamic | t/Qt=0.101+0.244t | 0.999 0 | Qe=4.09 mg·g-1, K2=0.59 | |
颗粒内扩散 diffusion particle | Qt =0.487t1/2 +2.572 | 0.714 3 | ||
D4020 | 一级动力学 First-order dynamic | In(Qe-Qt)=1.284-1.019t | 0.862 6 | Qe=4.03 mg·g-1, K2=0.42 |
二级动力学 Second-order dynamic | t/Qt=0.248+0.146t | 0.998 2 | ||
颗粒内扩散 diffusion particle | Qt=0.592t1/2 +2.147 | 0.729 8 | ||
NKA-9 | 一级动力学 First-order dynamic | In(Qe-Qt)=0.768-0.952t | 0.868 2 | Qe=3.25 mg·g-1, K2=0.71 |
二级动力学 Second-order dynamic | t/Qt=0.133+0.307t | 0.999 3 | ||
颗粒内扩散 diffusion particle | Qt=0.388t1/2 +2.325 | 0.759 0 |
温度Temperature/℃ | R2 | Langmuir 方程 Langmuir equation | Qm/(mg·g-1) | Ka |
---|---|---|---|---|
35 | 0.996 7 | Ce/ Qe =0.097 4Ce+0.058 3 | 17.15 | 0.59 |
30 | 0.996 6 | Ce/ Qe =0.083 2Ce+0.056 5 | 17.69 | 0.68 |
25 | 0.999 0 | Ce/ Qe= 0.073 0Ce+0.054 7 | 18.28 | 0.75 |
表4 Langmuir 等温吸附方程及相关参数
Table 4 Langmuir isothermal adsorption equation and related parameters
温度Temperature/℃ | R2 | Langmuir 方程 Langmuir equation | Qm/(mg·g-1) | Ka |
---|---|---|---|---|
35 | 0.996 7 | Ce/ Qe =0.097 4Ce+0.058 3 | 17.15 | 0.59 |
30 | 0.996 6 | Ce/ Qe =0.083 2Ce+0.056 5 | 17.69 | 0.68 |
25 | 0.999 0 | Ce/ Qe= 0.073 0Ce+0.054 7 | 18.28 | 0.75 |
温度Temperature/℃ | R2 | Freundlich 方程 Freundlich equation | 1/n | Kb |
---|---|---|---|---|
35 | 0.993 0 | InQe=0.695 0InCe+2.061 2 | 0.69 | 7.9 |
30 | 0.995 9 | InQe=0.715 1InCe+2.140 8 | 0.72 | 8.5 |
25 | 0.997 0 | InQe=0.738 8InCe+2.233 3 | 0.74 | 9.3 |
表5 Freundlich 等温吸附方程及相关参数
Table 5 Freundlich isothermal adsorption equation and related parameters
温度Temperature/℃ | R2 | Freundlich 方程 Freundlich equation | 1/n | Kb |
---|---|---|---|---|
35 | 0.993 0 | InQe=0.695 0InCe+2.061 2 | 0.69 | 7.9 |
30 | 0.995 9 | InQe=0.715 1InCe+2.140 8 | 0.72 | 8.5 |
25 | 0.997 0 | InQe=0.738 8InCe+2.233 3 | 0.74 | 9.3 |
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