中国农业科技导报 ›› 2024, Vol. 26 ›› Issue (5): 138-147.DOI: 10.13304/j.nykjdb.2023.0281
• 动植物健康 • 上一篇
张岚雄1,2(), 郑威2,3, 陈源桉2,3, 沈婧2,3, 邹双全2, 伍建榕1(
), 倪林2,3(
)
收稿日期:
2023-04-10
接受日期:
2023-06-14
出版日期:
2024-05-15
发布日期:
2024-05-14
通讯作者:
伍建榕,倪林
作者简介:
张岚雄 E-mail:Harlan_515@outlook.com
基金资助:
Lanxiong ZHANG1,2(), Wei ZHENG2,3, Yuan’an CHEN2,3, Jing SHEN2,3, Shuangquan ZOU2, Jianrong WU1(
), Lin NI2,3(
)
Received:
2023-04-10
Accepted:
2023-06-14
Online:
2024-05-15
Published:
2024-05-14
Contact:
Jianrong WU,Lin NI
摘要:
为研究樟叶总木脂素的最佳回流提取工艺及抑菌活性,以樟叶为研究对象,测定其芝麻素和9-羟基芝麻素含量,通过单因素联合Box-Behnken响应面法对樟叶总木脂素回流提取工艺进行优化,并采用带毒平板法对总木脂素的抑菌活性进行评价。结果表明,回流提取最佳工艺条件为提取温度86 ℃、料液比1∶8.39(g·mL-1)、提取时间43 min、提取溶剂为无水乙醇,在此条件下木脂素提取率达0.380 4%。各因素对提取率的影响表现为提取温度>提取时间>料液比。抑菌活性试验表明,木脂素含量为2 mg·mL-1时对链孢粘帚霉菌(Gliocladium catenulatum)、苹果黑腐皮壳菌(Valsa mali Miyabe
中图分类号:
张岚雄, 郑威, 陈源桉, 沈婧, 邹双全, 伍建榕, 倪林. 樟叶总木脂素提取工艺及其抑菌活性研究[J]. 中国农业科技导报, 2024, 26(5): 138-147.
Lanxiong ZHANG, Wei ZHENG, Yuan’an CHEN, Jing SHEN, Shuangquan ZOU, Jianrong WU, Lin NI. Study on Extraction Process and Antibacterial Activity of Lignans from Cinnamomum camphora Leaves[J]. Journal of Agricultural Science and Technology, 2024, 26(5): 138-147.
菌株Strain | 名称Name | 类别Taxonomy |
---|---|---|
F-3 | 西瓜尖孢镰孢菌Fusarium oxysporumf | 半知菌类Deuteromycetes |
G-1 | 链孢粘帚霉菌Gliocladium catenulatum | 半知菌类Deuteromycetes |
V | 苹果黑腐皮壳菌Valsa mali Miyabe et Yamada | 子囊菌门Ascomycota |
P-3 | 瓜果腐霉菌Pythium aphanidermatum | 卵菌门Oomycetes |
表1 供试菌种
Table 1 Tested fungal
菌株Strain | 名称Name | 类别Taxonomy |
---|---|---|
F-3 | 西瓜尖孢镰孢菌Fusarium oxysporumf | 半知菌类Deuteromycetes |
G-1 | 链孢粘帚霉菌Gliocladium catenulatum | 半知菌类Deuteromycetes |
V | 苹果黑腐皮壳菌Valsa mali Miyabe et Yamada | 子囊菌门Ascomycota |
P-3 | 瓜果腐霉菌Pythium aphanidermatum | 卵菌门Oomycetes |
水平level | 因素Factor | ||
---|---|---|---|
A:提取温度 Extraction temperature/℃ | B:料液比 Ratio of solid to liquid/(g·mL-1) | C:提取时间 Extraction time/min | |
-1 | 80 | 1∶6 | 30 |
0 | 85 | 1∶8 | 40 |
1 | 90 | 1∶10 | 50 |
表2 响应面试验因素水平
Table 2 Factor level of response surface
水平level | 因素Factor | ||
---|---|---|---|
A:提取温度 Extraction temperature/℃ | B:料液比 Ratio of solid to liquid/(g·mL-1) | C:提取时间 Extraction time/min | |
-1 | 80 | 1∶6 | 30 |
0 | 85 | 1∶8 | 40 |
1 | 90 | 1∶10 | 50 |
图3 不同处理下木脂素的提取率注:不同小写字母表示不同处理间在P<0.05水平差异显著。
Fig. 3 Extraction rate of lignan under different treatmentsNote: Different lowercase letters indicate significant differences between different treatments at P<0.05 level.
序号 Number | A:提取温度 Extraction temperature/℃ | B:料液比 Ratio of solid to liquid/(g·mL-1) | C:提取时间 Extraction time/min | 木脂素提取率 Extraction rate of lignans/% |
---|---|---|---|---|
1 | 80 | 1∶6 | 40 | 0.358 5 |
2 | 90 | 1∶6 | 40 | 0.362 0 |
3 | 80 | 1∶10 | 40 | 0.357 0 |
4 | 90 | 1∶10 | 40 | 0.370 9 |
5 | 80 | 1∶8 | 30 | 0.356 6 |
6 | 90 | 1∶8 | 30 | 0.365 8 |
7 | 80 | 1∶8 | 50 | 0.364 7 |
8 | 90 | 1∶8 | 50 | 0.369 9 |
9 | 85 | 1∶6 | 30 | 0.357 9 |
10 | 85 | 1∶10 | 30 | 0.367 0 |
11 | 85 | 1∶6 | 50 | 0.368 8 |
12 | 85 | 1∶10 | 50 | 0.373 4 |
13 | 85 | 1∶8 | 40 | 0.381 1 |
14 | 85 | 1∶8 | 40 | 0.379 6 |
15 | 85 | 1∶8 | 40 | 0.377 5 |
16 | 85 | 1∶8 | 40 | 0.378 3 |
17 | 85 | 1∶8 | 40 | 0.378 9 |
表3 响应面试验结果
Table 3 Experimental results of response surface test
序号 Number | A:提取温度 Extraction temperature/℃ | B:料液比 Ratio of solid to liquid/(g·mL-1) | C:提取时间 Extraction time/min | 木脂素提取率 Extraction rate of lignans/% |
---|---|---|---|---|
1 | 80 | 1∶6 | 40 | 0.358 5 |
2 | 90 | 1∶6 | 40 | 0.362 0 |
3 | 80 | 1∶10 | 40 | 0.357 0 |
4 | 90 | 1∶10 | 40 | 0.370 9 |
5 | 80 | 1∶8 | 30 | 0.356 6 |
6 | 90 | 1∶8 | 30 | 0.365 8 |
7 | 80 | 1∶8 | 50 | 0.364 7 |
8 | 90 | 1∶8 | 50 | 0.369 9 |
9 | 85 | 1∶6 | 30 | 0.357 9 |
10 | 85 | 1∶10 | 30 | 0.367 0 |
11 | 85 | 1∶6 | 50 | 0.368 8 |
12 | 85 | 1∶10 | 50 | 0.373 4 |
13 | 85 | 1∶8 | 40 | 0.381 1 |
14 | 85 | 1∶8 | 40 | 0.379 6 |
15 | 85 | 1∶8 | 40 | 0.377 5 |
16 | 85 | 1∶8 | 40 | 0.378 3 |
17 | 85 | 1∶8 | 40 | 0.378 9 |
方差来源 Source of variance | 平方和 Sum of squares | 自由度 df | 均方 Mean square | F值 F value | P值 P value |
---|---|---|---|---|---|
模型 Model | 1.136×10-3 | 9 | 1.262×10-4 | 52.72 | <0.000 1 |
A | 1.271×10-4 | 1 | 1.271×10-4 | 53.06 | 0.000 2 |
B | 5.544×10-5 | 1 | 5.544×10-5 | 23.15 | 0.001 9 |
C | 1.090×10-4 | 1 | 1.090×10-4 | 45.53 | 0.000 3 |
AB | 2.721×10-5 | 1 | 2.721×10-5 | 11.36 | 0.011 9 |
AC | 3.978×10-6 | 1 | 3.978×10-6 | 1.66 | 0.238 4 |
BC | 5.067×10-6 | 1 | 5.067×10-6 | 2.12 | 0.189 1 |
A2 | 4.009×10-4 | 1 | 4.009×10-4 | 167.42 | <0.000 1 |
B2 | 2.196×10-4 | 1 | 2.196×10-4 | 91.72 | <0.000 1 |
C2 | 1.096×10-4 | 1 | 1.096×10-4 | 45.79 | 0.000 3 |
残差 Residual error | 1.676×10-5 | 7 | 2.395×10-6 | ||
失拟项 Lack of fit | 9.321×10-6 | 3 | 3.107×10-6 | 1.67 | 0.309 2 |
纯误差 Pure error | 7.440×10-6 | 4 | 1.860×10-6 | ||
总和 Summation | 1.153×10-3 | 16 |
表4 木脂素含量作为响应值的回归模型方差分析
Table 4 ANOVA for regerssion model with lignanoids contents as response value
方差来源 Source of variance | 平方和 Sum of squares | 自由度 df | 均方 Mean square | F值 F value | P值 P value |
---|---|---|---|---|---|
模型 Model | 1.136×10-3 | 9 | 1.262×10-4 | 52.72 | <0.000 1 |
A | 1.271×10-4 | 1 | 1.271×10-4 | 53.06 | 0.000 2 |
B | 5.544×10-5 | 1 | 5.544×10-5 | 23.15 | 0.001 9 |
C | 1.090×10-4 | 1 | 1.090×10-4 | 45.53 | 0.000 3 |
AB | 2.721×10-5 | 1 | 2.721×10-5 | 11.36 | 0.011 9 |
AC | 3.978×10-6 | 1 | 3.978×10-6 | 1.66 | 0.238 4 |
BC | 5.067×10-6 | 1 | 5.067×10-6 | 2.12 | 0.189 1 |
A2 | 4.009×10-4 | 1 | 4.009×10-4 | 167.42 | <0.000 1 |
B2 | 2.196×10-4 | 1 | 2.196×10-4 | 91.72 | <0.000 1 |
C2 | 1.096×10-4 | 1 | 1.096×10-4 | 45.79 | 0.000 3 |
残差 Residual error | 1.676×10-5 | 7 | 2.395×10-6 | ||
失拟项 Lack of fit | 9.321×10-6 | 3 | 3.107×10-6 | 1.67 | 0.309 2 |
纯误差 Pure error | 7.440×10-6 | 4 | 1.860×10-6 | ||
总和 Summation | 1.153×10-3 | 16 |
图4 各因素对木脂素提取率影响的响应曲面图(左)和等高线图(右)
Fig. 4 Response surface diagram (left) and contour diagram (right) of the influence of different factors and extraction time on the extraction rate of lignans
序号 Number | 提取率 Extraction rate/% | 平均提取率 Average extraction rate/% | 相对误差 Relative error/% |
---|---|---|---|
1 | 0.381 7 | ||
2 | 0.380 2 | 0.380 5 | 0.015 6 |
3 | 0.379 5 |
表5 验证试验
Table 5 Verification experiment
序号 Number | 提取率 Extraction rate/% | 平均提取率 Average extraction rate/% | 相对误差 Relative error/% |
---|---|---|---|
1 | 0.381 7 | ||
2 | 0.380 2 | 0.380 5 | 0.015 6 |
3 | 0.379 5 |
菌株 Strain | 毒力回归方程 Virulence regression equation | 相关系数 Correlation coefficient | EC50/(mg·mL-1) | 95%置信限 95% confidence limit/(mg·mL-1) |
---|---|---|---|---|
G-1 | y=1.911x-0.177 | 0.886 | 1.238 | 0.672~1.925 |
V | y=2.093x-0.224 | 0.889 | 1.280 | 0.617~2.164 |
F-3 | y=1.988x-0.342 | 0.923 | 1.486 | 0.843~2.387 |
P-3 | y=1.767x+0.667 | 0.998 | 0.419 | 0.013~0.882 |
表6 樟木脂素对4种真菌的毒力测定
Table 6 Toxicity determination of C. camphora lignans to 4 fungi
菌株 Strain | 毒力回归方程 Virulence regression equation | 相关系数 Correlation coefficient | EC50/(mg·mL-1) | 95%置信限 95% confidence limit/(mg·mL-1) |
---|---|---|---|---|
G-1 | y=1.911x-0.177 | 0.886 | 1.238 | 0.672~1.925 |
V | y=2.093x-0.224 | 0.889 | 1.280 | 0.617~2.164 |
F-3 | y=1.988x-0.342 | 0.923 | 1.486 | 0.843~2.387 |
P-3 | y=1.767x+0.667 | 0.998 | 0.419 | 0.013~0.882 |
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