佛手果皮精油的提取工艺优化及其成分与抗氧化活性分析

王祎赫; 夏燕莉; 张欣; 杨航; 梅国富; 余波; 易建明; 薛慧玲

doi:10.13386/j.issn1002-0306.2022060055

佛手果皮精油的提取工艺优化及其成分与抗氧化活性分析

doi: 10.13386/j.issn1002-0306.2022060055

1.
成都大学食品与生物工程学院，四川成都 610106
2.
达州市达川区农业农村局，四川达州 635000
3.
广安市经济作物技术推广站，四川广安 638500

基金项目: 四川道地药材中药创新团队（SCCXTD-2020-19）。

详细信息

作者简介:
王祎赫（1998−），女，硕士研究生，研究方向：药食同源产品开发，E-mail：wyh980606@136.com

通讯作者:
薛慧玲（1981−），女，博士，副教授，研究方向：植物资源与新能源开发研究，E-mail：findlot@sohu.com

中图分类号: TS202.3
计量
- 文章访问数: 59
- HTML全文浏览量: 16
- PDF下载量: 7
- 被引次数: 0
出版历程
- 收稿日期: 2022-06-08
- 网络出版日期: 2022-12-15
- 刊出日期: 2023-01-17

Optimization of Extraction Process of Essential Oil from Bergamot Peel and Analysis of Its Components and Antioxidant Activity

1.
College of Food and Biological Engineering, Chengdu University, Chengdu 610106, China
2.
Dazhou Dachuan Agricultural and Rural Bureau, Dazhou 635000, China
3.
Guang'an Cash Crop Technology Extension Station, Guang'an 638500, China

摘要

摘要: 本研究采用纤维素酶辅助水蒸气蒸馏提取法提取佛手果皮精油，在以酶解pH、酶添加量、酶解温度及酶解时间作为单因素分析的基础上，通过Box-Behnken响应面设计法进行提取工艺优化。利用气相色谱-质谱（Gas Chromatography-Mass Spectrometry，GC-MS）法分析提取的精油的化学组成，最后以ABTS⁺·和DPPH·清除率为指标，评价佛手果皮精油的抗氧化活性。结果表明，佛手果皮精油最佳提取工艺为：酶解pH5.2、酶添加量0.7%、酶解温度52 ℃、酶解时间2.1 h，此条件下精油得率为3.11%。从提取的果皮精油中共鉴定出42种化合物，其中乙酸芳樟酯的相对含量最高（14.72%），其次为d-柠檬烯（14.58%）、芳樟醇（8.89%）。抗氧化活性研究结果显示：该法提取的佛手果皮精油在试验浓度范围内具有良好的抗氧化活性，并呈现明显量效关系。当精油浓度为40 mg/mL时，其对ABTS⁺·的清除率为91.20%；浓度达70 mg/mL时，其对DPPH·的清除率达93.19%。此优化工艺精油得率高，且佛手果皮精油其可作为天然抗氧化剂进行开发。
- 佛手果皮 /
- 精油提取 /
- 响应面优化 /
- 气相色谱-质谱法（GC-MS） /
- 抗氧化活性
Abstract: In this study, cellulase-assisted steam distillation was used to extract essential oil from bergamot peel. Based on the single factor analysis of enzymatic hydrolysis pH, enzymatic addition amount, enzymatic hydrolysis temperature and enzymatic hydrolysis time, the extraction process was optimized by using Box-Behnken response surface design method. Moreover, the chemical compositions of the extracted essential oil were analyzed by Gas chromatography-mass Spectrometry (GC-MS). Finally, the antioxidant activities of the essential oil was evaluated by the scavenging rates of ABTS⁺· and DPPH·. According to the research results, 3.11% yield of essential oil was achieved under the optimal extraction conditions: enzymatic hydrolysis of pH5.2, enzymatic dosage of 0.7%, enzymatic hydrolysis temperature of 52 ℃, and enzymatic hydrolysis time of 2.1 h. A total of 42 compounds were identified from the extracted peel essential oils. Among them, linalool acetate had the highest (14.72%) relative content, followed by d-limonene (14.58%) and linalool (8.89%). Besides, according to the results of antioxidant activity study, the essential oil extracted by this method presented good antioxidant activity in the experimental concentration range, and showed an obvious dose-effect relationship. The scavenging rate of ABTS⁺· reached 91.20% when the concentration of essential oil was 40 mg/mL; and the scavenging rate of DPPH· reached 93.19% when the concentration of essential oil was 70 mg/mL. Therefore, this optimized process has high yield of essential oil and can be developed as a natural antioxidant.
- bergamot peel /
- essential oil extraction /
- response surface optimization /
- gas chromatography-mass spectrometry (GC-MS) /
- antioxidant activity

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图 1 酶解pH对精油得率的影响

Figure 1. Effect of enzymatic hydrolysis pH on essential oil extraction rate

注：图中不同小写字母表示差异显著P<0.05，图2~图4同。

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图 2 酶添加量对精油得率的影响

Figure 2. Effect of enzyme addition on essential oil extraction rate

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图 3 酶解温度对精油得率的影响

Figure 3. Effect of enzymatic temperature on extraction rate of essential oil

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图 4 酶解时间对精油得率的影响

Figure 4. Effect of enzymatic hydrolysis time on essential oil extraction rate

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图 5 酶解pH与酶添加量的交互作用对精油得率的影响

Figure 5. Effect of the interaction between enzymatic hydrolysis pH and enzyme addition on the extraction rate

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图 6 酶解pH与酶解时间的交互作用对精油得率的影响

Figure 6. Effect of the interaction between pH and time of enzymatic hydrolysis on the extraction rate

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图 8 酶解温度与酶解时间的交互作用对精油得率的影响

Figure 8. Effect of the interaction between temperature and time of enzymatic hydrolysis on the extraction rate

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图 7 酶添加量与酶解时间的交互作用对精油得率的影响

Figure 7. Effect of interaction between enzyme addition and enzymatic hydrolysis time on extraction rate

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图 9 佛手果皮精油GC-MS总离子流色谱图

Figure 9. Total ion flow chromatogram of bergamot peel essential oil

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图 10 精油对ABTS⁺·的清除能力

Figure 10. Scavenging ability of essential oil to ABTS⁺· free radical

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图 11 精油对DPPH·的清除能力

Figure 11. Scavenging ability of essential oil to DPPH·

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表 1 Box-Behnken 设计因素与水平表

Table 1. Factors and levels of Box-Behnken experimental design

因素	水平
因素	−1	0	1
A（酶解pH）	4.5	5.0	5.5
B（酶添加量/%）	0.6	0.7	0.8
C（酶解温度/℃）	45	50	55
D（酶解时间/h）	1.5	2.0	2.5

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表 2 响应面试验设计及结果

Table 2. Response surface test design and results

试验编号	A	B	C	D	Y（精油得率（%））
1	−1	0	0	1	2.52
2	−1	−1	0	0	2.33
3	0	0	1	1	2.85
4	−1	1	0	0	2.65
5	0	0	1	−1	2.49
6	0	0	0	0	3.05
7	0	0	0	0	3.06
8	1	−1	0	0	2.78
9	0	0	−1	−1	2.22
10	0	0	−1	1	2.40
11	1	0	1	0	2.74
12	0	−1	0	−1	2.53
13	1	0	0	−1	2.51
14	−1	0	1	0	2.61
15	1	0	−1	0	2.44
16	0	0	0	0	3.02
17	1	0	0	1	2.85
18	0	0	0	0	3.06
19	−1	0	0	−1	2.4
20	0	1	0	1	2.92
21	0	0	0	0	3.04
22	0	−1	−1	0	2.33
23	−1	0	−1	0	2.26
24	0	1	0	−1	2.35
25	1	1	0	0	2.65
26	0	1	1	0	2.80
27	0	1	−1	0	2.38
28	0	−1	1	0	2.72
29	0	−1	0	1	2.63

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表 3 响应面二次回归方程模型方差分析

Table 3. Response surface quadratic regression equation model variance analysis

方差来源	平方和	自由度	均方	F值	P值	显著性
模型	1.93	14	0.14	126.42	<0.0001	**
A	0.12	1	0.12	110.32	<0.0001	**
B	0.015	1	0.015	14.17	0.0021	**
C	0.40	1	0.40	364.09	<0.0001	**
D	0.23	1	0.23	213.66	<0.0001	**
AB	0.051	1	0.051	46.54	<0.0001	**
AC	6.250E-004	1	6.250E-004	0.57	0.4610	NS
AD	0.012	1	0.012	11.12	0.0049	**
BC	2.250E-003	1	2.250E-003	0.21	0.6562	NS
BD	0.055	1	0.055	50.77	<0.0001	**
CD	8.100E-003	1	8.100E-003	7.45	0.0163	*
A²	0.36	1	0.36	335.42	<0.0001	**
B²	0.25	1	0.25	228.89	<0.0001	**
C²	0.58	1	0.58	535.50	<0.0001	**
D²	0.39	1	0.39	360.63	<0.0001	**
残差	0.015	14	1.088E-003
失拟检验	0.014	10	1.411E-004	5.04	0.0665	NS
纯误差	1.120E-003	4	2.800E-004
总误差	1.94	28
注：“*”差异极显著（P<0.01）；“”差异显著（（0.01<P<0.05）；“NS”不显著（P>0.05）。

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表 4 佛手果皮精油成分的GC-MS分析

Table 4. GC-MS analysis of essential oil components of bergamot peel

序号	留时间（min）	化合物名称	类别	分子式	CAS	相对含量（%）
1	9.454	（+）-α-蒎烯	萜烯烃	C₁₀H₁₆	7785-70-8	0.29
2	10.516	β-水芹烯	萜烯烃	C₁₀H₁₆	555-10-2	0.28
3	10.624	左旋-β-蒎烯	萜烯烃	C₁₀H₁₆	18172-67-3	1.04
4	10.959	β-月桂烯	萜烯烃	C₁₀H₁₆	123-35-3	0.91
5	11.830	邻-异丙基苯	芳香烃	C₁₀H₁₄	527-84-4	0.26
6	12.018	d-柠檬烯	萜烯烃	C₁₀H₁₆	5989-27-5	14.58
7	12.083	反式-β-罗勒烯	萜烯烃	C₁₀H₁₆	3779-61-1	0.92
8	12.619	γ-松油烯	萜烯烃	C₁₀H₁₆	99-85-4	5.18
9	13.608	芳樟醇	醇	C₁₀H₁₈O	78-70-6	8.89
10	14.249	1,2-二氢芳樟醇	醇	C₁₀H₂₀O	18479-51-1	0.18
11	15.201	（-）-4-萜品醇	醇	C₁₀H₁₈O	20126-76-5	0.50
12	15.515	α-松油醇	醇	C₁₀H₁₈O	10482-56-1	0.85
13	15.626	正癸醛	醛	C₁₀H₂₀O	112-31-2	0.30
14	16.105	橙花醇	醇	C₁₀H₁₈O	106-25-2	0.13
15	16.291	橙花醛	醛	C₁₀H₁₆O	106-26-3	0.79
16	16.532	乙酸芳樟酯	酯	C₁₂H₂₀O₂	115-95-7	14.72
17	16.817	牻牛儿醛	醛	C₁₀H₁₆O	141-27-5	0.96
18	17.935	叶绿醇	醇	C₂₀H₄₀O	150-86-7	0.15
19	18.104	9-十七烷醇	醇	C₁₇H₃₆O	624-08-8	1.17
20	18.338	乙酸橙花酯	酯	C₁₂H₂₀O₂	141-12-8	1.46
21	18.434	3,9-二乙基-6-羟基十三烷	醇	C₁₇H₃₆O	123-24-0	0.65
22	18.495	10-癸醇	醇	C₁₉H₄₀O	16840-84-9	0.46
23	18.673	乙酸香叶酯	酯	C₁₂H₂₀O₂	105-87-3	2.70
24	18.845	顺式-7-十六烯	脂肪烯烃	C₁₆H₃₂	35507-09-6	1.94
25	19.279	甲基丙烯酸十四酯	酯	C₁₈H₃₄O₂	2549-53-3	3.83
26	19.540	8-十六醇	醇	C₁₆H₃₄O	19781-83-0	2.78
27	19.724	己基癸醇	醇	C₁₆H₃₄O	2425-77-6	5.63
28	19.910	橙花叔醇	醇	C₁₅H₂₆O	7212-44-4	3.19
29	19.995	1,2-十六烷二醇	醇	C₁₆H₃₄O₂	6920-24-7	3.04
30	20.125	5-十二烷醇	醇	C₁₂H₂₆O	10203-33-5	2.24
31	20.265	2-异丙基-5-甲基-1-庚醇	醇	C₁₁H₂₄O	91337-07-4	2.66
32	20.415	11-甲基十二烷醇	醇	C₁₃H₂₈O	85763-57-1	2.68
33	20.810	2,6-二叔丁基-4-甲基苯酚	酚	C₁₅H₂₄O	128-37-0	1.76
34	20.928	7-表-反式-倍半桧烯-水合物	醇	C₁₅H₂₆O	58319-05-4	1.24
35	21.145	3,7,11-三甲基-1-十二烷醇	醇	C₁₅H₃₂O	6750-34-1	0.26
36	21.508	（E）-α-没药烯	萜烯烃	C₁₅H₂₄	25532-79-0	0.63
37	21.761	2-辛基十二烷酸	酸	C₂₀H₄₀O₂	40596-46-1	0.36
38	22.380	二十二碳烯酸丙酯	酯	C₂₅H₅₀O₂	26718-94-5	0.17
39	22.685	大根香叶烷	烷烃	C₁₅H₃₀	645-10-3	0.18
40	30.501	抗坏血酸二棕榈酸酯	酯	C₃₈H₆₈O₈	28474-90-0	0.21
41	33.778	共轭（9E,11E）-亚油酸	酸	C₁₈H₃₂O₂	544-71-8	1.33
42	33.877	反式-9-十八碳烯酸	酸	C₁₈H₃₄O₂	112-79-8	1.05

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