初榨橄榄油中多酚化合物的UPLC-FLD检测及其抗氧化活性研究

doi:10.3969/j.issn.1004-1524.2021.05.16

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (5): 907-915.DOI: 10.3969/j.issn.1004-1524.2021.05.16

初榨橄榄油中多酚化合物的UPLC-FLD检测及其抗氧化活性研究

李雪¹^,²^,³(), 张玉¹^,²^,³, 王君虹¹^,³, 朱作艺¹^,³, 孙素玲¹, 刘志¹^,⁴, 朱申龙³, 沈国新³, Agusti ROMERO⁵, 王伟¹^,³^,^*()

1.浙江省农业科学院农产品质量安全与营养研究所,浙江杭州 310021
2.农业农村部农产品信息溯源重点实验室,浙江杭州 310021
3.木本油料品质营养国际联合实验室,浙江杭州310021
4.湖南人文科技学院农业与生物技术学院,湖南娄底 417000
5.西班牙加泰罗尼亚农业与食品科技研究所, 西班牙加泰罗尼亚 08140

收稿日期:2020-07-07 出版日期:2021-05-25 发布日期:2021-05-25
通讯作者: 王伟
作者简介:*王伟,E-mail:wangwei5228345@126.com
李雪(1986—),女,吉林四平人,博士,助理研究员,主要从事木本油料营养品质与功能活性研究。E-mail:hei.semeng@163.com
基金资助:
浙江省农业科学院青年人才培养项目(2020);浙江省农业科学院B类学科建设项目;浙江省农业科学院国际合作项目(2019)

UPLC-FLD determination and antioxidant activity of polyphenols in virgin olive oil

LI Xue¹^,²^,³(), ZHANG Yu¹^,²^,³, WANG Junhong¹^,³, ZHU Zuoyi¹^,³, SUN Suling¹, LIU Zhi¹^,⁴, ZHU Shenlong³, SHEN Guoxin³, Agusti ROMERO⁵, WANG Wei¹^,³^,^*()

1. Institue of Agro-Product Safety and Nutrition, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2. Key Laboratory of Information Traceability for Agricultural Products, Ministry of Agriculture and Rural Affairs of China, Hangzhou 310021, China
3. International Joint Laboratory for Quality and Nutrition of Woody Oils, Hangzhou 310021, China
4. College of Agriculture and Biotechnology, Hunan University of Humanities, Science and Technology, Loudi 417000, China
5. Institute of Agriculture and Food Research and Technology, Catalonia 08140, Spain

Received:2020-07-07 Online:2021-05-25 Published:2021-05-25
Contact: WANG Wei

摘要/Abstract

摘要：

采用超高效液相色谱荧光检测法(UPLC-FLD)检测不同品种初榨橄榄油中6种多酚化合物的含量,并测定样品的总多酚含量及抗氧化活性,同时对多酚组分与抗氧化能力的相关性进行分析。结果表明:色谱条件为采用Cadenza色谱柱(150 mm×4.6 mm, 5 μm),柱温35 ℃,以0.1%甲酸-水(A)-乙腈(B)为流动相进行梯度洗脱,荧光光谱激发波长为278 nm,发射波长为339 nm。在此条件下,初榨橄榄油样品中6种多酚化合物可在17 min内得到较好分离,并在较宽的浓度范围内呈良好的线性关系(R²≥ 0.999 5),且精密度高,日内精密度相对标准偏差(RSD)≤ 4.91%,日间精密度RSD ≤5.41%,加标回收结果准确可靠,平均回收率93.42%~119.86%,RSD ≤ 6.26%。不同初榨橄榄油样品中羟基酪醇、酪醇、香草酸、松脂酚及总多酚含量差异较大,而橄榄苦苷和女贞苷含量最低,且样品间差异较小。初榨橄榄油多酚提取物具有良好的抗氧化活性,但不同品种间具有较大差异,ABTS法测定的总抗氧化能力为0.95~2.71 mmol·L^-1 Trolox,DPPH自由基清除率为7.22%~66.23%,抑制羟自由基能力为13.24~136.06 U·mL^-1,其中Coratina的抗氧化活性最强,总多酚含量与提取物总抗氧化能力呈显著正相关。

关键词: 初榨橄榄油, 多酚化合物, 超高效液相色谱荧光检测法, 抗氧化活性

Abstract:

An ultra high performance liquid chromatography (UPLC) coupled with fluorescence detector (FLD) was used to determine the contents of 6 selected phenolic compounds in different samples of virgin olive oil. The total polyphenols content and antioxidant activity of these samples were also detected. The chromatographic column used was Cadenza (150 mm×4.6 mm, 5 μm) and the mobile phase was composed of 0.1% formate-water (A) and acetonitrile (B). Column temperature was at 35℃. The excitation wavelength and emission wavelength were 278 nm and 339 nm. Under the conditions, 6 phenolic compounds in virgin olive oil could be separated well within 17 min, with a wider range of concentration, good linear relationship (R²≥ 0.999 5), high precision with the relative standard deviation (RSD) of intra-day precision≤ 4.91%, RSD of inter-day precision ≤ 5.41%, and reliable recovery with the average recovery rate of 93.42%-119.86%, RSD≤6.26%. The contents of hydroxytyrosol, tyrosol, vanillic acid, pinoresinol and total polyphenols in different virgin olive oil samples were significantly different, while the contents of oleuropein and ligstroside were the lowest and the differences between samples were relatively small. Polyphenols extracts of virgin olive oil have good antioxidant activity, but significant differences between different samples were detected. The total antioxidant capacities of the samples were from 0.95 to 2.71 mmol·L^-1 Trolox determined by ABTS method. DPPH free radical scavenging activities of samples were 7.22%-66.23%. Hydroxyl free radical clearance capabilities of samples were 13.24-136.06 U·mL^-1. Coratina showed the strongest antioxidant activity among samples. The content of total polyphenols was significantly correlated to total antioxidant capacity of the extracts of virgin olive oil.

Key words: virgin olive oil, polyphenol, UPLC-FLD, antioxidant activity

中图分类号:

S509

李雪, 张玉, 王君虹, 朱作艺, 孙素玲, 刘志, 朱申龙, 沈国新, Agusti ROMERO, 王伟. 初榨橄榄油中多酚化合物的UPLC-FLD检测及其抗氧化活性研究[J]. 浙江农业学报, 2021, 33(5): 907-915.

LI Xue, ZHANG Yu, WANG Junhong, ZHU Zuoyi, SUN Suling, LIU Zhi, ZHU Shenlong, SHEN Guoxin, Agusti ROMERO, WANG Wei. UPLC-FLD determination and antioxidant activity of polyphenols in virgin olive oil[J]. Acta Agriculturae Zhejiangensis, 2021, 33(5): 907-915.

图/表 5

参考文献 24

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多酚化合物 Polyphenols	荧光检测器FLD					紫外检测器UV
多酚化合物 Polyphenols	回归方程 Regress equation	相关系数 R²	线性范围 Linear range/ (μg·mL^-1)	定量限 LOQ/(μg· mL^-1)	检出限 LOD/(μg· mL^-1)	回归方程 Regress equation	相关系数 R²	线性范围 Linear range/ (μg·mL^-1)	定量限 LOQ/(μg· mL^-1)	检出限 LOD/(μg· mL^-1)
羟基酪醇 Hydroxytyrosol	y=2394141.98x -26214.42	1.0000	0.026-12.8	0.003	0.001	y=9007.84x -219.50	1.0000	0.128-12.8	0.128	0.039
酪醇 Tyrosol	y=2737496.85x -30373.56	1.0000	0.005-10.2	0.004	0.001	y=7006.96x -284.44	1.0000	0.102-10.2	0.102	0.034
香草酸 Vanillic acid	y=2638811.24x -21127.88	1.0000	0.022-11.0	0.006	0.002	y=12618.85x -485.26	1.0000	0.110-11.0	0.110	0.033
橄榄苦苷 Oleuropein	y=283557.60x -78095.20	0.9995	0.098-29.4	0.066	0.020	y=2634.68x -728.51	0.9999	0.490-29.4	0.490	0.148
女贞苷 Ligstroside	y=120011.15x -30187.66	0.9996	0.170-51.0	0.170	0.052	y=1503.57x -813.65	0.9998	0.850-51.0	0.850	0.258
松脂酚 Pinoresinol	y=6008326.90x -2178721.02	0.9995	0.0860-25.8	0.003	0.001	y=8179.26x -4051.88	0.9997	0.430-25.8	0.430	0.130

多酚化合物 Polyphenols	荧光检测器FLD					紫外检测器UV
多酚化合物 Polyphenols	回归方程 Regress equation	相关系数 R²	线性范围 Linear range/ (μg·mL^-1)	定量限 LOQ/(μg· mL^-1)	检出限 LOD/(μg· mL^-1)	回归方程 Regress equation	相关系数 R²	线性范围 Linear range/ (μg·mL^-1)	定量限 LOQ/(μg· mL^-1)	检出限 LOD/(μg· mL^-1)
羟基酪醇 Hydroxytyrosol	y=2394141.98x -26214.42	1.0000	0.026-12.8	0.003	0.001	y=9007.84x -219.50	1.0000	0.128-12.8	0.128	0.039
酪醇 Tyrosol	y=2737496.85x -30373.56	1.0000	0.005-10.2	0.004	0.001	y=7006.96x -284.44	1.0000	0.102-10.2	0.102	0.034
香草酸 Vanillic acid	y=2638811.24x -21127.88	1.0000	0.022-11.0	0.006	0.002	y=12618.85x -485.26	1.0000	0.110-11.0	0.110	0.033
橄榄苦苷 Oleuropein	y=283557.60x -78095.20	0.9995	0.098-29.4	0.066	0.020	y=2634.68x -728.51	0.9999	0.490-29.4	0.490	0.148
女贞苷 Ligstroside	y=120011.15x -30187.66	0.9996	0.170-51.0	0.170	0.052	y=1503.57x -813.65	0.9998	0.850-51.0	0.850	0.258
松脂酚 Pinoresinol	y=6008326.90x -2178721.02	0.9995	0.0860-25.8	0.003	0.001	y=8179.26x -4051.88	0.9997	0.430-25.8	0.430	0.130

多酚化合物 Polyphenols	样品含量 Concentration/ μg	低浓度Low concentration		高浓度High concentration
多酚化合物 Polyphenols	样品含量 Concentration/ μg	加标量 Concentration of addition standard/(μg·g^-1)	回收率 Recovery rates/%	加标量 Concentration of addition standard/(μg·g^-1)	回收率 Recovery rates/%
羟基酪醇Hydroxytyrosol	0	1.26	119.2±1.09	12.59	110.99±0.33
酪醇Tyrosol	0	1.00	99.75±0.39	10.03	106.53±0.62
香草酸Vanillic acid	0	0.96	93.42±0.31	9.64	116.68±0.34
橄榄苦苷Oleuropein	0	1.08	110.51±2.48	10.82	100.67±6.26
女贞苷Ligstroside	0	1.67	119.50±3.12	16.72	100.30±2.33
松脂酚Pinoresinol	0	0.85	119.86±1.10	8.46	101.69±0.67

多酚化合物 Polyphenols	样品含量 Concentration/ μg	低浓度Low concentration		高浓度High concentration
多酚化合物 Polyphenols	样品含量 Concentration/ μg	加标量 Concentration of addition standard/(μg·g^-1)	回收率 Recovery rates/%	加标量 Concentration of addition standard/(μg·g^-1)	回收率 Recovery rates/%
羟基酪醇Hydroxytyrosol	0	1.26	119.2±1.09	12.59	110.99±0.33
酪醇Tyrosol	0	1.00	99.75±0.39	10.03	106.53±0.62
香草酸Vanillic acid	0	0.96	93.42±0.31	9.64	116.68±0.34
橄榄苦苷Oleuropein	0	1.08	110.51±2.48	10.82	100.67±6.26
女贞苷Ligstroside	0	1.67	119.50±3.12	16.72	100.30±2.33
松脂酚Pinoresinol	0	0.85	119.86±1.10	8.46	101.69±0.67

品种 Variety	化合物含量 Content/(μg·g^-1)						总多酚含量 Total polyphenol content/ (μg·g^-1)	DPPH/%	ABTS/ (mmol· L^-1)	抑制羟自由基能力 Hydroxyl radical clearance capacity/ (U·mL^-1)
品种 Variety	羟基酪醇 Hydroxy- tyrosol	酪醇 Tyrosol	香草酸 Vanillic acid	橄榄苦苷 Oleuro- pein	女贞苷 Ligstro- side	松脂醇 Pinoresinol	总多酚含量 Total polyphenol content/ (μg·g^-1)	DPPH/%	ABTS/ (mmol· L^-1)	抑制羟自由基能力 Hydroxyl radical clearance capacity/ (U·mL^-1)
Coratina 科拉蒂	6.26± 0.16Ba	8.73± 0.17Aa	1.19± 1.23Dc	0.97± 0.94Ea	—	3.20± 1.85Cd	160.36± 4.26a	66.23± 3.11 a	2.71± 0.37 a	136.06± 16.52 a
Picual 皮瓜尔	4.13± 0.75 Bb	5.65± 0.37 Ac	1.58± 1.14 Da	0.96± 0.26Eb	—	4.05± 0.55Cb	70.99± 0.53c	11.47± 13.21 d	1.13± 3.19 c	44.19± 14.72 b
Arbosana 阿尔波萨纳	0.75± 3.53 Ed	3.45± 0.16 Bd	1.46± 0.30 Cb	0.96± 0.46Db	—	4.15± 0.45Aa	71.14± 1.31c	24.47± 6.44 b	1.38± 0.84 b	13.24± 4.49 c
Koroneiki 柯基	3.91± 1.56 Bc	6.63± 0.07 Ab	0.99± 0.87 De	0.95± 0.44 Db	—	3.12± 0.39 Cd	92.28± 0.82b	7.22± 7.64 d	1.40± 0.41 b	27.47± 6.46 bc
Arbequina 豆果	0.56± 2.17 Ee	2.83± 0.27 Be	1.12± 0.74 Cd	0.95± 0.19 Db	—	3.52± 0.46 Ac	45.82± 1.24d	19.54± 3.98 c	0.95± 4.76 d	39.59± 5.72 b

初榨橄榄油中多酚化合物的UPLC-FLD检测及其抗氧化活性研究

UPLC-FLD determination and antioxidant activity of polyphenols in virgin olive oil

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