特级初榨橄榄油中酚类化合物不同提取方法对比研究

采用液液萃取法(LLE)、固相萃取法(SPE)和液液微萃取法(LLME)分别对特级初榨橄榄油中酚类成分进行提取,应用超高效液相色谱-高分辨质谱法(UPLC-HRMS)及Folin-Ciocalteus比色法分别进行酚类化合物组成研究和总酚含量测定,比较3种方法的提取效率。结果表明:特级初榨橄榄油中共检测到40种酚类化合物,其中液液萃取样品检出33种,固相萃取样品检出20种,液液微萃取样品检出15种,液液萃取法提取酚类化合物种类显著多于其他两种方法;液液萃取法提取的总酚含量分别为固相萃取法及液液微萃取法的2. 9倍和3. 7倍,液液萃取法对酚类化合物的提取效率明显优于其他两种方法;但从实验过程角度考量,液液微萃取法具有操作简单、省时省力、有机溶剂用量少、对实验人员伤害小、环境污染小等优点,具有进一步优化的潜力。     

Determination of phenolic compounds in olive oil: New method based on liquid–liquid micro extraction and ultra high performance liquid chromatography-triple–quadrupole mass spectrometry

A novel analytical approach has been performed and evaluated for the identification and quantification of phenolic compounds (phenolic alcohols, secoiridoid derivates, lignans, flavonoids, phenolic acids and aldehydes) which can be found in both virgin olive oil (VOO) and extra virgin olive oil (EVOO). An improved liquid–liquid micro extraction (LLME) method combining with ultra high performance liquid chromatography (UHPLC) coupled to electrospray ionization source (ESI) and tandem mass spectrometry (MS/MS) in Dynamic Multiple Reaction Monitoring (DMRM) mode has been developed, reducing the amount of sample, reagents and time consumed. The proposed methodology was applied to standard solutions, and the quality control parameters obtained were compared with those achieved in spiked refined olive oil (SROO). Generally, these parameters showed lower values in SROO than in standard solutions. Matrix effect, recovery and process efficiency have been studied using SROO. The recoveries were around 90% and the process efficiency of the whole method was higher than 80% for most of the phenolic compounds studied. Examination of these parameters revealed that work straight in the olive oil matrix, which is closer to behave as a real sample, provides better results. The applicability and reliability of this methodology have been confirmed using real samples.     

添加新鲜橄榄叶和复合果胶酶压榨对油橄榄果出油率和初榨橄榄油品质的影响

为提高油橄榄果出油率、改善初榨橄榄油(VOO)品质,以甘肃陇南主栽的成熟度为7的莱星品种油橄榄鲜果为原料,考察压榨过程中新鲜橄榄叶(0、3%、5%)和复合果胶酶(0、0.01%、0.02%)添加量(以油橄榄果质量计)对油橄榄果出油率和VOO色泽、叶绿素含量、基本理化性质、总酚含量、脂肪酸组成及含量的影响。结果表明：添加适量的新鲜橄榄叶可提高出油率及总酚含量,降低酸值,但色泽加深,叶绿素含量和过氧化值升高;添加适量的复合果胶酶在提高出油率的同时,VOO的总酚含量上升,叶绿素含量和过氧化值降低,但酸值升高,色泽加深;压榨过程中添加新鲜橄榄叶和复合果胶酶对VOO脂肪酸组成没有影响,但对油酸、亚油酸、棕榈酸和棕榈烯酸等主要脂肪酸含量有一定影响。在压榨制取VOO时添加适量的新鲜橄榄叶与复合果胶酶可提高出油率,获得富含多酚的VOO。     

Solid-phase extraction and gas chromatographic analysis of phenolic compounds in virgin olive oil

A careful investigation on the potential application of solid phase extraction (SPE)-gas chromatography procedure in the analysis of phenol compounds in virgin olive oils was carried out. In doped refined olive oil samples a comparison between liquid/liquid and SPE extraction evidenced higher recovery when the C₁₈ sorbent phase was employed whereas, in the case of total suppression of residual sylanolic group (C_{18 EC}), only contradictory data was obtained. The same procedures were carried out on 10 genuine samples of virgin olive oil. As observed with the standards, C₁₈ sorbent phase gave higher values compared to C_{18 EC} and the liquid/liquid extraction procedures. Satisfactory results were obtained in the detection of ligstroside aglycon but for the oleuropein aglycon the quantitative is not completely reliable for the overlap of some unknown no-phenol compounds.     

Improvement of bioactive phenol content in virgin olive oil with an olive-vegetation water concentrate produced by membrane treatment

Olive-vegetation water (OVW) is very rich in phenols, which can cause pollution problems, due to their high hydrophilicity and antimicrobial activity. In this study, a three-phase membrane system was applied for the recovery of hydrophilic phenols from fresh OVW in an industrial plant, through a prior enzymatic treatment. This innovation yielded both a crude phenolic concentrate (CPC) and a very high reduction in OVW pollution. Furthermore, the CPC was utilised in a virgin olive oil (VOO) extraction process with the aim of improving VOO phenolic content. The results obtained with four different olive cultivars showed that the CPC extraction increased the phenolic content of the VOOs without any alteration of their aroma profiles.     

Monitoring the enrichment of virgin olive oil with natural antioxidants by using a new capillary electrophoresis method

The enrichment of virgin olive oil (VOO) with natural antioxidants contained in various herbs (rosemary, thyme and oregano) was studied. Three different enrichment procedures were used for the solid–liquid extraction of antioxidants present in the herbs to VOO. One involved simply bringing the herbs into contact with the VOO for 190 days; another keeping the herb–VOO mixture under stirring at room temperature (25 °C) for 11 days; and the third stirring at temperatures above room level (35–40 °C). The efficiency of each procedure was assessed by using a reproducible, efficient, reliable analytical capillary zone electrophoresis (CZE) method to separate and determine selected phenolic compounds (rosmarinic and caffeic acid) in the oil. Prior to electrophoretic separation, the studied antioxidants were isolated from the VOO matrix by using an optimised preconcentration procedure based on solid phase extraction (SPE). The CZE method was optimised and validated.     

Volatile phenols in virgin olive oils: Influence of olive variety on their formation during fruits storage

The potential significance as odorants and markers of olive fruits degradation has been recently pointed out for volatile phenols in virgin olive oil (VOO) and related to the appearance of VOO sensory defects. The few studies carried out in order to elucidate the factors affecting their formation in olive fruits or VOOs, indicated that they could be considered as analytical indices of olive fruits degradation during storage, likely reflecting the microbiological activity. In the present study, the effect of the olive variety (‘Arbequina’, ‘Arbosana’ and ‘Leccino’) on the production of volatile phenols during twelve days of storage in closed plastic bags was evaluated. The different resistance of each variety to the microbiological attach was observed during olive fruit storage, and it was reflected by the evolution of guaiacol, 4-ethylphenol and 4-ethylguaiacol, and related to free acidity values. On the contrary, a scarce dependence on the microbial growth or varietal factors was observed for 4-vinyl derivatives, which appeared more directly related to the time of olives storage. The evolution of volatile phenols found certain correspondence in the sensory characteristics of the resulting VOOs, while the rest of VOO chemical quality indices did not show major variations during fruits storage.     

Chlorophylls in olive and in olive oil: chemistry and occurrences

The chlorophylls are responsible for the characteristic green color of the olive fruits and their products. Virgin olive oil (VOO) is obtained from processing olives only by mechanical and physical means under conditions ensuring that the natural characteristics of the fruit composition are maintained as far as possible. In terms of the total chlorophyll content of oil, the extraction process entails a loss of chlorophyll of up to 80%. Many factors, both agronomical and technological, can affect the presence of green pigments in VOO. The analysis of green pigments in olives and/or oil requires an initial phase of extraction of these compounds from the solid and fluid matrix, followed by the selective separation and subsequent identification of the different components of the chlorophyll fraction. The aim of this review article is to summarize and critically analyze the available information about chlorophylls in VOO.     

Contribution of olive seed to the phenolic profile and related quality parameters of virgin olive oil

BACKGROUND: Conflicting results have been reported about the effect of fruit de‐stoning on the virgin olive oil (VOO) phenolic profile. The aim of the present study was to determine whether olive seed plays any role in the synthesis of this oil phenolic fraction. RESULTS: Increases of around 25% of total phenolic compounds were observed in oils obtained from de‐stoned olive fruits in three main Spanish cultivars. To investigate the involvement of olive seed in determining the phenolic profile of VOO, whole intact olive fruits were added with up to 400% olive stones. Excellent regression coefficients were found in general for the decrease of total phenolic compounds and, particularly, of o‐diphenolics in the resulting oils. On the other hand, it was found that olive seed contains a high level of peroxidase (POX) activity (72.4 U g^?1 FW), accounting for more than 98% of total POX activity in the whole fruit. This activity is able to modify VOO phenolics in vitro, similar to the effect of adding stones during VOO extraction. CONCLUSION: Olive seed plays an important role in determining VOO phenolic profile during the process to obtain an oil that seems to be associated with a high level of POX activity. Copyright © 2007 Society of Chemical Industry     

Determination of Polycyclic Aromatic Hydrocarbons (PAHs) in Olive and Refined Pomace Olive Oils with Modified Low Temperature and Ultrasound-Assisted Liquid–Liquid Extraction Method Followed by the HPLC/FLD

The cleanup method of modified low temperature was compared with the standardized method of modified ultrasound-assisted liquid–liquid (UALL) extraction for the analysis of 15 polycyclic aromatic hydrocarbons (PAHs) in olive oil and refined pomace olive oil. The modified UALL extraction consisted in purification on C₁₈ reversed-phase, Florisil-bonded-phase and NH₂ cartridges, and modified low-temperature extraction was followed by alumina-N and NH₂ solid-phase extraction (SPE) cartridges. Both methods are followed by reversed-phase high-performance liquid chromatography with fluorescence detection. The chromatograms of the final extracts showed lower interferences in both of the methods. The solvent consumption and cost for the modified UALL method were higher than those of the modified low temperature, and also, it needed more equipment, but its analysis time was less. The limit of detection and limit of quantitation of the modified UALL method were 0.16–0.97 and 0.57–2.93 μg kg^?1, respectively, and for the modified low temperature, they were 0.09–1.97 and 0.29–5.99 μg kg^?1, respectively. The PAH recoveries for the modified UALL extraction method ranged from 75.0 to 111.0 % (RSD?=?3–8 %), and for the modified low temperature, they ranged from 81.5 to 113.8 % (RSD?=?3–10 %).     

测定菜籽油中酚类化合物样品前处理方法的比较

采用液液萃取(LLE)、固相萃取(SPE)、QuEChERS 3种不同前处理方法提取菜籽油中酚类化合物,并采用超高效液相色谱串联质谱(LC-MS/MS)进行测定,分析比较了3种不同前处理方法对菜籽油中酚类化合物提取效果的影响。结果表明:在3种不同的前处理方法中,QuEChERS前处理方法的提取效果最好,对菜籽油中7种酚类化合物的回收率最高,可达85.16%～101.78%,基质共提物含量最低,为2.04 mg/g;LLE的回收率为66.04%～101.11%,基质共提物含量最高,为6.80 mg/g;SPE的回收率最低,仅为56.98%～83.79%,基质共提物含量为2.30 mg/g。     

High vacuum applied during malaxation in oil industrial plant: Influence on virgin olive oil extractability and quality

Vacuum technology was applied for the first time at industrial scale to the mechanical extraction processes of virgin olive oil (VOO). The study was carried out using an industrial plant with a working capacity of 4 tons of olives. The use of high vacuum conditions during the malaxation process induced an intense mass transfer into the olive paste that improved the coalescence phenomena of oil droplets with a significant increase in oil extractability for four different cultivars (Arbequina, Arbosana, Koroneiki and Picual). The evaluation of the quality characteristics of the Picual VOO showed an enhancement in the hydrophilic phenol fraction mainly related to the oleacein content. The low temperature set during the vacuum treatment (16 ± 1 °C for 30 min of malaxation) limited the stripping process of the main aldehydes, alcohols and esters. In contrast, other negative volatile compounds related to off-flavoured VOO (ethanol, 3-methyl-1-butanol, ethyl acetate and acetic acid) were significantly reduced.     

气相色谱-质谱检测葡萄酒中氨基甲酸乙酯预处理方法的比较研究

目的比较液液萃取法、固相萃取法和分散式固相萃取法作为预处理方法对葡萄酒中的氨基甲酸乙酯的处理效率。方法葡萄酒样品中的氨基甲酸乙酯分别用二氯甲烷进行液液萃取,用氨基甲酸乙酯专用柱进行固相萃取,用乙腈、石墨化炭黑和N-丙基乙二胺进行分散式固相萃取,再以气相色谱-质谱联用仪测定其含量。结果 3种预处理方法的平均回收率均在70%~110%之间,相对标准偏差(n=5)均小于6%。液液萃取法处理单个样品成本小于5元,富集倍数可达5倍,可用于低含量样品的预处理和进出口企业的产品合规性检测;固相萃取法操作简便,每人每工作日可处理40个样品,可用于企业的高通量检测;分散式固相萃取法成本适中、回收率最佳、综合预处理效率高,一次预处理可同时检测葡萄酒中农药残留和氨基甲酸乙酯含量,适用于葡萄酒中多种危害物质的筛查。结论三种方法各具优势,综合考虑处理效率和实验成本,分散式固相萃取法更适合葡萄酒生产企业的日常应用。该方法已用于葡萄酒中氨基甲酸乙酯的含量测定。     

Comparison of matrix solid-phase dispersion and liquid-liquid extraction for the chromatographic determination of fenthion and its metabolites in olives and olive oils

Matrix solid-phase dispersion (MSPD) methodology has been developed to extract fenthion and its metabolites from olives and olive oils, and the technique compared with conventional liquid-liquid extraction (LLE). The method was applied to olives and olive oil samples obtained from olive groves treated with fenthion. Pesticide residues were analysed by gas chromatography (GC) using a nitrogen-phosphorus detector. Both extraction methods gave a linear response over the concentration range assayed (0.03-1 mg kg^-1 for MSPD and 0.1-1 mg kg^-1 for LLE). Recoveries and RSD (n = 6) values in olives were 85-112% and 2-6% for MSPD, and 68-108% and 4-16% for LLE, respectively. In the case of olive oil, recoveries and RSD (n = 6) values were 67-98% and 5-11% for MSPD, and 63-115% and 6-14% for LLE, respectively. When compared to LLE, the newly developed MSPD method was twice as sensitive and required 10 times less sample weight.     

高压脉冲电场处理对初榨橄榄油得率和品质的影响

为开发一种能提高初榨橄榄油(VOO)得率、对品质无不利影响且适用于工业化生产的新工艺,以甘肃陇南产区莱星、鄂植8号品种油橄榄鲜果为原料,采用高压脉冲电场(PEF)处理辅助传统工艺提取VOO,通过与传统提取工艺比较,研究PEF处理对VOO得率、理化性质、总多酚含量和脂肪酸组成及相对含量的影响。结果表明：与传统提取工艺相比,采用PEF处理VOO得率平均增幅为15.38%,果渣含油率平均降幅为22.28%,对VOO的色泽、酸值、过氧化值、总多酚含量和脂肪酸组成及相对含量等总体无显著性影响。研究认为,在VOO传统提取工艺的基础上,鲜果破碎之后增加PEF处理环节,可显著提高VOO得率,而且对VOO品质无不利影响,该技术在VOO工业化生产中有良好的应用潜力。     

食用植物油中农药残留检测技术的研究进展

我国农产品中农药残留的中毒事件和进出口贸易中农药残留超标事件时有发生,已成为影响农业可持续发展的重要问题,因此农药残留检测技术也在迅速发展。文章综述了现阶段对植物油中农药残留检测技术的研究进展。主要从前处理和检测方法 2方面进行分析,根据前处理方法和检测方法的不同,分别介绍了各类前处理方法如液液萃取法(liquid-liquid extraction,LLE)、固相萃取法(solid phase extraction,SPE)、基质固相分散萃取法(matrix solid phase dispersion,MSPD)、分散固相萃取(disperse solid phase extraction,d SPE)法、凝胶渗透色谱法(gel permeation chromatography,GPC)等方法的原理和过程,以及各类检测方法如气相色谱法、气相色谱-质谱联用法(gas chromatography-mass spectrometry,GC-MS)、液相色谱法和液相色谱-串联质谱联用法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)等的特点和应用范围。文章总结各类方法的优缺点,并提出了食用植物油中农药残留分析方法的发展趋势。     

Recovery of Aromatic Aglycones from Grape Pomace Winemaking By-Products by Using Liquid-Liquid and Pressurized-Liquid Extraction

The potential of winemaking grape pomace by-products as a source of glycosidic aroma precursors that under enzymatic hydrolysis might release aroma compounds has been evaluated. Two different extraction methodologies, liquid–liquid and pressurized liquid extraction (LLE and PLE) were employed. Solid phase extraction (SPE)-GC-MS analysis of the hydrolyzed LLE glycosidic extract revealed 22 aroma compounds belonging to different chemical families (terpenes, C13 norisoprenoids, vanillines, etc.). Response surface methodology was employed to study the effect of the most significant PLE experimental variables (temperature and solvent composition) on the extraction of aromatic aglycones. The parameters of the model were estimated by multiple linear regressions. Most of the aroma compounds showed an adequate fit to the calculated model (18 compounds from 22 with R ²?>?0.8). The application of the optimized PLE conditions (50 % of ethanol in the hydroalcoholic solution) and 90 °C showed higher extraction yield of aglycones when comparing with the extraction yield obtained by LLE.     

Characterization of key aroma compounds in a representative aromatic extracts from citrus and astragalus honeys based on aroma extract dilution analyses

The aroma and aroma-active compounds of astragalus and citrus honeys were determined by GC-MS-olfactometry (GC-MS-O). The honey extracts were obtained by two different extraction techniques including liquid–liquid extraction (LLE) and solid phase extraction (SPE), and compared to determine the more representative odour aromatic honey extract. According to sensory analysis, the aromatic extract (40 g honey with 22 mL water) obtained by LLE was the most similar of honey odour. The amount of aroma compounds of citrus honey (14391.6 µg kg^?1) was higher than astragalus honey (4270.1 µg kg^?1). Terpenes and acids in citrus honey and esters and aldehydes in astragalus honey were the most dominant aroma compounds. Aroma extract dilution analysis (AEDA) was used to determine the aroma-active compounds of honey samples. By application of AEDA on the aromatic extracts isolated by LLE, 12 aroma-active compounds in astragalus honey and 23 in citrus honey could be detected within a flavour dilution (FD) factor range of 8–128. On the basis of the FD factor, phenylethyl alcohol in both honeys was the most powerful key compound and was described as the flowery odour.     

酶处理对初榨橄榄油品质及抗氧化活性的影响

将纤维素酶、果胶酶应用于橄榄油提取工艺,旨在生产具有较高总酚含量及较强抗氧化活性的高质量初榨橄榄油。随着果胶酶和纤维素酶添加量的提高,橄榄油的过氧化值及K₂₃₂均出现下降的趋势,油酸比例有一定程度提高,并在添加0.2%纤维素酶时油酸比例达到最高(65.85%)。结合主成分分析,确定了在油橄榄融合过程中添加0.5%纤维素酶得到的初榨橄榄油总酚含量和抗氧化活性最高。这是由于果胶酶和纤维素酶能有效降解橄榄细胞壁,减少亲水酚类物质与细胞壁多糖的络合,有助于橄榄果皮中的游离酚的释放,从而提高橄榄油中总酚含量及抗氧化活性。     

Direct analysis of glucuronidated metabolites of main olive oil phenols in human urine after dietary consumption of virgin olive oil

In the present study we report on a UPLC-MRM validated method for the simultaneous direct analysis of main glucuronidated metabolites of olive oil phenols: tyrosol, hydroxytyrosol and its O-methyl metabolite homovanillyl alcohol in human urine after dietary olive oil ingestion. The developed method was linear within the concentration range 20–2000 ng/mL with adequate recovery of analytes (>86%). Intra- and inter-day precision and accuracy were according to standard requirements for method validation criteria. Using the developed method, urinary concentrations and excretion rates of glucuronides of olive oil phenols were successfully estimated in an intervention study with 11 healthy volunteers supplemented with a dietary dose of virgin olive oil (VOO) (50 mL). Therefore, about 13% of the consumed olive oil polyphenols were recovered in 24-h urine, where 75% of them were in the form of glucuronides (3′- and 4′-O-hydroxytyrosol glucuronides, 4′-O-glucuronides of tyrosol and homovanillyl alcohol) and 25% as free compounds.