顶空固相微萃取与气相色谱-电子捕获技术联用检测软木塞中2,4,6-三氯苯甲醚

建立利用顶空固相微萃取与气相色谱电子捕获技术联用分析检测葡萄酒软木塞中2,4,6-三氯苯甲醚含量的方法。该方法测定软木塞中2,4,6-三氯苯甲醚的相关系数（R²）为0.999 9,精密度好（相对标准偏差不大于5.6%）,回收率为101%～108%,检出限为0.20 pg/L、定量限为0.66 pg/L。利用该方法测定了11 种不同类型的软木塞中2,4,6-三氯苯甲醚的含量,证实了所建方法适用于葡萄酒软木塞中痕量2,4,6-三氯苯甲醚含量的测定。     

气相色谱电子捕获技术分析检测葡萄酒及软木塞中TCA的研究进展

软木塞污染影响葡萄酒感官质量已成为葡萄酒行业最为关注的问题之一,主要原因在于软木塞中存在2,4,6-三氯苯甲醚(TCA).介绍了利用气相色谱电子捕获技术(GC-ECD)检测葡萄酒及软木塞中TCA的研究进展,同时对GC-ECD的工作原理及应用进行了阐述,为有效分析检测和控制葡萄酒及软木塞中的TCA含量提供参考.     

葡萄酒中2,4,6-三氯苯甲醚(TCA)检测新技术的研究进展

2,4,6-三氯苯甲醚(TCA)是危害葡萄酒感官品质的一种物质,通常来自于葡萄酒封装所用的软木塞,也可能来自葡萄原料或酿酒过程中的污染,它的存在使葡萄酒产生不良气味和口感。简介了最近十几年来检测葡萄酒中TCA的新方法和新仪器,包括葡萄酒中TCA的萃取技术和检测技术。并对这些新技术的特点、原理及应用优势进行了阐述。目的是为对葡萄酒中的TCA做到精确、低检出限和高灵敏性的定性及定量测定提供参考,从而对葡萄酒中的TCA进行准确的监测,最终提高葡萄酒的品质。     

超声波辅助-顶空固相微萃取-气相色谱法测定软木塞中痕量2,4,6-三氯苯甲醚

通过对软木塞样品提取过程的优化,建立了超声波法辅助顶空固相微萃取-气相色谱仪检测软木塞中痕量2,4,6-三氯苯甲醚(TCA)的方法。试验结果表明,该方法的线性范围0.5 ng/L~80.0 ng/L,R2为0.999 8,线性关系良好。方法检出限为0.2 ng/g,方法定量限为0.5 ng/g,精密度(RSD)在1.35%~5.47%之间,加标回收率为81.5%~97.8%。该方法操作简便、快速、易操作、准确度高、重现性好,适用于普通实验室对软木塞中痕量2,4,6-三氯苯甲醚残留的检测。     

法国消除软木塞污染新措施

最初,Dom P巖ignon为能够保留发酵过程中产生的气泡以酿造起泡葡萄酒,试图用新式软木塞替代传统的木制瓶塞。现在看来,如果不是他,人们也许永远也不会知道所谓的软木塞污染。到目前为止,软木塞在葡萄酒中的应用仍然相当广泛,但软木塞污染问题一直是困扰各葡萄酒厂和软木塞厂的问题。许多人认为软木塞中的2,4,6-三氯苯甲醚(TCA)是导致这一问题的罪魁祸首。当氯(常用来漂白)与软木塞中的霉菌接触就会产生这种具有湿报纸气味的物质,它具有巨大的危害力。加州大学戴维斯分校的酿酒专家Christian Butzke称,一汤匙容量的TCA就足以败坏美国全年…     

顶空-固相微萃取/气相色谱串联质谱法测定葡萄酒中多种氯酚及氯代茴香醚

样品中氯酚经乙酰化生成乙酰氯酚后,通过优化萃取头、离子强度、温度、时间等固相微萃取条件后,建立顶空-固相微萃取-气相色谱串联质谱法同时测定测定葡萄酒中的3种氯酚2,4,6-三氯酚(TCP)、2,3,4,6-四氯酚(TeCP)、五氯酚(PCP)以及两种氯代茴香醚2,4,6-三氯苯甲醚(TCA)、五氯苯甲醚(PCA)含量的方法。结果表明：该方法检出限为0.1ng/L,回收率为86.0%～105.9%,相对标准偏差小于16.5%。该方法前处理简单、高效、检测灵敏度高,可应用于进出口葡萄酒中木塞污染物的监控。     

山药酒中甾醇含量测定及清除DPPH自由基活性研究

采用固相微萃取（SPME）结合气相色谱-质谱联用（GC-MS）仪测定葡萄酒用软木塞中2,4,6-三氯苯甲醚的迁移量,对整个过程引入的不确定度进行分析评定。依据CNAS-GL006：2019《化学分析中不确定度的评估指南》和JJF 1059.1—2012《测量不确定度评定与表示》,构建了不确定度评定的数学模型,分析了不确定度来源,并对各分量加以量化和合成。评定结果表明,其不确定度主要来源于标准溶液系列配制过程,其次是内标物质添加过程;当木塞中2,4,6-三氯苯甲醚迁移量为2.04 ng/L时,其扩展不确定度为0.26 ng/L（k=2）。     

红葡萄酒中2,4,6-三氯苯甲醚(TCA)感官阈值的研究

2,4,6-三氯苯甲醚(TCA)是葡萄酒软木塞常见污染物之一,在酒中的含量极低(ng/L级别),但特殊的霉味会使消费者产生排斥.国外已对其进行了深入的仪器与感官分析研究,国内还未开展针对TCA的感官分析研究.本实验主要目的是测算多大浓度TCA污染的葡萄酒会使消费者产生排斥反应,通过成对比较及三点比较的检验方法,测定消费者排斥阈(CRT)及消费者觉察阈(DT),5%显著性结果分别为64ng/L和2ng/L,且两者间未呈现显著相关性.对比国外相关文献发现,国内外消费者察觉阈值比较接近,TCA的感官敏感性无较大差异;而国内消费者在对葡萄酒的喜好性评价中,相比国外消费者更不易受TCA的影响.     

不同软木塞中2,4,6-三氯苯甲醚迁移量的检测和评价分析

本文在建立了稳定的2,4,6-三氯苯甲醚(TCA)检测方法基础上,对不同类别的软木塞进行了TCA迁移量的检测,并对检测结果进行分析评价。结果表明,DIAM塞为TCA"零污染",天然塞和微颗粒塞次之,风险程度较低;1+1塞有较高的检出率,但TCA迁移量均在2 ng/L以下的可控范围内;而聚合塞不仅检出率高、合格率低,而且TCA检出值范围较宽,最大值达5.76 ng/L,对葡萄酒的污染风险程度最高。该研究为酿酒师木塞的选用提供数据支撑和指导作用。     

超声波辅助-顶空固相微萃取-气相色谱法测定葡萄酒中痕量2,4,6-三氯苯甲醚

建立了超声波-顶空固相微萃取-气相色谱检测葡萄酒中的痕量木塞污染物——2,4,6-三氯苯甲醚（TCA）的方法。试验结果表明,该方法有良好的线性关系,相关系数为0.999 89,方法定量限为0.5 ng/L,相对标准偏差RSD在1.98%～5.32%之间,加标回收率为89.0%～103.2%。该方法操作简便、快速、准确度高、重现性好,适用于普通实验室对葡萄酒中痕量TCA残留的检测。     

Determination of Chloroanisoles and Chlorophenols in Cork and Wine by using HS‐SPME and GC‐ECD Detection

Cork taint is an off‐flavor problem in wine, the main reason being the presence of 2,4,6‐trichloroanisole (TCA) in the cork stopper. In addition to the TCA, the presence of other chloroanisole and chlorophenol family compounds (the perception limits of which are very low) can also result in, or contribute to, cork taint problem. In this study, the levels of 2,4‐dichloroanisole (DCA), 2,4,6‐trichloroanisole (TCA), 2,3,4,6‐tetrachloroanisole (TeCA), pentachloroanisole (PCA), 2,4,6‐trichlorophenol (TCP), 2,3,4,6‐tetrachlorophenol (TeCP) and penthaclorophenol (PCP) were assayed in cork stoppers (natural, agglomerated and colmate) and in red wine samples from different wineries in Turkey using HS‐SPME and GC‐ECD detection. The performance parameters for all chloroanisole and chlorophenol compounds were as follows: recovery 92.48–102.53%, R 0.992‐0.996. The LOQ values were DCA (8.4 ng/L), TCA (0.8 ng/L), TeCa (0.6 ng/L), PCP (0.8 ng/L), TCP (0.8 ng/L), TeCP (1.2 ng/L), and PCP (1.1 ng/L) respectively. In cork stoppers, the amounts of 2,4,6 TCA ranged between 5.4–130.6 ng/g. The 2,3,4,6 TeCA ranged between 1.12‐8.2 ng/g and the PCA ranged between nd (not detected)‐11.01 ng/g. In the wine samples, 2,4,6 TCA ranged between 1.42–70.2 ng/L. The 2,3,4,6 TeCA ranged between nd‐15.1 ng/L and the PCA ranged from nd‐5.16 ng/L. The results indicated that there was a significant correlation between the TCA in wines and the TCA in cork stoppers.     

Migration of 2,4,6-trichloroanisole from cork stoppers to wine

The migration of 2,4,6-trichloroanisole (TCA) from cork stoppers to wine is studied under different experimental conditions. Corks that were either naturally contaminated or spiked with a TCA solution were immersed in an ethanol–water mixture (12% v/v) for 6 days or were used as a closure for bottled wines over periods of 1, 4 and 8 months. The TCA content was determined after the stated periods using headspace solid-phase microextraction and gas chromatography electron capture detection. Three different spiking procedures were tested: the injection of a TCA solution into the cork, the immersion of the cork in a TCA in hexane solution, and cutting off a thin slab to inject different quantities of a TCA solution into it before rejoining the two parts. Only the first of these procedures was subsequently used as the immersion technique failed to retain sufficient quantities of TCA and the slab technique retained too much. Corks spiked with 120 ng TCA gave 8% migration after 6 days for corks used in the immersion experiments and less than 1% migration for corked bottles kept for 1 month. When the contact time was 4 and 8 months, migration was 4 and 8% respectively for corks spiked with 1 g TCA. It was concluded that the contact surface, temperature and time all favour TCA migration. Under the same conditions, wine bottled with naturally contaminated cork stoppers showed TCA concentrations which varied depending on the characteristics of every individual cork stopper.     

Effect of wine style on the perception of 2,4,6-trichloroanisole,a compound related to cork taint in wine

2,4,6-Trichloroanisole (TCA) is a fungal metabolite that can contaminate cork stoppers and wines producing a very unpleasant mouldy odour (cork taint). Sensory control is very widely used for the detection of possible TCA presence in corks and wine, but the correct identification of TCA, especially at low concentrations, can be difficult. The aim of this study was to show how wine styles can affect the ability of the same panel to perceive TCA. For this purpose, a panel was selected, according to the panelists’ sensitivity to the TCA stimulus. The triangle test, a sensory difference test, was carried out by the panel with both white and red wines, using samples both free from TCA and spiked with known quantities of this contaminant. The results showed that the panelists identified the difference caused by the added TCA at different significance levels, depending on the wine style, so TCA detection was influenced by wine style for the selected panel.     

Presence of cork-taint responsible compounds in wines and their cork stoppers

 Chloroanisoles [2,4-dichloroanisole, 2,4,6-trichloroanisole (TCA), 2,3,4,6-tetrachloroanisole and pentachloroanisole], chlorophenols [2,4,6-trichlorophenol, 2,3,4,6-tetrachlorophenol and pentachlorophenol (PCP)] and guaiacol were detected in red and white cork-tainted bottled wines. These compounds were also found in the cork stoppers from those bottles. A significant correlation was found between TCA in wines and TCA in cork stoppers, and between TCA in wine and intensity of cork taint. At low levels of TCA, the presence of guaiacol or PCP were also found to influence in cork taint. Received: 29 November 1999     

The analysis of natural cork stoppers in transversal sections as an effective tool to determine the origin of the taint by 2,4,6-trichloroanisole

The distribution of several chlorophenols (CPs) and chloroanisoles (CAs), including 2,4,6-trichloroanisole (TCA), the main compound responsible for fungal taint of wines, was analyzed in four independent batches of natural cork stoppers that were dissected into three transversal slices. The contaminants were homogeneously distributed in the slices analyzed. All the stoppers were contaminated with at least one single CP or CA, although most of the corks contained several CPs and CAs. Pentachlorophenol was the more abundant contaminant, detected in 89.2% of the corks analyzed. The analysis of releasable CPs and CAs showed that most of the contaminants present in the stoppers cannot be released into wine. The same compounds were analyzed in a different batch of natural cork stoppers that had been used to close wine bottles in a winery whose facilities were contaminated with CPs and CAs. In this case, the highest amounts of contaminants were accumulated on the heads of the stoppers in contact with the winery environment. Bottled corks were able to efficiently absorb deuterium-labeled TCA (TCA-d ₅) and/or deuterium-labeled pentachloroanisole (PCA-d ₅) from an artificially tainted wine during 40 months. In this case, most of the TCA-d ₅ and PCA-d ₅ was located in the basal slices of the closures in direct contact with wine. These data, and also data from other authors, indicate that the distribution in transversal slices of the CPs and CAs contaminating cork stoppers is in direct relationship with the origin of the taint, and therefore that the analysis of cork stoppers by transversal sections could be an effective tool to clarify the origin of the taint.     

黄酒中9 种生物胺的高效液相色谱分析法

建立一种检测黄酒中生物胺的反相高效液相色谱法。用三氯乙酸提取黄酒中生物胺,丹磺酰氯作为衍生化试剂,1,7-二氨基庚烷作为内标定量。色谱条件为：Waters XBridge C18色谱柱（4.5 mm×250 mm,3.8 μm）作为分离柱,乙腈和水作为流动相进行梯度洗脱,流速1 mL/min,紫外检测波长254 nm。结果表明,9 种生物胺（盐酸吡哆胺、色胺、腐胺、尸胺、β-苯乙胺、组胺、酪胺、精胺和亚精胺）在40 min内能被很好分离。在1～50 mg/L质量浓度范围内,各种生物胺呈现良好的线性相关性（R2＞0.999）。各生物胺加标回收率为79.54%～89.55%,相对标准偏差为3.14%～6.35%,该法各组分检出限（RSN=3）为0.002～0.009 mg/L。应用柱前衍生化高效液相色谱-紫外检测法,实现了黄酒中生物胺的准确检测。     

Search for the contamination source of butyltin compounds in wine: agglomerated cork stoppers

A possible butyltin contamination source in wine was studied in this paper. Agglomerated cork stoppers, which were produced in Portugal, Spain, and Italy, used in wine bottles were examined. The domestic cork products, cork granules, and mucus used for cork products were also analyzed. The levels of mono- and dibutyltin compounds in corks were found in the range from <0.0024 to 3.3 and from <0.0029 to 6.7 microg of Sn/g, respectively. A low level of tributyltin contamination was also found in 2 of 31 tested samples. The presence of butyltin compounds in agglomerated cork stoppers was confirmed by GC-MS. Experimental results indicated that all overseas agglomerated cork stoppers studied contained mono- and/or dibutyltins. Butyltins were not detected in cork granules, mucus, most of the natural cork stoppers, and domestic agglomerated cork products. The concentrations of mono- and dibutyltins increased with the time in a 30-day experiment, showing that butyltin compounds can leach from agglomerated cork to the wine. When the butyltin concentrations in wine samples were compared with their levels in the corresponding agglomerated cork stoppers, a correlation was found. The potential harm of such food contamination was evaluated by the toxic research of butyltin compounds using Daphnia sp. as the experimental model.     

Matrix effect during the application of a rapid method using HS-SPME followed by GC–ECD for the analysis of 2,4,6-TCA in wine and cork soaks

An off-flavor in wine known as ‘cork taint’ is of concern in the wine and cork industry. Cork taint imparts a musty flavor to the wine and is primarily due to the presence of 2,4,6-trichloroanisole [2,4,6-TCA] in cork stoppers. During this study, an instrumental method for 2,4,6-TCA analysis was developed and evaluated using headspace solid-phase microextraction (HS-SPME) and gas chromatography coupled with an electron capture detector (GC–ECD). 2,3,6-Trichlorotoluene [2,3,6-TCT] was assayed as the internal standard. The method was developed in synthetic wine and was applied in commercial wine samples, as well as in cork soaks obtained by the extraction of TCA from cork stoppers and cork barks using synthetic wine. The method performance was evaluated through the estimation of its linearity (R² > 0.99), repeatability (RSD value = 5.72%) and sensitivity (recovery > 86%, LOD = 0.177–0.368 ng/L) in different types of samples. Due to the complexity of the samples used, the study has been especially focused on the matrix effects that were identified causing significant bias to the quantitative analysis of 2,4,6-TCA in cork soaks, where there is a lack of previous studies.     

不同种类葡萄酒软木塞的密封性能研究

对不同种类软木塞进行密封性能试验,并对封存的葡萄酒样品进行感官品评试验。结果表明,聚合塞适用于葡萄酒的短期储存;贴片1＋1塞适用于葡萄酒的中短期储存;天然软木塞适用于葡萄酒的长期储存。     

Migration of phenolic compounds from different cork stoppers to wine model solutions: antioxidant and biological relevance

Considering the enological interest of cork, this study aimed to identify and quantify the phenolic compounds able to migrate from different classes (natural cork stoppers “Flor” and “Third” quality and microagglomerate cork stopper) of cork stoppers into bottled wine model solutions. Another aim was to evaluate some antioxidant and biological features of cork phenolics that migrated into the wine model solutions. The main phenolic acids and aldehydes detected were as follows: gallic and protocatechuic acid detected both around 3.5 mg/L and vanillin and protocatechuic aldehyde detected around 2.5 and 1.5 mg/L after 27 months of bottling, respectively. Trace amounts of more complex polyphenols, namely hydrolysable tannins (castalagin/vescalagin and mongolicain A/B), were also detected. Two antioxidant features of the wine model solutions bottled with different wine cork stoppers were studied, namely the antiradical capacity and the reducing capacity, being the natural cork stoppers the ones with the higher activities. The intestinal absorption of the compounds in each wine model solution after 27 months in bottled was evaluated. The simpler phenolic compounds were able to cross Caco-2 cell model. The antiproliferative activity of the same wine model solutions was also evaluated against gastric and breast cancer cells. All samples were active against the two cell lines, which highlight the possible health outcomes of wine sealed with cork stoppers.