Volatility of patulin in apple juice

Patulin is a mycotoxin produced by certain fungi, such as those found commonly on apples. The patulin content of apple juice is a regulatory concern because patulin is a suspected carcinogen and mutagen. A simple model of the apple juice concentration process was carried out to examine the possible contamination of patulin in apple aroma, a distillate produced commercially in the concentration of apple juice. The results show no evidence for patulin volatility, and document a reduction in patulin content by at least a factor of 250 in the apple distillate obtained from apple juice. Furthermore, a survey of several commercial apple aroma samples found no evidence of patulin content.     

Rapid liquid chromatographic determination of patulin in apple juice

A rapid method is described for the quantitative determination of patulin in apple juice. The mycotoxin is extracted from the sample with ethyl acetate and the extract is cleaned up by extraction with a sodium carbonate solution. Patulin is determined by reverse phase liquid chromatography using a muBondapak C18 column and a 254 nm ultraviolet detector. The lower detection limit in patulin standard solution is 0.32 ng and recovery is greater than 75%.     

苹果汁中棒曲霉素臭氧脱毒设备的设计与应用

为了高效降解苹果汁中棒曲霉素,研制了一套利用臭氧技术降解棒曲霉素的设备。该设备利用臭氧的强氧化性,使其与苹果汁充分接触,从而使苹果汁中的棒曲霉素发生氧化分解。以人工污染的苹果汁为试验材料,研究了该设备的脱毒效果及对果汁主要品质指标的影响。研究结果表明:该臭氧脱毒设备能够有效降解苹果汁中的棒曲霉素,且臭氧浓度和臭氧处理时间均显著影响棒曲霉素的降解效果(P0.05)。在果汁棒曲霉素起始浓度约为201.60μg/L,pH值3.5和可溶性固形物质量分数为15%时,用12 mg/L的臭氧在3 L/min流速下处理15 min,可将棒曲霉素减少到约49.24μg/L,降解率达75.58%,达到世界卫生组织(WHO)和英国软饮料协会(British soft drinks association,BSDA)设定的最大允许限量50μg/L的要求。臭氧处理对果汁可溶性固形物含量、pH值和总酸度无显著影响(P0.05),但对果汁透光率、色值、苹果酸和总酚含量影响较大(P0.05)。该设备具有投资小、脱毒效率高、脱毒成本低、环保无污染、结构简单、易操作、适合各种规模的果汁企业等优势,具有良好的经济效益和社会效益。     

Comparison of metal oxide-based electronic nose and mass spectrometry-based electronic nose for the prediction of red wine spoilage

Taints caused by Brettanomyces sp. spoilage are of concern to winemakers and consumers. Typically the taints are described as "barnyard", "sweaty saddle", and "Band-aid" when present in red wine at concentrations of several hundred micrograms per liter or more. The two main components of the taint are 4-ethylphenol (4EP) and 4-ethylguaiacol (4EG), which are metabolites produced by Brettanomyces yeasts. There is a need for a rapid instrumental method to quantify these compounds in wines. In this paper are compared two techniques, the metal oxide sensor-based electronic nose (MOS-Enose) and the mass spectrometry-based electronic nose (MS-Enose). Gas chromatography-mass spectrometry (GC-MS) was used for quantification and prediction purposes. Following ethanol removal, the limits of detection of a MOS-Enose were determined as 44 microg L(-1) for 4EP and 91 microg L(-1) for 4EG, using the SY/gCT sensor. These values are significantly lower than the reported human sensory thresholds. Partial least-squares (PLS) regression of electronic nose signals against known levels of 4EP and 4EG in 46 Australian red wines showed that the MOS-Enose was unable to identify "brett" spoilage reliably because of the response of the gas sensors to intersample variation in volatile compounds other than ethylphenols. Conversely, the MS-Enose was capable of reliably estimating concentrations of 4EP higher than 20 microg L(-1). Correlations (r2) of 0.97 and 0.98 were obtained between estimates of 4EP and 4EG concentrations with the concentrations determined by conventional GC-MS. It is concluded that, following ethanol removal, existing metal oxide sensors are sufficiently sensitive to detect brett taints in wine but lack the selectivity needed to perform this task when the aroma volatile background varies.     

苹果汁冷冻浓缩工艺的研究

应用渐进冷冻浓缩原理对苹果汁冷冻浓缩特性进行研究,试验证明渐进浓缩苹果汁浓缩效果良好。影响冷冻浓缩的主要因素是搅拌速度。搅拌速度1200 r/min时浓缩效果最佳,溶质损失最少。冷冻浓缩对苹果汁中酸度和维生素C含量无影响,是提高浓缩品质量的较佳方法。浓缩产品感官质量均匀一致,保持了果汁的原有风味。     

Penicillium expansum: consistent production of patulin, chaetoglobosins, and other secondary metabolites in culture and their natural occurrence in fruit products

Penicillium expansum is known for its destructive rot and patulin production in apple juice. According to the literature, P. expansum can, among other compounds, produce citrinin, ochratoxin A, patulin, penitrem A, and rubratoxin B. In this study the qualitative production of metabolites was examined using TLC (260 isolates), HPLC (85 isolates), and MS (22 isolates). The results showed that none of the 260 isolates produced ochratoxin A, penitrem A, or rubratoxin B. However, chaetoglobosin A and communesin B were produced consistently by all 260 isolates. Patulin and roquefortine C were produced by 98% of the isolates. Expansolides A/B and citrinin were detected in 91 and 85% of the isolates, respectively. Chaetoglobosins and communesins were detected in naturally infected juices and potato pulp, whereas neither patulin nor citrinin was found. Because most P. expansum isolates produce patulin, citrinin, chaetoglobosins, communesins, roquefortine C, and expansolides A and B, foods contaminated with this fungus should ideally be examined for chaetoglobosin A as well as patulin.     

Analysis of patulin in apple juice by diphasic dialysis extraction with in situ acylation and mass spectrometric determination.

A procedure combining diphasic dialysis extraction with in situ acylation and gas chromatography/mass spectrometry (GC/MS) determination was developed for detection and quantification of the mycotoxin patulin in apple juice. Apple juice samples spiked with 4-N,N-dimethylaminopyridine were dialyzed using methane chloride and acetic anhydride inside dialysis tubing. Patulin was derivatized into its acetate and collected in the tubing after diphasic dialysis and was directly determined using GC/MS with the selective ion monitoring mode without further concentration and cleanup steps. Quantification was carried out by a calibration curve with an internal standard of correlation. The appropriate parameters of both dialysis and derivatization were examined. The linear range of the calibration curve was found to be 10-250 microg/L for patulin, and the limit of quantification was 10 microg/L. Levels of patulin ranging from 0 to 107.2 microg/L with 77-109% recovery were found in 10 apple samples. The technique combining diphasic dialysis extraction and acylation was demonstrated and showed potential for other applications.     

Micellar electrokinetic capillary electrophoresis for rapid analysis of patulin in apple cider

A micellar electrokinetic capillary chromatography (MECC) mode was applied to a capillary electrophoresis (CE) method, which was developed for detection and quantitation of patulin in apple ciders. This method used a small sample amount (2 mL) and consumed minimal organic solvent compared to the most commonly used HPLC methods. The sample preparation procedure of the CE method was also simpler than other chromatographic techniques developed for patulin analysis. Patulin was detected with a photodiode array detector at 273 nm. The standard curve was linear (r(2) = 0.9984) from 75 microgram/L to 121 microgram/mL with patulin working solutions corresponding to 3.8 microgram/L to 6.1 microgram/mL patulin in the sample. The linearity was better in a narrower range of concentrations (r(2) = 0.9999) from 75 microgram/L to 24.1 microgram/mL. The limit of detection of the method was 3.8 microgram/L. Patulin recoveries at 4 levels in spiked samples (10-121 microgram/L) ranged from 95.2 to 105.4%. The recoveries were 96. 9% and 99.2% for 2 levels (22.3 and 223 microgram/L, respectively) of patulin in infected apple samples. This method represents a unique alternative method for rapid and sensitive analysis of patulin in apple ciders.     

果汁护色剂JPX对苹果汁中棒曲霉素的降解作用及护色效果

为了考察果汁护色剂JPX在护色同时对苹果汁中棒曲霉素的降解作用与条件，该文研究处理温度、添加剂用量及处理时间对棒曲霉素降解率和果汁色值的影响。结果表明：在苹果浓缩汁生产过程中，用JPX控制棒曲霉素的较佳方法是，在榨汁阶段加入0.8g/L，酶解阶段加入1.6 g/L，棒曲霉素的降解率可以达到70%～80%，且对果汁有很好的护色效果。     

Activity and concentration of polyphenolic antioxidants in apple juice. 2. Effect of novel production methods

There is a great interest in food components that possess possible health-protecting properties, as is the case with flavonoids. Previous research showed that conventional apple juice processing resulted in juices poor in flavonoids and with a low antioxidant activity. This paper shows that it is possible to improve flavonoid content in juice and its antioxidant activity by applying an alcoholic extraction either on the pulp or on the pomace. The levels of flavonoids and chlorogenic acid in enriched juice were between 1.4 (chlorogenic acid) and 9 (quercetin glycosides) times higher than in conventional apple juice. In enriched juice the antioxidant activity was 5 times higher than in conventional apple juice, with 52% of the antioxidant activity of the originating fruits present. The novel processing method had similar effects for three apple cultivars tested (Elstar, Golden Delicious, and Jonagold). The taste and color of enriched juice were different from those of conventional juice.     

Activity and concentration of polyphenolic antioxidants in apple juice. 1. Effect of existing production methods

Apples are an important source of flavonoids in the human diet. The effect of processing apples into juice on polyphenolic antioxidant content and activity is described. Raw juice obtained from Jonagold apples by pulping and straight pressing or after pulp enzyming had an antioxidant activity that was only 10 and 3%, respectively, of the activity of the fresh apples. The levels of flavonoids and chlorogenic acid in the juice were reduced to between 50% (chlorogenic acid) and 3% (catechins). Most of the antioxidants were retained in the pomace rather than being transferred into the juice. Apparently, most of the antioxidant compounds are absorbed to the solid matter of the pomace. In apple juice, 45% of the total measured antioxidant activity could be ascribed to the analyzed antioxidants. For three apple cultivars tested (Elstar, Golden Delicious, and Jonagold), the processing methods had similar effects. The results indicate that processing can have a major impact on the bioactivity of products.     

Development of liquid chromatography-electrospray mass spectrometry for the determination of patulin in apple juice: investigation of its contamination levels in Japan

Patulin is a mycotoxin produced by mainly Penicillium species, for example, P. expansum, and Aspergillus species. There are several reports of patulin contamination in apple juice. Last year, the Ministry of Health, Labour and Welfare of Japan set the maximum allowable level of patulin in apple juice at 50 ppb and decided that the measurement of patulin levels in apple juice products should be conducted. To this end, a simple, accurate, and selective analytical method for the detection of patulin at levels lower than 5 ppb, the detection limit, is desired. This paper reports the development of an analytical method that employs solid-phase extraction-liquid chromatography-mass spectrometry (SPE-LC-MS). When MS measurements were conducted with the selected ion monitoring (SIM) mode, the pseudomolecular ions at m/z 153 and 156 were used to monitor patulin and (13)C(3)-labeled patulin, respectively. The detection limit (S/N = 3) and the quantification limit (S/N = 10) of patulin at injection levels into LC-MS were 12.5 and 25 pg, respectively. However, when the actual sample was applied for the analysis based on the developed method including the sample preparation, the detection limit (S/N = 3) and quantification limit (S/N = 10) were 2.5 and 5 pg in sample, respectively. The calibration curve obtained for concentrations ranging from 5 to 500 ppb showed good linearity with a coefficient of determination (r (2)) of 0.999. In addition, the recovery was >95% when an internal standard was used. The method was applied to the analysis of 76 apple juice samples from Japan, and as a result, patulin levels ranging from <1.0 to 45 ppb (detection frequency = 15/76) were detected. In this study, it was found that patulin was a greater contaminant in concentration/reduction than in "not from concentrate" apple juice.     

Characterization of Fuji apples from different harvest dates and storage conditions from measurements of volatiles by gas chromatography and electronic nose

Volatile compounds in Fuji apples harvested at two different maturities were measured at harvest and after 5 and 7 months of cold storage (1 degrees C) in four different atmospheres. When the samples were characterized by both chromatographic measurements of volatiles and responses of an electronic nose, the analyses showed a clear separation between fruits from different storage conditions (a normal cold atmosphere and three controlled atmospheres). During poststorage, the apples were left to ripen for 1, 5, and 10 days at 20 degrees C before analytical measurements were done involving headspace-gas chromatography methods and electronic nose type quartz crystal microbalances. Electronic nose responses registered by seven different sensors were used to classify the apples using principal component analysis. It was possible to identify the samples from different storage periods, days of shelf life, and harvest dates, but it was not possible to differentiate the fruits corresponding to different cold storage atmospheres.     

苹果浓缩汁二次混浊物的形态分析

通过苹果浓缩汁二次混浊物的形态分析以及浊度测定,初步确定混浊物的化学成分,为果汁生产、贮存中混浊的控制提供依据。将二次混浊果汁和后混浊果汁经乙酸双氧铀负染,观察其在透射式电镜下的形态并测定其浊度。结果表明：苹果浓缩汁经-18℃,冷冻8～10h处理后,苹果浓缩汁混浊减轻;再煮沸冷冻的果汁,混浊增加;二次混浊苹果浓缩汁形成的颗粒经乙酸双氧铀负染在透射式电镜下有蛋白质球状颗粒和链状的多酚类颗粒,或自身聚合或相互聚合形成大的颗粒;贮藏6个月的苹果浓缩汁后混浊比二次混浊严重。初步确定苹果浓缩汁的二次混浊物主要是蛋白质和酚类以及两者自身或相互聚合的产物。     

Activity and concentration of polyphenolic antioxidants in apple juice. 3. Stability during storage

Kinetic data are reported describing the stability of various classes of polyphenolic antioxidants in an apple juice enriched in these compounds as a function of storage temperature and oxygen concentration. The most thermally sensitive compounds were the various quercetin glycosides and epicatechin, whereas phloridzin and chlorogenic acid were more stable. The quercetin glycosides showed differences in their stability: quercetin galactoside approximately quercetin rhamnoside > quercetin glucoside/rutinoside > quercetin arabinoside. The effect of the presence of oxygen on the degradation rates was clear for only quercetin and to a lesser extent for epicatechin. Accelerated shelf-life testing of enriched apple juice during 4 days at 80 degrees C showed decreases in the antioxidant activity of 20-40%. The parameters obtained were used to predict the stability at different storage conditions. Calculations showed that polyphenolic antioxidants and antioxidant activity of enriched apple juice will be quite stable at ambient or refrigerated storage conditions up to half a year.     

基于电子鼻气味检测的苹果微波干燥方案优选

微波干燥中,过程参数设置不当会引起物料结构破坏和表面焦糊,期间往往伴随着特征气味的散发。该文以苹果为对象,采用60、70、80℃恒温、模糊逻辑控制和线性温度控制5种干燥方案,通过电子鼻采集的气味图谱,研究特征气味的散发规律,及优化干燥方案。根据恒温方案气味图谱中苹果风味峰和焦糊峰的变化规律,确定的模糊逻辑控制方案,可实现在线调节磁控管功率,保留风味、减少焦糊。将模糊逻辑控制方案中实测物料温度曲线进行优化,确定的线性温度控制方案,省去电子鼻检测环节,更适合工业应用。通过比较不同微波干燥方案下的产品品质,线性温度控制方案下干燥产品的品质高、耗时合理、干燥能力强,为优先方案。该研究为气味检测技术应用于苹果干燥提供了技术依据。     

High-pressure liquid chromatographic method for the determination of patulin in apple butter.

Patulin is extracted from apple butter samples with ethyl acetate and the extract is cleaned up on a silica gel column, using benzene-ethyl acetate (75+25) as the eluant. High-pressure liquid chromatography, using a 25 cm ZorbaxSil column, isooctane-ethyl ether-acetic acid (750+250+0.5) as the mobile solvent, and a 254 nm ultraviolet detector, is used for the determinative step. Under these conditions, patulin is eluted before 5-hydroxymethylfurfural, a component of apple butter which interferes with other liquid chromatographic and thin layer chromatographic methods. Recoveries of patulin added at levels of 34.6, 138.4, and 276.8 mug/kg ranged from 89.0 to 112.1%.