Fluorescence Analysis of Natural Organic Matter Fractionated by Ultrafiltration: Contrasting Between Urban-Impacted Water, and Radio-Contaminated Water from a Near-Pristine Site

Aqueous natural organic matter (NOM) impacted by two contrasting human impacts was analyzed using by multiresponse fluorescence, decoupled with the resolution routine PARAFAC. The first site is Chalk River, Ontario, Canada, near a pit formerly used to dispose low-level wastes. The second site is the Grand River in Cambridge, south-central Ontario, which is impacted by urban activities and agriculture. Our analysis included raw water, plus fractions from ultrafiltration and solid-phase extraction (SPE). The fluorescence spectra of the NOM, resolved with PARAFAC, showed three common features: humic-like components, at excitation/emission wavelengths 325?C350/450?C475 nm, fulvic-like components at 325/380?C420 nm and protein-like components, at 275/300 nm. Ultrafiltration revealed that most of the NOM comprised fine material below 5,000 Da cut-off (<4% of the total) in the urban-impacted sites and the clean site at Chalk River, but the colloidal fraction (larger than 5,000 Da) was substantially higher in the contaminated water, with ??18?C26% of the total. The protein-like components in the contaminated Chalk River water were affected by ultrafiltration, but less so in the clean Chalk River sample and the urban-impacted waters. SPE preferentially removed the protein-like component in the contaminated Chalk River water (typically 89?C95% signal decrease), but had a limited effect on humic- and fulvic-like components elsewhere. In conclusion, multiresponse fluorescence provided new information on the NOM quality from two contrasting sites, aided by ultrafiltration and SPE. These results are consistent with the in situ production of NOM in the Chalk River contaminated site, and natural production at the other sites.     

Monitoring chemical changes of dry-cured Parma ham during processing by surface autofluorescence spectroscopy

Parma hams at various processing stages were investigated by surface autofluorescence spectroscopy. Fluorescence "landscapes" of raw meat and salted (3 months), matured (11 and 12 months), and aged (15 and 18 months) Parma hams were obtained, and a three-dimensional data array (sample x emission x excitation) was used to develop a PARAFAC model including five components, which all exhibited characteristics of pure fluorophores regarding both excitation and emission spectra. The relative amount of each component related strongly to the processing stage, and sample age showed good correlation to fluorescence data (R = 0.98), with a relative error of prediction of approximately 1 month. Fluorescence measurements from samples of either semimembranosus or biceps femoris were used to predict chemical or sensory reference data, yielding good correlation for biceps femoris data, thereby enabling moisture content, sensory and instrumental color, and proteolysis value to be fairly well predicted. Overall, surface autofluorescence of Parma hams proved to hold relevant information, relating to major chemical/physical changes during processing. It is concluded that fluorescence spectroscopy has potential as an innovative method of quality control in dry-cured ham.     

Specific screening for color precursors and colorants in beet and cane sugar liquors in relation to model colorants using spectrofluorometry evaluated by HPLC and multiway data analysis

A comparison was made of the fluorophores in beet thick juice and cane final evaporator syrup, which are comparable in the production of cane and beet sugar; that is, both represent the final stage of syrup concentration prior to crystallization of sugar. To further elucidate the nature of the color components in cane and beet syrup, a series of model colorants was also prepared, consisting of mildly alkaline-degraded fructose and glucose and two Maillard type colorants, glucose--glycine and glucose--lysine. Fluorescence excitation--emission landscapes resolved into individual fluorescent components with PARAFAC modeling were used as a screening method for colorants, and the method was validated with size exclusion chromatography using a diode array UV--vis detector. Fluorophores from the model colorants were mainly located at visible wavelengths. An overall similarity in chromatograms and absorption spectra of the four model colorant samples indicated that the formation of darker color was the distinguishing characteristic, rather than different reaction products. The fluorophores obtained from the beet and cane syrups consisted of color precursor amino acids in the UV wavelength region. Tryptophan was found in both beet and cane syrups. Tyrosine as a fluorophore was resolved in only beet syrup, reflecting the higher levels of amino acids in beet processing. In the visible wavelength region, cane syrup colorant fluorophores were situated at higher wavelengths than those of beet syrup, indicating formation of darker colorants. A higher level of invert sugar in cane processing compared to beet processing was suggested as a possible explanation for the darker colorants.     

Fluorescence spectroscopy of aqueous pine litter extracts: effects of humification and aluminium complexation

Fluorescence excitation, emission, and synchronous-scan spectra were obtained for aqueous extracts of needle and litter layer (O-horizon) samples from ponderosa pine collected at a plantation site. The spectral lineshapes differed markedly between the needle and litter samples, and showed an increasing overall intensity with increasing extent of humification (increasing depth in the litter layer). At a dissolved organic C (DOC) concentration of 100 g m^?3, these effects were accompanied by a general shift in spectral density from lower to higher wavelength such that, in the excitation spectra, there was increasing prominence of a peak at 390 nm. When the DOC concentration was decreased from 100 to 25 g m^?3, the overall spectral intensity decreased and the peak at 390 nm in the excitation spectra of the litter samples gave way to a rising peak at 340 nm. Changes in pH from 4 to 5, characteristic of the undiluted litter extracts, produced little effect on the spectra. The addition of A1 at 40 mmol m^?3 generally produced enhancement of the fluorescence intensity in all three kinds of spectra for the needle and upper litter-layer extracts at 100 g C m^?3 and pH 4. Spectral density below 320 nm in the excitation and synchronous-scan spectra of the needle solution (possibly attributable to gentisic acid on the basis of model experiments) was, however, diminished by Al addition. For the lower litter-layer extracts, A1 addition decreased the fluorescence intensity in excitation and emission spectra, whereas it increased the intensity in synchronous-scan spectra. These trends indicated that the water-soluble fluorophores in the litter layer differed significantly from those in the fresh needles, and changed with the extent of humification. The 390-nm peak in the excitation spectrum, particularly its behaviour in the presence of added Al, may be useful as a spectral signature of products formed by litter humification processes.     

用于污染黄曲霉毒素花生分选的荧光信号研究

为在加工前将黄曲霉毒素超限的带衣花生米从原料中剔除,参照已有的色选系统,提出一种依据黄曲霉毒素含量超限带衣花生米的专属荧光信号进行逐粒分选的技术构想。采用Cary Eclipse荧光分光光度计测定100粒外观具有代表性的带衣花生米表面的紫外-荧光规律,通过与免疫亲和层析净化荧光光度法(GB/T18979-2003)检测结果对比,判定了黄曲霉毒素超限带衣花生米的荧光光谱特征;通过绘制450/490、460/490荧光强度比值的箱线图,评估了表面荧光法判断黄曲霉毒素超限带衣花生米的准确率;在搭建的荧光成像系统上,对黄曲霉毒素超限带衣花生米进行了荧光成像。检测发现,在365 nm波长激发下,黄曲霉毒素超限带衣花生米在420~460 nm处有荧光峰;以450/490荧光强度比值为依据剔除超限值带衣花生米的判断准确率为81%;a.u.40的带衣花生米可在图像中呈现亮蓝荧光光斑。表明表面荧光信号可作为带衣花生米在线、无损、逐粒分选的专属光学信号,用于黄曲霉毒素超限带衣花生米的剔除。     

Spectrophotometric determination of phosphine residues in rice

Phosphine residues were determined in 2 types of rice samples, raw polished and parboiled. The yellow chromophore due to phosphine from raw polished rice had 2 absorption maxima, a strong one at 370-380 nm and a weaker one at 400-410 nm. The chromophore from parboiled rice also had 2, one at 390-400 nm and the other at 410-420 nm. A closed system containing rice and a phosphine tablet or powder yielded 93-99% recovery of fortified phosphine. At any given dose of phosphine, parboiled rice retained a higher amount of residue after aeration than did raw polished rice.     

Application of fluorescence spectroscopy in the evaluation of light-induced oxidation in cheese

Light-induced oxidation of semihard cheese has been evaluated by fluorescence spectroscopy. The cheese was packaged in two packaging materials and exposed to different storage conditions, which included light/dark storage, oxygen availability, and storage time (0, 4, 7, 14, 21, 42, 70, or 84 days). Fluorescence excitation-emission matrices (EEM) were analyzed by PARAFAC, which gave an estimation of the pure excitation and emission spectra of the fluorophores and the concentrations of these. This analysis showed the presence of components such as tryptophan, tyrosine, vitamin A, fluorescent oxidation products, and riboflavin. Effects of packaging material, light or dark storage, and storage time were seen. However, there was no effect of the oxygen availability on the fluorescence measurements. The score values obtained by the PARAFAC models and chemical and physical measurements were analyzed together by principal component analysis (PCA). The loadings showed a separation of the variables into three groups; the first group was related to oxidation, the second group was related to the degradation of both riboflavin and vitamin A, and the third group was linked to the protein structure.     

Excitation-emission fluorescence spectroscopy combined with three-way methods of analysis as a complementary technique for olive oil characterization

This paper shows the potential of excitation-emission fluorescence spectroscopy (EEFS) and three-way methods of analysis [parallel factor analysis (PARAFAC) and multiway partial least-squares (N-PLS) regression] as a complementary technique for olive oil characterization. The fluorescence excitation-emission matrices of a set of Spanish extra virgin, virgin, pure, and olive pomace oils were measured, and the relationship between them and some of the quality parameters of olive oils (peroxide value, K232, and K270) was studied. N-PLS was found to be more suitable than PARAFAC combined with multiple linear regression for correlating fluorescence and quality parameters, yielding better fits and lower prediction errors. The best results were obtained for predicting K270. EEFS allowed detection of extra virgin olive oils highly degraded at early stages (with high peroxide value) and little oxidized pure olive oils (with low K270). The proposed methodology may be used as an aid to analyze doubtful samples.     

Fluorescence, browning index, and color in infant formulas during storage

Free and total fluorescent compounds, browning index, and color formation were measured in milk-based powdered infant formulas (IF) during 2 years of storage at 20 and 37 degrees C. The excitation spectra from 415 nm emission show three peaks (ex lambda1 = 270 nm, lambda2 = 325/315 nm, lambda3 = 350 nm) and from 347 nm excitation two emission peaks (415 and 520 nm), and no wavelength shifts were observed. Temperature and time of storage exert in general no significant effect on the development of fluorescence emission intensity and browning index. However, an important increase in pentodilysine was recorded-probably because of the iron and ascorbic acid contents of the samples-as well as in browning index in adapted IF. In both IF a color increase (deltaE) throughout storage was observed, this increase being greater in samples stored at 37 degrees C than in those stored at 20 degrees C. The increase in color with time fitted a linear regression model. Color appeared to be an indicator of sufficient sensitivity to measure the effect of temperature or storage time.     

Active photosensitizers in butter detected by fluorescence spectroscopy and multivariate curve resolution

In this study, fluorescence excitation and emission matrices and multivariate curve resolution (PARAFAC) were used to detect and characterize active photosensitizers spectrally in butter. Butter samples were packed under high (air) and low oxygen (<0.05%) atmospheres and exposed to violet, green, or red light. Six photosensitizers were found: riboflavin, protoporphyrin, hematoporphyrin, a chlorophyll a-like molecule, and two unidentified tetrapyrrols. By estimation of relative concentrations, we could follow how each sensitizer was photodegraded as function of wavelength, oxygen level, and time. The degradation rate of protoporphyrin, hematoporphyrin, chlorophyll a, and one of the tetrapyrrols correlated well (0.83-0.91) with the formation of sensory measured oxidation. The results suggest that mainly type I photoreactions were responsible for the degradation of photosensitizers in both high and low oxygen atmosphere. Type II photoreactions (generation of singlet oxygen) were involved in the oxidation of butter stored in air. The study shows that PARAFAC modeling of fluorescence landscapes is an excellent tool for studying photooxidation in complex systems.     

Fluorescence of vegetable oils: olive oils

Fluorescence spectra of undiluted extra virgin olive oil obtained with the traditional setup (right-angle fluorescence) show considerable artifacts and deformations due to self-absorption phenomena, even when the spectra are corrected for inner filter effects. On the other side, front-face fluorescence spectra are much less affected by self-absorption. Front-face fluorescence of native olive oil reveals the presence of different fluorophores and can provide information about their amount. From the intense emission at ca. 315-330 nm, it is possible to detect fluorescent polyphenols and pherols and to evaluate their overall content. Low-intensity emission bands at 350-600 nm are correlated to vitamins and other important molecules. Among them, the fluorescence of the riboflavin fluorophore can be used to evaluate its concentration. The intense emission of chlorophyll derivatives, measured in the 640-800 nm spectral region, can provide information on their concentration.     

Antioxidant properties of kilned and roasted malts

Compounds possessing antioxidant activity play a crucial role in delaying or preventing lipid oxidation in foods and beverages during processing and storage. Such reactions lead to loss of product quality, especially as a consequence of off-flavor formation. The aim of this study was to determine the antioxidant activity of kilned (standard) and roasted (speciality) malts in relation to phenolic compounds, sugars, amino acids, and color [assessed as European Brewing Convention units (degrees EBC) and absorbance at 420 nm]. The concentrations of sugars and amino acids decreased with the intensity of the applied heat treatment, and this was attributed to the extent of the Maillard reaction, as well as sugar caramelization, in the highly roasted malts. Proline, followed by glutamine, was the most abundant free amino/imino acid in the malt samples, except those that were highly roasted, and maltose was the most abundant sugar in all malts. Levels of total phenolic compounds decreased with heat treatment. Catechin and ferulic acid were the most abundant phenolic compounds in the majority of the malts, and amounts were highest in the kilned samples. In highly roasted malts, degradation products of ferulic acid were identified. Antioxidant activity increased with the intensity of heating, in parallel with color formation, and was significantly higher for roasted malts compared to kilned malts. In kilned malts, phenolic compounds were the main identified contributors to antioxidant activity, with Maillard reaction products also playing a role. In roasted malts, Maillard reaction products were responsible for the majority of the antioxidant activity.     

基于前表面荧光光谱鉴别新鲜与冻融大黄鱼

为研究反复冻融对水产品品质的影响,通过理化方法检测了不同冻融次数处理对大黄鱼解冻损失、pH 值、色泽、硫代巴比妥酸值、羰基含量等指标的影响,并采用前表面荧光光谱结合主成分分析（principal component analysis,PCA）和 Fisher 线性判别分析法（Fisher linear discriminant analysis,FLDA）对不同冻融次数的大黄鱼进行区分。结果显示随着冻融次数增加,大黄鱼的解冻损失显著增加（P<0.05）;pH 值呈先上升后下降的趋势;L*（亮度）值、b*（黄度）值均有不同程度的增加（P<0.05）,a*（红度）值下降（P<0.05）;羰基含量和硫代巴比妥酸反应物值（thiobarbituric acid reactive substances,TBARS）增加（P<0.05）,反复冻融导致大黄鱼的品质下降。色氨酸和烟酰胺腺嘌呤二核苷酸磷酸（nicotinamide adenine dinucleotide,NADH）的荧光光谱分别结合 PCA 和 FLDA 对不同冻融处理组进行分析,结果表明 FLDA 识别效果优于 PCA。通过 FLDA 建立了新鲜大黄鱼与冻融大黄鱼荧光光谱判别模型,发现色氨酸原始判别的准确率和交叉验证的准确率分别为68.3%和66.7%,NADH 原始判别的准确率和交叉验证的准确率均达到100%。由此可见,利用 NADH荧光光谱结合化学计量分析可以鉴别不同冻融处理的大黄鱼。研究结果为水产品新鲜度的快速评价提供参考。     

Native fluorescence from juvenile stages of common food storage insects

Insect infestation of stored foods has significant economic and health consequences; the development of novel methods of detection thus presents considerable opportunities. The fluorescence from nine species of storage insects (beetles and moths) was studied; the juvenile stages of all nine species exhibited fluorescence under long-wave (365 nm) UV light; none of the adult insects emit fluorescence, so the fluorophore(s) might be a compound(s) associated with the unsclerotized cuticle. The spectra of larval stages of Ephestia kuehniella, Oryzaephilus surinamensis, Corcyra cepahlonica, Tribolium castaneum, and Tribolium confusum exhibited excitation maxima in the range from 345 to 350 nm and emission maxima in the range from 421 to 427 nm, suggesting that fluorescence arises from a common chromophore; similarities in fluorescence properties implicate one of the many pteridine ring-containing compounds (pterins) commonly found in insects. Larvae and even eggs were readily imaged on foods using fluorescence under 365-nm excitation. Fluorescence thus appears to be ubiquitous in immature food storage insects, and fluorescence detection may be useful as a general method to detect insects in foods and agricultural commodities during storage or processing.     

Fluorescence Tracing of Diffuse Landfill Leachate Contamination in Rivers

Landfill leachates are composed of a complex mixture of degradation products which include a wide range of potentially fluorescent organic molecules and compounds. Here we investigate the use of fluorescence excitation–emission matrix (EEM) analysis in detecting diffuse landfill leachate contamination in rivers. Landfill leachates from three unlined landfill sites adjacent to our study river are characterised by intense fluorescence at excitation wavelength 220–230 nm, and emission wavelength 340–370 nm, which derives from fluorescent components of the xenobiotic organic matter fraction. Seven surface water sample sites on an adjacent polluted river system were analysed for fluorescence and water quality properties. The 220–230 nm excitation wavelength, 340–370 nm emission wavelength fluorescent centre was also detected in this river system at the sample locations downstream of the landfills, but not at upstream control sites, demonstrating its use as a tracer of landfill leachate contamination. Negative correlations are observed between this fluorescence centre and dissolved oxygen in the river water samples, demonstrating the water quality implications of leachate contamination at this study site. The fluorescence intensity at the 220–230 nm excitation wavelength, 340–370 nm emission wavelength fluorescent centre in landfill leachates is such that it remains detectable at dilutions of 10²–10³, and the fluorescence EEM technique is rapid and cost-effective for use by river managers and water quality regulators.     

Characterization of fresh and decomposed dissolved organic matter using excitation-emission matrix fluorescence spectroscopy and multiway analysis

Fresh and decomposed dissolved organic matter (DOM) derived from 13 plant biomass and animal manure sources was characterized using multidimensional fluorescence spectroscopy with parallel factor analysis (PARAFAC), high-performance size-exclusion chromatography, and UV-vis spectroscopy. The PARAFAC analysis modeled seven fluorescence components: tryptophan-like, tyrosine-like, and five humic substance-like components. For most of the plant-derived DOM solutions, decomposition significantly affected the concentration of three humic substance-like-associated components, increasing two and decreasing one. The effect of decomposition upon DOM derived from animal manures was dependent on the manure source. For a majority of the DOM extracts, the ratio of fluorescence intensity to absorptivity at 254 nm increased following decomposition, indicating that fluorescing DOM compounds were generally more resistant to biodegradation than nonfluorescing UV-absorbing compounds. Molar absorptivity, humification index (HIX), and apparent molecular weight (MWAP) increased by 38.0, 38.8, and 370%, respectively, following decomposition. Spearman correlation analysis showed a strong positive relationship between the humic substance-like components and the DOM MWAP, absorptivity, and HIX. The results of this study support the use of multidimensional fluorescence spectroscopy with PARAFAC as a method to monitor the decomposition of carbon-rich soil amendments such as crop residues, green manures, and animal manures.     

基于特征荧光信号的去囊衣带芯橘瓣分选

为实现去囊衣带芯橘瓣的机器视觉分选,给去囊衣带芯橘瓣的机器识别提供直接、准确的判别信号,采用荧光分光光度计对50颗去囊衣带芯橘瓣的橘芯和砂囊分别进行三维荧光光谱扫描,通过对橘芯、砂囊平均三维荧光光谱分析,确定了橘芯相对砂囊的特征荧光信号,据此对带芯橘瓣荧光图像识别的可行性进行了验证。检测发现,在370~390 nm紫外光激发下,橘芯和砂囊在440 nm处的荧光强度差异较大;橘芯与砂囊在370 nm紫外光激发下,440 nm处荧光强度分布的箱线图表明两者荧光强度分布存在明显差异,且以橘芯在440 nm处荧光强度的下四分位数(Q1)与砂囊在440 nm处荧光强度的上四分位数(Q3)的平均值作为分类标准,去囊衣带芯橘瓣检出准确率可达85%。对(370±2)nm激发下得到的单色(440±5)nm荧光图像进行二值化及形态学处理后,可在以砂囊为背景的橘瓣图形中显现橘芯亮斑。利用橘芯与砂囊的荧光特性差异进行机器视觉成像分析,可作为识别去囊衣带芯橘瓣的一种有效方法。     

Free radical scavenging capacity in the aging of selected red Spanish wines

Free radical scavenging capacity by the DPPH(*) method and main physicochemical properties, polyphenols content by HPLC, color by a tristimulus colorimeter, and UV-vis spectra in the aging of selected red Spanish wines, were studied. As the wines age, they become darker (lower lightness, L) and increase their hue angle (lower red color) as well as the ratio of absorbance at 420 nm to that at 520 nm. Main polyphenolics identified in the samples were tannic acid, oenin, and gallic acid. The antiradical efficiency of the samples increased during aging, which could be related to an increase in the tannic acid concentration shown by the following correlationship: EC(50) = 1/(0.18 + 0.0011[tannic acid](mg/L)) with a correlation coefficient of 0.744.     

航空施药雾滴沉积特性光谱分析检测系统研发与应用

为快速获取航空施药雾滴沉积的连续分布特性,弥补传统离散样点取样方式检测不足,提升航空施药雾滴沉积特性检测准确性,该文结合光谱分析和荧光激发技术设计研发了基于光谱分析的航空施药沉积特性检测系统。系统包括信息采集模块、采集装置模块和数据处理模块3部分。配置质量分数1.0%的荧光示踪剂溶液,采用农用植保无人机现场喷洒作业,同步放置雾滴获取介质和水敏试纸样本采集雾滴分布,系统采集雾滴获取介质的光谱特征曲线。与水敏试纸图像分析获取的雾滴沉积特性参数结果对比分析,结果表明:雾滴获取介质上的荧光示踪剂在450~460 nm和500~520 nm波段范围内产生显著荧光效应,其光谱平均值与雾滴沉积特性参数呈显著正相关。计算出450~460 nm和500~520 nm波段范围光谱平均值,建立雾滴沉积特性参数的检测多元线性回归模型,建模决定系数达0.80以上,验证决定系数达0.83以上,达到了较为理想的拟合结果。     

Characterization of Dissolved Organic Matter from Sewage Sludge using 3D-Fluorescence Spectroscopy and Chemometric Tools

The aim of the present article is to show the possibilities of chemometric tools and the parallel factor analysis (PARAFAC) model, as well as to understand the complexities of the fluorescence emission-excitation matrix (EEM) of water-soluble organic matter (WSOM) extracted from sewage sludge samples obtained with different origins and stabilization procedures. The variation in the composition of WSOM in the different sewage sludge samples could be correlated with the conditions of stabilization; therefore, the use of fluorescence spectroscopy to monitor these changes can be a useful tool to optimize the management of this residual flux, avoiding environmental pollution. However, the “contour density” EEM maps obtained with fluorescence present continuous spectra with different peaks, valleys, and shoulders where manual peak picking can be complicated and subjective, especially in complex matrixes such as sewage sludge. With the PARAFAC algorithm, it is possible to resolve the emission and excitation spectra of the different fluorescence components present in the samples and their contribution to the total fluorescence. Also, the contribution (scores) of the different fluorophors can be coupled with another chemometric tool to provide an effective classification method using the stabilization conditions of sewage sludge as main criterion. In this study, a wide survey of sewage sludge samples (287) was characterized by fluorescence emission-excitation matrix. The final molecular composition of the sewage sludge samples was not related to the stabilization (aerobic vs anaerobic) treatments. However, a sewage sludge classification based on a combined spectroscopic-chemometric approach was obtained, which could be used for the optimization of sewage sludge use in agriculture, minimizing the environmental risks.