Gas chromatographic analysis of histamine in mahi-mahi (Coryphaena hippurus)

Several authors have studied histamine using gas chromatography (GC) as a tool for quantitation, but the methods used were not always suitable depending on the kind of food. Problems frequently cited include incomplete histamine elution from the columns and peak tailing. Histamine is of interest because it is the factor common to all cases of scombroid poisoning, it has physiological and biological activity, and it is a chemical indicator of fish quality. In this study a modified GC method was used to quantify histamine in mahi-mahi (Coryphaena hippurus). Mean recovery was 67% for the GC method, compared with 90% for the AOAC fluorometric method. There was a 0.96 correlation of the GC histamine values with those of the AOAC fluorometric method. A temperature program, splitless/split injection, and analyte cleanup were essential for GC properties. Histamine retention time was 8.2 min. The method allowed peak height to be used for quantitation and simultaneous analysis of cadaverine and putrescine.     

Multiple compound quality index for cold-smoked salmon (Salmo salar) developed by multivariate regression of biogenic amines and pH

Production of biogenic amines during chill storage of 12 lots of cold-smoked salmon was studied. These data allowed for a multiple compound quality index to be developed by multivariate regression (partial least square regression). The quality index was based on concentrations of cadaverine, histamine, putrescine, and tyramine and pH and showed good correlation with sensory assessments. Biogenic amines were indicators of spoilage rather than casual agents of spoilage off-flavors. Four different biogenic amine profiles were found at the time of spoilage in cold-smoked salmon. These were the results of differences in the spoilage microflora. Histamine was detected above regulatory limits but below toxic levels. Measurements of salt and dry matter for calculation of water phase salt could be substituted by rapid water activity measurements.     

Occurrence of biogenic amines and polyamines in spinach and changes during storage under refrigeration

Biogenic amines and polyamines were studied in 18 market samples of spinach. Histamine and spermidine were detected in relatively high amounts in all samples within the ranges of 9.5-69.7 and 15.6-53.0 mg/kg, respectively. Other biologically active amines were either detected at low levels or not found at all. Changes in amine content during storage at 6 degrees C were studied. The content of most of the amines remained constant during storage, with the exception of spermidine and histamine. Spermidine showed a clear decreasing trend, whereas histamine significantly increased in all trials, but decreased at the end of the storage in two of the trials. Trials showing a decrease in histamine content also showed the highest spermidine decrease and recorded the highest pH values. Microbial loads throughout storage were also followed, with Pseudomonadaceae and Enterobacteriaceae being the predominant bacterial groups. Trials with higher microbial loads in initial samples also showed the highest histamine content in these samples. Potential explanations for both the formation and the degradation of histamine during storage are discussed.     

High-performance liquid chromatographic determination of biogenic amines in poultry carcasses

Biogenic amines, produced by bacterial decarboxylation of amino acids, have been associated with toxicological symptoms in broilers fed various poultry byproducts. A reversed-phase high-performance liquid chromatographic method is described for the quantitation of eight biogenic amines (tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine) in chicken carcasses. Amines were extracted with perchloric acid, derivatized with dansyl chloride, separated using gradient elution (methanol and water), and detected by fluorescence. Benzylamine was used as the internal standard. Linearity, repeatability, and recovery of the method were evaluated. The method was linear for all of the amines studied at concentrations ranging from 0.05 to 25 microg/mL. Average recoveries ranged from 92.6% to 96.8% for all amines except for histamine, which was 74.6%.     

HPLC method for analysis of free amino acids in fish using o-phthaldialdehyde precolumn derivatization

Precolumn derivatization applying o-phthaldialdehyde (OPA) was used to analyze free lysine, histidine, and ornithine, precursors of the respective biogenic amines cadaverine, histamine, and putrescine, which are considered indicators of fish quality and safety. This method uses 75% methanol to eliminate the use of strong acids as the extraction solution. Each analysis took 35 min, was reproducible, and allowed separation of primary amino acids in fish samples. A binary solvent delivery system coupled with a fluorescence detector and an Ultrasphere ODS column were utilized for HPLC separation. Linearity of the calibration curves was very good (r(2) = 0.99) for the amino acids of interest. Minimum concentrations of detection were 40 pmol/mL for histidine and lysine and 70 pmol/mL for ornithine. Average recoveries were 72% for lysine, 93% for histidine, and 98% for ornithine. This method used solvent gradient elution to study the levels of these analytes in mahi-mahi, bigeye tuna, and flounder.     

Freshness and quality criteria of iced farmed Senegalese sole (Solea senegalensis)

Senegalese sole (Solea senegalensis) is a high-value commercial species with increasing importance in aquaculture. The aim of this work was to study the quality changes of this species during chilled storage under refrigeration, through sensory and chemical methods. In particular, the optimization of a quality index method (QIM) scheme was proposed as well as the definition of sensory and chemical quality criteria. A shelf life of 15 days was reported, and a QIM scheme based on a total of 22 demerit points (dp) was proposed. Biogenic amines were never detected, and the usual spoilage indicators, such as TVB-N and TMA-N, were not significantly produced during the period of sensory-acceptable quality (15 days). On the basis of the significant correlations (p<0.001) between sensory data and Ki values, a quality index (QI) 19 dp and a Ki>40% indicate unacceptable quality of iced Senegalese sole.     

High-performance liquid chromatographic method for determination of biogenic amines in feedstuffs, complete feeds, and animal tissues

Numerous methods to analyze biogenic amines in biological materials have been described. A versatile and rapid methodology to analyze these compounds in feedstuffs, complete feeds, and animal tissues, however, has not been reported. The current method was developed to address this need. Biogenic amines in feedstuffs, complete animal feeds, and animal tissues were extracted with 10% trichloroacetic acid, reacted with O-phthaladehyde using high-performance liquid chromatographic employing a cation exchange column. Detection limits were 50 pmol/mL for tyramine, histamine, putrescine, and spermine; 40 pmol/mL for cadaverine; and 25 pmol/mL for spermidine. Extraction efficiency of biogenic amines in feedstuffs, duodenum, liver, ileum + jejunum, and whole shrimp and shrimp hepatopancreas ranged between 99-105, 93-135, 80-85, 65-102, 88-98, and 88-97%, respectively. It can be concluded that the current method can be applied to individual feedstuffs, complete feeds, and animal tissues for the rapid and accurate determination of concentration of biogenic amines.     

电子束辐照对真空包装章鱼生物胺的抑制作用研究

为分析电子束辐照处理对真空包装章鱼制品冷藏过程中生物胺形成的抑制作用,以真空包装的新鲜章鱼腕足段为研究对象,分别进行0、0.5、1.0、1.5、2.0和2.5 kGy剂量的电子束辐照处理,于4℃下冷藏63 d,每隔7 d进行感官品评和生物胺含量检测。结果表明,冷藏期间真空包装的章鱼中共检测出精胺、亚精胺、腐胺、尸胺、组胺和酪胺6种生物胺。电子束辐照处理能抑制章鱼冷藏过程中腐胺、尸胺、组胺和酪胺的生成,且抑制作用与辐照剂量成正比,其中对组胺生成的抑制效果最佳,仅在少数样品中被检出。此外,电子束辐照也抑制了章鱼中亚精胺和精胺在冷藏后期的分解。冷藏过程中的腐胺、尸胺和酪胺含量变化与样品感官品质表现出较好的一致性,高感官品质章鱼样品中的腐胺、尸胺和酪胺含量均≤10 mg·kg-1。综合电子束不同剂量辐照对章鱼感官品质和生物胺的抑制作用,1.0 kGy的电子束辐照剂量是真空包装章鱼4℃冷藏保鲜的最适前处理,其能延长高品质新鲜章鱼货架期至35 d。本研究结果为章鱼加工企业生产高品质章鱼产品提供了理论依据。     

Biogenic amines in low- and reduced-fat dry fermented sausages formulated with konjac gel

Biogenic amines in low- and reduced-fat dry fermented sausages made with konjac gel (KG) as pork backfat replacer were studied. An increase (P < 0.05) was observed in the microbial count during the fermentation process, reaching levels of over 8 Log cfu/g of total viable microorganisms and lactic acid bacteria. However, no significant differences were observed in the microbiota evolution as a function of the reformulation process (fat and konjac gel content). High levels of physiological amines (spermidine, spermine, and agmatine) were observed in the raw material. From day 2 of the fermentation process an increase (P < 0.05) was observed in tyramine and putrescine, which were the predominant amines at the end of the storage period. The increase in these amines was proportional to the presence of KG and fat reduction. This can also be seen for spermine, with agmatine showing the inverse. The biogenic amine levels in these products reformulated with KG are not considered to pose a health risk to consumers.     

Analysis of carbon monoxide in commercially treated tuna (Thunnus spp.) and mahi-mahi (Coryphaena hippurus) by gas chromatography/mass spectrometry

A simple and confirmative method for quantitative determination of carbon monoxide in tuna and mahi-mahi tissues using GC/MS, following chemical liberation of CO into headspace, is described. Carbon monoxide in recent years has been employed by the fishery industry to preserve fresh appearance in selected species of finfish during frozen storage, particularly in vacuum-packaged products. Indigenous CO contents of fresh Ahi tuna and mahi-mahi were examined using the method described in this study and found to be close to or less than 150 and 100 ng/g, respectively. Commercially CO-treated, vacuum-packaged tuna from multiple sources consistently showed CO level near or greater than 1 mug/g, while CO level in the only CO-treated frozen mahi-mahi sample was in the 500 ng/g range. The difference between untreated and treated specimens was in the range of 1 order of magnitude and thus suggested an easy quantitative and confirmative method of CO using widely available instrumentation that may be potentially useful for regulatory purpose in determining whether a commercially available product has been exposed to CO even if not labeled as such.     

Determination of biogenic amines in squid and white prawn by high-performance liquid chromatography with postcolumn derivatization

A simple method was developed for the determination of biogenic amines in aquatic food products using a reverse-phase high-performance liquid chromatography with postcolumn automatic o-phthalaldehyde derivatization and fluorescence detection. The linearity, repeatability, and recovery of the method for seven amines (tyramine, putrescine, cadaverine, histamine, agmatine, spermidine, and spermine) were evaluated. This optimized method was applied to detect biogenic amines in squid and white prawn during refrigerated storage. Sensory analysis, pH value, and total volatile base nitrogen value were combined to evaluate the freshness quality of these two raw materials. Agmatine and cadaverine in squid, cadaverine, and putrescine in white prawn were the most obviously changed amines during the storage at two different temperatures, and these biogenic amines could be the effective quality indicators for the freshness of the raw aquatic materials.     

Chemical and sensory properties of liquid beet sugar.

Sensory analysis and headspace gas chromatography-flame ionization detection (GC-FID) analysis of liquid sugars (sucrose) designated for food-industrial use were performed in order to elucidate which compounds are responsible for perceived off-odors and off-flavors in liquid sugar. The liquid sugar samples investigated had various kinds of off-odors and off-flavors that might influence the quality of any food product in which they might be used, such as a jam or a soft drink. The sensory scores for the different attributes and the levels of different volatile compounds as measured by GC were analyzed for correlation using partial least-squares (PLS) analysis. A link between sensory analysis and GC analysis was thereby established, and compounds critical for the odor and flavor were identified using MS (mass spectrometry). Approximately 10 compounds were found to be correlated with the defined sensory attributes. Among these compounds associated with the sensory attributes were dimethyl disulfide, 2,6-dimethylpyrazine, 4-methoxyphenol, and 2, 5-dimethylfuran.     

Chemical investigation of gamma-irradiated saffron (Crocus sativus L.)

Changes in aroma and coloring properties of saffron (Crocus sativus) after gamma-irradiation at doses of 2.5 and 5 kGy (necessary for microbial decontamination) were investigated. The volatile essential oil constituents responsible for aroma of the spice were isolated by steam distillation and then subsequently analyzed by gas chromatography/mass spectrometry (GC/MS). No significant qualitative changes were observed in these constituents upon irradiation, although a trained sensory panel could detect slight quality deterioration at a dose of 5 kGy. Carotene glucosides that impart color to the spice were isolated by solvent extraction and then subjected to thin-layer chromatography and high-performance liquid chromatography (HPLC). Fractionation of the above pigments into aglycon and glucosides was achieved by using ethyl acetate and n-butanol, respectively. Analysis of these fractions by HPLC revealed a decrease in glucosides and an increase in aglycon content in irradiated samples. The possibility of degradation of pigments during gamma irradiation is discussed.     

Characterization of volatile compounds in chilled cod (Gadus morhua) fillets by gas chromatography and detection of quality indicators by an electronic nose

Volatile compounds in cod fillets packed in Styrofoam boxes were analyzed during chilled storage (0.5 degrees C) by gas chromatography (GC)-mass spectrometry and GC-olfactometry to screen potential quality indicators present in concentrations high enough for detection by an electronic nose. Photobacterium phosphoreum dominated the spoilage bacteria on day 12 when the fillets were rejected by sensory analysis. Ketones, mainly 3-hydroxy-2-butanone, were detected in the highest level (33%) at sensory rejection, followed by amines (TMA) (29%), alcohols (15%), acids (4%), aldehydes (3%), and a low level of esters (<1%). The electronic nose's CO sensor showed an increasing response with storage time coinciding with the production of ethanol and 2-methyl-1-propanol that were produced early in the storage, followed by the production of 3-methyl-1-butanol, 3-methyl-butanal, 2,3-butandiol, and ethyl acetate. Lipid-derived aldehydes, like hexanal and decanal, were detected in similar levels throughout the storage time and contributed to the overall sweet odors of cod fillets in combination with other carbonyls (3-hydroxy-2-butanone, acetaldehyde, 2-butanone, 3-pentanone, and 6-methyl-5-heptene-2-one).     

以生物胺变化评价冷藏罗非鱼片腐败进程

为了探讨生物胺指标评价鱼肉腐败进程的可能性及其安全范围,应用反相高效液相色谱法测定冷藏罗非鱼片贮藏过程的生物胺变化,用以判断鱼片的腐败进程。结果表明:新鲜鱼片初始单胺和多胺总量较高,二胺总量很低。贮藏过程中单胺和多胺波动变化并呈下降趋势,尸胺、腐胺含量快速增加成为主要生物胺。含有尸胺+腐胺的二胺、生物胺指数、总生物胺等指标与其两者增长趋势相似。相关分析表明尸胺、腐胺、二胺、生物胺指数、总生物胺与贮藏时间、微生物数量和氨基态氮具有高度相关性。经回归分析表明假单胞菌、肠杆菌的增长与尸胺、腐胺具有重要的对应关系。尸胺、腐胺和二胺可简便有效评价鱼片腐败进程;但综合考虑组胺和酪胺的毒性,则生物胺指数(BAI)更适用,冷藏罗非鱼片生物胺指数的初步判别范围为:<20mg/kg,新鲜;20～40mg/kg,可接受;>40mg/kg,腐败。该研究为冷藏罗非鱼片生物胺的限量标准提供数据参考。     

贮藏温度对炝蟹品质的影响

为阐明不同贮藏温度下炝蟹品质的变化和食用安全性,本试验以三疣梭子蟹为材料制作炝蟹,探讨冻藏(-18℃)和冷藏(4℃)条件下炝蟹感官品质、菌落总数(TVC)、挥发性盐基氮(TVB-N)和生物胺的变化规律,并对其生物胺与其他品质指标进行相关性分析。结果表明,贮藏温度对炝蟹感官品质TVC、TVB-N等指标的劣变具有显著的影响。-18℃冻藏和4℃冷藏条件下,炝蟹感官品质分别于第10天和第6个月超出可接受限值;随着贮藏时间的延长,4℃冷藏条件下炝蟹TVC值显著增加,但至贮藏16 d时仍小于5 lg(CFU·g^-1),TVB-N值于贮藏12 d时超出一级鲜度临界值;-18℃冻藏过程中炝蟹TVC值略有增加,但变化不显著,TVB-N值于第6个月超出一级鲜度临界值。4℃冷藏过程中炝蟹中共检测出8种生物胺,而-18℃冻藏过程中共检测出5种生物胺。此外,色胺、腐胺和酪胺是炝蟹贮藏过程中产生的主要生物胺,且与其他品质均具有显著相关性。本研究结果为炝蟹在贮藏过程中的安全性和货架期预测提供了理论依据。     

Physicochemical and microbiological qualities of steamed and irradiated ground black pepper (Piper nigrum L.)

The effects of steam and irradiation treatments on the physicochemical properties (moisture content, pH, extractable yield, reducing sugar, soluble pigment, antioxidant activity, piperine, Hunter's color, and sensory attributes) and microbiological quality (total aerobic bacteria, coliforms, and yeasts and molds) of ground black pepper stored at refrigerated and room temperatures for 6 months were compared and evaluated. Irradiation resulted in a higher microbial reduction in pepper, with minimal effects on the proximate composition, functional components, color, and sensory attributes of the spice. Steamed peppers appeared darker, and a considerable decrease in the piperine content was observed after treatment and storage. This study illustrates that irradiation is a better decontamination method than steam treatment in eliminating microorganisms without apparently affecting the quality of the powdered spice. Storage at 4 degrees C enhanced the microbial quality and minimized the loss of piperine content in ground black peppers.     

High-performance liquid phase separation of glycosides. 5. Determination of individual glucosinolates in cabbage and rapeseed by laser-induced fluorescene capillary electrophoresis via the enzymatically released isothiocyanate aglycon.

A capillary electrophoresis (CE) method was developed for the profiling and determination of individual glucosinolates (GSs) via their isothiocyanate degradation products upon myrosinase digestion. The resulting isothiocyanates, the structures of which are reflective of the parent GS's, were then converted to their corresponding amines via base hydrolysis or reaction with 1, 2-benzenedithiol. Subsequently, the amines were fluorescently labeled to allow their sensitive detection by laser-induced fluorescence (LIF). The CE method involved the use of in situ charged micelles for the separation of isothiocyanates and their corresponding fluorescently labeled amines by micellar electrokinetic capillary chromatography (MECC). The term "in situ charged micelles" refers to micelles formed by complexing the polar hydroxyl groups of glycosidic surfactants with borate. The MECC method with on-column LIF detection was applied to the determination of GSs in white cabbage, rapeseed leaves, and rapeseed roots.     

Principal component analysis of biogenic amines and polyphenols in Hungarian wines

Biogenic amines, polyphenols, and resveratrol were analyzed quantitatively in 25 different Hungarian wines from the same wine-making region, harvest of 1998. Polyphenols were determined according to a spectrophotometric method, whereas other substrates were analyzed using overpressured-layer chromatography (OPLC). Principal component analyses (PCA) were performed on data matrices consisting of substrates (columns) and different sorts of wines (rows) from the region of Pécs (southern Hungary). It was found that four (unrotated) principal components account for >80% of the total variance in the data. The plots of component loadings showed significant groupings for concentrations of biogenic amines (and polyphenols). Similarly, the component scores grouped according to the different sorts of wines. The loading plots reveal that there is no need to measure all of the variables to achieve the same characterization. It is enough to measure one variable per group. Naturally, this conclusion is valid only within the limits of the present study; wines from other regions may behave differently.     

Relation of biogenic amines' formation with microbiological and sensory attributes in Lactobacillus-inoculated vacuum-packed rainbow trout (Oncorhynchus mykiss) fillets

The biogenic amine (BA) content of vacuum-packed filleted rainbow trout (Oncorhynchus mykiss) inoculated or not with two different Lactobacillus strains, individually or in combination, was monitored during refrigerated storage for 20 days and related to respective bacteriological and sensory changes occurring during the same period. Eight amines, namely putrescine, cadaverine, tyramine, tryptamine, beta-phenylethylamine, histamine, spermine, and spermidine, were determined, whereas agmatine was not detected in any of the samples. In all cases, BA concentration was higher (P < or = 0.05) in the controls compared to all inoculated treatments, whereas the trend with regard to the bacterial populations (Enterobacteriaceae, pseudomonads, and H2S-producing bacteria) and the off-odor scores was similar. Inoculation with Lactobacillus sakei CECT 4808 showed the best preservative effect among inoculated treatments. Concentrations of putrescine and cadaverine, the main BAs formed, correlated well with both spoilage bacterial counts and off-odor scores and can be useful indicators of shelf life. Spermine and spermidine contents decreased during storage, while levels of the other determined BAs remained below 10 mg/kg even after sensory rejection.