European Food Research and Technology

Published by Springer Nature

Online ISSN: 1438-2385


Print ISSN: 1438-2377


An automated HPLC determination of meticlorpindol in eggs with UV absorbance detection, using on-line dialysis and pre-concentration as sample clean-up; occurrence in and carry over to eggs
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  • Full-text available

January 1990


77 Reads

Ernst Maurits Mattern



H W van Gend
A fully automated HPLC determination of the coccidiostat meticlorpindol in whole egg, egg white and yolk is described. The sample homogenate is dialysed on-line against water. The dialysate is concentrated on-line on a short reversed-phase (RP) column. The contents of this column are transferred to the reversed-phase analytical column by means of the mobile phase. Meticlorpindol is detected using an absorbance detector at 270 nm. Linear calibration graphs are obtained in the range 40-900 ng/g in whole egg and egg white (detection limit 10 ng/g) and 80-1800 ng/g in yolk (detection limit 20 ng/g). Out of 111 commercially obtained egg samples 12 contained meticlorpindol with levels varying from 10 to 433 ng/g. A group of laying hens, kept in cages, received 10 mg/kg of Lerbek (meticlorpindol and methylbenzoquate; Dow Chemical) in the feed for 10 days. Meticlorpindol residues in the eggs rose to a level of 622 ng/g. Meticlorpindol was found in the eggs until 6 days after withdrawal of the medicated feed. Another group received 110 mg/kg in the feed. Meticlorpindol residues rose to levels of 4480 ng/g in the eggs, 5880 ng/g in the egg white and 2660 ng/g in the yolk. Meticlorpindol was found in the eggs and the egg white until 14 days and in the yolk until 8 days after withdrawal of the medicated feed.

Single nucleotide polymorphisms (SNPs) in key cytokines may modulate food allergy phenotypes

November 2012


91 Reads

Paula Brown


Bindukumar Nair


Supriya D Mahajan




Stanley A Schwartz
Single nucleotide polymorphisms (SNPs) can play a direct or indirect role in phenotypic expression in food allergy pathogenesis. Our goal was to quantitate the expression of SNPs in relevant cytokines that were expressed in food allergic patients. SNPs in cytokine genes IL-4 and IL-10 are known to be important in IgE generation and regulation. We examined IL-4 (C-590T), IL-4Rα (1652A/G) and IL-10 (C-627A) SNPs using real-time PCR followed by restriction fragment length polymorphism (RFLP) analysis. Our results show that the AA, AG and GG genotypes for IL-4Rα (1652A/G) polymorphisms were statistically different in radioallergosorbent test (RAST) positive versus negative patients, and although no statistically significant differences were observed between genotypes in the IL-4 (C-590T) and IL-10 (C-627A) SNPs, we observed a significant decrease in IL-4 (C-590T) gene expression and increase in IL-4Rα (1652A/G) and IL-10 (C-627A) gene expression between RAST(+) versus RAST(-) patients, respectively. We also observed significant modulation in the protein expression of IL-4 and IL-10 in the serum samples of the RAST(+) patients as compared to the RAST(-) patients indicating that changes in SNP expression resulted in altered phenotypic response in these patients.

Fig. 1 Continued
Fig. 3 Changes in (a) odor, (b) taste, and (c) texture of ozonated vs. control vacuum-packaged refrigerated (T=4±0.5 • C) trout. Points represent mean values of six determinations ± standard error (n=2×3). Error bars indicate the 95% confidence interval. Scoring scale: 0-10, with 7 being the lower limit of acceptability (development of first off-odor and/or off-taste)
Effects of ozonation on microbiological, chemical and sensory attributes of vacuum-packaged rainbow trout stored at 4±0.5°C

October 2005


166 Reads

The effects of ozone in aqueous solution on the shelf life of whole, vacuum-packaged rainbow trout, stored under refrigeration (4±0.5°C) were studied by monitoring the microbiological, chemical and sensory changes for a period of 15 days. Vacuum-packaged non-ozonated trout served as the control sample. Ozonation affected populations of bacteria namely, mesophilic aerobic bacteria, Pseudomonas spp. and H2S-producing bacteria until day 11 of storage, Brochothrix thermosphacta, lactic acid bacteria and Enterobacteriaceae until day 8 of storage. Trimethylamine (TMA) values of all rainbow trout samples remained low (<3mg N/100g) until day 11 of storage, and then increased to 12.2, 8.9 and 4.7mg N/100g for the control and the samples ozonated for 60 and 90min, respectively on day 15 of storage. Total volatile basic nitrogen (TVB-N) values remained relatively constant (20–25mg N/100g) until day 11 of storage, but increased to 61.1, 37.6 and 39.4mg N/100g flesh for the control and ozonated specimen for 60 and 90min, respectively on day 15 of storage. Thiobarbituric acid (TBA) values remained relatively constant (1–3mg MA/kg flesh) until day 12 of storage but increased to 8.4, 6.4 and 3.8mgMA/kg flesh on day 15 of storage for the control and the ozonated trout for 60 and 90min, respectively. Sensory evaluation (odor, taste and texture) of cooked rainbow trout showed a very good correlation with bacterial populations. On the basis of both sensory and microbiological data, a shelf life of 10 and 12 days was obtained for ozonated, vacuum-packaged and refrigerated rainbow trout at 60 and 90min, respectively.

Composition of some commercial grappas (grape marc spirit): The anomalous presence of 1,1-diethoxy-3-methylbutane: A case study

February 2009


118 Reads

A GC-MS study of the composition of nine different grappas (Italian grape marc spirit) was carried out. High-weight alcohols showed the greatest variations, while the lower congener concentrations generally were more uniform. Particularly interesting is the presence of 1,1-diethoxy-3-methylbutane (DMB), never detected before in grappa. DMB was present in all samples, ranging from 0.8 up to 30.6mgL−1. This latter concentration was the highest detected in an alcoholic beverage so far. A discussion on the other volatile congeners was also reported. Principal component analysis was applied to the data and explained >80% of the whole variability. Most grappas are grouped in a single cluster, while the other samples are completely separated. DMB, ethyl acetate, ethyl lactate, and n-butanol are some examples of constituent able to differentiate the tested products.

Occurrence of 3-chloropropane-1,2-diol and its esters in coffee

August 2005


172 Reads

Fifteen coffee samples comprising green, roasted, decaffeinated and instant coffees were analysed for their content of free and bound 3-chloropropane-1,2-diol (3-MCPD). Green coffees contained only traces of the free 3-MCPD. 3-MCPD in roasted coffees was found at the level of 10.1–18.5g/kg. The highest 3-MCPD level of 18.5g/kg was found in one instant coffee sample and in coffees with very long time application during roasting. The final colour of the roasted coffee beans was directly linked to the 3-MCPD formed, the darker beans having the greatest concentration, and arabica coffees contained lower 3-MCPD levels than robusta coffees. The level of bound 3-MCPD varied between 6g/kg (soluble coffees) and 390g/kg (decaffeinated coffee) and exceeded the free 3-MCPD level 8–33 times. Arabica coffees contained higher levels of the bound 3-MCPD than robusta coffees. The recognised precursors of 3-MCPD (salt, glycerol, lipids) were also determined and their influence on the formation of the free and bound 3-MCPD was discussed.

Kinetics of 3-chloropropane-1,2-diol (3-MCPD) degradation in high temperature model systems

July 2002


166 Reads

A study of the decay kinetics of 3-chloropropane-1,2-diol (3-MCPD) at high temperatures in aqueous model systems is reported. Data obtained over the pH range 5–6.5 and temperatures of 80–142 °C agreed well with a predicative model derived from earlier kinetic data. The results showed that 3-MCPD was unstable in aqueous solutions approaching infinite dilution at temperatures above 80 °C. Comparison of the experimental decay of 3-MCPD with that based on the kinetic parameters A (=107.73347+0.83775pHs–1) and E a (=119.2 kJ mol–1) was good to within 20% over the temperature and pH ranged studied. A putative mechanism for the degradation reaction is discussed together with the implications for the decay and formation of 3-MCPD in food systems.

Studies on the occurrence and formation of 1,2-dicarbonyls in honey

January 2004


222 Reads

The 1,2-dicarbonyl compounds 3-deoxyglucosulose (3-DG), methylglyoxal (MGO) and glyoxal (GO) were measured for the first time in 21 honey samples as the corresponding quinoxalines after derivatization with orthophenylenediamine using RP-HPLC and UV-detection. Compared to 5-hydroxymethylfural (HMF), which was also quantified, and ranged between 0.6 and 44mg/kg, up to 100-fold higher amounts of 3-DG were found, ranging from 79 to 1,266mg/kg. During storage of honey at 35C and 45C, a linear increase of 3-DG was observed. Values for GO and MGO were in the ranges 0.2–2.7mg/kg and 0.4–5.4mg/kg respectively, and were not affected by storage. Using semi-preparative RP-HPLC, glucosone, a 1,2-dicarbonyl compound previously unknown to occur in foods was isolated from a honey sample and characterized by LC-MS and NMR spectroscopy.

Formation of 3-chloropropane-1,2-diol in systems simulating processed foods

May 2004


159 Reads

Formation of 3-chloropropane-1,2-diol (3-MCPD) from the precursors glycerol, triolein and soy lecithin in the presence of sodium chloride was studied. The precursors were reacted with sodium chloride in an emulsion stabilised with an emulsifier under conditions which modelled the thermal treatment of foods during processing. Three sets of experiments were carried out aimed to monitor the influence of various factors (NaCl, water content and temperature) on the yield of 3-MCPD. The formation of 3-MCPD strongly depended on the concentration of NaCl and reached a maximum level at about 4–7% NaCl. The highest amount of 3-MCPD was formed in media containing approximately 13–17% water. The amount of 3-MCPD increased with increasing temperature over the range 100–230C and reached its highest value at 230C. The production of 3-MCPD was also followed in models very closely related to selected foods which had been shown to have a high potential to yield 3-MCPD during processing (salami, beefburgers, processed cheese, biscuits, crackers, doughnuts). The highest levels of 3-MCPD were formed in models simulating salami as they had the highest content of both fat and salt of all the samples. The lowest amount of 3-MCPD was formed in the models simulating biscuits and crackers as they had a low salt content and, at the same time, their water content was below the optimum level.

Table 2 Analyzed products of infant and baby foods and their basic characteristics 
Table 4 Diesters of 3-MCPD found in product 1 
Occurrence of 3-chloropropane-1,2-diol fatty acid esters in infant and baby foods

February 2009


639 Reads

A series of fourteen starter formula, follow-up formula, and growing-up milk produced by three manufacturers was analyzed by for the chloropropanols 1,3-dichloropropan-2-ol (1,3-DCP), 3-chloropropane-1,2-diol (3-MCPD) and their fatty acid esters. While none of the products contained free chloropropanols at levels exceeding the method detections limits, all of them contained relatively high amounts of 3-MCPD esters. The latter concentrations ranged from 62 to 588μg of 3-MCPD esters (bound 3-MCPD)/kg on the product basis and <300 (limit of quantification)—2,060μg/kg expressed on the fat basis. These concentrations were proportional to the fat content in products as refined vegetable oils used by the manufacturers contain elevated levels of 3-MCPD esters depending on their origin and other factors. Regular intake of these products can lead to exceeding the tolerable daily intake that has been set for free 3-MCPD. The major types of 3-MCPD esters were the symmetric diesters with palmitic and oleic acids and asymmetric diesters with palmitic/oleic acid among which 1,2-palmitoyl, oleoyl-3-MCPD prevailed.

Optimised quantification method for yeast-derived 1,3-β-D-glucan and α-D-mannan

January 2005


469 Reads

The polysaccharides 1,3--d-glucan and -d-mannan show numerous beneficial effects for the health of humans and animals. For several years, an increasing number of glucan- and mannan-containing products intended for food and feed applications are commercially available. For the determination of glucan and mannan contents, however, widely accepted methods have not yet been established. We have developed a practicable and reliable quantification method characterised by acidic hydrolysis with trifluoroacetic acid and subsequent determination of released d-glucose and d-mannose. The unavoidable loss of the monosaccharides due to the acidic conditions was minimised by optimisation of the hydrolysis parameters. The best conditions found were compared with literature methods in order to demonstrate the suitability. Finally, glucan and mannan contents of various commercial products were determined and compared to the specifications given by the manufacturers.

A new imidazolinone resulting from the reaction of peptide-bound arginine and oligosacccharides with 1,4-glycosidic linkages

February 2004


54 Reads

After heating N--hippuryl-L-arginine (Hip-Arg) with varying amounts of lactose for 1–4h at 100C, a previously unknown arginine derivative could be detected by RP-HPLC and UV-detection. Following semi-preparative isolation, the compound was unequivocally identified as 2-(2-benzoylamino-acetylamino)-5-[5-(3-hydroxypropyl)-4-oxo-imidazolon-2-yl]-L-ornithine (Hippuryl-PIO, Hip-PIO) by electrospray-time of flight-mass spectroscopy as well as one- and two-dimensional 1H- and 13C-nuclear magnetic resonance. Hip-PIO was exclusively formed during incubation of Hip-Arg with disaccharides containing a 1,4-glycosidic linkage. As reference for amino acid analysis, free PIO was synthesized starting from N-(tert-butoxycarbonyl)-L-arginine (t-Boc-Arg) via t-Boc-PIO and final hydrolysis with acetic acid. Preliminary evidence for the formation of protein-bound PIO, which proved to be acid-labile, was obtained using amino acid analysis with ninhydrin detection for enzymatic hydrolysates of casein samples which had been heated in the presence of lactose at 100C. The ornithinoimidazolinone PIO represents a new type of post-translational protein modification formed during food processing, which might be responsible for the major part of arginine derivatisation in disaccharide-containing foods like milk.

Isolation and identification of 3,4-dideoxypentosulose as specific degradation product of oligosaccharides with 1,4-glycosidic linkages

October 2006


96 Reads

Although it is well known that arginine side chains of proteins become extensively modified in the course of the advanced Maillard reaction, to date only few possible arginine derivatives are known. Recently, N-δ-[5-(3-hydroxypropyl)-4-oxo-imidazolon-2-yl]-l-ornithine (PIO) was identified as a new arginine derivative, formed exclusively from the side-chain of peptide-bound arginine and degradation products of oligosaccharides with 1,4-glycosidic linkages. In the present paper, a previously unknown C5 dicarbonyl compound, namely 3,4-dideoxypentosulose (3,4-DDPS), was isolated by means of semi-preparative RP-HPLC as the corresponding chinoxaline after heating lactose followed by trapping the 1,2-dicarbonyl compound with o-phenylenediamine. Identification was achieved using LC–MS as well as 1H- and 13C-NMR. Synthesis of 3,4-DDPS as the corresponding 2-hydroxy-dihydro-pyran-3-one proved the structure analysis. During heating of N-α-hippuryl arginine (Hip-Arg) with 2-hydroxy-dihydro-pyran-3-one, Hip-PIO was formed as the only arginine derivative. The formation of 3,4-DDPS from oligosaccharides with 1,4-glycosidic linkages follows a new and quantitatively important pathway of carbohydrate degradation in foods.

Performance of Lactobacillus sanfranciscensis TMW 1.392 and its levansucrase deletion mutant in wheat dough and comparison of their impact on bread quality

June 2007


106 Reads

The performance of Lactobacillus sanfranciscensis TMW 1.392 and its levansucrase deletion mutant TMW 1.392Δlev in wheat dough was compared. The effects of both strains on dough and bread characteristics were determined in order to find benchmarks for in situ production of exopolysaccharides (EPS). Growth and acidification were lower in doughs prepared with the Δlev mutant than in those employing the wild type. Extensogram resistance of the dough was reduced and extensibility increased with the addition of L.sanfranciscensis levan. Added EPS positively influenced water absorption, bread volume and firming of the crumb. In situ production of EPS was not sufficient to achieve the same positive effects of EPS, as they partially overlapped with effects resulting from enhanced acidification. Control doughs were made to separate effects of predough, EPS and different metabolism/acidification. High acetic acid levels decreased extensibility and volume. High lactic acid levels negatively influenced crumb hardness and firming kinetics. The use of knock out mutants proved helpful to judge overall performance of a strain, although the interpretation of specific effects must consider all changes in its metabolism.

Sourdough fermented by Lactobacillus plantarum FST 1.7 improves the quality and shelf life of gluten-free bread

April 2008


375 Reads

Lactobacillus plantarum FST1.7 has been recently shown to produce antifungal compounds, which improve the shelf life of wheat bread. In the present study, this strain was investigated for its ability to improve the quality and shelf life of gluten-free bread. Effects of incorporation of sourdough fermented by strain FST1.7 into a gluten-free bread mixture were compared to those obtained with sourdough fermented by the non-antifungal strain Lactobacillus sanfranciscensis TMW1.52 as well as to those obtained with chemically or non-acidified batters. Fundamental rheological tests revealed that the addition of sourdough to the gluten-free mix led to an increase in firmness and increase in elasticity overtime (P<0.05). Bread characteristics such as pH, total titratable acidity, and crumb hardness (5-day storage) were evaluated. Results showed that the biologically acidified gluten-free breads were softer after 5days than the chemically acidified gluten-free breads (P<0.001). Antifungal challenge tests employing conidial suspensions of Fusarium culmorum were carried out using the sourdough, non-acidified batter and bread. The rate of mould growth for the fungal species used was retarded by L.plantarum FST 1.7 when compared to the controls. In conclusion, the results of this study indicate that L.plantarum FST1.7 can be used to produce gluten-free bread with increased quality and shelf life.

Exclusive quantification of methyl-branched fatty acids and minor 18:1-isomers in foodstuff by GC/MS in the SIM mode using 10,11-dichloroundecanoic acid and fatty acid ethyl esters as internal standards

March 2007


137 Reads

Minor fatty acids (iso- and anteiso-fatty acids, vaccenic acid, elaidic acid) in foodstuff (seafood, milk, and dairy products) were analyzed by gas chromatography with electron ionisation mass spectrometry in the selected ion monitoring mode (GC/EI-MS-SIM). For this purpose, lipids were obtained by accelerated solvent extraction (ASE) and the fatty acid constituents were converted into methyl esters. Instead of the determination of the relative contribution of the minor fatty acids to the sum of all fatty acids detected (the so-called 100% method), we exclusively quantified the minor fatty acids, which was possible by using two types of internal standards (IS-1 and IS-2). For recovery checks during the extraction and/or the transesterification step we added the novel 10,11-dichloroundecanoic acid (DC-11:0) as IS-1. DC-11:0, which has never been detected in foodstuff, was synthesized by electrophilic addition of chlorine to 10-undecenoic acid (11:1n-10). The novel IS eluted in the range of 23:0 from the polar GC column used and showed the same properties as fatty acids in foodstuff during sample preparation. Recovery rate of DC-11:0 was generally >96% in the various samples analyzed. Ethyl esters (FAEE) of a12:0, a14:0, a15:0, a16:0, a17:0, and a18:0 (IS-2) were added to both the external standard (a quantitative mixture of methyl esters of methyl-branched fatty acids and 18:1n-9trans) in order to determine their response factors relative to FAEE and to the food samples. With this technique, (only) methyl-branched fatty acids (MBFAs) as well as vaccenic acid (18:1n-7) and elaidic acid (18:1n-9trans) were quantified in a range of dairy products (including twelve cheeses) and seafood. All samples were analyzed in triplicates, and good standard deviations (concentrations 0.002–5g/100g; standard deviations 0.00–0.03) were obtained in all cases. MBFAs were detected in all samples analyzed. The highest content of MBFAs (3.0g/100g) was determined in red-smear of romadur cheese. In all except two cheeses, i17:0 was the most abundant MBFA. The highest amount of 18:1n-9trans was found in feta (2.84g/100g) whereas 0.03g/100g in big eye snapper (Pricanthus tayenus) marked the lowest record of this minor fatty acid. Seal oil contained the highest amount of 18:1n-7 with 5.00g/100g, whereas emmental cheese was the sample with the lowest content of this monoenoic fatty acid. The combination of suitable IS and a sensitive GC/EI-MS-SIM method proved to be well suited for the quantification of minor fatty acids in foodstuff. When only a set of fatty acids is going to be analyzed, this method is less time consuming compared to “100% methods” and less prone to false results due to the higher selectivity of GC/MS compared to GC in combination with flame ionisation detection (GC/FID).

α 67-106 of bovine hemoglobin: A new family of antimicrobial and angiotensin I-converting enzyme inhibitory peptides

April 2011


65 Reads

Protein hydrolysates are of a significant interest, due to their potential application as a source of bioactive peptides in nutraceutical and pharmaceutical domains. The present study was focused on bovine hemoglobin hydrolysate obtained with pig pepsin in the presence of 30% ethanol. This hydrolysate was fractioned by reversed-phase high-performance liquid chromatography (RP-HPLC) into 12 major fractions (F1–F12). All fractions were analyzed by ESI/MS and ESI/MS/MS, in order to characterize and identify the peptides in these fractions. This hydrolysis permitted to generate a new serial of bioactive peptides with both antimicrobial and ACE inhibitory activities. Identified peptides were TKAVEHLDDLPGALSELSDLHAHKLRVDPVNFKLLSHSLL, LDDLPGALSELSDLHAHKLRVDPVNFKLLSHSL, KLLSHSL, and LLSHSL corresponding respectively to the 67-106, 73-105, 99-105, and 100-105 fragments of the α chain of bovine hemoglobin. They were the first found from bovine hemoglobin. These purified peptides have an antibacterial activity against four bacteria strains: Kocuria luteus A270, Listeria innocua, Escherichia coli, and Staphylococcus aureus with a MIC between 187.1 and 35.2μM. On the other hand, these peptides displayed at the same time ACE inhibitory activity with an IC50 range from 42.55 to 1,095μM. KeywordsBovine hemoglobin–Hydrolysis–Antimicrobial peptides–ACE inhibitory activity

Purification and characterization of a new 11S globulin-like protein from grape (Vitis vinifera L.) seeds

March 2010


178 Reads

Around 10 million tons of grape pomace containing 38–52% (on a dry matter basis) seeds was produced each year in the world, so how utilize these residual matters has been receiving great attention. In this study, 11S globulin-like protein (GSG) was isolated and purified from grape (Vitis vinifera L.) seeds by two consecutive cation exchange and size exclusion chromatography. A yield of 10mg the protein from 1kg seeds was obtained. The apparent molecular mass of the native GSG was found to be about 300kDa. The protein consisted of two subunits with molecular masses of 25.5 and 40.0kDa, respectively. MADLI-TOF–MS result showed that they were distinct from each other. N-terminal amino acid sequence of the 40.0kDa subunit is RQQTSRQQKE. Amino acid composition analyses indicated that GSG contained all eight essential amino acids, while lysine was the first limiting amino acids. To the best of our knowledge, this is the first report on the purification of 11S globulin-like protein from Vitis vinifera L. seeds.

Effect of wheat puroindoline alleles on functional properties of starch. Eur Food Res and Technol 226: 1205-1212

February 2008


220 Reads

Puroindoline a and b (Pina, Pinb) form the molecular basis of bread wheat grain hardness. Varieties with a softer endosperm and a wild genotype, in which both Pina and Pinb were present, seemed to produce less damaged starch flour than hard varieties, where Pin mutations occurred and changed the starch rheological properties. The functional property of starch samples extracted from wheat varieties with different Pin alleles was evaluated. Starch morphology was characterized by scanning electron microscopy and laser light scattering. Thermal properties were evaluated by differential scanning calorimetry. Amylose content, starch damage and rapid visco-analyser (RVA) parameters were also determined. Significant variations (P<0.05) were identified between different Pin variants for the distribution pattern of starch granule volume, amylose content, starch damage, RVA viscosity breakdown and retrogradation, gelatinisation transition temperatures and enthalpies. Hard genotypes presented higher medium diameter granules and lower enthalpic values. However, the differences detected are more evident among varieties that present both Pina and Pinb, than among those presenting only one of the two (a or b).

Online determination of 2H/ 1H and 13C/ 12C isotope ratios of cinnamaldehyde from different sources using gas chromatography isotope ratio mass spectrometry

November 2003


120 Reads

The combination of gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) and gas chromatography-pyrolysis-isotope ratio mass spectrometry (GC-P-IRMS) is applied to the authenticity assessment of cinnamaldehyde from various sources. For that reason, cinnamon oils were self-prepared by steam distillation from three different varieties of cinnamon bark on the market, C. ceylanicum (ceylon), C. cassia (cassia) and C. burmanii (cassia vera). Furthermore, the so-called wood cinnamon was investigated, which is produced from the outer bark of older branches of cinnamon of minor quality. Self-prepared oils were analysed from commercial cinnamon powder. In addition several commercial samples of cinnamon oil and cinnamaldehyde, some of them declared to be natural, were investigated. 2V-SMOW and 13CV-PDB values of cinnamaldehyde were determined and characteristic authenticity ranges were deduced, allowing the differentiation between synthetic and natural samples. By correlation of both the 2V-SMOW and 13CV-PDB values, characteristic authenticity ranges were defined for ceylon, cassia and wood cinnamon. The 2V-SMOW and 13CV-PDB values of cassia vera samples are in the range of cassia. By comparing the 2V-SMOW values of different self-prepared samples (ground bark, distillate) of cinnamon determined by TC/EA-IRMS with the corresponding GC-IRMS values, online GC-IRMS methods are proved to be essential in the authentication of complex natural products.

Determination of the 13C/12C ratios of ethanol and higher alcohols in wine by GC-C-IRMS analysis

June 2010


292 Reads

More than 50 authentic EU data bank wines from Germany (vintages: 2003, 2004 and 2006) have been extracted by liquid–liquid extraction with a mixture of cyclohexane and 2-methyl-2-propanol. The δ13C values of the extracted wine ethanol and other co-extracted higher alcohols have been determined by gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). The stable isotope ratios of wine ethanol obtained by liquid–liquid extraction were compared with those obtained after distillation according to the official EU method. No significant differences of the isotope ratios of ethanol measured by the two compared methods were found (R 2 = 0.972), thus it was concluded that no significant isotope effect during sample preparation occurred. Additionally, the 13C/12C isotope ratios of the major co-extracted alcohols, e.g. 2-methylpropan-1-ol, 2- and 3-methylbutan-1-ol, butan-2,3-diol and 2-phenyl-1-ethanol were determined and set in relation to the δ13C values of the corresponding extracted wine ethanol. Also, high correlations between their isotope ratios (e.g. 2,3-methylbutan-1-ol R 2 = 0.829) were found. The key benefits of the introduced liquid–liquid extraction are the time- and cost-saving determination of δ13C values of wine ethanol, the small amount of sample needed for the extraction and additionally the potential analysis of the co-extracted higher alcohols which could be valuable for the detection of fraud.

Authenticity assessment of 2- and 3-methylbutanol using enantioselective and/or 13C/12C isotope ratio analysis

January 1999


15 Reads

 Enantioselective gas chromatography and/or 13C/12C isotope ratio analysis are suitable tools for the authenticity assessment of the fusel alcohols, 2- and 3-methylbutanol (1 and 2, respectively). The chiral compound, 1 is biosynthesised almost completely as the (S)-enantiomer, regardless of which carbohydrate source is used for fermentation. The type of CO2 fixation and some plant-specific influences were of prime importance to the 13C/12C isotope ratios of the starting materials in alcoholic fermentation, and the δ13C values of 1 and 2 differed significantly. In general, the δ13C values of 2 were about 4–5ö lower than those of 1 produced via the same fermentation process. 13C/12C isotope ratio analysis results can be used to determine between fusel alcohols produced from different sources, and provides a new and valuable method of authenticity assessment.

Fig. 1 Electrophoretic analyses of PCR products generated after DNA amplification of mitochondrial 12S ( left ) and 16S ( right ) rRNA genes from the gadoid species studied. Numbers on top indicate fish species as follows: 1, Trisopterus minutus minutus ; 2, Micromesistius poutassou ; 3, Molva elongata ; 4, Phycis blennoides ; 5, Theragra calchogramma ; 6, Trisopterus minutus capelanus ; 7, Gadus morhua . Lane C represents an example of negative control (amplification without DNA) and lane L indicates 100 bp DNA ladder (Invitrogen) 
Fig. 3 Alignment by ClustalW method of DNA sequences from a portion of the 16S rRNA gene amplified by PCR using DNASTAR software. Numbers on the right side indicate fish species as follows: 1, Trisopterus minutus minutus ; 2, Micromesistius poutassou ; 3, Molva elongata ; 4, Phycis blennoides ; 5, Theragra calchogramma ; 6, Trisopterus minutus capelanus ; 7, Gadus morhua . Sequence #7 (GenBank accession no. X99772) [40] has been used as reference to align the remaining sequences. Boxes indicate residues that do not match the consensus (sequence #7, Gadus morhua ). Dotted lines indicate the position of primers 16Sar and 16Sbr 
Fig. 4 
Identification of gadoid species (Pisces, Gadidae) by sequencing and PCR–RFLP analysis of mitochondrial 12S and 16S rRNA gene fragments

July 2007


455 Reads

We applied PCR–RFLP (Polymerase Chain Reaction – Restriction Fragment Length Polymorphism) analysis to identify seven gadoid species of different biogeographical origin and commercial relevance, namely Gadus morhua (Atlantic ocean); Trisopterus minutus capelanus, Trisopterus minutus minutus, Molva elongata, Phycis blennoides, Micromesistius poutassou (Atlantic ocean and Mediterranean sea); Theragra chalcogramma (Pacific ocean). Two DNA fragments belonging to mitochondrial 12S and 16S rRNA genes of about 430 and 630bp, respectively, were isolated by PCR amplification. Their direct sequencing showed a significant genotypic diversity among gadoid species, useful for species identification. Digestion of 16S rRNA gene PCR fragment with MvaI or Bsh1285I restriction enzymes, followed by agarose gel electrophoresis of the cleaved products, yielded specific restriction profiles that enabled direct, visual identification of the species analyzed. This PCR–RFLP method allowed a clear and rapid discrimination of the gadoid species studied.

Structural modification of soy protein by 13-hydroperoxyoctadecadienoic acid

September 2009


41 Reads

To reveal the role of primary products of lipid peroxidation during soy protein oxidation process, oxidative modification of soy protein by 13-hydroperoxyoctadecadienoic acid (13-HPODE) generated by lipoxygenase-catalyzed oxidation of linoleic acid was investigated in this article. Incubation of soy protein with increasing concentration of 13-HPODE resulted in generation of protein carbonyl derivatives and loss of protein sulfhydryl groups. Circular dichroism spectra indicated that exposure of soy protein to 13-HPODE led to loss of α-helix structure. Effect of oxidation on tertiary structure was demonstrated by surface hydrophobicity and tryptophan fluorescence. Surface hydrophobicity gradually decreased, accompanied by loss and burial of some tryptophan residues. The results of surface hydrophobicity and tryptophan fluorescence implied that aggregation was induced by oxidation. Size exclusion chromatogram indicated that the extent of aggregation was increased in a 13-HPODE dose-dependent manner. Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated that non-disulfide linkages were involved in aggregate formation, and β-conglycinin was more vulnerable to 13-HPODE than glycinin.

Application of 13C NMR to the selection of the thermal processing conditions of canned fatty fish

January 2000


14 Reads

  13C nuclear magnetic resonance spectroscopy has been applied to select the processing conditions for tuna sterilisation. The work aimed to determine the heating times and retort temperatures required to guarantee a particular degree of microbiological sterility, but also to provide maximum nutrients and/or the retention of quality factors. Quality assessment was made on the basis of lipid degradation, hydrolysis and oxidation, and the estimation of the nutritional value (n-3 fatty acid content). Treatments involving the highest temperatures for the shortest times led to a final food with the highest n-3 fatty acid content and the lowest lipid degradation.

Determination of the Enantiomeric Ratios of Chiral PCB 95 and 149 in Human Milk Samples by Multidimensional Gas Chromatography with ECD and MS(SIM) Detection

August 1999


16 Reads

 The enantiomeric ratios of the chiral polychlorinated biphenyls 2, 2′, 3, 5′, 6-pentachlorobiphenyl (PCB 95) and 2, 2′, 3, 4′, 5′, 6-hexachlorobiphenyl (PCB 149) were determined in human milk samples by multidimensional gas chromatography, using two different achiral columns for preseparation and a chiral column for enantiomeric separation. Detection was carried out by electron capture detection and by mass spectrometry in the single ion monitoring mode. For PCB 95 a slight enrichment of the first eluted peak was determined whereas PCB 149 is present as racemate.

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