Evidence of 4-ethylcatechol as one of the main phenolic off-flavour markers in French ciders

A new HPLC method using a diode array detector was developed and validated to quantify 4-ethylphenol, 4-vinylphenol, 4-ethylguaiacol, 4-vinylguaiacol and 4-ethylcatechol in cider. The procedure was linear up to 150 mg/l for each of the five volatile phenols, precise (RSD < 2.9%) and sensitive, with limits of detection between 0.03 and 0.10 mg/l; moreover, it did not require any sample preparation. This method was applied to 11 phenolic off-flavour defective ciders. In these ciders, the main volatile phenol corresponded to 4-ethylcatechol. Moreover, the observed concentrations (maximum of 164 mg/l) indicated, for the first time, that this compound is an important phenolic off -flavour marker in cider. Then, volatile phenols concentrations were determined for 47 French commercial ciders and showed mean quantities of 3.2 (4-EC), 0.8 (4-EP), 0.1 (4-EG), 0.2 (4-VP) and 0.3 mg/l (4-VG). The majority of the tested commercial ciders presented low volatile phenol levels.     

Development,validation and application of a specific method for the quantitative determination of wine esters by headspace-solid-phase microextraction-gas chromatography–mass spectrometry

A method for specifically quantifying 32 apolar esters in wine is reported that employs head space solid phase microextraction (HS-SPME) followed by gas chromatography–mass spectrometry (GC–MS). A dozen esters were studied for the first time in wines, among them methyl trans-geranate, never reported in red wines until now. The target esters were apolar but bore a broad range of functional groups and have concentrations ranging from mg/l to ng/l levels; polar esters being effectively extracted by dichloromethane. Extraction and desorption conditions were optimised to obtain the best compromise for the simultaneous analysis of all 32 studied esters. To provide specificity for the method, deuterated ethyl esters were used as internal standards. These were ethyl-d₅ butyrate, hexanoate, octanoate and cinnamate, synthesised from the appropriate acyl chloride and ethanol-d₆. The esters were quantified in the wines with satisfactory repeatability (1.8% < RSD < 11.2%), reproducibility (1.5% < RSD < 15%), sensitivity (0.4 ng/l < LOQ < 4 μg/l), accuracy and specificity. The validation was carried out with several wine types as matrices (red, dry and sweet white). The optimised method was applied to 19 French wines and the results confirmed some well established oenological principles and opened prospects for further study on wine esters that had not been previously measured.     

Acrylamide in Caribbean foods – Residual levels and their relation to reducing sugar and asparagine content

The acrylamide levels in commercial and homemade Caribbean foods were determined by pre-derivatisation of acrylamide to 2-bromopropenamide and analysed by gas chromatography with mass spectrometric (GC/MS) detection. Over 100 Caribbean food samples were analysed for the presence of acrylamide. These samples include: biscuits, breakfast cereals, banana chips and home-prepared foods: breadfruit; Artocarpus altilis, banana fritters, and dumplings. The limit of detection (LOD) for the GC/MS method was found to be dependent on the type of column used for the GC/MS analysis. The DB-1701 and the DB-VRX columns gave LODs of 20 and 4 μg/kg, respectively. Acrylamide has not been found in raw foods or foods which have been cooked by boiling. Its content in all other foods had concentrations in the range, 65–3640 μg/kg. The relationship between acrylamide levels and precursor concentration as well as the health implications of our findings are discussed.     

Influence of the distillation processes from Rio de Janeiro in the ethyl carbamate formation in Brazilian sugar cane spirits

Ethyl carbamate (EC) or urethane, a potentially carcinogenic substance found in significant amounts in distilled fermented beverages, has become the main technical obstacle to the exportation of Brazilian sugar cane spirits (cachaças). In this evaluation, the EC levels were measured in samples of recently distilled beverages and of finished products, collected at 28 production plants using different distillation systems, from various regions of the state of Rio de Janeiro. The results obtained through GC–MS, using selective monitoring of ions with m/z 62 and 74, showed that approximately 45% of the products and raw materials are above the maximum allowed value of 150 μg/l, with an EC average value of 160 μg/l. The lower values were those for samples from alembics using low temperature and high reflux rate distillation, which emphasized the role of these two variables on the EC levels.     

Ethyl carbamate in cachaça (Brazilian sugarcane spirit): Extended survey confirms simple mitigation approaches in pot still distillation

In 2009, we reported an association between low levels of ethyl carbamate (EC) in pot still cachaças from Paraíba State, Brazil, and distillation in copper pot stills equipped with cooled columns. To strengthen these observations, we extended our study to Pernambuco State and assessed 13 pot still and 20 column still cachaça brands. An EC range from <40 to 532 μg/l was found; 18 brands exceeded the Brazilian limit (150 μg/l), 89% of which were column still types. Mean EC concentration of pot still cachaças was very low (64 μg/l), and was well below the Paraíba study (220 μg/l). An on-site investigation of pot still distilleries associated with <40 μg/l brands showed a connection to differences in the distillation apparatus. Maximising distillation reflux ratios in the ascending parts and minimising exposure to copper in the descending parts (through the use of stainless steel) can reduce EC, and also avoid copper contamination.     

Inactivation of spoilage microorganisms in apple cider using a continuous flow pulsed electric field system

The effect of pulsed electric field (PEF) in inactivating naturally occurring microorganisms (yeast and molds) in freshly squeezed apple cider was investigated in a continuous flow system. The microbial count decreased with an increase of applied pulses (17.6-58.7 total) and treatment temperature (45-50 °C), and a decrease of flow rate (3-10 l/h). At field strength of 27-33 kV/cm (3 mm electrode gap in a concentric chamber), 200 pulses/s, 3 l/h flow rate, and 50 °C process temperature, there was a 3.10 log reduction in microbial counts. By PEF treatment in the presence of a mixture of nisin and lysozyme (27.5 U/ml for nisin and 690 U/ml for lysozyme), there was an increase in the microbial count from 1.12 to 1.78 log reductions for 10 l/h flow. When cider samples were treated in the presence of clove oil (3 or 5 ml/100 ml), an additive reduction of 1.99 log cycles in microbial counts was observed.     

Ethyl carbamate in pot still cachaças (Brazilian sugar cane spirits): Influence of distillation and storage conditions

Ethyl carbamate (EC), a known genotoxic carcinogen, was studied in 25 brands of pot still cachaças from 19 distilleries in Paraíba State, Brazil. A concentration range of 55–700 μg/l was found with most brands (∼70%) exceeding the international EC limit for spirits (150 μg/l). Brand characteristics (colour, distillation [single or double], and bottle colouration) showed no consistent connection with EC levels. However, when EC levels of yellowish (cask matured) and colourless single-distiled cachaças from the same distillery were compared, the yellowish type was much more heavily contaminated. Eleven distilleries were visited and information regarding the distillation scale, kettle heating system, kettle’s shape, and cooling system of the column was collected. A close connection between EC levels and cooling system was found, with the non-cooled and cooled columns dominating the brands with high (200–700 μg/l range) and low (55–100 μg/l range) contamination levels, respectively.     

Concentrations of medium-chain 2- and 3-hydroxy fatty acids in foodstuffs

Medium chain 2-hydroxy fatty acids and 3-hydroxy fatty acids (2-OH-FAs and 3-OH-FAs with 8–20 carbons) are widespread in nature but little was known about the quantities of these minor fatty acids in food. For this reason, the concentrations and composition of 2- and 3-OH-FAs in several foodstuffs (milk and dairy products, animal brains, suet, vegetable oils) and two non-food samples (wool wax and vernix caseosa) were determined by gas chromatography with electron-capture negative ion mass spectrometry in the selected ion monitoring mode. Lipids were isolated from samples by accelerated solvent extraction. After transesterification the obtained fatty acid methyl esters (FAMEs) were separated into OH-FAMEs and non-OH-FAMEs. The OH-FAMEs were converted into pentafluorobenzoyl (PFBO) derivatives (PFBO-O-FAMEs). 2- and 3-OH-FAs were detected in all samples analysed. Chain-length ranged from 8–20 carbons. In general, 3-OH-FAs dominated in milk and dairy products (maximum concentration 17.7 mg/100 g l.w. in goat milk) whereas 2-OH-FAs were more abundant in animal brains and suet. The lowest concentrations were determined in vegetable oils (0.21–2.42 mg/100 g l.w.) whereas the highest concentrations of 2-OH-FAs were determined in wool wax (1180 mg/100 g l.w.) and vernix caseosa (34.5 mg/100 g l.w.).     

Browning on the surface of cut lettuce slices inhibited by short term exposure to nitric oxide (NO)

Freshly cut lettuce slices (Latuca sativa L.) were fumigated with nitric oxide (NO) gas at concentrations between 5 and 1000 μl/l in air at 20 °C for 1–4 h or dipped in an aqueous solution of the NO-donor compound, 2,2′-(hydroxynitrosohydrazino)-bisethanamine (DETANO) at concentrations between 10 and 1000 mg/l for 15 s to 60 min at 20 °C. Development of browning on the cut surfaces was inhibited during subsequent storage at 0 °C. The most effective treatments for extending postharvest life of lettuce slices were fumigation with 500 μl/l NO for 1 h, and dipping in 500 mg/l DETANO for 5 min. Dipping in DETANO solution was, however, more effective as it generated a 100% increase in postharvest life compared with a 70% increase due to NO gas. Solutions of DETANO in water were found to be relatively stable as the same extension in postharvest life was obtained for five batches of lettuce sequentially dipped in the same solution.     

Development and application of a method for the analysis of two trichothecenes: deoxynivalenol and T-2 toxin in meat in China by HPLC-MS/MS

A reliable and sensitive method was developed and successfully applied for the determination of deoxynivalenol and T-2 toxin simultaneously in pig dorsal muscle, pig back fat and chicken muscle by high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS) analysis. Limit of detection of deoxynivalenol and T-2 was 0.02 μg/kg and 0.007 μg/kg, and limit of quantification of deoxynivalenol and T-2 was 0.07 μg/kg and 0.02 μg/kg, respectively. Sixty-six meat samples were analyzed and deoxynivalenol was detected in the samples of pig back fat, with concentrations lower than 0.5 μg/kg, and T-2 toxin was detected in the samples of pig dorsal muscle, pig back fat and chicken muscle, with concentrations lower than 0.5 μg/kg. The results of sample analysis show that only trace residues of deoxynivalenol and T-2 toxin were detected in the samples analyzed.     

Ethyl-carbamate determination by gas chromatography–mass spectrometry at different stages of production of a traditional Brazilian spirit

Ethyl carbamate (EC), which is probably carcinogenic to humans, can be produced during the alcoholic fermentation of sugar-cane juice to give cachaça. The stages to produce cachaça are obtainment of sugar-cane juice, sugar-cane fermentation to wine, and obtainment of distilled fractions and residue. In order to investigate the presence of EC in the wine and in the fractions of the distillation process, as well as in the vinasse (the residue left after distillation), gas chromatography–mass spectrometry was employed. After the fermentation phase, the wine showed an average content of 122 mg L⁻¹ of EC. Average EC content in distilled fractions was 59.7 mg L⁻¹ for head, 52 μg·L⁻¹ for heart and 1.57 mg L⁻¹ for tail. EC content was 53.1 mg L⁻¹ for vinasse. The results showed that it is essential to separate the head and tail fractions to ensure cachaça quality, with respect to EC content.     

Formation of the highly stable pyranoanthocyanins (vitisins A and B) in red wines by the addition of pyruvic acid and acetaldehyde

Pyruvic acid (500 mg/l) and acetaldehyde (200 mg/l) were added, either as a large single dose or as smaller weekly doses over a 10 week period, to a young red wine (Vitis vinifera L. cv Tempranillo) in order to study the formation of vitisin A and B, and p-coumaroylvitisin A and B. In a further trial, pyruvic acid and acetaldehyde were added simultaneously as a single administration to test for any synergistic effect on vitisin formation. The addition of pyruvic acid led to the production of higher concentrations of vitisin A (4.08 ± 0.86 mg/l; 2.03% of the total anthocyanin content), while additions of acetaldehyde increased the concentration of vitisin B (2.47 ± 0.09 mg/l; 1.35%). The single, large dose administrations led to greater vitisin formation than did the smaller, weekly doses. Different patterns of formation were seen for vitisin A and B: the highest vitisin A content was achieved during the latter half of the 10 week study period while the highest vitisin B concentration was achieved early. The addition of acetaldehyde produced a greater reduction in monomeric anthocyanins than did the addition of pyruvic acid (loss of total anthocyanins 81.5%). The simultaneous addition of pyruvic acid and acetaldehyde led to less vitisin formation than did the addition of the reagents separately. p-Coumaroylvitisin A reached a maximum concentration of 0.86 ± 0.15 mg/l when the single dose of pyruvic acid was added, while the maximum recorded for p-coumaroylvitisin B was 0.66 ± 0.05 mg/l when the single dose of acetaldehyde was added. All anthocyanins were identified using HPLC/DAD and HPLC/ESI–MS.     

Influence of technological processing on apple aroma analysed by high resolution gas chromatography–mass spectrometry and on-line gas chromatography-combustion/pyrolysis-isotope ratio mass spectrometry

Extracts obtained by simultaneous distillation extraction (SDE) from industrial raw materials, namely single strength apple juices, and concentrates and aromas made thereof (each n = 31, from one production line; origin Poland, Germany, Turkey, Romania and China), as well as commercially available juices (n = 27), were analysed by standard controlled capillary gas chromatography–mass spectrometry (HRGC–MS). During the technological processing from juice to the aroma, no qualitative changes in the apple aroma profile were observed. Major constituents of the juices and aromas under study were found to be 1-hexanol (juice, 0.06–5.9 mg/l; aroma, 47–685 mg/l), 1-butanol (juice, 0.1–4.7 mg/l; aroma, 17–370 mg/l); E-2-hexenol (juice, 0.01–3.4 mg/l; aroma, 12–300 mg/l); E-2-hexenal (juice, 0–3.0 mg/l; aroma 0–470 mg/l), and butyl acetate (juice, 0–1.7 mg/l; aroma, 0–165 mg/l). By far the major component of the apple juice concentrates under study was furfural (2.4–56 mg/kg). The observed occurrence of 3-methyl-1-butanol (juice, 0.01–2.1 mg/l; aroma, 1.5–134 mg/l) and, in part, its acetate (juice, 0–0.3 mg/l; aroma, 0–3.3 mg/l), both known not to be genuine apple constituents, was obviously caused by fermentative effects in the course of industrial juice production. In addition, on-line capillary gas chromatography–isotope ratio mass spectrometry was used in the combustion (C) and the pyrolysis (P) modes (HRGC–C/P–IRMS) for the determination of δ¹³C_V-PDB and δ²H_V-SMOW values of selected apple flavour constituents to check potential isotope discrimination during distillative aroma production. As shown by means of the representative examples of E-2-hexenal, 1-hexanol and E-2-hexenol, their δ²H_V-SMOW values were slightly depleted. However, authenticity assessment by stable IRMS will not be influenced by this effect.     

Beta-carotene,lycopene, and alpha-tocopherol contents of selected Thai fruits

A total of 37 varieties of fresh fruits obtained from six representative markets in Bangkok, Thailand, were determined for their beta-carotene, lycopene, and alpha-tocopherol contents using high performance liquid chromatography. Beta-carotene content ranged from undetectable up to 616 μg/100 g of edible portion, lycopene content from undetectable up to 6693 μg/100 g and vitamin E content from not undetectable up to 1.43 mg/100 g. Red watermelon, Citruluss vulgalis (“jin-trarah” variety) was the richest source of dietary beta-carotene (1040 μg/serving) and lycopene (11,378 μg/serving), whilst the highest alpha-tocopherol content was found in unripe mango, Mangifera indica (“keosawoei” variety) with approximately 0.90 mg/75 g of edible portion, providing 9% of the Thai recommended daily intake of vitamin E.     

Toxicology and risk assessment of acrolein in food

Acrolein is an α,β-unsaturated aldehyde formed by thermal treatment of animal and vegetable fats, carbohydrates and amino acids. In addition it is generated endogenously. As an electrophile, acrolein forms adducts with gluthathione and other cellular components and is therefore cytotoxic. Mutagenicity was shown in some in vitro tests. Acrolein forms different DNA adducts in vivo, but mutagenic and cancerogenous effects have not been demonstrated for oral exposure. In subchronic oral studies, local lesions were detected in the stomach of rats. Systemic effects have not been reported from basic studies. A WHO working group established a tolerable oral acrolein intake of 7.5?μg/kg body weight/day. Acrolein exposure via food cannot be assessed due to analytical difficulties and the lack of reliable content measurements. Human biomonitoring of an acrolein urinary metabolite allows rough estimates of acrolein exposure in the range of a few μg/kg body weight/day. High exposure could be ten times higher after the consumption of certain foods. Although the estimation of the dietary acrolein exposure is associated with uncertainties, it is concluded that a health risk seems to be unlikely.     

Measurement of anthocyanins and other phytochemicals in purple wheat

The major anthocyanin composition of normal purple wheat and heat stressed purple wheat were measured using HPLC, LC–MS/MS and the pH differential method. The lignan secoisolariciresinol diglucoside (SDG) and melatonin content were also measured. Total anthocyanin profile of normal purple wheat (491.3 mg/kg) was significantly (P < 0.05) lower than that of the heat stressed purple wheat (522.7 mg/kg). Thirteen major anthocyanins were isolated and cyanidin 3-glucoside was the predominant anthocyanin in purple wheat. Using the pH differential method, the total anthocyanin content of normal (500.6 mg/kg) and heat stressed (526.0 mg/kg) purple wheat were similar to those observed using HPLC. The SDG content of normal and heat stressed purple wheat were 770 and 520 μg/kg, while melatonin content was 4 and 2 μg/kg, respectively. The presence of SDG and melatonin in addition to anthocyanins may contribute to the health benefits associated with consumption of coloured cereal grains.     

Essential oil composition,antimicrobial and antioxidant properties of Mosla chinensis Maxim

The essential oil of Mosla chinensis Maxim was analysed by gas chromatography/mass spectrometry, and its main components are carvacrol (57.08%), p-cymene (13.61%), thymol acetate (12.68%), thymol (6.67%), and γ-terpinene (2.46%). The essential oil exhibited great potential antimicrobial activity against all eight bacterial and nine fungal strains. Antioxidant activity was also tested, the essential oil showing significantly higher antioxidant activity than that of the methanol extract. In addition, the amounts of total phenol components in the plant methanol extract (47.3 ± 0.4 μg/mg) and the oil (80.7 ± 0.5 μg/mg) were determined. The results presented here indicate that the essential oil of M. chinensis has antimicrobial and antioxidant properties, and is therefore a potential source of antimicrobial and antioxidant agents for the food and pharmaceutical industries.     

Validation of antibiotics in catfish by on-line solid phase extraction coupled to liquid chromatography tandem mass spectrometry

For the first time automated on-line solid phase extraction coupled to liquid chromatography tandem mass spectrometry was developed for the simultaneous determination of 13 antibiotics (sulfonamides and tetracyclines) in catfish. The method proposed was validated according to Commission Decision 2002/657/EC, showing good linearity between 2 and 350 μg kg⁻¹, high recovery (80–99%) and reproducibility (13–20%) values, lower detection limits than 0.1 μg kg⁻¹, and quantification limits under 2.4 μg kg⁻¹ (between 39 and 84 times lower than the MRL fixed by the EU). Moreover, the proposed method was also used to determine sulfonamides and tetracyclines in 16 out of 107 samples, all previously analysed by microbiological screening that gave positive results. Five out of 13 antibiotics were found, having tetracycline the higher occurrence (10 samples); in all cases the concentrations were lower than the MRL established.     

Essential oil composition,antimicrobial and antioxidant activities of Satureja cuneifolia Ten.

Satureja cuneifolia Ten. is a well-known aromatic plant which is frequently used as a spice and herbal tea in Anatolia. S. cuneifolia oil was analyzed by gas chromatography/mass spectrometry (GC/MS). The major components of S. cuneifolia oil were carvacrol (44.99%) and p-cymene (21.61%). The essential oil of S. cuneifolia exhibited antimicrobial activity against all of the tested foodborne and spoilage bacteria. The minimum inhibitory concentration (MIC) values for test bacteria which were sensitive to the essential oil of S. cuneifolia were in the range of 600–1400 μg/ml. Antioxidant activities of the essential oil and the methanolic extract from S. cuneifolia were evaluated by using DPPH radical scavenging, β-carotene–linoleic acid bleaching and metal chelating activity assays. In addition, the amounts of total phenol components in the plant methanolic extract (222.5 ± 0.5 μg/mg) and the oil (185.5 ± 0.5 μg/mg) were determined.