Ionic liquid-based ultrasound-assisted surfactant-emulsified microextraction for simultaneous determination of three important flavoring compounds in plant extracts and urine samples

For the first time, a novel, efficient and environmentally friendly method termed ionic liquid-based ultrasound-assisted surfactant-emulsified microextraction (IL-UA-SE-ME) was developed and compared with normal-, reversed-, surfactant assisted-dispersive liquid–liquid microextraction and ultrasound-assisted surfactant-based emulsification microextraction methods for the analysis of three bioactive and flavoring compounds (para-anisaldehyde, trans-anethole and its isomer estragole) in some fresh plants (fennel and basil), their extracts and urine samples. The results showed that IL-UA-SE-ME is a much more effective method and under the optimum conditions (including extraction solvent: 90 μL of [C₆MIM][PF₆]; disperser: 5 mg of N-dodecylbenzenesulfonic acid sodium salt; sample pH: 3; sonication time: 5 min; and centrifuging time: 5 min), limits of detection, limits of quantification, linear ranges, recoveries, and enrichment factors were in the range of 16–22 ng mL^− 1, 49–67 ng mL^− 1, 0.04–90 μg mL^− 1, 94.3%–101.1%, and 118–127, respectively.     

Comparison of palladium–magnesium nitrate and ammonium dihydrogenphosphate modifiers for lead determination in honey by electrothermal atomic absorption spectrometry

The aim of the present work was to develop and optimize two procedures for the determination of Pb content in honeys by electrothermal atomic absorption spectrometry without any sample pretreatment. Palladium–magnesium nitrate and ammonium dihydrogenphosphate were used as chemical modifiers. Honey was diluted in water, and hydrogen peroxide, nitric acid, and Triton X-100 were added to minimize the matrix effect. RSD (lower than 10%) and the analytical recovery (98–101%) were acceptable for both methods. Pd–Mg(NO₃)₂ was the method selected for further direct Pb determinations in honey samples because it presented the best limit of detection (LOD = 1.6 ng g⁻¹). Moreover, this method allowed Pb determination using a calibration graph, instead of an addition graph, which is an important advantage. The direct proposed methods have been applied to the determination of Pb content in representative honey samples from Galicia (NW Spain). The lead concentrations found in the analyzed samples were in the range 1.71–75.0 ng g⁻¹.     

Immunoglobulins,growth factors and growth hormone in bovine colostrum and the effects of processing

In colostrum collected 0–80 h postpartum the contents of immunoglobulins (Igs), transforming growth factor beta-2 (TGF-β2), insulin-like growth factor-1 (IGF-1) and growth hormone (GH) were analysed. Colostrum initially contained 90 mg mL⁻¹ IgG1, 2.8 mg mL⁻¹ IgG2, 1.6 mg mL⁻¹ IgA, 4.5 mg mL⁻¹ IgM, and these concentrations declined by 92%, 87%, 93% and 84%, respectively, in the samples collected later. Of the growth factors, colostrum initially contained 289–310 ng mL⁻¹ TGF-β2 and the concentration diminished to 66 ng mL⁻¹. The content of IGF-1 and GH postpartum decreased from 870 to 150 ng mL⁻¹, and from 0.17 to <0.03 ng mL⁻¹, respectively. Heat treatment and freeze-drying of colostral whey decreased the content of Igs to 75%, while the contents of IGF-1 and TGF-β2 were unaffected. A similar processing, including filtration steps reduced also the IGF-1 and TGF-β2 by 25%. IgM seems to be the most sensitive of the Igs to processing.     

Modeling the effect of natamycin,pine-resin and environmental factors on the growth and OTA production by Aspergillus carbonarius using response surface methodology

The effect of two antifungal compounds (natamycin, pine-resin), temperature and water activity, on the growth rate, lag phase duration and Ochratoxin A (OTA) production by three Aspergillus carbonarius isolates (Ac-28, Ac-29, and Ac-33), was studied by means of Response Surface Methodology (RSM) based on a Central Composite Design (CCD). Two different experimental designs were performed as a function of temperature (16.6–33.4 °C), water activity (0.90–0.97 a_w), natamycin (0–1000 ng ml^− 1) or pine-resin (0–2.61%, w/v) on a Synthetic Grape-juice Medium (SGM). OTA production was analyzed after 5, 10 and 15 days of incubation. A second-order polynomial model was fitted to each response parameter to assess the growth and OTA potential of all fungal isolates. Results showed that natamycin, a_w and temperature had significant effects on the lag phase duration of all isolates, as well as on OTA accumulation after 10 days of incubation for Ac-29 and 15 days for Ac-28 and Ac-33 isolates. The same results were obtained for OTA production after treatment with pine-resin. However, fungal growth rates were not statistically significant in both experiments, with the exception of Ac-29 and Ac-33 after treatment with pine-resin. Overall, high natamycin concentrations (800 and 1000 ng ml^− 1) delayed fungal growth depending on the environmental factors assayed. Moreover, treatment with pine-resin at 16.6 °C/0.94 a_w/1.1% w/v, as well as at 25 °C/0.90 a_w/1.1% w/v, completely inhibited fungal growth up to 15 days of incubation.     

Isolation and characterisation of a novel antibacterial peptide from bovine αS1-casein

Bovine casein was hydrolysed with a range of proteolytic enzymes including pepsin, trypsin, α-chymotrypsin and β-chymotrypsin, and assessed for antibacterial activity. The pepsin digest of bovine casein, which showed antibacterial activity, was fractionated using reverse phase high performance liquid chromatography and the antibacterial peptides isolated were characterised using electrospray ionisation mass spectrometry. Two antibacterial peptides were identified, a novel peptide (Cp1) which corresponded to residues 99–109 of bovine α_S1-casein and a previously reported peptide (Cp2) which corresponded to residues 183–207 of bovine α_S2-casein. The minimum inhibitory concentration (MIC) of Cp1 and Cp2 were determined against a range of bacterial cultures. Cp1 exhibited an MIC of 125 μg mL⁻¹ against all Gram-positive bacteria tested, and MIC ranging between 125 and >1000 μg mL⁻¹ against the Gram-negative bacteria tested. Cp2 was generally far more potent against the Gram-positive bacteria, exhibiting an MIC of 21 μg mL⁻¹, compared to MICs ranging from 332 to >664 μg mL⁻¹ against most of the Gram-negative bacteria tested.     

Biochemical changes throughout the ripening of a traditional Spanish goat cheese variety (Babia-Laciana)

The physico-chemical characteristics, proteolysis (classical nitrogen fractions, caseins and their degradation products and free amino acids), and lipolysis (fat acidity and free fatty acids) were studied throughout the ripening of three batches of Babia-Laciana cheese, a Spanish traditional variety made from raw goats’ milk. The main compositional characteristics of this cheese at the end of the ripening are its high content of total solids (TS) (78.0±2.4 g 100 g⁻¹ of cheese) and fat (61.1±1.2 g 100 g⁻¹ of TS), the presence of residual lactose (1.6±0.8 g 100 g⁻¹ of TS) and its low content of sodium chloride (1.1±0.7 g 100 g⁻¹ of TS) and ash (2.8±0.5 g 100 g⁻¹ of TS). Its pH values (4.44±0.72) are extraordinarily low. The evolution and final values of the different nitrogen fractions show that this cheese undergoes a very slight proteolysis, a fact which was corroborated when the caseins and their degradation products were quantified: β-casein did not undergo any modification throughout ripening, while only 21% of the α_s-caseins were degraded. Free amino acids content increased by a factor of about 7 throughout ripening, resulting in a high content of γ-amino butyric acid and a low content of glutamic acid at the end of the process. Fat acidity increased very slightly, approximately 4.5 times, during ripening, reaching final values of 3.5±2.2 mg KOH g⁻¹ of fat. The total free fatty acids content showed a similar evolution to fat acidity. At the end of the ripening process, the main free fatty acid was C_18:1, followed by C₁₆ and C₁₀.     

Aflatoxin M1 in milk: Reliability of the immunoenzymatic assay

To evaluate the reliability of ELISA in aflatoxin M₁ (AFM₁) determination in milk, more than 15,000 samples were tested and results from 600 of them were compared with those of the HPLC reference method. Extraction efficiency by immunoaffinity columns was checked under various conditions. ELISA and HPLC assays of spiked samples gave same precision (coefficient of variation), recovery, and regression coefficient R² values (0.9–8%, 96.8–108%, 0.993, respectively) for samples containing less than 70 ng L⁻¹ of AFM₁. At higher concentrations up to 100 ng L⁻¹, ELISA gave a slight overestimation, with CV 7–21% and R²=0.876. This overestimation was confirmed (R²=0.788) for the 600 comparison samples. Data from this study support ELISA as a reliable routine quality control assay for AFM₁ in milk, quantifying exactly its content for values above, but close to, the legal limit. Regulatory rules require HPLC confirmation of positive samples, but by using ELISA, no cases of false-negative determination should occur.     

Determination of lactoferrin in bovine milk,colostrum and infant formulas by optical biosensor analysis

An automated, rapid, sensitive and label-free biosensor-based immunoassay for lactoferrin in bovine milk utilising surface plasmon resonance (SPR) optical detection is described. Lactoferrin content is estimated from its specific interaction with an anti-bovine lactoferrin antibody immobilised on the sensor surface in a direct- binding assay format. Samples are prepared for analysis by direct dilution into buffer. Analysis conditions, including ligand immobilisation, flow-rate, contact time and regeneration have been defined and non-specific binding considerations evaluated. Performance parameters include a working range of 0–1000 ng mL⁻¹, a method-detection limit of 19.9 μg mL⁻¹ in undiluted milk, overall instrument response RSD_R of 3.50%, a mean inter-assay RSD_R of 10.8% for milk and surface stability over ca 500 samples. The technique was applied to the measurement of lactoferrin content of consumer milks, colostrum and infant formulas, and temporal change during early bovine lactation.     

In-vitro mineral binding capacity of five fiber sources and their insoluble components for magnesium and calcium1

In-vitro binding of calcium (Ca) and magnesium (Mg) by total dietary fiber, hemicellulose A (HCL A), lignocellulose (LCL), cellulose (CL), and lignin (L) fractions isolated from rice bran (RB), wheat bran (WB), oat fiber (OF), apple fiber (AF) and tomato fiber (TF) was evaluated. At pH 6.8, significant amounts of Ca were bound by whole fibers, ranging from 800 μg g⁻¹ for RB to 10 097 μg g⁻¹ for TF. Mg bound by whole fibers varied from 496 μg g⁻¹ for OF to 2177 μg g⁻¹ for WB. Re-acid washing (pH<2.0) released 95–99% of the Ca and Mg bound to the fibers. Fibers with the highest endogenous Ca and Mg concentrations bound significantly (P<0.05) the highest amounts of the minerals studied. The Ca bound by HCL A varied from 9753 μg g⁻¹ for RB to 11 337 mg g⁻¹ for TF, whereas Mg bound varied from 1151 μg g⁻¹ for OF to 5626  μg g⁻¹ for TF hemicellulose fractions, respectively. Among the fiber components, Mg binding decreased in the order HCL A>LCL>L>CL, whereas Ca bound was in the order HCL A>LCL>CL>L. A relatively strong correlation was observed between the combined effects of protein content, hemicellulose, and lignin vs total Ca and Mg bound. 1998 Elsevier Science Ltd. All rights reserved     

Thermal and light stabilities and antioxidant activity of carotenoids from tomatoes extracted using an ultrasound-assisted completely solvent-free method

The use of petroleum-derived solvents, particularly volatile organic compounds (VOCs), in the chemical industry has increased the contamination and residual effects of these solvents. Ionic liquids (ILs) can potentially replace VOCs, thereby reducing the risks of environmental contamination and toxicity. In this context, the objectives of this study were as follows: 1 — to obtain an ionic liquid for use in extracting carotenoids from tomatoes with ultrasound assistance; and 2 — to determine the stability and antioxidant activity of tomato carotenoid extracts. Ultrasound can also efficiently extract carotenoid compounds with ionic liquids in comparison with conventional VOC solvents (obtained from an all-trans-lycopene 7.5–8.0 μg·g^− 1 tomato sample by IL and 6.2–7.7 μg·g^− 1 by acetone). Similarly, the activation energies (E_a) in aqueous medium were obtained for the IL carotenoid extract (10.8 kJ·mol^− 1) and acetone carotenoid extract (9.4 kJ·mol^− 1). The antioxidant activities of the tomato carotenoid extract were 7.4 and 12.4 relative to α-tocopherol for the ionic liquid extract and acetone extract, respectively. The combination of chromatographic analysis and degradation kinetics provided data for positive assessment similarity of thermal and light stabilities of tomato carotenoids extracted by IL and extracted by acetone.     

Purification of angiotensin I-converting enzyme inhibitory peptides and antihypertensive effect of milk produced by protease-facilitated lactic fermentation

Fresh low-fat milk was fermented with five mixed lactic acid bacteria for up to 30 h at 42 °C. A protease, prozyme 6, was added 5 h after the beginning of fermentation. The whey was separated from the fermented milk and freeze-dried. As the fermentation time extended to 30 h, soluble protein content increased from 30.9 to 195.9 mg g⁻¹, free amino acid content increased from 2.8 to 192.8 mg g⁻¹, peptide content increased from 6.4 to 402.8 mg g⁻¹ and γ-aminobutyric acid (GABA) increased from 0 to 80.6 mg 100 g⁻¹, while inhibition of angiotensin I-converting enzyme (ACE) increased as indicated by a decrease of IC₅₀ from 1.18 to 0.24 mg mL⁻¹, respectively. The amino acid sequences of two ACE inhibitory peptides were Gly–Thr–Trp and Gly–Val–Trp, of which the IC₅₀ values were 464.4 and 240.0 μm, respectively. The systolic blood pressure and diastolic blood pressure of spontaneously hypertensive rat (SHR) were reduced 22 and 21.5 mm Hg, respectively, after 8 weeks of oral administration of diluted whey (peptide concentration 5 mg mL⁻¹) from the 30 h fermentation.     

Antimicrobial agent detection in ewes’ milk by the microbial inhibitor test brilliant black reduction test—BRT AiM®

The detection limits of 24 antimicrobial agents were determined in ewes’ milk by one commercially available version of brilliant black reduction test, BRT Inhibitor Test with prediffusion AiM^® (BRT AiM^®). For each drug, eight concentrations were tested on 20 milk samples from individual ewes. The detection limits of the BRT AIM^® method were determined by means of logistic regression models: 6 μg kg⁻¹ amoxycillin, 6 μg kg⁻¹ ampicillin, 51 μg kg⁻¹ cloxacillin, 2 μg kg⁻¹ penicillin “G”, 230 μg kg⁻¹ cefadroxil, 1330 μg kg⁻¹ cephalosporin “C”; 270 μg kg⁻¹ cephalexin, 92 μg kg⁻¹ cefoperazone, 120 μg kg⁻¹ ceftiofur, 69 μg kg⁻¹ cefuroxime, 6000 μg kg⁻¹ streptomycin, 1200 μg kg⁻¹ gentamycin, 3700 μg kg⁻¹ neomycin, 630 μg kg⁻¹ erythromycin, 120 μg kg⁻¹ tylosin, 390 μg kg⁻¹ doxycycline, 5500 μg kg⁻¹ oxytetracycline, 6200 μg kg⁻¹ tetracycline, 5400 μg kg⁻¹ sulfadiazine, 3200 μg kg⁻¹ sulfamethoxazole, 6500 μg kg⁻¹ sulfamethoxypyridazine, 6200 μg kg⁻¹ sulfaquinoxaline, 22000 μg kg⁻¹ chloramphenicol and 4100 μg kg⁻¹ trimethoprim. The BRT AiM^® method presents detection limits for β-lactam antibiotics that are similar to those obtained as Maximum Residue Limits (MRLs) according to Regulation 2377/90 EEC as set out by the European Union. However, for other antimicrobial agents the estimated limits were higher than those of the EU-MRLs. It is therefore advisable to enhance the sensitivity of the method for the detection of the different antimicrobial groups or to develop a combined system of different microbiological inhibitor tests that would enable the detection of a greater number of antimicrobial agents.     

Effect of combinations of high-pressure treatment and bacteriocin-producing lactic acid bacteria on the survival of Listeria monocytogenes in raw milk cheese

The combined effect of high-pressure (HP) treatment and bacteriocin-producing lactic acid bacteria (BP-LAB) on the survival of Listeria monocytogenes Scott A in cheeses made from raw milk that was inoculated with the pathogen at 4.80 log cfu mL⁻¹, a commercial starter and one of seven strains of BP-LAB was investigated. On day 3, the counts of L. monocytogenes were 7.03 log cfu g⁻¹ in a control cheese (without BP-LAB, not HP treated), 6.06–6.74 log cfu g⁻¹ in cheeses with BP-LAB, 6.13 log cfu g⁻¹ in a cheese without BP-LAB and treated on day 2 at 300 MPa, 2.01 log cfu g⁻¹ in a cheese without BP-LAB and treated on day 2 at 500 MPa, 3.83–5.43 log cfu g⁻¹ in cheeses with BP-LAB and treated on day 2 at 300 MPa, and 1.81 log cfu g⁻¹ or less in cheeses with BP-LAB and treated on day 2 at 500 MPa. HP treatment was more effective on day 51 than on day 2.     

Enterobacteriaceae in beef products from retail outlets in the Republic of Ireland and comparison of the presence and counts of E. coli O157:H7 in these products

This study investigated the prevalence and numbers of Enterobacteriaceae in minced beef and beef burgers purchased from supermarkets and butcher shops in the Republic of Ireland (RoI). Samples (n=1303) collected between June 2001 and April 2002 from every county in the RoI (∼60 per county) were examined for the presence of Enterobacteriaceae using method BS 5763. Overall, in the 43 beef products in which E. coli O157:H7 was present the Enterobacteriaceae counts ranged from 0.52 to 6.98 log₁₀ cfu g⁻¹. There was no correlation between the number of Enterobacteriaceae and the presence of E. coli O157:H7. There were no significant differences between Enterobacteriaceae numbers in fresh, unpackaged, minced beef samples from butcher shops and supermarkets, or in fresh, unpackaged, beef burgers from butcher shops and supermarkets. However, there were significant differences among the numbers of Enterobacteriaceae detected in different minced beef products. The numbers of Enterobacteriaceae in fresh, unpackaged, minced beef (6.54–6.98 log₁₀ cfu g⁻¹) were considerably higher than in preprepared or prepackaged minced beef (2.95–3.62 log₁₀ cfu g⁻¹).     

Mercury speciation in the muscle of two commercially important fish,hake (Merluccius merluccius) and striped mullet (Mullus barbatus) from the Mediterranean sea: estimated weekly intake

Total mercury and methylmercury concentrations were measured in the muscle tissue of two fish species from the Ionian and Adriatic seas. Higher total mercury and methylmercury concentrations were detected in striped mullet (Mullus barbatus), a benthic species (Ionian sea: Hg=0.40 μg g⁻¹ wet wt, MeHg=0.40 μg g⁻¹ wet wt; Adriatic sea: Hg=0.49 μg g⁻¹ wet wt, MeHg=0.44 μg g⁻¹ wet wt), than in hake (Merluccius merluccius), a pelagic species (Ionian sea: Hg=0.09 μg g⁻¹ wet wt, MeHg=0.09 μg g⁻¹ wet wt; Adriatic sea: Hg=0.18 μg g⁻¹ wet wt; MeHg=0.16 μg g⁻¹ wet wt). Total mercury residues were determined in all samples of both species from the Adriatic sea, while levels below the limit of detection were registered in 25% and 11%, respectively, of striped mullet and hake samples from the Ionian sea. In 18.8% and 22.2% of striped mullet samples from the Ionian and Adriatic seas, respectively, total mercury concentrations exceeded the maximum level fixed by the European Commission Decision (Hg=0.5 μg g⁻¹ wet wt). In the two different species, mercury was present almost completely in the methylated form with mean percentages between 60% and 100%. The estimated weekly intake for total mercury was below the established the provisional tolerable weekly intake (PTWI) for both species, though their consumption provides a methylmercury intake above the WHO safety limit.     

Survival of Listeria monocytogenes in Galotyri,a traditional Greek soft acid-curd cheese,stored aerobically at 4°C and 12°C

Galotyri is a traditional Greek soft acid-curd cheese, which is made from ewes’ or goats’ milk and is consumed fresh. Because cheese processing may allow Listeria monocytogenes post-process contamination, this study evaluated survival of the pathogen in fresh cheese during storage. Portions (0.5 kg) of two commercial types (<2% salt) of Galotyri, one artisan (pH 4.0±0.1) and the other industrial (pH 3.8±0.1), were inoculated with ca. 3 or 7 log cfu g⁻¹ of a five-strain cocktail of L. monocytogenes and stored aerobically at 4°C and 12°C. After 3 days, average declines of pathogen's populations (PALCAM agar) were 1.3–1.6 and 3.7–4.6 log cfu g⁻¹ in cheese samples for the low and high inocula, respectively. These declines were independent (P>0.05) of the cheese type or the storage temperature. From day 3, however, declines shifted to small or minimal to result in 1.4–1.8 log cfu g⁻¹ of survivors at 28 days of storage of all cheeses at 4°C, indicating a strong “tailing” independent of initial level of contamination. Low (1.2–1.7 log cfu g⁻¹) survival of L. monocytogenes also occurred in cheeses at 12°C for 14 days, which were prone to surface yeast spoilage. When ca. 3 log cfu g⁻¹ of L. monocytogenes were inoculated in laboratory scale prepared Galotyri of pH ≅4.4 and ≅3% salt, the pathogen died off at 14 and 21 days at 12°C and 4°C, respectively, in artisan type cheeses fermented with the natural starter. In contrast, the pathogen survived for 28 days in cheeses fermented with the industrial starter. These results indicate that L. monocytogenes cannot grow but may survive during retail storage of Galotyri despite its low pH of or slightly below 4.0. Although contamination of Galotyri with L. monocytogenes may be expected low (<100 cfu g⁻¹) in practice, that long-term survival of the pathogen in commercial cheeses was shown to be unaffected by the artificial contamination level (3 or 7 logs) and the storage temperature (4°C or 12°C), which should be a concern.     

Production of volatile aroma compounds by kefir starter cultures

Production of carbonyl compounds by single-strain cultures, kefir starter (Lactobacillus delbrueckii subsp. bulgaricus HP1+Lb. helveticus MP12+Lactococcus lactis subsp. lactis C15+Streptococcus thermophilus T15+Saccharomyces cerevisiae A13) and kefir grains during fermentation and storage of kefir was studied. The content of carbonyl compounds produced by kefir starter was greater than that produced by kefir grains. The maximum acetaldehyde concentration (18.3 μg g⁻¹) in kefir with starter culture was mainly due to the metabolic activity of Lb. delbrueckii subsp. bulgaricus HP1 isolated from kefir grains. The highest diacetyl production activity was recorded in the starter culture (1.87 μg g⁻¹) and the single-strain culture St. thermophilus T15 (1.62 μg g⁻¹), followed by Lb. helveticus MP12 (0.85 μg g⁻¹) and Lc. lactis subsp. lactis C15 (0.42 μg g⁻¹). The lactobacilli Lb. delbrueckii subsp. bulgaricus HP1 and Lb. helveticus MP12 produced acetone, which was not found in the cocci cultures. The presence of 2-butanone was related to the production ability of Lb. helveticus MP12. In comparison, Lc. lactis subsp. lactis C15 synthesized ethyl acetate more actively than the other single-strain cultures included in the starter. S. cerevisiae A13 produced ethanol and CO₂ in amounts (3975 μg g⁻¹; 1.80 g L⁻¹) that lent cultured kefir distinctive flavour and aroma characteristic of authentic kefir.     

Impact of relative humidity and temperature on visible fungal growth and OTA production of ochratoxigenic Aspergillus ochraceus isolates on grapes

Aspergillus ochraceus is an ochratoxin A (OTA) producer mould found in grapes and this may contribute to OTA contamination in wines and grape juices. The influence of relative humidity (R.H.; 80, 90 and 100%) and temperature (10, 20 and 30 °C) on visible mould growth on grapes and OTA accumulation after 14 days of incubation by this fungal species has been studied using a full factorial design with three replicates.The two abiotic factors and their interaction (R.H.×temperature) affected significantly the A. ochraceus growth in berries, which was maximum at 90–100% R.H. levels. With regard to the optimum temperature level, it occurred at 30 °C at 80 and 90% R.H., whereas no significant differences were detected at 20 and 30 °C when R.H. was 100%.OTA production by A. ochraceus on grapes was not significantly modified by the assayed levels of temperature and R.H, with a mean value of 3.53 ng g⁻¹.Predictive models of percentage of grapes with visible growth of A. ochraceus isolates under different relative humidity and temperature are presented.     

Enrichment of the conjugated linoleic acid content of bovine milk fat by dry fractionation

This study investigated the effect of fat fractionation on the conjugated linoleic acid (cis-9, trans-11-C_18 : 2) content of bovine milk fat. Anhydrous milk fat was fractionated into hard and soft fractions using controlled cooling and agitation. Fractionation of milk fat pre-melted at 60°C using a temperature programme of 33–10°C and a cooling rate of 0.58°C h⁻¹ yielded a soft fraction containing 63.2% more conjugated linoleic acid (2.22 g 100 g⁻¹ FAME), which was also enriched in polyunsaturated fatty acids and vaccenic acid (trans-11-C_18 : 1) compared with the parent fat. Agitation following fractionation was found to have a negative effect on the conjugated linoleic acid content of the soft fraction. Refractionation of the soft fraction did not increase the yield of conjugated linoleic acid. The conjugated linoleic acid and trans fatty acid content of 26 selected food products ranging in milk fat content from 0 to 100% is reported. Conjugated linoleic acid concentrations ranged from 0 to 16.2 mg g⁻¹ fat and were generally lower than the trans fatty acid content which ranged from 0 to 155.7 mg g⁻¹ fat. Spreads containing vegetable oils contained higher trans fatty acid and lower conjugated linoleic acid contents than milk fat-containing products. This study highlights that a milk fat fraction enriched in conjugated linoleic acid may be achieved by dry fractionation.     

Multicommutation hydride generation atomic fluorescence determination of inorganic tellurium species in milk

A multicommutated flow system has been developed for hydride generation, atomic fluorescence (HG-AFS) determination of tellurate (Te^VI) and tellurite (Te^IV) in milk samples. After a batch leaching of Te by sonication at room temperature for 10 min with aqua regia, sample slurries in acidic medium were merged with sodium borohydride and HCl to obtain data on Te^IV. Another portion of the acidic slurry was mixed with KBr and passed through a reaction coil introduced inside a microwave oven to reduce quantitatively Te^VI to Te^IV which was analyzed by HG-AFS. The detection limit was 0.57 ng g⁻¹ in the original samples. The linear range obtained was till 4 ng ml⁻¹ and the average recovery of different amounts of Te^VI and Te^IV added to real milk samples were 98 ± 4% and 98 ± 2%, respectively, indicating the absence of analyte losses or contaminations and original species modification. Average relative standard deviation of 6.3% was found for Te determination in a series of commercially available milk samples containing from 1.0 to 10.1 ng ml⁻¹ total Te. The proposed method provided a high sampling frequency of 24 h⁻¹ for the determination of both, free Te^IV and total Te, in a same sample with a two times reduced waste generation and a four times reduced reagent consumption as compared with the continuous hydride generation. Additionally, the method developed requires a minimum operator attention and sample manipulation.