Determination of polycyclic aromatic hydrocarbons in smoked fish samples by a new microextraction technique and method optimisation using response surface methodology.

Microwave-assisted extraction coupled with dispersive liquid-liquid microextraction as a recently introduced method was applied to determine polycyclic aromatic hydrocarbons in Iranian smoked fish. The results showed that the interaction of hydrolysing solution volume with ethanol ratio, volumes of extracting and disperser solvent is significant in the obtained model. Optimized conditions were: a hydrolysing solution volume 10 ml with 50% ethanol, a pH of 5, and extracting and disperser solvent volumes of 150 and 500 µl respectively. The level of 16 polycyclic aromatic hydrocarbons (PAHs) was determined in 80 smoked fish consisting of four species. The contamination of benzo[a]pyrene in all samples except three was below the European Commission's maximum level of 2 µg kg(-1) for smoked fish, while the ∑4 PAHs (benzo[a]pyrene, chrysene, benzo[a]anthracene and benzo[b]fluoranthene) were between 3 and 12 µg kg(-1) wet weight in all samples. Of the species examined, Hypophthalmichthys molitrix had the highest PAHs (∑16).

Microwave-assisted extraction and dispersive liquid-liquid microextraction followed by gas chromatography-mass spectrometry for isolation and determination of polycyclic aromatic hydrocarbons in smoked fish.

A simple and efficient method was developed using microwave-assisted extraction (MAE) and dispersive liquid-liquid microextraction (DLLME) coupled with gas chromatography-mass spectrometry (GC-MS) for the extraction and quantification of 16 polycyclic aromatic hydrocarbons (PAHs) in smoked fish. Benzo[a]pyrene, chrysene and pyrene were employed as model compounds and spiked to smoked fish to assess the extraction procedure. Several parameters, including the nature and volume of hydrolysis, extracting and disperser solvents, microwave time and pH, were optimized. In the optimum condition for MAE, 1g of fish sample was extracted in 12 mL KOH (2M) and ethanol with a 50:50 ratio in a closed-vessel system. For DLLME, 500 µL of acetone (disperser solvent) containing 100 µL of ethylene tetrachloride (extraction solvent) was rapidly injected by syringe into 12 mL of the sample extract solution (previously adjusted to pH 6.5), thereby forming a cloudy solution. Phase separation was performed by centrifugation and a volume of 1.5 µL of the sedimented phase was analyzed by GC-MS in select ion monitoring (SIM) mode. Satisfactory results were achieved when this method was applied to analyze the PAHs in smoked fish samples. The MAE-DLLME method coupled with GC-MS provided excellent enrichment factors (in the range of 244-373 for 16 PAHs) and good repeatability (with a relative standard deviation between 2.8 and 9%) for spiked smoked fish. The calibration graphs were linear in the range of 1-200 ng g(-1), with the square of the correlation coefficient (R(2))>0.981 and detection limits between 0.11 and 0.43 ng g(-1). The recoveries of those compounds in smoked fish were from 82.1% to 105.5%. A comparison of this method with previous methods demonstrated that the proposed method is an accurate, rapid and reliable sample-pretreatment method that gives very good enrichment factors and detection limits for extracting and determining PAHs from smoked fish.

Self-assembly of ß-lactoglobulin and the soluble fraction of gum tragacanth in aqueous medium.

Spectrophotometric and light scattering measurements, along with optical microscopy, were used to follow the complexation and coacervation process that occur when ß-lactoglobulin (BLG)/tragacanthin (T) mixed dispersions (0.3 wt.% total concentration; BLG:T ratio of 2:1) were brought from pH 6 to pH 2. In addition, the coupling of slow in situ acidification of the mixture and rheometry was utilised to gain deeper insights into pH-induced structural transitions during the assembly process. The results obtained by this multi-methodological approach allowed the associative phase separation process to be parameterised in terms of a set of characteristic pH values (~5.3, ~4.8, ~4.5, ~4.15, ~4, ~3.8, ~2.5) at which critical structural changes took place. Investigation of the absorbance profiles of complexed/coacervated systems as a function of time revealed that several transitions could occur at different time scales. Morphological changes in the assemblies and the subsequent formation of some flocculant substances during the late stage of process were clearly visualised using microscopy.