Development of a membraneless extraction module for the extraction of volatile compounds: application in the chromatographic analysis of vicinal diketones in beer.
A membraneless extraction module (MLEM) for the sample preparation of volatile compounds and its use for the chromatographic analysis of vicinal diketones in beer are reported. The extraction process is based on the same principles of gas diffusion (GD) and pervaporation (PV); however it does not use a membrane. This module has a lower chamber where the sample continuously flows, while volatile compounds evaporate to the headspace. Inside the module there is a suspended small reactor, where a small volume of a suitable acceptor solution is placed. This extraction module was tested in the determination of vicinal diketones (VDKs) in beer (CV=5%; LOD=4 microg L(-1)), showing applicability with real samples. Several parameters of the extraction process, such as temperature, sample flow and extraction time, were studied and optimized. This module proved to be a good tool for the sampling of volatile compounds, since the extraction is made without using a membrane avoiding all the robustness problems related with its use.
Development and validation of a high-performance liquid chromatography method for the determination of diacetyl in beer using 4-nitro-o-phenylenediamine as the derivatization reagent.
Diacetyl is a natural byproduct of fermentation and known to be an important flavor compound in many food products. Because of the potential undesirable effects of diacetyl on health safety and beer flavor, determination of its concentration in beer samples is essential and its analytical methods have attracted close attention recently. The aim of the present work is to develop and validate a novel high-performance liquid chromatography method for the quantification of diacetyl in beer based on the derivatization reaction of diacetyl with 4-nitro-o-phenylenediamine (NPDA). After the derivatization with NPDA in pH 3.0 at 45 °C for 20 min, diacetyl was separated on a kromasil C(18) column at room temperature in the form of the resulting 6-nitro-2,3-dimethylquinoxaline and detected by the ultraviolet detector at 257 nm. The results showed that the correlation coefficient for the method was 0.9992 in the range of 0.0050-10.0 mg L(-1) and the limit of detection was 0.0008 mg L(-1) at a signal-to-noise ratio of 3. The applicability of the proposed method was evaluated in the analysis of beer samples with the recovery range of 94.0-99.0% and relative standard deviation range of 1.20-3.10%. The concentration levels of diacetyl detected in beer samples from 12 brands ranged from 0.034 to 0.110 mg L(-1). The proposed method showed efficient chromatographic separation, excellent linearity, and good repeatability that can be applied to quantification of diacetyl in beer samples.
Intentional poisoning cases of animals with anticholinesterase pesticide-carbofuran in Sri Lanka.
Carbofuran is a broad spectrum insecticide and nematicide which inhibits acetyl cholinesterase. Several intentional poisoning cases of animals and birds including crows, dogs, cow, and elephant, using carbofuran were reported in Sri Lanka. Qualitative analysis of carbofuran in biological specimens was carried out using T.L.C and GC-MS. The quantitative analysis was carried out by HPLC using Zorbax Eclips XDB-C18 (150 x 4.6 mm I.D x 5 microm particle size) column with acetonitrile: water 25:75 v/v mobile phase and UV detection at 210 nm. The liquid-liquid extraction with chloroform was reproducible and sensitive. The procedure was validated in terms of linearity (0.996<r<0.999) for concentrations ranging from 1 to 200 microg/ml, repeatability (RSD 4.4% at 100 microg/ml, n=10) extraction recovery (91.7% at 100 microg/g, 77.3% at 20 microg/g), limit of detection (S/N ratio=3, 0.7 microg/ml) and limit of quantification (S/N ratio=10, 2.4 microg/ml). The method was found satisfactory and successfully applied in fatal cases of animals and birds due to carbofuran poisoning. The fatal carbofuran levels detected were in the range of 42-910 microg/g in crow's gizzard and contents, 50-800 microg/g in dog's stomach contents, 13-20 microg/g in dog's liver, 0.9 microg/g in cow's stomach and contents, 35 microg/g in elephant's stomach and contents.