One-step sample preparation technique for broad spectrum gas chromatographic/mass spectrometric determination of organic priority pollutants in water

A rapid and cost-effective sample preparation technique is described for the qualitative and quantitative determination of a broad spectrum of organic contaminants in water. The technique involves simultaneous-in-situ acetylation of phenols with acetic anhydride and one-step extraction of phenol acetates and basic and neutral organic compounds from water. The extract is concentrated and analyzed by gas chromatography/mass spectrometry. The recovery and precision data of selected base, neutral, and phenolic compounds are given. The advantages and disadvantages of the in-situ acetylation/extraction technique are discussed.     

Isolation of antioxidant compounds from orange juice by using countercurrent supercritical fluid extraction (CC-SFE).

Antioxidants from orange juice are isolated by the use of countercurrent supercritical fluid extraction (CC-SFE) and characterized by reversed-phase liquid chromatography (RPLC) coupled to mass spectrometry (MS) and diode-array detection (DAD). A pilot-scale SFE plant equipped with a packed column has been employed for countercurrent extraction and fractionation of raw orange juice with carbon dioxide. Several experiments have been performed in order to study the effect of the countercurrent conditions on the content of antioxidative compounds. In this study, the main variable that has been considered is the solvent-to-feed ratio (S/F) because it plays an essential role in the extraction efficiency. The values tested covered a wide range of sample and solvent (CO(2)) flow rates. In each experimental run, two different extracted fractions and the residual nonextracted juice were obtained and characterized. Different flavonoids have been identified in the fractions obtained after CC-SFE. The possibility of using this process for antioxidant compounds enrichment is discussed.     

Quantitative confirmation of dimetridazole and ipronidazole in swine feed by capillary gas chromatography/mass spectrometry with multiple ion detection

Extracts from 4 types of swine feed containing 0.11 ppm each of dimetridazole (DMZ) and ipronidazole (IPR) were analyzed by capillary gas chromatography/mass spectrometry (GC/MS) using multiple ion detection (MID) techniques. We demonstrate in this paper that the quantitative results obtained by capillary GC/MS with MID are comparable for both compounds to results obtained by liquid chromatography and have a lower coefficient of variation for DMZ. Moreover, consistency in the ion ratios (5 ions in DMZ and 6 ions in IPR) permits identification of these compounds by electron ionization MS.     

Optimization of multiple headspace solid-phase microextraction for the quantification of volatile compounds in dry fermented sausages

Multiple headspace solid-phase microextraction (HS-SPME) is a stepwise method that eliminates the influence of the matrix sample on the quantitative analysis of solid samples. The process was optimized for the analysis of volatile compounds in dry fermented sausages by gas chromatography and mass spectrometry. Different amounts of fermented sausages and different vial volumes were studied to obtain the theoretical exponential decay of the peak area of the four successive extractions in order to calculate the total area in the sausage. The highest number of volatile compounds analyzed by multiple HS-SPME in dry fermented sausages was obtained in a 10 mL headspace vial with 0.07 g of sample in the presence of water, 0.75 mg butylated hydroxytoluene, and 0.5 g sodium chloride. Finally, the method was characterized in terms of linearity and detection limits and applied to analyze the volatile compounds present in fermented sausages manufactured with either nitrate or nitrite.     

Quantitative determination of linoleic acid in infant formulas by gas chromatography

A method has been developed for the quantitative determination of linoleic acid in infant formulas by gas chromatography (GC). A known amount of triheptadecanoin was spiked into the sample. Total lipid was extracted from the product by an ethyl ether-petroleum ether-ethanol system in a Mojonnier flask. The sample was saponified by methanolic KOH after the solvents were evaporated. Methyl esters of the fatty acids were prepared by boron trifluoride (BF3) in methanol and analyzed by gas chromatography. A glass column packed with 10% SP-2340 (75% cyanopropyl silicone) was used to separate and identify the methyl linoleate and the methyl heptadecanoate. The quantity of methyl linoleate was calculated by comparing the integrated peak areas of these 2 fatty acid methyl esters. This method was satisfactory for both milk protein-based and soy protein-based matrixes. The results obtained by this method are comparable to those obtained by the AOAC spectrophotometric method 28.082-28.085.     

Gas chromatographic determination of N-nitrosamines in beverages following automatic solid-phase extraction

A semiautomatic method for the determination of seven N-nitrosamines in beverages by gas chromatography with nitrogen-phosphorus detection is proposed. Beverage samples are aspirated into a solid-phase extraction module for preconcentration and cleanup. The influence of the experimental conditions was examined by using various sorbents among which LiChrolut EN was found to provide quantitative elution and the highest preconcentration factors of all. The proposed method is sensitive, with limits of detection between 7 and 33 ng/kg, and precise, with relative standard deviations from 4.3% to 6.0%. The recoveries of N-nitrosamines from beverage samples spiked with 0.5 or 1 microg/kg concentrations of these compounds ranged from 95% to 102%. The method was successfully applied to the determination of residues of the studied N-nitrosamines in beverages including beer, wine, liquor, whisky, cognac, rum, vodka, grape juice, cider, tonic water, and soft drinks. The analytes were only detected in beer samples, positives being confirmed by gas chromatography coupled with impact ionization mass spectrometry.     

Truffle aroma analysis by headspace solid phase microextraction

An experimental design has been used to optimize the extraction of volatile compounds from summer truffle aroma (Tuber aestivum) by using headspace solid phase microextraction. The extracted compounds have been analyzed by gas chromatography with a flame ionization detector and by gas chromatography-mass spectrometry (GC-MS). In an attempt to develop an objective method to fully characterize truffle aroma, a fiber of medium polarity (for flavors) was used to avoid discrimination toward very nonpolar and polar volatile compounds. To optimize the extraction conditions, a response surface experimental design was applied considering three factors such as extraction temperature, equilibrium time, and extraction time. From the statistical analysis of the experimental design, it was possible to determine that the most important factor influencing the abundance of aroma compounds was the extraction temperature. Optimal extraction temperature was established at approximately 50 degrees C. By using GC-MS, it was possible to identify 37 compounds, most of them previously described as responsible for truffle aroma.     

Identification of impact odorants contributing to fresh mushroom off-flavor in wines: incidence of their reactivity with nitrogen compounds on the decrease of the olfactory defect

Analysis of wines from different grape varieties marked by sometimes intense aromatic nuances of fresh mushroom was performed by gas chromatography coupled with olfactometry. This analysis has led to the identification of several odoriferous zones, which were recalling a fresh mushroom odor. Two trace compounds responsible for these odoriferous zones, 1-nonen-3-one and 1-octen-3-one, have been identified and their content has been determined by using either a multidimensional gas chromatography technique coupled to olfactometry and mass spectrometry detection (in the case of 1-nonen-3-one) or the preparation of the derivative with O-2,3,4,5,6-pentafluorobenzylhydroxylamine hydrochloride in the presence of the deuterated form, as the internal standard (in the case of 1-octen-3-one), then gas chromatography coupled to mass spectrometry detection. The assays allowed the quantification of these compounds at concentration levels sometimes well above their detection and recognition olfactory threshold. We show that adding nitrogen compounds to the altered wines, such as an amino acid (glycine) or a tripeptide (glutathione), led to lower concentrations of 1-octen-3-one in wines and diminished smell of fresh mushrooms. The study of the reaction in a model medium, whose composition is close to wine, by liquid chromatography coupled to mass spectrometry demonstrated the formation of adducts between 1-octen-3-one and glycine, and 1-octen-3-one and glutathione characterized by NMR.     

Optimization of microwave-assisted extraction for the characterization of olive leaf phenolic compounds by using HPLC-ESI-TOF-MS/IT-MS(2)

In the present work, a simple and rapid method for the extraction of phenolic compounds from olive leaves, using microwave-assisted extraction (MAE) technique, has been developed. The experimental variables that affect the MAE process, such as the solvent type and composition, microwave temperature, and extraction time, were optimized using a univariate method. The obtained extracts were analyzed by using high-performance liquid chromatography (HPLC) coupled to electrospray time-of-flight mass spectrometry (ESI-TOF-MS) and electrospray ion trap tandem mass spectrometry (ESI-IT-MS(2)) to prove the MAE extraction efficiency. The optimal MAE conditions were methanol:water (80:20, v/v) as extracting solvent, at a temperature equal to 80 °C for 6 min. Under these conditions, several phenolic compounds could be characterized by HPLC-ESI-MS/MS(2). As compared to the conventional method, MAE can be used as an alternative extraction method for the characterization of phenolic compounds from olive leaves due to its efficiency and speed.     

A rapid method for the quantitative determination of short-chain free volatile fatty acids from cheese

The determination of free volatile fatty acids (FVFA) is of interest in the analysis of cheeses. As these compounds are components of taste and flavor, they give indications on metabolic reactions taking place during cheese ripening and can provide an evaluation of cheese defects and their causes. One of the most widely used methods for the determination of FVFA in cheese involves preliminary recovery from the matrix by steam distillation, followed by gas chromatography separation. Relatively high distillate volumes must be collected to achieve a quantitative yield of all the compounds of interest, so that, as a result, the solution is too diluted to achieve good instrumental sensitivity. In this paper, an alternative method for the determination of C2-C6 free carboxylic acids in cheeses involving the use of a Nukol capillary column and crotonic acid as internal standard is described. This method is quick and cheap, as the sample preparation is a simple extraction with water. The underivatized FVFA are then directly separated by gas chromatography. Using this method, all FVFA in cheeses can be quantified with good repeatability and excellent recovery.     

Method for the quantitative extraction of resveratrol and piceid isomers in grape berry skins. Effect of powdery mildew on the stilbene content

A simple method for the quantitative extraction of resveratrol and its glycosides from grape berry skins has been developed. Optimal conditions for the extraction were investigated. Type of solvent, time, and temperature assayed influenced resveratrol and piceid yield. Adequate extraction was attained with ethanol/water (80:20 v/v) maintained at 60 degrees C for 30 min. Recovery (>96%) and reproducibility (6.83-15.13%) were satisfactory. After extraction, resveratrol and piceid isomers were quantified by high-performance liquid chromatography coupled to a ultraviolet-visible diode array detector. The amounts detected in 13 samples of 7 different varieties analyzed were, on average, 92.33 microg/g of dry skin for cis-piceid, 42.19 microg/g of dry skin for trans-piceid, and 24.06 microg/g of dry skin for trans-resveratrol. cis-Resveratrol was not detected in any sample. In grape berries infected by powdery mildew the contentw of these compounds were considerably increased and the degree of infection was positively related to their stilbene content.     

Preparative HPLC method for the purification of sulforaphane and sulforaphane nitrile from Brassica oleracea

An extraction and preparative HPLC method has been devised to simultaneously purify sulforaphane and sulforaphane nitrile from the seed of Brassica oleracea var. italica cv. Brigadier. The seed was defatted with hexane, dried, and hydrolyzed in deionized water (1:9) for 8 h. The hydrolyzed seed meal was salted and extracted with methylene chloride. The dried residue was redissolved in a 5% acetonitrile solution and washed with excess hexane to remove nonpolar contaminants. The aqueous phase was filtered through a 0.22-microm cellulose filter and separated by HPLC using a Waters Prep Nova-Pak HR C-18 reverse-phase column. Refractive index was used to detect sulforaphane nitrile, and absorbance at 254 nm was used to detect sulforaphane. Peak identification was confirmed using gas chromatography and electron-impact mass spectrometry. Each kilogram of extracted seed yielded approximately 4.8 g of sulforaphane and 3.8 g of sulforaphane nitrile. Standard curves were developed using the purified compounds to allow quantification of sulforaphane and sulforaphane nitrile in broccoli tissue using a rapid GC method. The methodology was used to compare sulforaphane and sulforaphane nitrile content of autolyzed samples of several broccoli varieties.     

Determination of tri-n-butyltin and di-n-butyltin compounds in fish by gas chromatography with flame photometric detection

An analytical method is described for the simultaneous quantitative determination of tri-n-butyltin and di-n-butyltin compounds in fish. The sample was extracted with 0.5N HCl-methanol, and the methanol solution was extracted with hexane. The extract was purified by gel permeation chromatography and treated with Grignard reagent to yield the methyl derivatives, which were determined by gas chromatography with flame photometric detection operated in the tin mode (610 nm). Recoveries of tri-n-butyltin chloride (Bu3SnCl) and di-n-butyltin dichloride (Bu2SnCl2) spiked to fish at the levels of 0.2 and 1.0 ppm ranged from 80 to 105%. Detection limits were 0.02 micrograms/g for both compounds. Tri-n-butyltin compounds equivalent to Bu3SnCl levels of 0.07-2.0 ppm and di-n-butyltin compounds equivalent to Bu2SnCl2 levels of 0.02-0.11 ppm were found in reared yellowtails, and these values showed good agreement with the results from gas chromatographic-mass spectrometric analysis.     

Determination of daminozide in apples by gas chromatography/chemical ionization mass spectrometry

A method using gas chromatography/chemical ionization mass spectrometry (GC/CIMS) for the determination of daminozide residues in apples has been developed. Daminozide was separated from the sample matrix by water extraction and cation exchange, converted to the methyl ester by treatment with HCl-methanol, and determined by GC/CIMS using succinonitrile as an internal standard. The detection level was 0.05 ppm. Recoveries were 92-104% from apples spiked at the 0.05-0.5 ppm levels. Of the 25 apple samples analyzed, only 2 were positive for daminozide (1.04 and 0.32 ppm).     

Gas chromatographic profile analysis of basic nitrogen-containing aromatic compounds (azaarenes) in high protein foods

A method is described for the determination of basic nitrogen-containing polycyclic aromatic compounds (N-PACs, azaarenes) in meat. The enrichment procedure includes liquid-liquid partition (dimethylformamide-water-cyclohexane), extraction of N-PACs by sulfuric acid, reextraction after neutralization by cyclohexane or, alternatively, by nonadsorbing ion exchange chromatography. Further purification is performed by column chromatography on Sephadex LH 20 using a closed system to avoid sample contamination by laboratory pollutants. N-PACs are analyzed by capillary gas chromatography and measured by comparing to the corresponding peak areas of an internal standard (e.g., 10-azabenzo(a)pyrene). The limit of detection of this method ranges from 0.1 to 0.4 ng for benzacridines, dibenzacridines, and their methyl derivatives. The results of a collaborative study, stimulated by IUPAC, are reported: Coefficients of variation for the various azaarenes were 4.0-13.6% for the check analysis and 10.4-25.4% for a spiked ham sample. Consequently, IUPAC suggests this procedure as a recommended method.     

Development of an enzyme-linked immunosorbent assay for quantitative determination of quizalofop-p-ethyl

Accurate quantification of quizalofop-p-ethyl is essential for it may do harm to humans and animals through both water and food. Currently, detection of quizalofop-p-ethyl mainly relies on methods such as gas chromatography, high performance liquid chromatography, and gas chromatography-mass spectrometry. Although these techniques are reliable, they are relatively expensive and time-consuming because of multistep sample cleanup. To address this, we developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) with a polyclonal antibody against quizalofop-p-ethyl that was generated in our lab. The IC(50) of detection was 0.03495 microg/mL, and the lowest detection limit reached 0.00192 microg/mL. Furthermore, the method had high specificity for it did not cross-react with other structure-related compounds. When water and soil samples that were fortified with quizalofop-p-ethyl were analyzed by this ELISA, recoveries were in the range of 89-110% from water and 81-108% from soil. Good correlations between this immunoassay and gas chromatography data were obtained for residues of quizalofop-p-ethyl in water and soil. Our data indicate that this method is a convenient analytical technique for monitoring quizalofop-p-ethyl in waters without extraction and the extra cleanup step and in soil without the cleanup step.     

Comparison of methods for the exhaustive extraction of hypericins, flavonoids, and hyperforin from Hypericum perforatum L

Renewed interest in plant-derived drugs has led to an increased need for efficient extraction methods. Hypericum perforatum L. contains several groups of bioactive compounds with noteworthy pharmacological activities. Direct sonication of H. perforatum was investigated and compared with conventional maceration, indirect sonication, Soxhlet extraction, and accelerated solvent extraction (ASE). Highly selective liquid chromatography/tandem mass spectrometry analysis showed that the content of six investigated active compounds (hypericin, pseudohypericin, hyperoside, rutin, quercitrin, and hyperforin) in extracts obtained by direct sonication was significantly higher than in extracts obtained by the other methods. The active compound contents increased on increasing the ultrasonic power from 40 to 60 W when using direct sonication. Conventional maceration gave the lowest amount of analyzed active compounds. Soxhlet extraction gave better results than ASE or indirect sonication.     

Residue methodology for AMDRO fire ant insecticide (AC 217,300) in pasture grass and crops

Residue methodology is described for the determination of AC 217,300 residues in pasture grass and crop samples. After extraction and subsequent cleanup on an XAD-2 column, residues of AC 217,300 are determined by liquid chromatography (LC), using a reverse phase paired-ion chromatographic system and detection at 300 nm. The method has a validated limit of sensitivity of 0.05 ppm with corresponding control values for the commodities analyzed of less than 0.01 ppm. Apparent residues over 0.05 ppm can be confirmed by either gas chromatography with an electron capture detector (GC-EC) or gas chromatography-negative ion chemical ionization mass spectrometry (GC-NICI). The direct GC-NICI method circumvents the need for sample cleanup on the XAD-2 column, and offers a greatly simplified procedure that is useful for screening samples. Recoveries of AC 217,300 from the commodities analyzed have been satisfactory with all methods of analysis.     

Corn Gluten Meal Odorants and Volatiles After Treatment to Improve Flavor

Production of corn gluten meal (CGM), a high‐protein coproduct from wet milling of corn, is increasing as production of fuel ethanol from corn increases. Unpleasant taste and odor have limited the use of CGM in human food. Adjustment of pH and extraction with water have been reported to reduce the off‐flavor of CGM but the improvement is not enough for substantial addition of CGM to the human diet. More study of CGM is needed. In this study, volatile compounds released under different conditions of pH, water extraction, and temperature were identified and compared using solid‐phase microextraction‐gas chromatography‐mass spectrometry (SPME‐GC‐MS). The water‐extractable portion, which improves the taste of CGM by its absence, was dried and analyzed by SPME‐GC‐MS. In addition, materials extractable from CGM with methylene chloride were identified by gas chromatography‐mass spectrometry (GC‐MS). Further, the spontaneous generation of a CGM‐like odor accompanied by a change in physical appearance of the CGM sample was described. Flavors and odors known to be associated with the identified CGM compounds were listed. Some possible origins of the volatiles, from degradation of corn constituents or as fermentation products of the corn steeping process, were noted.     

Rapid and sensitive determination of 4-nitrophenol, 3-methyl-4-nitrophenol, 4,6-dinitro-o-cresol, parathion-methyl, fenitrothion, and parathion-ethyl by liquid chromatography with electrochemical detection

Liquid chromatography with electrochemical detection has been used to determine various nitropesticides, DNOC, fenitrothion, and parathion (methyl and ethyl), and some of their main metabolites, 4-nitrophenol for parathion (methyl and ethyl) and 3-methyl-4-nitrophenol for fenitrothion, by using indirect detection. Analysis of them in river water samples has been performed without a preconcentration step. The recovery efficiencies of the tested compounds yielded values between 96 and 112% at the fortification level of 0.5 ppb in a river water sample, and their relative standard deviations were between 1 and 15%. The detection limits of these compounds ranged between 0.05 and 0.14 ppb.