Liquid chromatography-electrospray ionization-tandem mass spectrometry method for the determination of epoxidized soybean oil in food products

Epoxidized soybean oil (ESBO) is widely used as a plasticizer and stabilizer in such polymers [poly(vinyl chloride) in particular] commonly adopted for manufacturing of gaskets of the lids for glass jars and plastic films for food packaging. Human exposure to ESBO and its derivatives is likely to occur over a lifetime with a significant variation according to life stage. A reversed phase liquid chromatography interfaced with electrospray ion trap tandem mass spectrometry method for the determination of ESBO in foods was developed. A simple sample treatment procedure entailing the use of an extraction step with dichloromethane without any further cleanup was proved. Chromatographic separation was performed using two C18 columns with an aqueous acetic acid-acetone-acetonitrile mixture as the mobile phase under gradient conditions. The method was validated in terms of detection limits (4 mg kg(-1)), quantitation limits, linearity (established over 2 orders of magnitude), recovery (good mean recoveries, higher than 90% for all of the signals detected), precision (RSD% < 8), and trueness. The applicability of the method to the determination of ESBO in different food matrices (in particular those rich in edible oil) was demonstrated, and the performances were compared to those reachable by the commonly well-known gas chromatography-mass spectrometry procedure.     

Micellar electrokinetic capillary electrophoresis for rapid analysis of patulin in apple cider

A micellar electrokinetic capillary chromatography (MECC) mode was applied to a capillary electrophoresis (CE) method, which was developed for detection and quantitation of patulin in apple ciders. This method used a small sample amount (2 mL) and consumed minimal organic solvent compared to the most commonly used HPLC methods. The sample preparation procedure of the CE method was also simpler than other chromatographic techniques developed for patulin analysis. Patulin was detected with a photodiode array detector at 273 nm. The standard curve was linear (r(2) = 0.9984) from 75 microgram/L to 121 microgram/mL with patulin working solutions corresponding to 3.8 microgram/L to 6.1 microgram/mL patulin in the sample. The linearity was better in a narrower range of concentrations (r(2) = 0.9999) from 75 microgram/L to 24.1 microgram/mL. The limit of detection of the method was 3.8 microgram/L. Patulin recoveries at 4 levels in spiked samples (10-121 microgram/L) ranged from 95.2 to 105.4%. The recoveries were 96. 9% and 99.2% for 2 levels (22.3 and 223 microgram/L, respectively) of patulin in infected apple samples. This method represents a unique alternative method for rapid and sensitive analysis of patulin in apple ciders.     

Ochratoxin a determination in beer by solid-phase microextraction coupled to liquid chromatography with fluorescence detection: a fast and sensitive method for assessment of noncompliance to legal limits

A solid-phase microextraction-liquid chromatography-fluorescence detection (SPME-LC-FD) method for the determination of ochratoxin A (OTA) in commercial beer samples was developed for the first time using a 60 microm thick poly(dimethylsiloxane)/divinylbenzene (PDMS/DVB) fiber. The procedure required a very simple sample pretreatment, an isocratic elution, and provides a selective extraction. All of the factors influencing fiber adsorption (extraction time, temperature, pH, and salt addition) and desorption of the analyte (desorption and injection time and desorption solvent mixture composition) have been investigated. The linear range investigated in beer was 0.03-2 ng/mL; within-day and between-days relative standard deviation in beer were 4.3 and 5.9%, respectively. The limit of quantification in spiked beer was 53 pg mL(-)(1), well below all European regulatory levels.     

Liquid chromatographic determination of morphine sulfate and some contaminants in injections and bulk drug material

A liquid chromatographic (LC) procedure is described for the assay of morphine sulfate in bulk drug material and injection solutions. The bulk drug and injection samples are prepared by direct dilution with LC mobile solvent. The average bulk drug purity (5 manufacturers) determined by the LC method was 99.9% with a difference of 0.1% from the average purity (anhydrous) found by the official USP XX procedure. The average LC recovery (19 studies) of morphine sulfate added to injection samples was 99.4% with a coefficient of variation (CV) of 1.14%. Morphine sulfate content was determined in triplicate for 53 injection samples (1-15 mg morphine sulfate/mL) formulated by 6 manufacturers, using the proposed LC procedure. Individual sample CV (n = 3) averaged 1.14%. The LC method is simple and specific for morphine sulfate. Major degradation products, preservatives, and some contaminants and related compounds are separated during LC.     

Use of anion-exchange membrane extraction for the high-performance liquid chromatographic analysis of mustard seed glucosinolates

A new one-step extraction using anion-exchange membranes for the HPLC determination of glucosinolates in mustard seeds is reported. The exchange of glucosinolates on the membranes was studied using sinigrin in solutions and sinigrin added as an internal standard to seeds of yellow mustard. By varying time of extraction, membrane size, and sample size, the optimal conditions for maximum glucosinolate recovery were determined and the following procedure was adopted: 0.2 g of ground mustard seeds are heated in 20 mL of boiling water for 5 min. After cooling, samples are transferred to plastic centrifuge tubes, 9-cm(2) membranes are added, and suspensions are shaken on a mechanical shaker for 2.5 h. Glucosinolates are then eluted from the membranes with 25 mL of 1 N KCl by shaking again for 2.5 h. Using this procedure, the sinigrin extraction from solutions and from mustard seeds was linear with 80% recovery. Seeds of yellow, brown, oriental, and Indian mustard were analyzed by this procedure; excellent reproducibility, with coefficients of variation in the range 1.0-4.3% were obtained. This method offers a simple and inexpensive alternative to complicated and tedious procedures for glucosinolate isolation/purification required for chromatographic determinations.     

Determination of reserpine and hydrochlorothiazide in commercial tablets by liquid chromatography with fluorescence and UV absorption detectors in series

A procedure is presented for the determination of reserpine and hydrochlorothiazide in commercial tablets by liquid chromatography (LC). Reference and sample solutions are prepared in methanol. For LC, a normal phase column is used, methanol is the eluting solvent, and 2 detectors are arranged in series. A fluorescence detector set at an excitation wavelength of 280 nm and emission wavelength of 360 nm quantitates reserpine, and a UV absorption detector set at 345 nm determines hydrochlorothiazide. Several synthetic mixtures containing the 2 ingredients in the amounts approximately present in commercial tablets were analyzed by the proposed method. Two samples of commercial tablets were also analyzed; for each sample, 5 determinations were made on a ground composite of 20 tablets; 10 individual tablets were also analyzed. For comparison, some of the solutions were analyzed for each ingredient by an alternative procedure.     

FTIR approaches for diuron determination in commercial pesticide formulations

Two strategies have been developed for Diuron determination by FTIR spectrometry, an off-line extraction and stopped-flow determination and a fully mechanized procedure, based on the on-line extraction of Diuron and FIA-FTIR measurement of the extracts. The aforementioned procedures have been compared with a reference chromatographic method. The off-line FTIR spectra were obtained at a nominal resolution of 4 cm(-1) from 4000 to 900 cm(-1) by accumulating 25 scans. Diuron was determined using peak height measurements at 1582 cm(-1) corrected using a baseline defined between 1562 and 1614 cm(-1). The waste generation of the off-line procedure was 3.4 mL chloroform for each sample, and the method provided a LOD of 40 microg g(-1), corresponding to 0.8% (w/w) Diuron in the original sample. The fully mechanized FIA method provided a LOD of 35 microg g(-1), which corresponds to 0.7% (w/w) in the solid sample and a maximum sampling frequency of the whole procedure of 30 h(-1), with a waste generation of 9.3 mL per sample, taking into account the volume of CHCl(3) required for sample dissolution and that need as a carrier. All those methods consume less organic solvent than a HPLC method, which involves the use of 39 mL of acetonitrile per sample and a sampling frequency of 12 h(-1).     

Speciation analysis of mercury in cereals by liquid chromatography chemical vapor generation inductively coupled plasma-mass spectrometry

A simple and rapid procedure for the separation and determination of inorganic, methyl, and ethyl mercury compounds was described using liquid chromatography (LC) followed by vapor generation inductively coupled plasma-mass spectrometry (VG-ICP-MS). Well resolved chromatograms were obtained within 5 min by reversed-phase liquid chromatography with a C8 column as the stationary phase and a pH 4.7 solution containing 0.5% v/v 2-mercaptoethanol and 5% v/v methanol as the mobile phase. The separated mercury compounds were converted to mercury vapors by an in situ nebulizer/vapor generation system for their introduction into ICP. The concentrations of NaBH4 and HNO3 required for vapor generation were also optimized. The method was applied for the speciation of mercury in reference materials NIST SRM 1568a Rice Flour and NIST SRM 1567a Wheat Flour and also rice flour and wheat flour samples purchased locally. The accuracy of the procedure was verified by analyzing the certified reference material NRCC DOLT-3 Dogfish Liver for methyl mercury. Precision between sample replicates was better than 13% for all the determinations. The detection limits of the mercury compounds studied were in the range 0.003-0.006 ng Hg mL(-1) in the injected solutions, which correspond to 0.02-0.06 ng g(-1) in original flour samples. A microwave-assisted extraction procedure was adopted for the extraction of mercury compounds from rice flour, wheat flour, and fish samples using a mobile phase solution.     

Metallspeziierung in Bodenlösungen mittels Dialysen- und Ionenaustauscherverfahren

Metal speciation in soil solution by dialysis-and ion exchange resin procedures A graduated separation procedure was applied for determining ionic and organic species of main and transition elements in soil solutions. Dialysis of the samples against pure water was found to be an appropriate method for analyzing weak complexes. Ionic forms could be differentiated from soluble strong organic complexes by varying electrolyte concentrations and cation exchange resin. The results of experiments which cation exchange resin in direct contact with the sample (column-mode and batch- mode) were compared with those from dialysis. Thus, three types of element forms could be distinguished: such forms, which could be characterized as inorganically bound ones (K, Na), such elements, which were only weakly bound to organic compounds in order to maintain electrical neutrality (Ca, Mg, Mn, Zn, Cd) and such elements, which were strongly complexed by organic molecules to a high extent (Cu, Cr, Pb, Fe, Al).     

Simplified automated procedure for measurement of certain catecholamine drugs delivered from aerosol inhalation units

An automated colorimetric method is described for the measurement of certain catecholamine drugs, such as isoproterenol and epinephrine, in sample solutions derived from 2 metered doses delivered from the mouthpiece of aerosol inhalation units. The procedure, applicable to the expressed product, involves oxidizing the catecholamine with potassium ferricyanide at pH 6 to produce an aminochrome. This method is similar to the well known trihydroxyindole reaction procedure but differs in that the formed aminochrome is measured spectrophotometrically at 495 nm instead of being further derivatized by alkaline rearrangement to the trihydroxyindole species followed by either spectrophotometric or fluorometric measurement.     

Evaluation and validation of a commercially available enzyme-linked immunosorbent assay for the neonicotinoid insecticide imidacloprid in agricultural samples

The performance of a commercially available microtiter plate ELISA kit for the determination of the neonicotinoid insecticide imidacloprid was evaluated for sensitivity, selectivity, influence of organic solvent used for extraction procedure, matrix interference originated from agricultural sample, accuracy, and method comparison with conventional HPLC analysis. The limit of detection for the kit (0.1 or 0.5 ng/mL) was determined. The working range (1-39 ng/mL) experimentally calculated on the basis of a criterion, which is determined as the range from I(20) to I(80), was comparable to that established by the manufacturer (1-50 ng/mL). The linearity of the standard curve based on the kit-assembled standard solutions agreed with the one based on the self-made standard solutions. Specificity studies indicate that the imidacloprid monoclonal antibody can readily distinguish the target compound from other structurally related neonicotinoid analogues and some metabolites, with the exception of clothianidin, the cross-reactivity of which was approximately 12%. To extract imidacloprid from an agricultural sample (apple) as simply and rapidly as possible, some extraction methods were examined. Consequently, the extraction method with hand-shaking for 5 min was the best among the examined methods. For the analysis of imidacloprid in apple samples, it was extracted directly with methanol and the extracts were diluted 10-fold (100-fold in the well) with water prior to ELISA analysis. No significant matrix interference was observed with the dilution factor. Recoveries of imidacloprid from fortified apple samples ranged from 87.7 to 112.0%. The results obtained with the ELISA kit correlated well with those by the reference method (conventional HPLC analysis) for apple samples (r > 0.998). These findings strongly indicate that the ELISA kit may be employed routinely for an on-site imidacloprid residue analysis of apple samples.     

Gas chromatographic-mass spectrometric determination of adipate-based polymeric plasticizers in foods

A method for the quantitative determination of adipate-based polymeric plasticizers in foods is described. The procedure involves extraction from the food and transmethylation of the polymeric plasticizer to form dimethyladipate (DMA). The derivative is cleaned up by size-exclusion chromatography and determined by capillary gas chromatography-mass spectrometry with selected ion monitoring. The use of a deuterated internal standard at the extraction stage enables quantitation by stable isotope dilution. A detection limit of 0.1 mg/kg of the polymeric plasticizer in foods and a relative standard deviation of 4% have been achieved routinely. The method has been applied successfully to the analysis of cheese, sandwiches, meat, biscuits, and cake that have been in contact with polymeric plasticized poly(vinyl chloride) films.     

Gas chromatographic determination with electron capture detection of residual ethylene oxide in intraocular lenses

A sensitive method is described to determine trace quantities of ethylene oxide (EO) in EO-sterilized intraocular lenses (IOLs). An IOL is dipped in ethanol containing 0.25 ppm propylene oxide (PO) in a 4 mL vial, 2 drops of freshly distilled hydrobromic acid is added through a septum, and the mixture is warmed at 50 degrees C for 24 h. It is then neutralized by vigorous shaking with sodium bicarbonate, dehydrated with anhydrous sodium sulfate, and filtered. The filtrate is injected into a gas chromatograph with electron-capture detection, and the peak height ratio of ethylene bromohydrin/propylene bromohydrin is measured. EO residue is calculated from the calibration curve obtained through a similar procedure with the standard EO/PO solutions. The limit of determination is 0.04 microgram/lens (ca 2.0 ppm). When EO residue levels were determined for IOLs sampled at 3 different aeration periods after sterilization, we found that 9 days of aeration was necessary to meet the U.S. Food and Drug Administration proposed limit for EO residue in IOLs.     

Determination of sulfite in food by flow injection analysis

A method is described for the determination of sulfite levels in food products by flow injection analysis (FIA). The method is based on the decolorization of malachite green by SO2, which is isolated from the flowing sample stream by means of a gas diffusion cell. The FIA method has a detection limit in food sample extracts of 0.1 ppm SO2 (3 times peak height of blank), which corresponds to 1-10 ppm SO2 in a food product, depending on the extraction procedure used. At the 5 ppm SO2 level in a food extract, the precision of replicate injections is +/- 1-2%. The method was tested on a variety of both sulfite-treated and untreated food products and the results compared favorably with those obtained by the Monier-Williams, colorimetric (pararosaniline), and enzymatic (sulfite oxidase) methods. The average differences from the FIA results were 19, 11, and 12%, respectively, for those samples (n = 12) above 50 ppm SO2. At lower levels the results were somewhat more erratic due to inaccuracies of the various methods at low concentrations.     

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.     

Analyzing capillary‐rise method settings for contact‐angle determination of granular media§

Water repellency is a relevant topic in soil‐science research due to its effects on vegetation growth, occurrence of surface runoff, infiltration, and erosion. Different methods have been adapted for the assessment of soil wettability by measuring the contact angle (CA), like the capillary‐rise method (CRM), where the liquid penetration dynamics into a dry sample is analyzed. However, questions related to sample preparation and the use of a suitable reference liquid in order to improve the reproducibility in such heterogeneous materials are still open. Different methods use ethanol as a reference liquid to quantify the degree of water repellency, like the molarity of ethanol droplets (MED), whereas other methods (CRM) suggest in addition n‐hexane as reference. To date, no generally accepted protocol has been invented to apply the CRM to soil particles. By using model porous materials with defined and stable levels of water repellency (silt, sand, and glass beads), CA results were compared for different initial settings of the sample. The main objective was to prove the CRM as a reliable and reproducible method to characterize soil wettability and to specify general guidelines for its application for granular materials in terms of sample size, sample‐packing procedure, and reference liquid. Results showed that a sample weight of ≈ 2 g has a relatively lower CA variation between replications. The packing procedure showed erratic results in CA, proving to be a critical factor in reproducibility. A uniform criterion of samples packing is recommended regarding application of CRM in soils. Regarding the reference liquid, n‐hexane should be preferred instead of ethanol for dynamic CA determination, because ethanol increased the tendency of CRM to overestimate angles due to dynamic effects, especially in finer‐textured materials (i.e., silt).     

Headspace gas chromatographic determination of residual 1,3-butadiene in rubber-modified plastics and its migration from plastic containers into selected foods

A headspace gas chromatographic procedure has been developed for the determination of 1,3-butadiene in rubber-modified plastics and in some foods. Polymer solutions or foods are equilibrated in sealed vials at 90 degrees C, and headspace samples are injected into a gas chromatograph. 1,3-Butadiene residues are measured using a flame ionization detector and are quantitated by the method of standard additions or an external calibration curve. Refrigerator tubs, vegetable oil bottles, chewing gum, and foods in contact with this type of packaging were analyzed. Limits of quantitation varied with the matrix, ranging from 2 ng/g (ppb) in chewing gum to 20 ng/g in polymers. 1,3-Butadiene was found in one polymer at 53 ng/g with an 8% coefficient of variation. The procedure yields "apparent" trace levels of 1,3-butadiene, and confirmation by a complementary technique is required.     

Determination of morantel-related residues in bovine milk by electron capture gas chromatography

A gas chromatographic assay was developed to determine major residues of morantel in bovine milk over a range that is suitable for monitoring residues of the drug. The method is based on hydrolysis of the N-methyl-tetrahydropyrimidine portion of morantel and its metabolites to N-methyl-1,3-propanediamine, and converting the diamine to an N,N-bis-(2-nitro-4-trifluoromethylphenyl) derivative. The addition of an internal standard, the N-desmethyl-N-ethyl homolog of pyrantel, to the milk sample circumvents any potential problem that could arise from variable reaction yields, and eliminates the true recovery as a factor affecting the accuracy and precision of the procedure. The concentrations of the derivatives are determined by pulsed electron capture gas chromatography over a linear dynamic range that is equivalent to 12.5-50 ppb morantel. The method was evaluated at the 0, 12.5, 25, and 50 ppb levels in fortified bovine milk, and in a withdrawal sample containing physiologically incurred morantel residues. Mean values of 14 +/- 1.7, 24 +/- 3.7, and 47 +/- 6.9 were found for the fortified samples, approximately 3 ppb for control milk, and 16 +/- 1.7 ppb for the withdrawal sample.     

Quantitative determination of plant opal content in soils, using a combined method of heavy liquid separation and alkali dissolution

The aim of this study was to improve the quantitative determination of the plant opal content (i.e. phytoliths) in soils. The proposed method is based on: (i) the separation of plant opal from the silt and sand fractions of the soil, using heavy liquid flotation (aqueous solution of ZnBr₂, density = 1.92 g cm^?3); (ii) the subsequent determination of alkali-soluble silicon by atomic absorption spectrometry. Extraction and analytical procedures were tested on a broad sample of temperate and tropical soils with very different phytolith contents. Our investigations lead to the following conclusions: (i) a selective dissolution of opal in alkaline solutions (e.g. hot 0.5 m NaOH as proposed by Jones, 1969) is inaccurate so that a sink-float method must be used before any dissolution procedure; (ii) to dissolve opal completely, a 0.5 M NaOH dissolution treatment at 150°C can be easily and successfully carried out in steel PTFE-lined pressure vessels; (iii) the reproducibility of the determination is satisfactory for a step-by-step procedure (mean coefficient of variation = 13.4%). The comparison of this new method of quantitative assessment of soil opal with two other methods (gravimetric and phytolith-counting methods), shows very highly significant correlations (P<0.001). Therefore, this procedure is a useful tool in studies connected with pedological and environmental history.     

Improved Method for Isolating and Quantitating α‐Amino Nitrogen as Nonprotein,True Protein,Salt‐Soluble Proteins,Zeins, and True Glutelins in Maize Endosperm

The conventional Landry‐Moureaux method for selective extraction of maize proteins was modified by reducing the contact time of meal with extractants and by removing 55% 2‐propanol as extractant. The new procedure, coupled with a method for quantitating protein at microgram level, was used for assessing the nitrogen distribution of four soluble protein fractions present in 100‐mg samples of endosperm originating from six maize inbreds and opaque‐2 versions. Proteins extracted with 55% 2‐propanol plus reductant were made up of α‐, β‐, γ‐, and δ‐zeins. Proteins extracted subsequently with salt plus reductant were minor and poor in lysine (1 mol%).They were associated with zeins. Comparison of present data with those available in the literature showed a close similarity for a given genotype between the percentage of total α‐amino nitrogen extracted by 2‐propanol plus reductant than by salt plus reductant under conditions of the modified procedure and that of total Kjeldhal nitrogen extracted by 2‐propanol with and without reductant, and by salt plus reductant, using the conventional procedure. A simplified protocol was described and tested for isolating and quantitating α‐amino nitrogen as nonprotein, true protein, salt‐soluble proteins, zeins, and true glutelins in any sample of maize endosperm.