Rapid

Dichloroacetic acid (DCA) is a compound found in chlorinated drinking water. In addition, the compound is a metabolite of several halogenated solvents, including trichloroethylene (TCE) and perchloroethylene (PCE). Exposure to DCA is of concern because high doses of the compound have been shown to cause cancer in laboratory animals. Dosages of TCE administered to animals in cancer studies are designed to elicit maximal DCA formation in vivo, whereas levels of DCA to which individuals are exposed in drinking water are very low. Analysis of DCA in biological samples has been quite challenging. Derivatizing reagents commonly used to convert DCA into a more volatile form for analysis by gas chromatography (GC) have been found to convert trichloroacetic acid (TCA), a major metabolite of TCE and PCE, into DCA. High-performance liquid chromatography (HPLC) analysis does not require derivatization of DCA and can thus avoid this problem. However, the most popular stationary phases in HPLC columns do not retain small, polar compounds such as DCA well. The liquid chromatography/tandem mass spectrometry (LC/MS/MS) method described in this paper uses hydrophilic interaction liquid chromatography (HILIC), a type of chromatography that is able to retain these small, polar compounds. Method validation was performed using the United States Food and Drug Administration (USFDA) and International Conference on Harmonziation (ICH) Guidance for Industry: Bioanalytical Method Validation as a guide. Levels of DCA found in rats dosed with 2 g/kg TCE were 17.2 ng/mL (liver), 262.4 ng/mL (kidney), 175.1 ng/mL (lung), and 39.5 ng/mL (blood). Copyright © 2005 John Wiley & Sons, Ltd.

Following previous studies on counterfeit of wines with synthetic ingredients, the possibility of frauds by natural external L-tartaric acid has also been investigated. The aim of this research was to map the stable isotope ratios of L-tartaric acid coming from botanical species containing large amounts of this compound: grape and tamarind. Samples of L-tartaric acid were extracted from the pulp of tamarind fruits originating from several countries and from grape must. δ13C and δ18O were measured for all samples. Additional δ2H measurements were performed as a complementary analysis to help discrimination of the botanical origin. Different isotopic patterns were observed for the different botanical origins. The multivariate statistical analysis of the data shows clear discrimination among the different botanical and synthetic sources. This approach could be a complementary tool for the control of L-tartaric acid used in oenology. Copyright © 2007 John Wiley & Sons, Ltd.

We report on the development and validation under ISO 17025 criteria of a multi-residue confirmatory method to identify and quantify 17 widely chemically different pesticides (insecticides: Carbofuran, Methiocarb, Pirimicarb, Dimethoate, Fipronil, Imidacloprid; herbicides: Amidosulfuron, Rimsulfuron, Atrazine, Simazine, Chloroturon, Linuron, Isoxaflutole, Metosulam; fungicides: Diethofencarb) and 2 metabolites (Methiocarb sulfoxide and 2-Hydroxytertbutylazine) in honey. This method is based on an on-column liquid–liquid extraction (OCLLE) using diatomaceous earth as inert solid support and liquid chromatography (LC) coupled to mass spectrometry (MS) operating in tandem mode (MS/MS). Method specificity is ensured by checking retention time and theoretical ratio between two transitions from a single precursor ion. Linearity is demonstrated all along the range of concentration that was investigated, from 0.1 to 20 ng g−1 raw honey, with correlation coefficients ranging from 0.921 to 0.999, depending on chemicals. Recovery rates obtained on home-made quality control samples are between 71 and 90%, well above the range defined by the EC/657/2002 document, but in the range we had fixed to ensure proper quantification, as levels found in real samples could not be corrected for recovery rates. Reproducibility is found to be between 8 and 27%. Calculated CCα and CCβ (0.0002–0.943 ng g−1 for CCα, and 0.0002–1.232 ng g−1 for CCβ) show the good sensitivity attained by this multi-residue analytical method. The robustness of the method has been tested in analyzing more than 100 raw honey samples collected from different areas in Belgium, as well as some wax and bee samples, with a slightly adapted procedure.

A method for the determination of several household biocides in water by liquid chromatography/electrospray ionisation tandem mass spectrometry (LC/ESI-MS/MS) is presented. It permits the simultaneous determination of triclosan (TCS), triclocarban (TCC) and seven parabens, including the distinction between branched and linear isomers of propyl (i-PrP and n-PrP) and butyl parabens (i-BuP and n-BuP). Prior to LC/MS/MS, analytes are preconcentrated by solid-phase extraction (SPE) on Oasis HLB (60 mg) cartridges at natural sample pH and subsequently eluted with 4 mL of methanol. This simple SPE procedure provides extraction recoveries above 85% except for raw wastewater, where it falls to 65% for TCC. The performance of the method was tested with two triple-quadrupole LC/MS instruments from a low/mid and mid/high market range: a Varian 1200L and an API-4000. The latter system provided between 3 and 80 times lower limits of quantification (LOQs) than the first one, in the 0.08–0.44 ng/L range for surface water. Moreover, a comparison of matrix effects on both instruments showed a very different behaviour, particularly in the case of parabens. For these compounds signal suppression was observed in the 1200L instrument and signal enhancement with the 4000 instrument. As a result, different calibration approaches were chosen for them and this pointed to the need of matrix effect re-evaluation in method transfer between different LC/MS systems. The application of the method to real samples showed the ubiquity of methyl paraben (MeP) and n-PrP (at the 1–6 µg/L in raw wastewater) and the coexistence of i-BuP and n-BuP at similar levels (ca. 100–200 ng/L in raw wastewater). Copyright © 2009 John Wiley & Sons, Ltd.

The production of recombinant therapeutic proteins is one of the fastest growing sectors of the pharmaceutical industry, particularly monoclonal antibodies and Fc-fusion proteins. Currently, mammalian cells are the dominant production system for these proteins because they can perform complex post-translational modifications that are often required for efficient secretion, drug efficacy, and stability. These protein modifications include misfolding and aggregation, oxidation of methionine, deamidation of asparagine and glutamine, variable glycosylation, and proteolysis. Such modifications not only pose challenges for accurate and consistent bioprocessing, but also may have consequences for the patient in that incorrect modifications and aggregation can lead to an immune response to the therapeutic protein. This mini-review describes examples analytical and preventative advances in the fields of protein oxidation, deamidation, misfolding and aggregation (glycosylation is covered in other articles in this issue). The feasibility of partially replacing traditional analytical methods such as peptide mapping with high-throughput screens and their use in clone and media selection are evaluated. This review also discusses how further technical advances could improve the manufacturability, potency, and safety of biotherapeutics.

The determination of the positions of double bonds after dimethyl disulfide (DMDS) addition is presented for linear or branched di-unsaturated esters and a di-substituted γ lactone. Different DMDS derivatives are studied according to the number, n, of methylene groups separating the double bonds. For n=0, a mono derivative is obtained following treatment with DMDS; for n = 1, 2 or 3, a cyclic thioether is synthesized. For n=0, only one double bond is located after a classical cleavage. For n = 1, 2 and 3, a systematic fragmentation pathway is observed allowing the determination of the positions of the double bonds and the molecular structures of the compounds.

ABSTRACTThis study aimed to compare Petrifilm Aerobic Count (AC) plates and the conventional pour plate methodology using the de Man–Rogosa–Sharpe (MRS) agar for the enumeration of lactic acid bacteria (LAB) in fermented milks (FMs), with different starter cultures added. FM samples (n = 66) were collected and plated on both methodologies, with incubation under anaerobic conditions at 35C for 48 h. The count results were compared by analysis of variance (P ≤ 0.05) and regression analysis. No differences between the mean counts obtained by both methodologies were observed, even when distinct FMs were compared. Considering all samples, a high correlation level was obtained between Petrifilm AC and MRS agar (r = 0.92), but these indexes were lower in FMs with Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (r = 0.90) and Lactobacillus fortis (r = 0.81). Despite some slight interferences, Petrifilm AC has proven to be a convenient methodology on enumerating LAB in FM.This study aimed to compare Petrifilm Aerobic Count (AC) plates and the conventional pour plate methodology using the de Man–Rogosa–Sharpe (MRS) agar for the enumeration of lactic acid bacteria (LAB) in fermented milks (FMs), with different starter cultures added. FM samples (n = 66) were collected and plated on both methodologies, with incubation under anaerobic conditions at 35C for 48 h. The count results were compared by analysis of variance (P ≤ 0.05) and regression analysis. No differences between the mean counts obtained by both methodologies were observed, even when distinct FMs were compared. Considering all samples, a high correlation level was obtained between Petrifilm AC and MRS agar (r = 0.92), but these indexes were lower in FMs with Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (r = 0.90) and Lactobacillus fortis (r = 0.81). Despite some slight interferences, Petrifilm AC has proven to be a convenient methodology on enumerating LAB in FM.PRACTICAL APPLICATIONSThis study focused on the evaluation of an important tool for monitoring the microbiologic quality in the food industry. Petrifilm plates are widely used in the food industry, and their application for several microorganisms groups and foods must be properly evaluated for convenient application. Petrifilm Aerobic Count (AC) plates, associated to the de Man–Rogosa–Sharpe broth, were already described as adequate for the lactic acid bacteria (LAB) enumeration in foods. However, A previous study was conducted only with experimental inoculation of LAB in sterile milk and naturally occurring LAB in raw milk (Nero et al. 2006). This article is a complement of the first one, and we show the convenience of Petrifilm AC plates on LAB enumeration in fermented milks, considering different types of LAB starter cultures on the performance of this methodology.This study focused on the evaluation of an important tool for monitoring the microbiologic quality in the food industry. Petrifilm plates are widely used in the food industry, and their application for several microorganisms groups and foods must be properly evaluated for convenient application. Petrifilm Aerobic Count (AC) plates, associated to the de Man–Rogosa–Sharpe broth, were already described as adequate for the lactic acid bacteria (LAB) enumeration in foods. However, A previous study was conducted only with experimental inoculation of LAB in sterile milk and naturally occurring LAB in raw milk (Nero et al. 2006). This article is a complement of the first one, and we show the convenience of Petrifilm AC plates on LAB enumeration in fermented milks, considering different types of LAB starter cultures on the performance of this methodology.

All four regioisomers of feruloyl quinic acid and isoferuloyl quinic acid were synthesized and a liquid chromatography/tandem mass spectrometry (LC/MS/MS) method developed that resolves all eight regioisomers. All eight regioisomers can be readily distinguished by their MS/MS spectra in the negative ion mode, illustrating the power of tandem mass spectrometry to elucidate the structures of regioisomeric compounds. Compound identification is possible, either by direct comparison of spectral fingerprints or by rational probing of diagnostic fragment ions, thus allowing the identification of these important classes of natural products and potential human metabolites. Copyright © 2010 John Wiley & Sons, Ltd.