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Development of a UHPLC-MS/MS method for the determination of quercetin in milk and its application to a pharmacokinetic study
2019
Gbylik-Sikorska, Małgorzata | Gajda, Anna | Burmańczuk, Artur | Grabowski, Tomasz | Posyniak, Andrzej
Introduction: Quercetin is a polyphenolic flavonoid which has been used in traditional Chinese medicine as a natural therapeutic agent with a broad spectrum of activities (antioxidant, anticancer, neuroprotective, anti-inflammatory, antiviral and antibacterial). The aim of this study was to develop and validate a rapid and simple ultra-high-performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) method for the determination of quercetin in milk. Material and Methods: Sample preparation was based on a liquid-liquid extraction with 0.5% formic acid in acetonitrile. The chromatographic separation was performed on a ZORBAX SB-C18 column with methanol and 0.5% formic acid as a mobile phase. Results: The procedure was successfully validated. The mean recovery of the analyte was 98%, with the corresponding intra- and inter-day variation less than 10% and 15%, respectively, and the repeatability and reproducibility were in the range of 3%–7.2% and 6.1%–12%, respectively. The lowest level of quantification was 1.0 μg/kg. Conclusion: The proposed method was successfully applied in evaluating the pharmacokinetics of quercetin in milk obtained from dairy cows with clinical mastitis after intramammary administration.
Afficher plus [+] Moins [-]Electrochemical reduction of azo dyes mimicking their biotransformation to more toxic products
2019
Pietruk, Konrad | Piątkowska, Marta | Olejnik, Małgorzata
Some azo dyes, including Sudans I–IV and Para Red, are genotoxic and may be biotransformed to cancerogenic aromatic amines. They are banned as food and feed additives, but their presence has been detected in food. Aromatic amines are also considered potentially toxic. Online EC–MS is a promising tool to study the transformation mechanisms of xenobiotics such as azo dyes. The aim of the study was to investigate emulation of how azo dyes are enzymatically transformed to amines with EC–MS. The reduction reactions of five azo dyes (Sudans I–IV and Para Red) were conducted using a glassy carbon working electrode and 0.1% formic acid in acetonitrile. Reduction results were compared with the literature and in silico to select preliminary candidates for metabolites. The LC-MS/MS method was used to confirm results obtained by electrochemical reactor. A limited number of pre-selected compounds were confirmed as azo dyes metabolites – aniline for Sudan I, aniline and 4-aminoazobenzene for Sudan III, o-toluidine for Sudan IV, and 4-nitroaniline for Para Red. No metabolites were found for Sudan II. Electrochemistry–mass spectrometry was successfully applied to azo dyes. This approach may be used to mimic the metabolism of azo dyes, and therefore predict products of biotransformation.
Afficher plus [+] Moins [-]Histologic assessment of age-related changes in the temporomandibular joints of horses
2019
Smyth, Travis T. | Allen, Andrew L. | Carmalt, James L.
OBJECTIVE To describe histologic changes in the temporomandibular joints (TMJs) of horses of various ages. SAMPLE 22 TMJs from cadavers of 11 horses. PROCEDURES Horses were categorized into 3 age groups (group 1, 2 to 10 years old [n = 3]; group 2, 11 to 20 years old [3]; and group 3, > 20 years old [5]). Each TMJ was sectioned into 5-mm slices, preserved in formalin, decalcified in formic acid, and routinely processed for histologic analysis. Joints were systematically assessed by use of previously described methods. Multilevel mixed-effects models were used to examine the data. RESULTS The number of changes was significantly fewer and degree of changes was significantly less within the TMJs of group 1 horses, compared with those of group 3 horses. Comparison among groups revealed that the combination of temporal and mandibular scores for group 1 was significantly lower than for groups 2 or 3. Disk score did not differ significantly between groups 1 and 2, but disk scores of groups 1 and 2 were significantly lower than the disk score of group 3. CONCLUSIONS AND CLINICAL RELEVANCE The assessed lesions were associated with osteoarthritis, and they accumulated in the TMJs as horses aged. In the absence of signs of pain manifested as changes in mastication, behavior, or performance, it would be difficult to determine the point at which accrued pathological changes represented the onset of clinically important osteoarthritis of the TMJs.
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