Analytical method for fast screening and confirmation of multi-class veterinary drug residues in fish and shrimp by LC-MS/MS

A multi-class, multi-residue analytical method based on LC-MS/MS detection was developed for the screening and confirmation of 28 veterinary drug and metabolite residues in flatfish, shrimp and eel. The chosen veterinary drugs are prohibited or unauthorised compounds in Korea, which were categorised into various chemical classes including nitroimidazoles, benzimidazoles, sulfones, quinolones, macrolides, phenothiazines, pyrethroids and others. To achieve fast and simultaneous extraction of various analytes, a simple and generic liquid extraction procedure using EDTA-ammonium acetate buffer and acetonitrile, without further clean-up steps, was applied to sample preparation. The final extracts were analysed by ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). The method was validated for each compound in each matrix at three different concentrations (5, 10 and 20 ng g^–1) in accordance with Codex guidelines (CAC/GL 71-2009). For most compounds, the recoveries were in the range of 60–110%, and precision, expressed as the relative standard deviation (RSD), was in the range of 5–15%. The detection capabilities (CCβs) were below or equal to 5 ng g^–1, which indicates that the developed method is sufficient to detect illegal fishery products containing the target compounds above the residue limit (10 ng g^–1) of the new regulatory system (Positive List System – PLS).     

Application of Hyphenated Chromatography–Mass Spectrometry Techniques to Plant Allelopathy Research

Plant allelopathy offers hope as an additional means of weed control in modern agriculture. Its mechanisms and molecular basis are not yet well understood. Research on the chemical basis for allelopathy has often been hindered by the complexity of plant and soil matrices, making it difficult to track active compounds. Recent improvements in the cost and capabilities of bench-top chromatography–mass spectrometry instruments make these tools more powerful and more widely available to assist with molecular studies conducted in today's expanding field. Such instrumental techniques are herein recommended as economically efficient means of advancing the rigor of allelopathy research and assisting the development of a better understanding of the chemical basis for the allelopathy phenomenon.     

Identification of Human UDP-Glucuronosyltransferase 1A4 as the Major Isozyme Responsible for the Glucuronidation of 20(S)-Protopanaxadiol in Human Liver Microsomes

20(S)-protopanaxadiol (PPD), one of the representative aglycones of ginsenosides, has a broad spectrum of pharmacological activities. Although phase I metabolism has been investigated extensively, information regarding phase II metabolism of this compound remains to be elucidated. Here, a glucuronidated metabolite of PPD in human liver microsomes (HLMs) and rat liver microsomes (RLMs) was unambiguously identified as PPD-3-O-β-d-glucuronide by nuclear magnetic resonance spectroscopy and high resolution mass spectrometry. The chemical inhibition and recombinant human UDP-Glucuronosyltransferase (UGT) isoforms assay showed that the PPD glucuronidation was mainly catalyzed by UGT1A4 in HLM, whereas UGT1A3 showed weak catalytic activity. In conclusion, PPD-3-O-β-d-glucuronide was first identified as the principal glucuronidation metabolite of PPD in HLMs, which was catalyzed by UGT1A4.     

Chemical Profile and Biological Activities of Two Edible Plants: Chemical Investigation and Quantitative Analysis Using Liquid Chromatography Tandem Mass Spectrometry and Gas Chromatography Mass Spectrometry

The objectives of this study were to define the phenolic and fatty acid profiles, anticholinesterase, antioxidant, antimicrobial activities, and total phenolic-flavonoid contents of Lycopsis orientalis and Tragopogon latifolius var. angustifolius which have been used as food source and food supplement in Anatolia and have never been examined before. Rosmarinic and quinic acids (21.11 and 11.46 mg g^–1 extract, respectively) were found to be the most abundant constituents in L. orientalis and T. latifolius var. angustifolius among the studied 27 compounds by liquid chromatography tandem mass spectrometry. In the fatty acid compositions of L. orientalis and T. latifolius var. angustifolius that were determined by gas chromatography mass spectrometry, oleic (29.1%) and palmitic (28.7%) acids were identified as the major components, respectively. The high antioxidant activity of the methanol extract of L. orientalis shows parallelism to its rosmarinic acid content. Besides, this extract showed medium anticholinesterase activity. The results of the present study proves that the L. orientalis might also be used as a food source due to its high phenolic acid content and strong antioxidant property.     

Investigating the chlorination of acidic pharmaceuticals and by-product formation aided by an experimental design methodology

The degradation of seven acidic drugs and two metabolites during chlorination was investigated by liquid chromatography-mass spectrometry (LC-MS). A triple-quadrupole (QqQ) system was used to follow the time course of the pharmaceuticals and by-products, while a quadrupole time-of-flight (Q-TOF) system was also used for the identification of the by-products. Under strong chlorination conditions (10 mg/L Cl₂, 24 h), only four of the target compounds were significantly degraded: salicylic acid, naproxen, diclofenac and indomethacine. The degradation kinetics of these four compounds were investigated at different concentrations of chlorine, bromide and pH by means of a Box-Behnken experimental design. Depending on these factors, measured pseudo-first order half-lives were in the ranges: 23-573 h for salicylic acid, 13-446 min for naproxen, 5-328 min for diclofenac and 0.4-13.4 min for indomethacine. Also, it was observed that chlorine concentration was the overall most significant factor, followed by the bromide concentration (except for indomethacine), resulting in increased degradation kinetics as they are increased. The degradation path of salicylic acid, naproxen and diclofenac consisted of aromatic substitution of one or two hydrogens by chlorine and/or bromide. Moreover, for diclofenac, two other by-products corresponding to a decarboxylation/hydroxylation pathway from the monohalogenated products were also identified. On the other hand, indomethacine degradation did not lead to halogenation products but to oxidation ones. The investigation of these by-products in real samples by LC-MS/MS (QqQ) showed that the halogenated derivates of salicylic acid occurred in all the drinking water and wastewater samples analysed.     

Degradation Products of Tryptophan in Cell Culture Media: Contribution to Color and Toxicity

Biomanufacturing processes may be optimized by storing cell culture media at room temperature, but this is currently limited by their instability and change in color upon long-term storage. This study demonstrates that one of the critical contributing factors toward media browning is tryptophan. LC-MS technology was utilized to identify tryptophan degradation products, which are likely formed primarily from oxidation reactions. Several of the identified compounds were shown to contribute significantly to color in solutions but also to exhibit toxicity against CHO cells. A cell-culture-compatible antioxidant, a-ketoglutaric acid, was found to be an efficient cell culture media additive for stabilizing components against degradation, inhibiting the browning of media formulations, and decreasing ammonia production, thus providing a viable method for developing room-temperature stable cell culture media.     

An improved LC-MS/MS method for the quantitation of acrylamide in processed foods

An improved liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of acrylamide in processed foods. The homogenized samples, spiked with ¹³C₃-acrylamide as an internal standard, were extracted with water and centrifuged. D₅-3-chloropropanediol as a recovery standard was added to 1-ml aliquots, and the sample was purified with a C₁₈-cartridge column. The extract was directly analyzed using LC-MS/MS without derivatization. The ion transitions of 72–55 m/z (acrylamide), 75–58 m/z (¹³C₃-acrylamide), and 116–98 m/z (d₅-3-chloropropanediol) were found to be the most reliable for the identification and quantification of acrylamide in multiple reaction monitoring. The limit of quantification for acrylamide, defined as a signal-to-noise ratio of 10:1, was 2 μg/kg. The use of d₅-3-chloropropanediol minimized the effects of variation in the sample matrixes and increased the quality of analysis. This method could be applied to the quantification of acrylamide in processed foods.     

大米中环已烯酮类除草剂残留量的测定

建立了大米中环己烯酮类除草剂残留量同时测定的液相色谱-质谱/质谱法。试样中残留的环己烯酮类除草剂用酸性乙腈高速匀浆提取,提取液经N-丙基乙二胺（PSA）、十八烷基硅烷（ODS）和石墨化炭黑净化,用液相色谱-质谱／质谱仪检测和确证,外标法定量。环己烯酮类除草剂的浓度在0．0025～0．1000μg·mL^-1范围内时,线性关系良好,8种环己烯酮类除草剂的相关系数为0．9947～0．9992。在0．005～0．050mg·kg^-1浓度范围内,样品平均加标回收率在73．2％～107．0％之间,相对标准偏差为5．14％～10．15％。8种环己烯酮类除草剂的最低检出限均为0．005mg·kg^-1。     

Antioxidant activity and polyphenol content of Turkish thyme (Thymus vulgaris) monitored by liquid chromatography and tandem mass spectrometry

Like tea, the leaves of Turkish thyme (Thymus vulgaris) can be boiled in water to produce an extract. This is widely used as syrup for the treatment of coughs and bronchitis at alternative medicine clinics in many parts of the world. In the current study, we assessed the phenolic content and antioxidant activity of thyme. The antioxidant activities of both ethanol and aqueous extracts of thyme were determined using various in vitro methods. The total phenolic and total flavonoid contents were determined to be a gallic acid equivalent and a quercetin equivalent, respectively. Finally, the quantities of the phenolic compounds were detected using high-performance liquid chromatography and tandem mass spectrometry. The total phenolic compounds in the aqueous extract and ethanol extracts of Turkish thyme were 256.0 μg gallic acid equivalent/mg dried extract and 158.0 μg gallic acid equivalent/mg dried extract, respectively. Conversely, the total flavonoid compounds in both extracts were 44.2 μg and 36.6 μg quercetin equivalent/mg dried extract, respectively. For the first time, we determined phenolic contents and investigated the antioxidant potential of thyme. The results indicate that Turkish thyme is a good dietary source with phenolic properties.     

Determination of testosterone esters and nortestosterone esters in animal blood serum by LC-MS/MS

A sensitive and robust confirmatory method for determination of steroid esters in blood serum is essential for reliable monitoring of possible illegal use of steroid hormones as growth promoters in meat production. A previously used sample preparation methodology was improved. The procedure consists of protein precipitation and removal of phospholipids by dispersive SPE Supel? QuE Z-Sep (Sigma-Aldrich) followed by clean-up on alumina column and LC-MS/MS measurement. The modified method has been validated according to Commission Decision 2002/657/EC. Validation parameters for determination of six testosterone esters and five nortestosterone esters in bovine and porcine blood serum are presented in this article. Decision limits for all analytes were observed in the range 10–20 pg mL^?1. The method described is considerably robust for bovine and porcine serum analyses and can be applied both for screening and confirmatory determination in routine residue monitoring.