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Willemijn J Comuth, Linda Ø Henriksen, Daan van de Kerkhof, Steen E Husted, Steen D Kristensen, Moniek P M de Maat, Anna-Marie B Münster
BACKGROUND: Issues with laboratory measurement of dabigatran include: 1. Do coagulation assays reflect dabigatran plasma concentrations? 2. Do samples from patients treated with dabigatran have the same coagulability as dabigatran-spiked samples from healthy volunteers? 3. What is the long-term stability of dabigatran after storage at -80 °C? This study aims to evaluate these questions. MATERIALS AND METHODS: Ecarin chromogenic assay (ECA), a laboratory-developed diluted thrombin time (LD-dTT), prothrombin time (PT) and activated partial thromboplastin time (APTT) and ROTEM® were used to measure dabigatran anticoagulant activity and liquid chromatography-tandem mass spectrometry (LC-MS/MS) to measure dabigatran plasma concentrations...
February 17, 2018: Thrombosis Research
Berit P Jensen, Rajneeta Saraf, Jing Ma, Sarah Berry, Cameron C Grant, Carlos A Camargo, Christiaan W Sies
BACKGROUND: Demand for measurement of 25-hydroxyvitamin D (25OHD) is growing and dried blood spot (DBS) sampling is attractive as samples are easier to collect, transport and store. METHODS: A 2D LC-MS/MS assay without derivatization was developed. DBS punches (3.2 mm) were ultrasonicated with d6 -25OHD3 in 70% methanol followed by hexane extraction, dry-down and reconstitution. The assay was validated and applied to two studies comparing whole blood adult DBS with serum samples (n = 40) and neonatal whole blood DBS with cord serum samples (n = 80)...
February 20, 2018: Clinica Chimica Acta; International Journal of Clinical Chemistry
Huaidong Zhang, Qin Li, Lina Wang, Yan Chen
BACKGROUND: Alcohol dehydrogenases (ADHs) catalyze the reversible oxidation of alcohol using NAD+ or NADP+ as cofactor. Three ADH homologues have been identified in Komagataella phaffii GS115 (also named Pichia pastoris GS115), ADH1, ADH2 and ADH3, among which adh3 is the only gene responsible for consumption of ethanol in Komagataella phaffii GS115. However, the relationship between structure and function of mitochondrial alcohol dehydrogenase isozyme III from Komagataella phaffii GS115 (KpADH3) is still not clear yet...
February 20, 2018: Biochimica et Biophysica Acta
R Siva Kumar, K V Sravan Kumar, L Kondareddy, K R Yogeshwara, Gangrade Manish, Jayachandran Jeenet, Kanyawar Nitesh
A new sensitive and stability indicating reverse phase high-performance liquid chromatography method has been developed for the quantitative determination of potential impurities of Ledipasvir, active pharmaceutical ingredient. Efficient chromatographic separation was achieved on waters × select column contains C18 stationary phase in gradient mode, quantitation at wavelength of 325 nm and mobile phase A is 50 mM ammonium formate pH 3.5 and mobile phase B is acetonitrile with flow rate 1.2 mL. The method was validated according to International Council for Hormonisation guideline, linearity studies has shown correlation coefficient value greater than 0...
February 21, 2018: Journal of Chromatographic Science
Claire Pressiat, Huu-Hien Huynh, Alain Plé, Hélène Sauvageon, Isabelle Madelaine, Cécile Chougnet, Christine Le Maignan, Samia Mourah, Lauriane Goldwirt
BACKGROUND: A simple, rapid, and sensitive LC-MS/MS method has been developed and validated for the quantification of ruxolitinib, olaparib, vismodegib, and pazopanib in human plasma. METHODS: After a simple protein precipitation of plasma samples, the chromatographic separation was performed using an UPLC system coupled with MS/MS in a positive ionization mode. The mobile phase consisted of a gradient elution of 10 mmol/L formate ammonium buffer containing 0.1% (v/v) formic acid (phase A) and acetonitrile with 0...
February 22, 2018: Therapeutic Drug Monitoring
Zhenfang Cheng, Fuyou Du, Qun Qin, Lingshun Sun, Qiulian Zeng, Guihua Ruan, Jianping Li
In this work, an easy, effective and sensitive method, based on graphene oxide@silica@magnetite composites as adsorbent of magnetic solid phase extraction combined with liquid chromatography with tandem mass spectrometry, was established and validated for the trace analysis of cytokinins in different plants. The prepared magnetic composite was characterized by infrared spectroscopy, transmission electron microscopy, Brunauer-Emmett-Teller analysis and magnetic hysteresis. Under the optimized conditions, good linearities in the range of 0...
February 23, 2018: Journal of Separation Science
Can-Jun Ruan, Wei Guo, Miao Zhou, Gui-Xin Guo, Chuan-Yue Wang, Wen-Biao Li, Jose de Leon
A recent guideline recommends therapeutic drug monitoring (TDM) for risperidone, paliperidone and olanzapine, which are frequently-used second-generation antipsychotics. We developed a simple high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) coupled with an online solid-phase extraction (SPE) method that can be used to measure risperidone, paliperidone and olanzapine using small (40 μL) samples. The analytes were extracted from serum samples automatically pre-concentrated and purified by C8 (5 μm, 2...
February 23, 2018: Biomedical Chromatography: BMC
Alexandra M Koenig, Alexander Karabatsiakis, Thomas Stoll, Sarah Wilker, Thomas Hennessy, Michelle M Hill, Iris-Tatjana Kolassa
Childhood maltreatment (CM) can increase the risk of adverse health consequences in adulthood. A deeper insight in underlying biological pathways would be of high clinical relevance for early detection and intervention. The untargeted investigation of all detectable metabolites and lipids in biological samples represents a promising new avenue to identify so far unknown biological pathways associated with CM. Using an untargeted approach, liquid chromatography-mass spectrometry (LC-MS) was performed on peripheral blood serum samples collected three months postpartum from 105 women with varying degrees of CM exposure...
February 22, 2018: Scientific Reports
Melissa M Goggin, Stephanie D Gozum, Anna Miller, Gregory C Janis
The recent increase in illicit opioids sold on the black market, cut into heroin and masqueraded as prescription pills prompts a significant public health concern. Most designer opioids possess unknown potencies and unknown pharmacokinetics and their unregulated, variable dosages lead to rashes of overdoses. Additionally, many of the designer opioids, especially the fentanyl analogs are significantly more potent than heroin. High-profile cases involving overdoses of U-47700 and carfentanil have been reported in the media; however, the true prevalence of these and other designer opioids is unknown...
February 17, 2018: Journal of Analytical Toxicology
Bingjun Qian, Lirong Yin, Xiaomin Yao, Yaoguang Zhong, Juan Gui, Feifeng Lu, Fuming Zhang, Jianhua Zhang
The saponins, as components of tea seed meal (TSM), are undesirable hemolytic components, and should be degraded for reducing their hemolytic activity in order to be used in animal feed. In this study, β-glucuronidase was verified to be a potent hydrolase of tea seed saponins to reduce their hemolytic activity, and a β-glucuronidase-producing Lactobacillus crustorum (L. crustorum) strain was screened from raw bovine milk. Next, solid-state fermentation with the isolated L. crustorum and a Bacillus subtilis natto strain, which can produce cellulase and hence improve the fermentation performance of tea seed meal, was processed for detoxification of tea seed meal...
February 20, 2018: FEMS Microbiology Letters
Yu Dai, Amanda Li Cheng Tan, Huan Chen, Pei Shi Ong, Xiaoqiang Xiang, Jinzhu Wu, Hai-Shu Lin
Desoxyrhapontigenin (DRG, 4-methoxyresveratrol or trans-3,5-dihydroxy-4'-methoxystilbene) is a naturally occurring resveratrol (RES) derivative with a variety of biological activities. To facilitate its further medicinal exploration, a reliable LC-MS/MS method was developed and validated for the quantification of DRG in rat plasma using heavy isotope labelled RES as an internal standard. The ESI was operated in its negative ion mode while DRG and RES were determined by multiple reaction monitoring (MRM) using precursor-to-product ion transitions of m/z 241...
February 13, 2018: Journal of Pharmaceutical and Biomedical Analysis
Nadia Gagnon, Isabelle Fréchette, Pierre-Luc Mallet, Jean Dubé, Ghislaine Houde, Guy D Fink
OBJECTIVES: To determine salivary cortisol reference intervals in a healthy adult population, at 6 different time points during a 24-hour (h) period. METHODS: In a prospective study, salivary cortisol concentrations were measured upon waking, one-hour post-waking and at specific times of the day: at 12 h00, 16 h00, 20 h00 and midnight. Samples were analyzed by the first and second-generation electrochemiluminescence assays (ECLIA) from Roche Cobas Cortisol®...
February 19, 2018: Clinical Biochemistry
Hai H Bui, Phillip E Sanders, Diane Bodenmiller, Ming Shang Kuo, Gregory P Donoho, Anthony S Fischl
Phosphatidylinositol (3,4,5) trisphosphate (PIP3 ) is a biologically active membrane phospholipid that is essential for the growth and survival of all eukaryotic cells. We describe a new method that directly measures PIP3 and describe the HPLC separation and measurement of the positional isomers of phosphatidylinositol bisphosphate, PI(3,5)P2 , PI(3,4)P2 and PI(4,5)P2 . Mass spectrometric analyses were performed online using ultra-high performance liquid chromatography (UHPLC)-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) in the negative multiple-reaction monitoring (MRM) modes...
February 19, 2018: Analytical Biochemistry
Cuiyun Wang, Fang Yue, Guangfeng Ai, Jie Yang
A simple and sensitive liquid chromatography tandem mass spectrometry method was validated for simultaneous quantification of evodiamine and its metabolites 10-hydroxyevodiamine (M1), 18-hydroxyevodiamine (M2), 10-hydroxyevodiamine-glucuronide (M3) and 18-hydroxy- evodiamine-glucuronide (M4) in rat plasma for the first time. The analytes were extracted with acetonitrile and separated on a C18 column within 3 min. The detection was achieved in positive selected reaction monitoring mode with precursor-to-product transitions at m/z 304...
February 22, 2018: Biomedical Chromatography: BMC
Arindam Ghatak, Palak Chaturvedi, Wolfram Weckwerth
Metabolomics is an essential technology for functional genomics and systems biology. It plays a key role in functional annotation of genes and understanding towards cellular and molecular, biotic and abiotic stress responses. Different analytical techniques are used to extend the coverage of a full metabolome. The commonly used techniques are NMR, CE-MS, LC-MS, and GC-MS. The choice of a suitable technique depends on the speed, sensitivity, and accuracy. This chapter provides insight into plant metabolomic techniques, databases used in the analysis, data mining and processing, compound identification, and limitations in metabolomics...
February 23, 2018: Advances in Biochemical Engineering/biotechnology
Christina Lund Kidholm, Hans Christian Beck, Julie Bukh Madsen, Nikolai Bjødstrup Palstrøm, Jes Sanddal Lindholt, Lars Melholt Rasmussen
OBJECTIVE: The pathogenesis of abdominal aortic aneurysms (AAA) involves a disturbed balance of breakdown and buildup of arterial proteins. We envision that individuals with AAA carry generalized arterial protein alterations either because of effects of genetically or environmental AAA risk factors or because of compensatory changes due to signaling molecules released from the affected aneurysmal tissue. APPROACH: Protein extraction and quantitative proteome analysis by LC-MS/MS (liquid chromatography-mass spectrometry) was done on individual samples from the internal mammary artery from 11 individuals with AAA and 33 sex- and age-matched controls without AAA...
2018: PloS One
Andreas E Steinhilber, Felix F Schmidt, Wael Naboulsi, Hannes Planatscher, Alicia Niedzwiecka, Jutta Zagon, Albert Braeuning, Alfonso Lampen, Thomas O Joos, Oliver Poetz
The ban of processed animal proteins (PAPs) in feed for farmed animals introduced in 2001 was one of the main EU measures to control the bovine spongiform encephalopathy (BSE) crisis. Currently, microscopy and polymerase chain reaction (PCR), are the official methods for the detection of illegal PAPs in feed. However, the progressive release of the feed ban, recently with the legalization of non-ruminant PAPs for the use in aquaculture, requires the development of alternative methods to determine the species origin and the source (legal or not)...
February 22, 2018: Analytical Chemistry
Neeloo Singh, Renu Goel, Ekta Jain
PURPOSE: Although in trypanosomatids, monoxeny (Leptomonas) is ancestral to dixeny (Leishmania), however clinical cases of visceral leishmanisis with Leptomonas co-infection are increasingly being reported from India. Using a proteogenomic approach, we carried out a detailed proteome analysis of these two kinetoplastid parasites viz. Leishmania and its sister Leptomonas, to catalog the key proteins associated with and therefore possibly responsible for phenotype changes in Leptomonas evolution and domestication as co-infection with Leishmania...
February 22, 2018: Proteomics. Clinical Applications
Xiaoyan Guan, Le Zhang, Jette Wypych
Disulfide linkage is critical to protein folding and structural stability. The location of disulfide linkages for antibodies is routinely discovered by comparing the chromatograms of the reduced and non-reduced peptide mapping with location identification confirmed by collision-induced dissociation (CID) mass spectrometry (MS)/MS. However, CID product spectra of disulfide-linked peptides can be difficult to interpret, and provide limited information on the backbone region within the disulfide loop. Here, we applied an electron-transfer dissociation (ETD)/CID combined fragmentation method that identifies the disulfide linkage without intensive LC comparison, and yet maps the disulfide location accurately...
February 22, 2018: MAbs
Mehmet Ali Tetik, Onur Sevindik, Hasim Kelebek, Serkan Selli
The principal purpose of the present work is to characterize the aroma, aroma-active, and anthocyanin profiles of Okuzgozu wines and to observe the effect of the pomace pressing technique on these parameters. A total of 58 and 59 volatile compounds were identified and quantified in free-run juice wine (FRW) and pressed pomace wine (PW). Alcohols were found as the most dominant group among aroma compounds followed by esters and acids. However, among all these compounds only 11 and 13 of them could be considered as key odorants in aromatic extracts of FRW and PW, respectively...
February 22, 2018: Journal of Mass Spectrometry: JMS
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