Separation and identification of a complex flurbiprofen-polyethylene glycol mono- and diester mixture via a hyphenated HPLC-DAD-HRMS/SPE NMR system.

We report the use of a hyphenated HPLC-DAD(diode array detector)-HRMS/SPE NMR system for the separation and isolation of a complex mixture of esters, containing substances with very similar LC retention times. The literature known mono- and diesters of the drug Flurbiprofen and polyethylene glycol, which form a large number of substances with varying chain lengths, were chosen for this study. We demonstrate the use of this hyphenated system to quickly and effectively isolate sixteen of these highly similar individual esters in an automated fashion, demonstrating its applicability in standard pharmaceutical analysis and quality control of drugs. Both, synthetic solutions of these esters and extracts from Flurbiprofen lozenges were used for this purpose. By the sole use of this system, the individual compounds were isolated and UV, HRMS and 1D and 2D NMR data could be collected, enabling the identification and differentiation of the individual esters.

Isolation, Identification and Structural Verification of a Methylene-Bridged Naloxone "Dimer" Formed by Formaldehyde.

We report the isolation and characterization of a methylene bridged "dimer" of the opioid antagonist Naloxone, previously detected in experimental Buprenorphine-Naloxone oral films. This compound was found to form via an aldol addition followed by a condensation reaction under acidic conditions between two units of Naloxone and one unit of formaldehyde. HPLC-UV-HRMS analysis revealed the formation of three individual stereoisomers during this reaction, which were separately isolated using solid-phase extraction. These isomers were shown to freely react into one another in solvent, forming an equilibrium. The structure of the unknown compound was determined via HRMS spectrometry and 1D and 2D NMR spectroscopy.

Formation of a Thiol-Ene Addition Product of Asthma Medication Montelukast Caused by a Widespread Tin-Based Thermal Stabilizer.

We report the formation and characterization of two diastereomeric thiol-ene addition products of the asthma medication Montelukast within chewing tablets. Widespread tin-based thermal stabilizers dioctyltin bis(2-ethylhexyl thioglycolate) and monooctyltin tris(2-ethylhexyl thioglycolate), used in the manufacturing process of the medication's forming foil, were identified as the source of the thiol reactant, showing that these stabilizers may play a part in the degradation of Montelukast and APIs with functionalities similar to those of Montelukast, which should be considered during development of medication. The isolation and analysis of these impurities was performed by HPLC and UV-vis spectroscopy. HRMS and NMR data were collected to characterize and determine the structures of these compounds.

Beyond Verb Meaning: Experimental Evidence for Incremental Processing of Semantic Roles and Event Structure.

We present an event-related potentials (ERP) study that addresses the question of how pieces of information pertaining to semantic roles and event structure interact with each other and with the verb's meaning. Specifically, our study investigates German verb-final clauses with verbs of motion such as fliegen 'fly' and schweben 'float, hover,' which are indeterminate with respect to agentivity and event structure. Agentivity was tested by manipulating the animacy of the subject noun phrase and event structure by selecting a goal adverbial, which makes the event telic, or a locative adverbial, which leads to an atelic reading. On the clause-initial subject, inanimates evoked an N400 effect vis-à-vis animates. On the adverbial phrase in the atelic (locative) condition, inanimates showed an N400 in comparison to animates. The telic (goal) condition exhibited a similar amplitude like the inanimate-atelic condition. Finally, at the verbal lexeme, the inanimate condition elicited an N400 effect against the animate condition in the telic (goal) contexts. In the atelic (locative) condition, items with animates evoked an N400 effect compared to inanimates. The combined set of findings suggest that clause-initial animacy is not sufficient for agent identification in German, which seems to be completed only at the verbal lexeme in our experiment. Here non-agents (inanimates) changing their location in a goal-directed way and agents (animates) lacking this property are dispreferred and this challenges the assumption that change of (locational) state is generally a defining characteristic of the patient role. Besides this main finding that sheds new light on role prototypicality, our data seem to indicate effects that, in our view, are related to complexity, i.e., minimality. Inanimate subjects or goal arguments increase processing costs since they have role or event structure restrictions that animate subjects or locative modifiers lack.

Age-Related Changes in Predictive Capacity Versus Internal Model Adaptability: Electrophysiological Evidence that Individual Differences Outweigh Effects of Age.

Hierarchical predictive coding has been identified as a possible unifying principle of brain function, and recent work in cognitive neuroscience has examined how it may be affected by age-related changes. Using language comprehension as a test case, the present study aimed to dissociate age-related changes in prediction generation versus internal model adaptation following a prediction error. Event-related brain potentials (ERPs) were measured in a group of older adults (60-81 years; n = 40) as they read sentences of the form "The opposite of black is white/yellow/nice." Replicating previous work in young adults, results showed a target-related P300 for the expected antonym ("white"; an effect assumed to reflect a prediction match), and a graded N400 effect for the two incongruous conditions (i.e. a larger N400 amplitude for the incongruous continuation not related to the expected antonym, "nice," versus the incongruous associated condition, "yellow"). These effects were followed by a late positivity, again with a larger amplitude in the incongruous non-associated versus incongruous associated condition. Analyses using linear mixed-effects models showed that the target-related P300 effect and the N400 effect for the incongruous non-associated condition were both modulated by age, thus suggesting that age-related changes affect both prediction generation and model adaptation. However, effects of age were outweighed by the interindividual variability of ERP responses, as reflected in the high proportion of variance captured by the inclusion of by-condition random slopes for participants and items. We thus argue that - at both a neurophysiological and a functional level - the notion of general differences between language processing in young and older adults may only be of limited use, and that future research should seek to better understand the causes of interindividual variability in the ERP responses of older adults and its relation to cognitive performance.

Think globally: cross-linguistic variation in electrophysiological activity during sentence comprehension.

This paper demonstrates systematic cross-linguistic differences in the electrophysiological correlates of conflicts between form and meaning ("semantic reversal anomalies"). These engender P600 effects in English and Dutch (e.g. Kolk et al., 2003; Kuperberg et al., 2003), but a biphasic N400 - late positivity pattern in German (Schlesewsky and Bornkessel-Schlesewsky, 2009), and monophasic N400 effects in Turkish (Experiment 1) and Mandarin Chinese (Experiment 2). Experiment 3 revealed that, in Icelandic, semantic reversal anomalies show the English pattern with verbs requiring a position-based identification of argument roles, but the German pattern with verbs requiring a case-based identification of argument roles. The overall pattern of results reveals two separate dimensions of cross-linguistic variation: (i) the presence vs. absence of an N400, which we attribute to cross-linguistic differences with regard to the sequence-dependence of the form-to-meaning mapping and (ii) the presence vs. absence of a late positivity, which we interpret as an instance of a categorisation-related late P300, and which is observable when the language under consideration allows for a binary well-formedness categorisation of reversal anomalies. We conclude that, rather than reflecting linguistic domains such as syntax and semantics, the late positivity vs. N400 distinction is better understood in terms of the strategies that serve to optimise the form-to-meaning mapping in a given language.

The role of animacy in the real time comprehension of Mandarin Chinese: Evidence from auditory event-related brain potentials.

Two auditory ERP studies examined the role of animacy in sentence comprehension in Mandarin Chinese by comparing active and passive sentences in simple verb-final (Experiment 1) and relative clause constructions (Experiment 2). In addition to the voice manipulation (which modulated the assignment of actor and undergoer roles to the arguments), both arguments were either animate or inanimate. This allowed us to examine the interplay of animacy with thematic interpretation. In Experiment 1, we observed no effect of animacy at NP1, but N400 effects for inanimate actor arguments in second position. This result mirrors previous findings in German, thus suggesting that an initial undergoer universally leads to the prediction of an ideal (animate) actor. We also observed an N400 effect for passive sentences with an inanimate initial (undergoer) argument. We attribute this effect to a language-specific property of the passive construction in Chinese, namely that the first argument is negatively affected by the event described (i.e. bears an experiencer role). Experiment 2 showed that both of these effects can also be observed in sentence constructions of another type, in which the critical information sources become available in a different order. These findings provide the first demonstration that the N400 is not only sensitive to general (universal) aspects of thematic processing (i.e. "who is acting on whom") but also to the interaction between thematic interpretation and language-specific pragmatic principles.