ALBERT

Description: Ginkgo biloba extract (GbE) is a dietary supplement derived from an ethanolic extract of Ginkgo biloba leaves. Unfinished bulk GbE is used to make finished products that are sold as dietary supplements. The variable, complex composition of GbE makes it difficult to obtain consistent toxicological assessments of potential risk. The National Toxicology Program (NTP) observed hepatotoxicity in its rodent studies of a commercially available, unfinished GbE product, but the application of these results to the broader GbE supplement market is unclear. Here, we use a combination of non-targeted and targeted chromatographic and spectrophotometric methods to obtain profiles of 24 commercially available finished GbE products and unfinished standardized and unstandardized extracts with and without hydrolysis, then used principal component analysis to group unfinished products according to their similarity to each other and to National Institute of Standards and Technology (NIST) standard reference materials (SRM), and the finished products. Unfinished products were grouped into those that were characteristic and uncharacteristic of standardized GbE. Our work demonstrates that different analytical approaches produced similar classifications of characteristic and uncharacteristic products in unhydrolyzed samples, but the distinctions largely disappeared once the samples were hydrolyzed. Using our approach, the NTP GbE was most similar to two unfinished GbE products classified as characteristic, finished products, and the NIST GbE SRM. We propose that a simple analysis for the presence, absence, or amounts of compounds unique to GbE in unhydrolyzed samples could be sufficient to determine a sample’s authenticity. Graphical abstract

Description: Impacts of spectral nudging on simulations of Arctic climate in coupled simulations have been investigated in a set of simulations with a regional climate model (RCM). The dominantly circumpolar circulation in the Arctic lead to weak constraints on the lateral boundary conditions (LBCs) for the RCM, which causes large internal variability with strong deviations from the driving model. When coupled to an ocean and sea ice model, this results in sea ice concentrations that deviate from the observed spatial distribution. Here, a method of spectral nudging is applied to the atmospheric model RCA4 in order to assess the potentials for improving results for the sea ice concentrations when coupled to the RCO ocean-sea ice model. The spectral nudging applied to reanalysis driven simulations significantly improves the generated sea ice regarding its temporal evolution, extent and inter-annual trends, compared to simulations with standard LBC nesting. The method is furthermore evaluated with driving data from two CMIP5 GCM simulations for current and future conditions. The GCM biases are similar to the RCA4 biases with ERA-Interim, however, the spectral nudging still improves the surface winds enough to show improvements in the simulated sea ice. For both GCM downscalings, the spectrally nudged version retains a larger sea ice extent in September further into the future. Depending on the sea ice formulation in the GCM, the temporal evolution of the regional sea ice model can deviate strongly.

Description: The increased rate of Tropical Indian Ocean (TIO) surface warming has gained a lot of attention in the recent years mainly due to its regional climatic impacts. The processes associated with this increased surface warming is highly complex and none of the mechanisms in the past studies could comprehend the important features associated with this warming such as the negative trends in surface net heat fluxes and the decreasing temperature trends at thermocline level. In this work we studied a previously unexplored aspect, the changes in large scale surface circulation pattern modulating the surface warming pattern over TIO. We use ocean reanalysis datasets and a suit of Ocean General Circulation Model (OGCM) experiments to address this problem. Both reanalysis and OGCM reveal strengthening large scale surface circulation pattern in the recent years. The most striking feature is the intensification of cyclonic gyre circulation around the thermocline ridge in the southwestern TIO. The surface circulation change in TIO is mainly associated with the surface wind changes and the geostrophic response to sea surface height decrease in the western/southwestern TIO. The surface wind trends closely correspond to SST warming pattern. The strengthening mean westerlies over the equatorial region are conducive to convergence in the central and divergence in the western equatorial Indian Ocean (IO) resulting central warming and western cooling. The resulting east west SST gradient further enhances the equatorial westerlies. This positive feedback mechanism supports strengthening of the observed SST trends in the equatorial Indian Ocean. The cooling induced by the enhanced upwelling in the west is compensated to a large extent by warming due to reduction in mixed layer depth, thereby keeping the surface temperature trends in the west to weak positive values. The OGCM experiments showed that the wind induced circulation changes redistribute the excess heat received in the western TIO to central and east thereby enhancing warming in the central equatorial IO. The increased surface warming in central TIO increases the latent heat loss, and keeps the net heat flux trends negative. Model sensitivity experiments reveal that the subsurface cooling at thermocline level in TIO is contributed by variability in Pacific via Indonesian Through Flow whereas the surface warming trend is mainly induced by the changes in the local forcing. The long term changes in IO Rossby waves are not induced by local atmospheric forcing but are forced by Pacific. The thermocline shoaling in the west is therefore amplified by the remote influence of Pacific via wave transmission through Indonesian archipelago.

Description: Stable isotope dilution assays (SIDAs) are becoming ever commoner in mycotoxin analysis, and the number of synthesized or commercially available isotopically labelled compounds has greatly increased in the 7 years since our last review dealing with this topic. Thus, this review is conceived as an update for new applications or improvements of SIDAs for compounds discussed earlier, but the main focus is on newly introduced labelled substances and the development of SIDAs for, for example, fusarin C, moniliformin or the enniatins. Mycotoxin research has concentrated on the emerging group of Alternaria toxins in recent years, and a series of SIDAs have been developed, including ones for tenuazonic acid, alternariol, altertoxins and tentoxin that are discussed in detail in this review. Information about synthetic routes, isotopic purity and mass-spectrometric characterization of labelled compounds is given, as well as about the development and validation of SIDAs and their application to foods, feeds or biological samples. As the number of commercially available labelled standards is increasing continuously, a general tendency for the use of analytical methods based on liquid chromatography coupled with mass spectrometry capable of identifying a series of mycotoxins simultaneously (“multimethods”) and using one or more labelled internal standards can be observed. An overview of these applications is given, thus demonstrating that SIDAs are increasingly being used in routine analysis.

Description: Synchrotron radiation nanoscale computed tomography (SR nano-CT) is a powerful analysis tool and can be used to perform chemical identification, mapping, or speciation of carbon and other elements together with X-ray fluorescence and X-ray absorption near edge structure (XANES) imaging. In practical applications, there are often challenges for SR nano-CT due to the misaligned geometry caused by the sample stage axial vibration. It occurs quite frequently because of experimental constraints from the mechanical error of manufacturing and assembly and the thermal expansion during the time-consuming scanning. The axial vibration will lead to the structure overlap among neighboring layers and degrade imaging results by imposing artifacts into the nano-CT images. It becomes worse for samples with complicated axial structure. In this work, we analyze the influence of axial vibration on nano-CT image by partial derivative. Then, an axial vibration calibration method for SR nano-CT is developed and investigated. It is based on the cross correlation of plane integral curves of the sample at different view angles. This work comprises a numerical study of the method and its experimental verification using a dataset measured with the full-field transmission X-ray microscope nano-CT setup at the beamline 4W1A of the Beijing Synchrotron Radiation Facility. The results demonstrate that the presented method can handle the stage axial vibration. It can work for random axial vibration and needs neither calibration phantom nor additional calibration scanning. It will be helpful for the development and application of synchrotron radiation nano-CT systems.

Description: An on-line, ensemble-based data assimilation (DA) method is performed to reconstruct the climate for 1750–1850 AD, and the performance is evaluated on large and small spatial scales. We use a low-resolution version of the Max Planck Institute for Meteorology MPI-ESM model and assimilate the PAGES 2K continental mean temperature reconstructions for the Northern Hemisphere (NH). The ensembles are generated sequentially for sub-periods based on the analysis of previous sub-periods. The assimilation has good skill for large-scale temperatures, but there is no agreement between the DA analysis and proxy-based reconstructions for small-scale temperature patterns within Europe or with reconstructions for the North Atlantic Oscillation (NAO) index. To explain the lack of added value in small spatial scales, a maximum covariance analysis (MCA) of links between NH temperature and sea level pressure is performed based on a control simulation with MPI-ESM. For annual values, winter and spring the Northern Annular Mode (NAM) is the pattern that is most closely linked to the NH continental temperatures, while for summer and autumn it is a wave-like pattern. This link is reproduced in the DA for winter, spring and annual means, providing potential for constraining the NAM/NAO phase and in turn regional temperature variability. It is shown that the lack of actual small-scale skill is likely due to the fact that the link might be too weak, as the NH continental mean temperatures are not the best predictors for large-scale circulation anomalies, or that the PAGES 2K temperatures include noise. Both factors can lead to circulation anomalies in the DA analysis that are substantially different from reality, leading to unrealistic representation of small-scale temperature variability. Moreover, we show that even if the true amplitudes of the leading MCA circulation patterns were known, there is still a large amount of unexplained local temperature variance. Based on these results, we argue that assimilating temperature reconstructions with a higher spatial resolution might improve the DA performance.

Description: We analyse the impact of migration and strength of Southern Hemisphere westerly winds on the ocean carbon cycle in a systematic sensitivity study with the University of Victoria Earth System Climate Model. We find that changes in the biological pump are mainly driven by changes in ocean residence times while changes in export production are negligible. Changes in the biological and physical pumps are always of opposite sign; with the physical pump being dominant for southward shifts and the biological pump being dominant for northward shifts. Furthermore, changes in the Pacific Ocean carbon budget dictate the overall changes in global marine and atmospheric carbon. Overall, atmospheric \(\hbox {CO}_2\) increases (and \(\Delta ^{14}\hbox {C}\) decreases) for northward shifts or a strengthening in wind forcing. The opposite is true for a southward shift or a weakening in wind forcing. Combining forcings (shift and intensity change) results in a combination of their impacts with the direction of the shift being the first order forcing. The terrestrial carbon reservoir absorbs (releases) 50–70 % of the net oceanic carbon loss (increase), counterbalancing the effect on atmospheric \(\hbox {CO}_2\) .

Description: Effects of wind and fresh water on the Atlantic meridional overturning circulation (AMOC) are investigated using a fully coupled climate model. The AMOC can change significantly when perturbed by either wind stress or freshwater flux in the North Atlantic. This study focuses on wind stress effect. Our model results show that the wind forcing is crucial in maintaining the AMOC. Reducing wind forcing over the ocean can cause immediately weakening of the vertical salinity diffusion and convection in the mid-high latitudes Atlantic, resulting in an enhancement of vertical salinity stratification that restrains the deep water formation there, triggering a slowdown of the thermohaline circulation. As the thermohaline circulation weakens, the sea ice expands southward and melts, providing the upper ocean with fresh water that weakens the thermohaline circulation further. The wind perturbation experiments suggest a positive feedback between sea-ice and thermohaline circulation strength, which can eventually result in a complete shutdown of the AMOC. This study also suggests that sea-ice variability may be also important to the natural AMOC variability on decadal and longer timescales.

Description: The relationship between the large-scale circulation dynamics and regional precipitation regime in the Tibetan Plateau (TP) has so far not been well understood. In this study, we classify the circulation types using the self-organizing maps based on the daily field of 500 hPa geopotential height and link them to the precipitation climatology in the eastern and central TP. By virtue of an objective determining method, 18 circulation types are quantified. The results show that the large amount of precipitation in summer is closely related to the circulation types in which the enhanced and northward shifted subtropical high (SH) over the northwest Pacific and the obvious cyclconic circulation anomaly over the Bay of Bengal are helpful for the Indian summer monsoon and East Asian summer monsoon to take abundant low-latitude moisture to the eastern and southern TP. On the contrary, the dry winter in the central and eastern Tibet corresponds to the circulation types with divergence over the central and eastern TP and the water vapor transportations of East Asian winter monsoon and mid-latitude westerly are very weak. Some circulation types are associated with some well-known circulation patterns/monsoons influencing the TP (e.g. East Atlantic Pattern, El Niño Southern Oscillation, Indian Summer Monsoon and the mid-latitude westerly), and exhibit an overall good potential for explaining the variability of regional seasonal precipitation. Moreover, the climate shift signals in the late 1970s over the eastern Pacific/North Pacific Oceans could also be reflected by both the variability of some circulation types and their correspondingly composite precipitations. This study extends our understandings for the large-scale atmospheric dynamics and their linkages with regional precipitation and is beneficial for the climate change projection and related adaptation activities in the highest and largest plateau in the world.

Description: Based on the historical and RCP8.5 experiments from 25 Coupled Model Intercomparison Project phase 5 (CMIP5) models, the impacts of sea surface temperature (SST) warming in the tropical Indian Ocean (IO) on the projected change in summer rainfall over Central Asia (CA) are investigated. The analysis is designed to answer three questions: (1) Can CMIP5 models reproduce the observed influence of the IO sea surface temperatures (SSTs) on the CA rainfall variations and the associated dynamical processes? (2) How well do the models agree on their projected rainfall changes over CA under warmed climate? (3) How much of the uncertainty in such rainfall projections is due to different impacts of IO SSTs in these models? The historical experiments show that in most models summer rainfall over CA are positively correlated to the SSTs in the IO. Furthermore, for models with higher rainfall-SSTs correlations, the dynamical processes accountable for such impacts are much closer to what have been revealed in observational data: warmer SSTs tend to favor the development of anti-cyclonic circulation patterns at low troposphere over north and northwest of the Arabian Sea and the Bay of Bengal. These anomalous circulation patterns correspond to significantly enhanced southerly flow which carries warm and moisture air mass from the IO region up to the northeast. At the same time, there is a cyclonic flow over the central and eastern part of the CA which further brings the tropical moisture into the CA and provides essential moist conditions for its rainfall generation. In the second half of twenty-first century, although all the 25 models simulate warmed SSTs, significant uncertainty exists in their projected rainfall changes over CA: half of them suggest summer rainfall increases, but the other half project rainfall decreases. However, when we select seven models out of the 25 based on their skills in capturing the dynamical processes as observed, then the model projected changes are much closer. Five out of the seven models predicted more rainfall over CA. Such a result is helpful for allowing us to attribute part of the observed upward rainfall trend in the CA region in the last several decades to the IO SST warming.

Description: A selenosugar (selenosugar 1 , methyl-2-acetamido-2-deoxy-1-seleno-β-D-galactopyranoside) was identified in aqueous extracts of muscle tissue of three marine fish species, mackerel ( Scomber scombrus ), sardine ( Sardina pilchardus ), and tuna ( Thunnus albacares ), by high-performance liquid chromatography coupled to elemental and high-resolution molecular mass spectrometry. Selenoneine (2-selenyl-N α , N α , N α -trimethyl-L-histidine), a known selenium compound in fish, was the major form of selenium in the aqueous extracts, and the methylated derivative of selenoneine, namely Se-methylselenoneine, was also identified as a minor natural constituent in the fish. Selenosugar 1 , a major urinary excretion product of selenium often found in organs and body fluids related to selenium excretion, has so far not been reported in muscle tissue. Se-methylselenoneine has been proposed as the main urinary metabolite from selenoneine. This first report of selenosugar 1 and Se-methylselenoneine as natural constituents of fish muscle tissue opens up a new perspective on the role of these compounds in selenium metabolism and is relevant to selenium supplementation studies.

Description: Cyclonic windstorms are one of the most important natural hazards for Europe, but robust climate projections of the position and the strength of the North Atlantic storm track are not yet possible, bearing significant risks to European societies and the (re)insurance industry. Previous studies addressing the problem of climate model uncertainty through statistical comparisons of simulations of the current climate with (re-)analysis data show large disagreement between different climate models, different ensemble members of the same model and observed climatologies of intense cyclones. One weakness of such evaluations lies in the difficulty to separate influences of the climate model’s basic state from the influence of fast processes on the development of the most intense storms, which could create compensating effects and therefore suggest higher reliability than there really is. This work aims to shed new light into this problem through a cost-effective “seamless” approach of hindcasting 20 historical severe storms with the two global climate models, ECHAM6 and GA4 configuration of the Met Office Unified Model, run in a numerical weather prediction mode using different lead times, and horizontal and vertical resolutions. These runs are then compared to re-analysis data. The main conclusions from this work are: (a) objectively identified cyclone tracks are represented satisfactorily by most hindcasts; (b) sensitivity to vertical resolution is low; (c) cyclone depth is systematically under-predicted for a coarse resolution of T63 by both climate models; (d) no systematic bias is found for the higher resolution of T127 out to about three days, demonstrating that climate models are in fact able to represent the complex dynamics of explosively deepening cyclones well, if given the correct initial conditions; (e) an analysis using a recently developed diagnostic tool based on the surface pressure tendency equation points to too weak diabatic processes, mainly latent heating, as the main source for the under-prediction in the coarse-resolution runs. Finally, an interesting implication of these results is that the too low number of deep cyclones in many free-running climate simulations may therefore be related to an insufficient number of storm-prone initial conditions. This question will be addressed in future work.

Description: We present a novel strategy based on data-independent acquisition coupled to targeted data extraction for the detection and identification of site-specific modifications of targeted peptides in a completely unbiased manner. This method requires prior knowledge of the site of the modification along the peptide backbone from the protein of interest, but not the mass of the modification. The procedure, named multiplex adduct peptide profiling (MAPP), consists of three steps: 1) A fragment-ion tag is extracted from the data, consisting of the b-type and y-type ion series from the N and C-terminus, respectively, up to the amino-acid position that is believed to be modified; 2) MS1 features are matched to the fragment-ion tag in retention-time space, using the isolation window as a pre-filter to enable calculation of the mass of the modification; and 3) modified fragment ions are overlaid with the unmodified fragment ions to verify the mass calculated in step 2. We discuss the development, applications, and limitations of this new method for detection of unknown peptide modifications. We present an application of the method in profiling adducted peptides derived from abundant proteins in biological fluids with the ultimate objective of detecting biomarkers of exposure to reactive species.

Description: Circulating cell-free DNA (ccfDNA) has great potential for non-invasive diagnostics, and prediction and monitoring of treatment response, but its amount is usually limited. Therefore, the choice of methods to extract and characterize ccfDNA is crucial. In the current study, we performed the most comprehensive comparison of methods for ccfDNA extraction (11 methods), quantification (3 methods), and estimation of the integrity index (2 methods) from small quantities of different kinds of plasma. The QIAamp® Circulating Nucleic Acid Kit and the Norgen Plasma/Serum Circulating DNA Purification Mini Kit showed the best accuracy and reproducibility, but the Norgen kit allowed to extract a higher amount of ccfDNA. This workflow provides a reliable protocol for the multiple applications of ccfDNA in biomedicine. Graphical Abstract Workflow for the evaluation of methods to isolate, quantify and characterize circulating cell-free DNA from small volumes of plasma

Description: After internal contamination, uranium rapidly distributes in the body; up to 20 % of the initial dose is retained in the skeleton, where it remains for years. Several studies suggest that uranium has a deleterious effect on the bone cell system, but little is known regarding the mechanisms leading to accumulation of uranium in bone tissue. We have performed synchrotron radiation-based micro-X-ray fluorescence (SR μ-XRF) studies to assess the initial distribution of uranium within cortical and trabecular bones in contaminated rats’ femurs at the micrometer scale. This sensitive technique with high spatial resolution is the only method available that can be successfully applied, given the small amount of uranium in bone tissue. Uranium was found preferentially located in calcifying zones in exposed rats and rapidly accumulates in the endosteal and periosteal area of femoral metaphyses, in calcifying cartilage and in recently formed bone tissue along trabecular bone. Furthermore, specific localized areas with high accumulation of uranium were observed in regions identified as micro-vessels and on bone trabeculae. These observations are of high importance in the study of the accumulation of uranium in bone tissue, as the generally proposed passive chemical sorption on the surface of the inorganic part (apatite) of bone tissue cannot account for these results. Our study opens original perspectives in the field of exogenous metal bio-mineralization.

Description: Extreme precipitation and flood episodes in the Himalayas are oftentimes traced to synoptic situations involving connections between equatorward advancing upper level extratropical circulations and moisture-laden tropical monsoon circulation. While previous studies have documented precipitation characteristics in the Himalayan region during severe storm cases, a comprehensive understanding of circulation dynamics of extreme precipitation mechanisms is still warranted. In this study, a detailed analysis is performed using rainfall observations and reanalysis circulation products to understand the evolution of monsoon-extratropical circulation features and their interactions based on 34 extreme precipitation events which occurred in the Western Himalayas (WEH) during the period 1979–2013. Our results provide evidence for a common large-scale circulation pattern connecting the extratropics and the South Asian monsoon region, which is favorable for extreme precipitation occurrences in the WEH region. This background upper level large-scale circulation pattern consists of a deep southward penetrating midlatitude westerly trough, a blocking high over western Eurasia and an intensifying Tibetan anticyclone. It is further seen from our analysis that the key elements of monsoon-midlatitude interactions, responsible for extreme precipitation events over the WEH region, are: (1) midlatitude Rossby wave breaking, (2) west-northwest propagation of monsoon low-pressure system from the Bay of Bengal across the Indian subcontinent, (3) eddy shedding of the Tibetan anticyclone, (4) ageostrophic motions and transverse circulation across the Himalayas, and (5) strong moist convection over the Himalayan foothills. Furthermore, high-resolution numerical simulations indicate that diabatic heating and mesoscale ageostrophic effects can additionally amplify the convective motions and precipitation in the WEH region.

Description: This paper examines the difference in the Atlantic Meridional Overturning Circulation (AMOC) mean state between free and assimilative simulations of a common ocean model using a common interannual atmospheric forcing. In the assimilative simulation, the reproduction of cold cores in the Nordic Seas, which is absent in the free simulation, enhances the overflow to the North Atlantic and improves AMOC with enhanced transport of the deeper part of the southward return flow. This improvement also induces an enhanced supply of North Atlantic Deep Water (NADW) and causes better representation of the Atlantic deep layer despite the fact that correction by the data assimilation is applied only to temperature and salinity above a depth of 1750 m. It also affects Circumpolar Deep Water in the Southern Ocean. Although the earliest influence of the improvement propagated by coastal waves reaches the Southern Ocean in 10–15 years, substantial influence associated with the arrival of the renewed NADW propagates across the Atlantic Basin in several decades. Although the result demonstrates that data assimilation is able to improve the deep ocean state even if there is no data there, it also indicates that long-term integration is required to reproduce variability in the deep ocean originating from variations in the upper ocean. This study thus provides insights on the reliability of AMOC and the ocean state in the Atlantic deep layer reproduced by data assimilation systems.

Description: The southeast region of Brazil experienced in austral summer 2014 a major drought event leading to a number of impacts in water availability for human consumption, agricultural irrigation and hydropower production. This study aims to perform a diagnostic analysis of the observed climate conditions during this event, including an inspection of the occurred precipitation anomalies in the context of previous years, and an investigation of possible relationships with sea surface temperatures and atmospheric circulation patterns. The sea surface temperature analysis revealed that the southwestern South Atlantic Ocean region near the coast of southeast Brazil showed strong negative association with precipitation over southeast Brazil, indicating that increased sea temperatures in this ocean region are consistent with reduced precipitation as observed in summer 2014. The circulation analysis revealed prevailing anti-cyclonic anomalies at lower levels (850 hPa) with northerly anomalies to the west of southeast Brazil, channeling moisture from the Amazon towards Paraguay, northern Argentina and southern Brazil, and drier than normal air from the South Atlantic Ocean towards the southeast region of Brazil. This circulation pattern was found to be part of a large-scale teleconnection wave train linked with the subsidence branch of the Walker circulation in the tropical east Pacific, which in turn was generated by an anomalous tropical heat source in north/northeastern Australia. A regional Hadley circulation with an ascending branch to the south of the subsidence branch of the Walker circulation in the tropical east Pacific was identified as an important component connecting the tropical and extratropical circulation. The ascending branch of this Hadley circulation in the south Pacific coincided with an identified Rossby wave source region, which contributed to establishing the extratropical component of the large-scale wave train connecting the south Pacific and the Atlantic region surrounding southeast Brazil. This connection between the Pacific and the Atlantic was confirmed with Rossby ray tracing analyses. The local circulation response was associated to downward air motion (subsidence) over Southeast Brazil, contributing to the expressive negative precipitation anomalies observed during summer 2014, and leading to a major drought event in the historical context. The analysis of atmospheric and oceanic patterns of this event helped defining a schematic framework leading to the observed drought conditions in southeast Brazil, including the involved teleconnections, blocking high pressure, radiative and humidity transport effects.

Description: The spectroscopic approaches of FTIR imaging and Raman mapping were applied to the characterisation of a new carbon hydroxyapatite/β-glucan composite developed for bone tissue engineering. The composite is an artificial bone material with an apatite-forming ability for the bone repair process. Rabbit bone samples were tested with an implanted bioactive material for a period of several months. Using spectroscopic and chemometric methods, we were able to determine the presence of amides and phosphates and the distribution of lipid-rich domains in the bone tissue, providing an assessment of the composite’s bioactivity. Samples were also imaged in transmission using an infrared microscope combined with a focal plane array detector. CaF 2 lenses were also used on the infrared microscope to improve spectral quality by reducing scattering artefacts, improving chemometric analysis. The presence of collagen and lipids at the bone/composite interface confirmed biocompatibility and demonstrate the suitability of FTIR microscopic imaging with lenses in studying these samples. It confirmed that the composite is a very good background for collagen growth and increases collagen maturity with the time of the bone growth process. The results indicate the bioactive and biocompatible properties of this composite and demonstrate how Raman and FTIR spectroscopic imaging have been used as an effective tool for tissue characterisation.

Description: Cervical cancer is the fourth most common cancer in women worldwide and mainly affects younger women. The mortality associated with cervical cancer can be reduced if the disease is detected at the pre-cancer stage. Current best-practice methods include cytopathology, HPV testing, and histopathology, but these methods are limited in terms of subjectivity, cost, and time. There is an unmet clinical need for new methods to aid clinicians in the early detection of cervical pre-cancer. These methods should be objective and rapid and require minimal sample preparation. Raman spectroscopy is a vibrational spectroscopic technique by which incident radiation is used to induce vibrations in the molecules of a sample and the scattered radiation may be used to characterise the sample in a rapid and non-destructive manner. Raman spectroscopy is sensitive to subtle biochemical changes occurring at the molecular level, enabling spectral variations corresponding to disease onset to be detected. Over the past 15 years, there have been numerous reports revealing the potential of Raman spectroscopy together with multivariate statistical analysis for the detection of a variety of cancers. This paper discusses the recent advances and challenges for cervical-cancer screening and diagnosis and offers some perspectives for the future.

Description: The existence of cut-off lows (COLs) over South Pacific and South America is often associated with adverse weather events such as intense precipitation over the central region of South America, frost episodes in southern Brazil and the development of Andes lee cyclones and intense cyclones over the southern coast of Brazil. Despite this importance, the formation and maintenance mechanisms of the COLs are not well understood. To detail the significant variability in terms of the eddy kinetic energy equation for fifty cases of COLs that formed over the southeastern Pacific Ocean is the aim of this study. Only the cases of COLs that formed over the ocean and remained there during most of their life were chosen. The main terms of the equation [ageostrophic flux convergence (AFC), baroclinic conversion (BRC) and barotropic conversion (BRT)] were calculated using the 6-hourly gridded data from the National Centers for Environmental Prediction/Department of Energy reanalysis. The formation mechanism of the COLs was associated with BRC and AFC. During the midlife period, the BRC term converted eddy kinetic energy to eddy potential energy and the AFC had a positive contribution until 6 h after the midlife point. In the dissipation phase, the BRC term remained positive and AFC became negative. The BRT extracted kinetic energy from the COL during the entire life cycle. The AFC term was the most important in all phases of the cut-off lifetime, and it was the responsible for extending the cut-off lifetime while the others terms were negatives.

Description: The present study examines the ability of high resolution (T382) National Centers for Environmental Prediction coupled atmosphere–ocean climate forecast system version 2 (CFS T382) in simulating the salient spatio-temporal characteristics of the boreal summertime mean climate and the intraseasonal variability. The shortcomings of the model are identified based on the observation and compared with earlier reported biases of the coarser resolution of CFS (CFS T126). It is found that the CFS T382 reasonably mimics the observed features of basic state climate during boreal summer. But some prominent biases are noted in simulating the precipitation, tropospheric temperature (TT) and sea surface temperature (SST) over the global tropics. Although CFS T382 primarily reproduces the observed distribution of the intraseasonal variability over the Indian summer monsoon region, some difficulty remains in simulating the boreal summer intraseasonal oscillation (BSISO) characteristics. The simulated eastward propagation of BSISO decays rapidly across the Maritime Continent, while the northward propagation appears to be slightly slower than observation. However, the northward propagating BSISO convection propagates smoothly from the equatorial region to the northern latitudes with observed magnitude. Moreover, the observed northwest-southeast tilted rain band is not well reproduced in CFS T382. The warm mean SST bias and inadequate simulation of high frequency modes appear to be responsible for the weak simulation of eastward propagating BSISO. Unlike CFS T126, the simulated mean SST and TT exhibit warm biases, although the mean precipitation and simulated BSISO characteristics are largely similar in both the resolutions of CFS. Further analysis of the convectively coupled equatorial waves (CCEWs) indicates that model overestimates the gravest equatorial Rossby waves and underestimates the Kelvin and mixed Rossby-gravity waves. Based on analysis of CCEWs, the study further explains the possible reasons behind the realistic simulation of northward propagating BSISO in CFS T382, even though the model shows substantial biases in simulating mean state and other BSISO modes.

Description: The TAO/TRITON array is the cornerstone of the tropical Pacific and ENSO observing system. Motivated by the recent rapid decline of the TAO/TRITON array, the potential utility of TAO/TRITON was assessed for ENSO monitoring and prediction. The analysis focused on the period when observations from Argo floats were also available. We coordinated observing system experiments (OSEs) using the global ocean data assimilation system (GODAS) from the National Centers for Environmental Prediction and the ensemble coupled data assimilation (ECDA) from the Geophysical Fluid Dynamics Laboratory for the period 2004–2011. Four OSE simulations were conducted with inclusion of different subsets of in situ profiles: all profiles (XBT, moorings, Argo), all except the moorings, all except the Argo and no profiles. For evaluation of the OSE simulations, we examined the mean bias, standard deviation difference, root-mean-square difference (RMSD) and anomaly correlation against observations and objective analyses. Without assimilation of in situ observations, both GODAS and ECDA had large mean biases and RMSD in all variables. Assimilation of all in situ data significantly reduced mean biases and RMSD in all variables except zonal current at the equator. For GODAS, the mooring data is critical in constraining temperature in the eastern and northwestern tropical Pacific, while for ECDA both the mooring and Argo data is needed in constraining temperature in the western tropical Pacific. The Argo data is critical in constraining temperature in off-equatorial regions for both GODAS and ECDA. For constraining salinity, sea surface height and surface current analysis, the influence of Argo data was more pronounced. In addition, the salinity data from the TRITON buoys played an important role in constraining salinity in the western Pacific. GODAS was more sensitive to withholding Argo data in off-equatorial regions than ECDA because it relied on local observations to correct model biases and there were few XBT profiles in those regions. The results suggest that multiple ocean data assimilation systems should be used to assess sensitivity of ocean analyses to changes in the distribution of ocean observations to get more robust results that can guide the design of future tropical Pacific observing systems.

Description: Large arrays of femtoliter-sized chambers are important tools for single molecule research as well as bioanalytical applications. We have optimized the design and fabrication of two array types consisting of 250 × 250 (62 500) femtoliter chambers either by surface etching of fused silica slides or by polydimethylsiloxane (PDMS) molding. Highly diluted solutions of β-galactosidase were enclosed in such arrays to monitor the fluorogenic reactions of hundreds of individual enzyme molecules in parallel by wide-field fluorescence microscopy. An efficient mechanical sealing procedure was developed to prevent diffusion of the fluorescent reaction product out of the chambers. Different approaches for minimizing non-specific surface adsorption were explored. The signal acquisition was optimized to grant both a large field of view and an efficient signal acquisition from each femtoliter chamber. The optimized femtoliter array has enabled a three-in-one enzyme assay system: First, the concentration of active enzyme can be determined in a digital way by counting fluorescent chambers in the array. Second, the activity of the enzyme bulk solution is given by averaging many individual substrate turnover rates without the need for knowing the exact enzyme concentration. Third—unlike conventional enzyme assays—the distribution of individual substrate turnover rates yields insight into the conformational heterogeneity in an enzyme population. The substrate turnover rates of single β-galactosidase molecules were found to be broadly distributed and independent of the type of femtoliter array. In general, both types of femtoliter arrays are highly sensitive platforms for enzyme analysis at the single molecule level and yield consistent results. Graphical Abstract Isolation and analysis of individual enzyme molecules in large arrays of femtoliter-sized chambers

Description: We present a comprehensive discussion of the role that microchip electrophoresis (ME) instrumentation could play in future NASA missions of exploration, as well as the current barriers that must be overcome to make this type of chemical investigation possible. We describe how ME would be able to fill fundamental gaps in our knowledge of the potential for past, present, or future life beyond Earth. Despite the great promise of ME for ultrasensitive portable chemical analysis, to date, it has never been used on a robotic mission of exploration to another world. We provide a current snapshot of the technology readiness level (TRL) of ME instrumentation, where the TRL is the NASA systems engineering metric used to evaluate the maturity of technology, and its fitness for implementation on missions. We explain how the NASA flight implementation process would apply specifically to ME instrumentation, and outline the scientific and technology development issues that must be addressed for ME analyses to be performed successfully on another world. We also outline research demonstrations that could be accomplished by independent researchers to help advance the TRL of ME instrumentation for future exploration missions. The overall approach described here for system development could be readily applied to a wide range of other instrumentation development efforts having broad societal and commercial impact.

Description: Vanadium speciation in the fungus Phycomyces blakesleeanus was examined by X-ray absorption near-edge structure (XANES) spectroscopy, enabling assessment of oxidation states and related molecular symmetries of this transition element in the fungus. The exposure of P. blakesleeanus to two physiologically important vanadium species (V 5+ and V 4+ ) resulted in the accumulation of this metal in central compartments of 24 h old mycelia, most probably in vacuoles. Tetrahedral V 5+ , octahedral V 4+ , and proposed intracellular complexes of V 5+ were detected simultaneously after addition of a physiologically relevant concentration of V 5+ to the mycelium. A substantial fraction of the externally added V 4+ remained mostly in its original form. However, observable variations in the pre-edge-peak intensities in the XANES spectra indicated intracellular complexation and corresponding changes in the molecular coordination symmetry. Vanadate complexation was confirmed by 51 V NMR and Raman spectroscopy, and potential binding compounds including cell-wall constituents (chitosan and/or chitin), (poly)phosphates, DNA, and proteins are proposed. The evidenced vanadate complexation and reduction could also explain the resistance of P. blakesleeanus to high extracellular concentrations of vanadium.

Description: Proton transfer reaction mass spectrometry (PTR-MS) is a well-established technique for real-time analysis of volatile organic compounds (VOCs). Although it is extremely sensitive (with sensitivities of up to 4500 cps/ppbv, limits of detection 〈1 pptv and the response times of approximately 100 ms), the selectivity of PTR-MS is still somewhat limited, as isomers cannot be separated. Recently, selectivity-enhancing measures, such as manipulation of drift tube parameters (reduced electric field strength) and using primary ions other than H 3 O + , such as NO + and O 2 + , have been introduced. However, monoterpenes, which belong to the most important plant VOCs, still cannot be distinguished so more traditional technologies, such as gas chromatography mass spectrometry (GC-MS), have to be utilised. GC-MS is very time consuming (up to 1 h) and cannot be used for real-time analysis. Here, we introduce a sensitive, near-to-real-time method for plant monoterpene research—PTR-MS coupled with fastGC. We successfully separated and identified six of the most abundant monoterpenes in plant studies (α- and β-pinenes, limonene, 3-carene, camphene and myrcene) in less than 80 s, using both standards and conifer branch enclosures (Norway spruce, Scots pine and black pine). Five monoterpenes usually present in Norway spruce samples with a high abundance were separated even when the compound concentrations were diluted to 20 ppbv. Thus, fastGC-PTR-ToF-MS was shown to be an adequate one-instrument solution for plant monoterpene research.

Description: S -Nitrosoglutathione (GSNO) is a very important biomolecule that has crucial functions in many physiological and physiopathological processes. GSNO acts as NO donor and is a candidate for future medicines. This work describes, for the first time, the separation and the detection of GSNO and its decomposition products using capillary electrophoresis coupled to mass spectrometry (CE–MS). The separation was performed in slightly alkaline medium (pH 8.5) under positive-ionization MS detection. The identification of three byproducts of GSNO was formally performed for the first time: oxidized glutathione (GSSG), glutathione sulfinic acid (GSO 2 H), and glutathione sulfonic acid (GSO 3 H). GSO 2 H and GSO 3 H are known to have important biological activity, including inhibition of the glutathione transferase family of enzymes which are responsible for the elimination of many mutagenic, carcinogenic, and pharmacologically active molecules. We observed, after the ageing of GSNO in the solid state, that the proportion of both GSSG and GSO 3 H increases whereas that of GSO 2 H decreases. These results enabled us to propose an oxidation scheme explaining the formation of such products.

Description: A novel method was developed for d / l -isomeric separation of aldopentoses and aldohexoses as their ( S )-(+)-4-( N , N -dimethylaminosulfonyl)-7-(3-aminopyrrolidin-1-yl)-2,1,3-benzoxadiazole derivatives using phenylboronate buffer containing sodium dodecyl sulfate as a background electrolyte. The combination of derivatization with a chiral labeling reagent and micellar electrokinetic chromatography with phenylboronate made possible the efficient separation of d / l isomers as well as epimeric isomers of aldopentoses and aldohexoses. Laser-induced fluorescence detection permitted the micromolar-level determination of monosaccharide derivatives. The limit of detection was 105 amol (300 nM), and the repeatabilities of the migration times and peak area responses were 0.8 % and 7.9 % (relative standard deviation; n  = 6), respectively. The method was applied to the determination of d / l - galactose in red seaweed.

Description: The chemical additives used in fracturing fluids can be used as tracers of water contamination caused by hydraulic fracturing operations. For this purpose, a complete chemical characterization is necessary using advanced analytical techniques. Liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC/Q-TOF-MS) was used to identify chemical additives present in flowback and produced waters. Accurate mass measurements of main ions and fragments were used to characterize the major components of fracking fluids. Sodium adducts turned out to be the main molecular adduct ions detected for some additives due to oxygen-rich structures. Among the classes of chemical components analyzed by mass spectrometry include gels (guar gum), biocides (glutaraldehyde and alkyl dimethyl benzyl ammonium chloride), and surfactants (cocamidopropyl dimethylamines, cocamidopropyl hydroxysultaines, and cocamidopropyl derivatives). The capabilities of accurate mass and MS-MS fragmentation are explored for the unequivocal identification of these compounds. A special emphasis is given to the mass spectrometry elucidation approaches used to identify a major class of hydraulic fracturing compounds, surfactants.

Description: Enzyme immunoassays are currently the methods of choice for gluten control in foods labelled as gluten free , providing a mechanism for assessing food safety for consumption by coeliac and other allergic patients. However, their limitations, many of them associated to the reactivity of the different antibodies used and their degree of specificity, have prevented the establishment of a standardised method of analysis. We explore new methods for quantitatively determining gluten content in foods based on the use of two recently described aptamers, raised against a 33-mer peptide recognised as the immunodominant fragment from α2-gliadin. The assays use the target peptide immobilised onto streptavidin-coated magnetic beads in combination with a limited amount of biotin-aptamer in a competitive format, followed by streptavidin-peroxidase labelling of the aptamer that remains bound to the magnetic beads. The enzyme activity onto the beads, measured by chronoamperometry in disposable screen-printed electrodes, is inversely related to the target concentration in the test solution. We find that while the assay using the aptamer with the highest affinity towards the target (Gli 4) achieves low detection limits (~0.5 ppm) and excellent analytical performance, when challenged in samples containing the intact protein, gliadin, it fails in detecting the peptide in solution. This problem is circumvented by employing another aptamer (Gli 1), the most abundant one in the SELEX pool, as a receptor. The proposed assays allow the convenient detection of the allergen in different kinds of food samples, including heat-treated and hydrolysed ones. The obtained results correlate with those of commercially available antibody-based assays, providing an alternative for ensuring the safety and quality of nominally gluten-free foods. Graphical Abstract Electrochemical magnetoassay for gluten determination using biotin-aptamers as receptors

Description: Application of biochar-based fertilizers is increasingly being considered for its potential agronomic and environmental benefits. However, biochar may contain residues of polycyclic aromatic hydrocarbons (PAHs) as a result of its production by pyrolysis. The strong adsorption of PAHs to biochar makes extraction and analysis of biochar-based fertilizers difficult. This study optimizes the extraction of PAHs in biochar-based fertilizer samples by using an ultrasonic bath for quantification by gas chromatography–mass spectrometry. Among 12 solvents, acetone–cyclohexane (1:1) mixture was selected as the optimum solvent for extraction. Three variables affecting the extraction were studied by Box–Behnken design. The optimum conditions were 57 °C extraction temperature, 81 min extraction time, and two extraction cycles, which were validated by assessing the linearity of analysis, LOD, LOQ, recovery, and levels of PAHs in real biochar-based fertilizer samples. Results revealed that the 16 U.S. EPA PAHs had good linearity, with squared correlation coefficients greater than 0.99. LODs were low, ranging from 2.2 ng g −1 (acenaphthene) to 23.55 ng g −1 (indeno[1,2,3- cd ]perylene), and LOQs varied from 7.51 ng g −1 to 78.49 ng g −1 . The recoveries of 16 individual PAHs from the three biochar-based fertilizer samples were 81.8–109.4 %. Graphical Abstract Use of RSM to optimize UAE for extraction of the PAHs in biochar-based fertilizer

Description: Preparative temperature rising elution fractionation (prepTREF) is the standard technique for the preparative fractionation of polyolefins according to crystallisability. For olefin copolymers such as linear low-density polyethylene (LLDPE), it was believed that the TREF elution temperature correlates directly with the copolymer composition. For copolymers having different bulk comonomer contents, the prepTREF fractions of different samples collected at a given temperature were assumed to have the same chemical composition. It was acknowledged quite early that co-crystallisation effects may disturb TREF fractionation and fractions are obtained that are not completely homogeneous. This, however, has not been investigated quantitatively so far. The fundamental statement of prepTREF is challenged for the first time quantitatively using advanced analytical techniques including high-temperature high-performance liquid chromatography (HPLC). Ethylene-1-octene copolymers having bulk comonomer contents ranging from 0.3 to 6.4 mol% were fractionated by prepTREF, and the fractions were analysed by high-temperature size exclusion chromatography, crystallisation analysis fractionation, differential scanning calorimetry and high-temperature HPLC. All analytical results prove that the TREF fractions collected from different samples at the same elution temperature have different chemical compositions. The chemical compositions of the fractions correlate with the compositions of the bulk samples in that the comonomer contents of similar TREF fractions increase with an increase of the comonomer content of the bulk samples. These results are in clear contrast to the previous assumption that the TREF fraction composition is independent of the bulk copolymer composition for a given copolymer type. Graphical abstract Advanced analysis of LLDPE by combination of HT-HPLC, DSC and CRYSTAF

Description: The exploration of new chemical entities from herbal medicines may provide candidates for the in silico screening of drug leads. However, this significant work is hindered by the presence of multiple classes of plant metabolites and many re-discovered structures. This study presents an integrated strategy that uses ultrahigh-performance liquid chromatography/linear ion-trap quadrupole/Orbitrap mass spectrometry (UHPLC/LTQ-Orbitrap-MS) coupled with in-house library data for the systematic characterization and discovery of new potentially bioactive molecules. Exploration of the indole alkaloids from Uncaria rhynchophylla (UR) is presented as a model study. Initially, the primary characterization of alkaloids was achieved using mass defect filtering and neutral loss filtering. Subsequently, phytochemical isolation obtained 14 alkaloid compounds as reference standards, including a new one identified as 16,17-dihydro- O -demethylhirsuteine by NMR analyses. The direct-infusion fragmentation behaviors of these isolated alkaloids were studied to provide diagnostic structural information facilitating the rapid differentiation and characterization of four different alkaloid subtypes. Ultimately, after combining the experimental results with a survey of an in-house library containing 129 alkaloids isolated from the Uncaria genus, a total of 92 alkaloids (60 free alkaloids and 32 alkaloid O -glycosides) were identified or tentatively characterized, 56 of which are potential new alkaloids for the Uncaria genus. Hydroxylation on ring A, broad variations in the C-15 side chain, new N -oxides, and numerous O -glycosides, represent the novel features of the newly discovered indole alkaloid structures. These results greatly expand our knowledge of UR chemistry and are useful for the computational screening of potentially bioactive molecules from indole alkaloids. Graphical Abstract A four-step integrated strategy for the systematic characterization and efficient discovery of new indole alkaloids from Uncaria rhynchophylla

Description: Recent numerical studies in stratospheric dynamics and its variability as well as climate, have highlighted the need of more observational analyses to improve simulation of the West African monsoon (WAM). In this paper, activity and spectral characteristics of short-scale vertical waves (wavelengths 〈4 km) are analysed in equatorial coastal and tropical lower stratosphere during the WAM. A first detailed description of such waves over West Africa is derived from high-resolution vertical profiles of temperature and horizontal wind obtained during Intensive Observation Period of the African Monsoon Multidisciplinary Analyses (AMMA) Campaign 2006. Monthly variation of wave energy density is revealed to trace the progression of the inter-tropical convergence zone (ITCZ) over West Africa. Mesoscale inertia gravity-waves structures with vertical and horizontal wavelengths of 1.5–2.5 and 400–1100 km respectively and intrinsic frequencies of 1.1–2.2 f or periods 〈2 days are observed in the tropical LS with intense activity during July and August when the WAM is installed over the tropical West Africa. Over equatorial region, gravity waves with intrinsic frequencies of 1.4–4 f or periods 〈5.2 days, vertical wavelength of 2.1 km and long horizontal wavelengths of 1300 km are intense during the WAM coastal phase. From July to October, gravity waves with intrinsic frequencies of 1.2–3.8 f or periods 〈6 days, vertical wavelength of 2.1 km and horizontal wavelengths of 1650 km are less intense during the WAM Sahelian phase of the WAM, March–June. Unlike potential energy density, kinetic energy density is observed to be a good proxy for the activity of short-scale vertical waves during the WAM because quasi-inertial waves are dominant. Long-term wave activity variation from January 2001 to December 2009, highlights strong year-to-year variation superimposed on convective activity and quasi-biennial oscillation-like variations especially above tropical stations.

Description: Satellite based top-of-atmosphere (TOA) and surface radiation budget observations are combined with mass corrected vertically integrated atmospheric energy divergence and tendency from reanalysis to infer the regional distribution of the TOA, atmospheric and surface energy budget terms over the globe. Hemispheric contrasts in the energy budget terms are used to determine the radiative and combined sensible and latent heat contributions to the cross-equatorial heat transports in the atmosphere (AHT EQ ) and ocean (OHT EQ ). The contrast in net atmospheric radiation implies an AHT EQ from the northern hemisphere (NH) to the southern hemisphere (SH) (0.75 PW), while the hemispheric difference in sensible and latent heat implies an AHT EQ in the opposite direction (0.51 PW), resulting in a net NH to SH AHT EQ (0.24 PW). At the surface, the hemispheric contrast in the radiative component (0.95 PW) dominates, implying a 0.44 PW SH to NH OHT EQ . Coupled model intercomparison project phase 5 (CMIP5) models with excessive net downward surface radiation and surface-to-atmosphere sensible and latent heat transport in the SH relative to the NH exhibit anomalous northward AHT EQ and overestimate SH tropical precipitation. The hemispheric bias in net surface radiative flux is due to too much longwave surface radiative cooling in the NH tropics in both clear and all-sky conditions and excessive shortwave surface radiation in the SH subtropics and extratropics due to an underestimation in reflection by clouds.

Description: Mineral dust aerosols are an essential component of climate over West Africa, however, little work has been performed to investigate their contributions to potential climate change. A set of regional climate model experiments with and without mineral dust processes and land cover changes is performed to evaluate their climatic effects under the Representative Concentration Pathway 8.5 for two global climate models. Results suggest surface warming to be in the range of 4–8 °C by the end of the century (2081–2100) over West Africa with respect to the present day (1981–2000). The presence of mineral dusts dampens the warming by 0.1–1 °C in all seasons. Accounting for changes in land cover enhances the warming over the north of Sahel and dampens it to the south in spring and summer; however, the magnitudes are smaller than those resulting from dusts. Overall dust loadings are projected to increase, with the greatest increase occurring over the Sahara and Sahel in summer. Accounting for land cover changes tends to reduce dust loadings over the southern Sahel. Future precipitation is projected to decrease by 5–40 % in the western Sahara and Sahel and increase by 10–150 % over the eastern Sahel and Guinea Coast in JJA. A dipole pattern of future precipitation changes is attributed to dust effects, with decrease in the north by 5–20 % and increase by 5–20 % in the south. Future changes in land cover result in a noisy non-significant response with a tendency for slight wetting in MAM, JJA, and SON and drying in DJF.

Description: Phenylethanolamine A (PA) is a β-adrenergic agonist, which was first used in animal husbandry as a growth promoter in China in 2010. In this study, a monoclonal-antibody (mAb)-based indirect competitive enzyme-linked immunosorbent assay (icELISA) and lateral-flow immunoassay (LFA) for the detection of PA in swine urine and pork were developed. The immunogen was prepared by linking PA hapten with carrier protein via a diazotization method. The IC 50 value of the optimized icELISA was 0.44 ng mL −1 . The limits of detection of the icELISA for PA in swine urine and pork were 0.13 ng mL −1 and 0.39 ng g −1 , respectively. The recoveries of PA from spiked swine urine and pork were in the range 82.0–107.4 % and 81.8–113.3 %, respectively, with the coefficients of variation in the range 4.1–16.2 % and 1.2–6.3 %, respectively. The mAbs had negligible cross reactivity with 10 other β-agonists. In contrast, the LFA had a cut-off level of 5 ng mL −1 (g) in swine urine and pork, and the results could be achieved within 5 min. Ten blind samples of swine urine were analyzed simultaneously by icELISA, LFA, and ultra-high-performance liquid chromatography–tandem mass spectrometry, and the results of the three methods agreed well. Therefore, the combination of two immunoassays provides an effective and rapid screening method for detection of PA residues in biological samples. Graphical Abstract Determination of phenylethanolamine A in biological samples by immunological methods

Description: In a previous study Pons et al. (Clim Res 54(3):197–207, 2010 . doi: 10.3354/cr01117g ) reported a significant decreasing trend of snowfall occurrence in the Northern Iberian Peninsula since the mid 70s. The study was based on observations of annual snowfall frequency (measured as the annual number of snowfall days NSD) from a network of 33 stations ranging from 60 to 1350 m. In the present work we analyze the skill of Regional Climate Models (RCMs) to reproduce this trend for the period 1961–2000 (using both reanalysis- and historical GCM-driven boundary conditions) and the trend and the associated uncertainty of the regional future projections obtained under the A1B scenario for the first half of the twenty-first century. In particular, we consider the regional simulation dataset from the EU-funded ENSEMBLES project, consisting of thirteen state-of-the-art RCMs run at 25 km resolution over Europe. While ERA40 severely underestimates both the mean NSD and its observed trend (−2.2 days/decade), the corresponding RCM simulations driven by the reanalysis appropriately capture the interannual variability and trends of the observed NSD (trends ranging from −3.4 to −0.7, −2.1 days/decade for the ensemble mean). The results driven by the GCM historical runs are quite variable, with trends ranging from −8.5 to 0.2 days/decade (−1.5 days/decade for the ensemble mean), and the greatest uncertainty by far being associated with the particular GCM used. Finally, the trends for the future 2011–2050 A1B runs are more consistent and significant, ranging in this case from −3.7 to −0.5 days/decade (−2.0 days/decade for the ensemble mean), indicating a future significant decreasing trend. These trends are mainly determined by the increasing temperatures, as indicated by the interannual correlation between temperature and NSD (−0.63 in the observations), which is preserved in both ERA40- and GCM-driven simulations.

Description: The interannual-decadal variability of the wintertime mixed layer depths (MLDs) over the North Pacific is investigated from an empirical orthogonal function (EOF) analysis of an ensemble of global ocean reanalyses. The first leading EOF mode represents the interannual MLD anomalies centered in the eastern part of the central mode water formation region in phase opposition with those in the eastern subtropics and the central Alaskan Gyre. This first EOF mode is highly correlated with the Pacific decadal oscillation index on both the interannual and decadal time scales. The second leading EOF mode represents the MLD variability in the subtropical mode water (STMW) formation region and has a good correlation with the wintertime West Pacific (WP) index with time lag of 3 years, suggesting the importance of the oceanic dynamical response to the change in the surface wind field associated with the meridional shifts of the Aleutian Low. The above MLD variabilities are in basic agreement with previous observational and modeling findings. Moreover the reanalysis ensemble provides uncertainty estimates. The interannual MLD anomalies in the first and second EOF modes are consistently represented by the individual reanalyses and the amplitudes of the variabilities generally exceed the ensemble spread of the reanalyses. Besides, the resulting MLD variability indices, spanning the 1948–2012 period, should be helpful for characterizing the North Pacific climate variability. In particular, a 6-year oscillation including the WP teleconnection pattern in the atmosphere and the oceanic MLD variability in the STMW formation region is first detected.

Description: Real-time feedback about dissected tissue during the neurosurgical procedure is strongly requested. A novel direct ionization mass spectrometric method for identifying pathological differences in tissues is proposed. The method is based on simultaneous extraction of tissue lipids and electrospray ionization which allows mass spectrometric data to be obtained directly from soft tissues. The advantage of this method is the stable flow of solvent, which leads to stable time-dependent spectra. The tissues included necrotized tissues and tumor tissues in different combinations. Capability for direct analysis of samples of dissected tissues during the neurosurgical procedure is demonstrated. Data validation is conducted by compound identification using precise masses from the MS profile, MS/MS, and isotopic distribution structure analysis. The method can be upgraded and applied for real-time identification of tissues during surgery. This paper describes the technique and its application perspective. For these purposes, other methods were compared with the investigated one and the results were shown to be reproducible. Differences in lipid profiles were observed even in tissues from one patient where distinctions between different samples could be poor. The paper presents a proof of concept for the method to be applied in neurosurgery particularly and in tissue analysis generically. The paper also contains preliminary results proving the possibility of observing differences in mass spectra of different tumors.

Description: Immunoglobulin Y (IgY) is derived from egg yolk and has been identified as a cheap and high-yield immunoreagent. The application of IgY in immunoassays for the detection of chemical contaminants in food samples has rarely been reported. In this work, we describe a rapid and sensitive fluorescence polarization immunoassay (FPIA) for valnemulin (VAL) using IgY which was produced using a previously prepared immunogen. Three fluorophore-labeled VAL tracers were synthesized and the sensitivity of the best tracer (VAL–DTAF) in the optimized FPIA with antibody IgY 100 demonstrated an IC 50 value of 12 ng mL −1 in buffer. After evaluation of several extraction procedures, acidified acetonitrile was selected to extract VAL from swine tissue. The recoveries of VAL in spiked swine tissue at three levels (50, 100, and 200 μg kg −1 ) were higher than 79 % with coefficients of variation (CVs) lower than 12 %. The limit of detection (LOD) of the FPIA in swine tissue was 26 μg kg −1 and was lower than the maximum residue limit (MRL) of VAL set by the European Union. The study showed that IgY could be a good substitute for IgG when developing a high-throughput assay for chemical residues. Graphical Abstract FPIA based on IgY for rapid detection of Valnemulin in swine tissue

Description: In this paper, decadal variability of the Pacific-South America (PSA) mode is examined from year 1871 to 2008 based on the newly developed ocean and atmosphere reanalysis products. The PSA mode, mirroring the Pacific-North America mode in the Northern Hemisphere, emerges as the second EOF mode of 500 mb geopotential height anomalies. The mode displays substantial interannual-decadal variability with distinct timescales between 3–8 and 10–18 years, respectively. The decadal variability of the PSA mode is found to be associated with the coupled ocean–atmosphere interaction over the subtropical South and tropical Pacific. The subduction of the subtropical temperature anomalies in the South Pacific in conjunction with the tropical–subtropical atmospheric teleconnection plays important role in the decadal variability of the PSA mode.

Description: The National Center for Atmospheric Research Community Earth System Model is used to study the “spring predictability barrier” (SPB) problem for El Niño events from the perspective of initial error growth. By conducting perfect model predictability experiments, we obtain two types of initial sea temperature errors, which often exhibit obvious season-dependent evolution and cause a significant SPB when predicting the onset of El Niño events bestriding spring. One type of initial errors possesses a sea surface temperature anomaly (SSTA) pattern with negative anomalies in the central–eastern equatorial Pacific, plus a basin-wide dipolar subsurface temperature anomaly pattern with negative anomalies in the upper layers of the eastern equatorial Pacific and positive anomalies in the lower layers of the western equatorial Pacific. The other type consists of an SSTA component with positive anomalies over the southeastern equatorial Pacific, plus a large-scale zonal dipole pattern of the subsurface temperature anomaly with positive anomalies in the upper layers of the eastern equatorial Pacific and negative anomalies in the lower layers of the central–western equatorial Pacific. Both exhibit a La Niña-like evolving mode and cause an under-prediction for Niño-3 SSTA of El Niño events. For the former initial error type, the resultant prediction errors grow in a manner similar to the behavior of the growth phase of La Niña; while for the latter initial error type, they experience a process that is similar to El Niño decay and transition to a La Niña growth phase. Both two types of initial errors cause negative prediction errors of Niño-3 SSTA for El Niño events. The prediction errors for Niño-3 SSTA are mainly due to the contribution of initial sea temperature errors in the large-error-related regions in the upper layers of the eastern tropical Pacific and/or in the lower layers of the western tropical Pacific. These regions may represent ‘‘sensitive areas’’ for El Niño–Southern Oscillation (ENSO) predictions, thereby providing information for target observations to improve the forecasting skill of ENSO.

Description: Using Raman and infrared spectroscopy, we monitored spectral changes occurring in the blood plasma of patients with Alzheimer’s disease (AD) in relation to healthy controls. The protein secondary structure as reflected by amide I band involves β-sheet enrichment, which may be attributable to Aβ peptide formation and to increasing proportion of the globulins that are β-sheet rich. Likewise, the behavior of the infrared 1200–1000–cm −1 region and the Raman 980–910- and 450–400-cm −1 regions can be explained in terms of the said plasma composition change. Further, the 744-cm −1 Raman band from healthy control plasma shows frequency upshifting in the course of AD, which may be generated by the platelets collected in blood plasma. Linear discrimination analysis and receiver operating characteristic (ROC) analysis have been used to distinguish between patients with AD and age-matched healthy controls with a diagnostic accuracy of about 94 %. Graphical Abstract ROC assessment of sample classifications by Raman spectroscopy ( dashed line ) and linear combination of infrared and Raman spectroscopies ( solid line )

Description: Molecular imprinting is a very powerful synthetic method for preparation of robust materials with pre-designed molecular selectivity. Molecularly imprinted polymers (MIPs) are attractive substitutes for antibodies in many analytical and bioanalytical applications, e.g. for development of biosensors and for drug assays. In addition to selective molecular binding, new functions are being added to MIPs to make the synthetic materials responsive to different environmental conditions, making it possible to modulate the binding and release of different molecular targets and to simplify affinity separation. Introduction of signal-transduction functionality into MIPs also brings in new, more easily operated chemical sensors for detection and quantification of important analytical targets. Graphical Abstract Multi-functional molecularly imprinted polymers

Description: In the last few decades, MALDI-TOF MS has become a useful technique not only in proteomics, but also as a fast and specific tool for whole cell analysis through intact cell mass spectrometry (IC-MS). The present study evaluated IC-MS as a novel tool for the detection of distinct patterns that can be observed after exposure to a certain toxin or concentration by utilizing the eukaryotic fish cell line RTL-W1. Two different viability assays were performed to define the range for IC-MS investigations, each of which employing copper sulfate, acridine, and β-naphthoflavone (BNF) as model compounds for several classes of environmental toxins. The IC-MS of RTL-W1 cells revealed not only specific spectral patterns for the various toxins, but also that the concentration used had an effect on RTL-W1 profiles. After the exposure with copper sulfate and acridine, the spectra of RTL-W1 showed a significant increase of certain peaks in the higher mass range ( m / z 〉7000), which is probably attributed to the apoptosis of RTL-W1. On the contrary, exposure to BNF showed a distinct change of ion abundances only in the lower mass range ( m / z 〈7000). Furthermore, a set of mass peaks could be identified as a specific biomarker for a single toxin treatment, so IC-MS demonstrates a new method for the distinction of toxic effects in fish cells. Due to fast sample preparation and high throughput, IC-MS offers great potential for ecotoxicological studies to investigate cellular effects of different substances and complex environmental samples. Graphical Abstract Use of intact cell MALDI-TOF mass spectrometry (IC-MS) to detect and differentiate toxic effects of environmental toxins in rainbow trout liver cell line RTL-W1

Description: In this contribution, we investigated whether surface-enhanced Raman scattering (SERS) of serum can be a candidate method for detecting “luminal A” breast cancer (BC) at different stages. We selected three groups of participants aged over 50 years: 20 healthy women, 20 women with early localized small BC, and 20 women affected by BC with lymph node involvement. SERS revealed clear spectral differences between these three groups. A predictive model using principal component analysis (PCA) and linear discriminant analysis (LDA) was developed based on spectral data, and its performance was estimated with cross-validation. PCA-LDA of SERS spectra could distinguish healthy from BC subjects (sensitivity, 92 %; specificity, 85 %), as well as subjects with BC at different stages, with a promising diagnostic performance (sensitivity and specificity, ≥80 %; overall accuracy, 84 %). Our data suggest that SERS spectroscopy of serum, combined with multivariate data analysis, represents a minimally invasive, easy to use, and fast approach to discriminate healthy from BC subjects and even to distinguish BC at different clinical stages.

Description: In this study, the relationship between the El Niño-Southern Oscillation (ENSO) and precipitation variability over the Korean Peninsula is investigated. In contrast to the previously-known positive correlation between them during an El Niño developing summer and winter, we found a considerably significant negative correlation in September between Niño3 Sea Surface Temperature and Korean precipitation during ENSO developing phase. The northerly wind is only seen during El Nino developing phase and is part of the cyclonic flow over the subtropical North Pacific. The cyclonic flow over the subtropical North Pacific is induced by the subtropical diabatic heating, which is a peculiar feature during El Niño developing phase. In addition, it is demonstrated that the negative correlation is partly attributed to the tropical cyclone (TC), particularly during La Niña phase. That is, TC tends to pass through Korean Peninsula more frequently during La Niña years, which leads to more precipitation over the Korean Peninsula.

Description: Explosive cyclones are rapidly intensifying low pressure systems generating severe wind speeds and heavy precipitation primarily in coastal and marine environments. This study presents the first analysis on how explosive cyclones respond to climate change in the extratropics of the Northern Hemisphere. An objective-feature tracking algorithm is used to identify and track cyclones from 23 CMIP5 climate models for the recent past (1981–1999) and future (2081–2099). Explosive cyclones are projected to shift northwards by about \(2.2^\circ\) latitude on average in the northern Pacific, with fewer and weaker events south of \(45^\circ \hbox {N}\) , and more frequent and stronger events north of this latitude. This shift is correlated with a poleward shift of the jet stream in the inter-model spread ( \(R=0.56\) ). In the Atlantic, the total number of explosive cyclones is projected to decrease by about 17 % when averaging across models, with the largest changes occurring along North America’s East Coast. This reduction is correlated with a decline in the lower-tropospheric Eady growth rate ( \(R=0.51\) ), and is stronger for models with smaller frequency biases ( \(R=-0.65\) ). The same region is also projected to experience a small intensification of explosive cyclones, with larger vorticity values for models that predict stronger increases in the speed of the jet stream ( \(R=0.58\) ). This strengthening of the jet stream is correlated with an enhanced sea surface temperature gradient in the North Atlantic ( \(R=-0.63\) ). The inverse relationship between model bias and projection, and the role of model resolution are discussed.

Description: We identify the dynamical drivers of systematic changes in persistent quasi-stationary states (regimes) of the Southern Hemisphere troposphere and their secular trends. We apply a purely data-driven approach, whereby a multiscale approximation to nonstationary dynamical processes is achieved through optimal sequences of locally stationary fast vector autoregressive factor processes, to examine a high resolution atmospheric reanalysis over the period encompassing 1958–2013. This approach identifies regimes and their secular trends in terms of the predictability of the flow and is Granger causal. A comprehensive set of diagnostics on both isentropic and isobaric surfaces is employed to examine teleconnections over the full hemisphere and for a set of regional domains. Composite states for the hemisphere obtained from nonstationary nonparametric cluster analysis reveal patterns consistent with a circumglobal wave 3 (polar)–wave 5 (subtropical) pattern, while regional composites reveal the Pacific South American pattern and blocking modes. The respective roles of potential vorticity sources, stationary Rossby waves and baroclinic instability on the dynamics of these circulation modes are shown to be reflected by the seasonal variations of the waveguides, where Rossby wave sources and baroclinic disturbances are largely contained within the waveguides and with little direct evidence of sustained remote tropical influences on persistent synoptic features. Warm surface temperature anomalies are strongly connected with regions of upper level divergence and anticyclonic Rossby wave sources. The persistent states identified reveal significant variability on interannual to decadal time scales with large secular trends identified in all sectors apart from a region close to South America.

Description: The presented work reports a generic fluorescent aptasensing design employing carboxy-modified fluorescent particles as a signal-generating probe and magnetic particles as a solid separation support. Carboxy-modified fluorescent particles were used to modify the aptamer and act as a signal-generating probe. Magnetic beads were used as an immobilization surface to perform the function of a solid separation support. As a proof of concept, the assay was used to detect ochratoxin A (OTA). Fluorescent detection based on the displacement and competition format was performed, and the obtained results were compared. The competition-based assays were characterized with improved analytical characteristics as compared to those based on the displacement principle. The competitive fluorescent assays showed a high sensitivity where the detection limit and IC 50 were 0.005 and 7.2 nM respectively. The aptasensing platform was finally demonstrated for the detection of OTA in a beer sample. However, this is a generic approach that can be very easily extended to other matrixes to determine OTA. Additionally, the proposed concept of fluorescent particles as a signal-generating probe in combination with magnetic particles can also be integrated to other fluorescent-based affinity assays.

Description: Over the past decades, several in vitro methods have been tested for their ability to predict either human intestinal absorption (HIA) or penetration across the blood–brain barrier (BBB) of drugs. Micellar liquid chromatography (MLC) has been a successful approach for retention time measurements of drugs to establish models together with other molecular descriptors. Thus far, MLC approaches have only made use of commercial surfactants such as sodium dodecyl sulfate (SDS) and polyoxyethylene (23) lauryl ether (Brij35), which are not representative for the phospholipids present in human membranes. Miltefosine, a phosphocholine-based lipid, is presented here as an alternative surfactant for MLC measurements. By using the obtained retention factors and several computed descriptors for a set of 48 compounds, two models were constructed: one for the prediction of HIA and another for the prediction of penetration across the BBB expressed as log BB. All data were correlated to experimental HIA and log BB values, and the performance of the models was evaluated. Log BB prediction performed better than HIA prediction, although HIA prediction was also improved a lot (from 0.5530 to 0.7175) compared to in silico predicted HIA values.

Description: Aldosterone is a mineralocorticoid steroid hormone, the measurement of which in the clinical laboratory is principally performed for the investigation of primary hyperaldosteronism. Primary hyperaldosteronism is a specifically treatable and potentially curable form of hypertension, which typically presents as drug-resistant hypertension and, in up to 37 % of cases, hypokalemia. Accurate measurement of aldosterone concentration is essential for correct diagnosis. The serum concentrations of aldosterone are in the picomolar range and therefore sensitive aldosterone assays are required. With the advancement in instrumentation of LC-MS/MS, the picomolar range of aldosterone can be easily measured by the newer models, but for those with a less sensitive instrument, special technique for sample preparation to enhance assay sensitivity is required. This work described the use of charge-tagging for the picomolar measurement of serum aldosterone in a less sensitive LC-MS/MS instrument. The assay was linear up to 3000 pmol/L with lower limit of quantitation at 80 pmol/L. The mean relative recovery was 96.5 % with a range of 89.3–101.6 % for aqueous calibrators and the mean relative recovery was 94.8 % with a range of 87.5–101.4 % for serum calibrators. Intra-assay CVs range from 8.2 % to 11.3 %, and inter-assay CVs ranged from 8.5 % to 13.5 % at concentration range from 229 to 1720 pmol/L. The LC-MS/MS method compared well (y = 1.04x + 8.97) with the in-use radioimmunoassay method. There was no significant difference found ( p  = 0.7135) between results determined by LC-MS/MS and radioimmunoassay method.

Description: Molecular heterogeneity of cancer is a major obstacle in tumor diagnosis and treatment. To deal with this heterogeneity, a multidisciplinary combination of different analysis techniques is of urgent need because a combination enables the creation of a multimodal image of a tumor. Here, we develop a computational workflow in order to combine matrix-assisted laser desorption/ionization mass spectrometric (MALDI-MS) imaging and Raman microspectroscopic imaging for tissue based studies. The computational workflow can be used to confirm a spectral histopathology (SHP) based on one technique with another technique. In this contribution, we confirmed a Raman spectroscopic based SHP with MALDI-imaging. Owing to this combination, we could demonstrate, for a larynx carcinoma sample, that tissue types and different metabolic states could be extracted from the Raman spectra. Further investigations with the help of MALDI spectra yield a better characterization of variable epithelial differentiation and a better understanding of ongoing dysplastic alterations.

Description: A fuzzy chromatography mass spectrometric (FCMS) fingerprinting method combined with chemometric analysis has been established for rapid discrimination of whole-grain flour (WF) from refined wheat flour (RF). Bran, germ, endosperm, and WF from three local cultivars or purchased from a grocery store were studied. The state of refinement (whole vs. refined) of wheat flour was differentiated successfully by use of principal-components analysis (PCA) and soft independent modeling of class analogy (SIMCA), despite potential confounding introduced by wheat class (red vs. white; hard vs. soft) or resources (different brands). Twelve discriminatory variables were putatively identified. Among these, dihexoside, trihexoside, apigenin glycosides, and citric acid had the highest peak intensity for germ. Variable line plots indicated phospholipids were more abundant in endosperm. Samples of RF and WF from three cultivars (Hard Red, Hard White, and Soft White) were physically mixed to furnish 20, 40, 60, and 80 % WF of each cultivar. SIMCA was able to discriminate between 100 %, 80 %, 60 %, 40 %, and 20 % WF and 100 % RF. Partial least-squares (PLS) regression was used for prediction of RF-to-WF ratios in the mixed samples. When PLS models were used the relative prediction errors for RF-to-WF ratios were less than 6 %. Graphical Abstract Workflow of targeting discriminatory compounds by use of FCMS and chemometric analysis

Description: Ammonia in gastric juice is considered a potential biomarker for Helicobacter pylori infection and as a factor contributing to gastric mucosal injury. High ammonia concentrations are also found in patients with chronic renal failure, peptic ulcer disease, and chronic gastritis. Rapid and specific methods for ammonia detection are urgently required by the medical community. Here we present a method to detect ammonia directly in gastric juice based on Fourier transform infrared spectroscopy. The ammonia dissolved in biological liquid samples as ammonium ion was released in air as a gas by the shifting of the pH equilibrium of the ammonium/ammonia reaction and was detected in line by a Fourier transform infrared spectroscopy system equipped with a gas cell for the quantification. The method developed provided high sensitivity and selectivity in ammonia detection both in pure standard solutions and in a simulated gastric juice matrix over the range of diagnostic concentrations tested. Preliminary analyses were also performed on real gastric juice samples from patients with gastric mucosal injury and with symptoms of H. pylori infection, and the results were in agreement with the clinicopathology information. The whole analysis, performed in less than 10 min, can be directly applied on the sample without extraction procedures and it ensures high specificity of detection because of the ammonia fingerprint absorption bands in the infrared spectrum. This method could be easily used with endoscopy instrumentation to provide information in real time and would enable the endoscopist to improve and integrate gastroscopic examinations.

Description: Analytical methods allowing sensitive determination of reduced homocysteine (rHcy), one of the so-called biothiols, in human serum is a topic of growing interest due to its close relation to several human disorders, mainly cardiovascular diseases. Although most widely used analytical strategies to determine total Hcy involve derivatization by means of fluorescent labels, this work proposes the use of ebselen, a Se-containing labelling agent to derivatize the reactive sulfhydryl group of the Hcy molecule in its “free” reduced form, which is more likely to play different roles in disease pathogenesis. Optimization of the derivatization and separation conditions by high-performance liquid chromatography (HPLC) to isolate the excess of derivatizing reagent is carried out here using UV/VIS detection. Further, the study of the Se labelling reaction by electrospray ionization tandem mass spectrometry (ESI-MS/MS) provides a stoichiometry of the derivative of 1:1. The main advantage of using ebselen as a labelling agent is the presence of the Se atom in the molecule that allows the use of inductively coupled plasma mass spectrometry (ICP-MS) as a sensitive and selective Se detector. The coupling of HPLC with ICP-MS provided excellent features for the determination of Se-derivatized rHcy (detection limit of 9.6 nM) in real samples. Quantification was accomplished by using post-column isotope dilution (ID) of Se in serum samples, after precipitation of the main serum proteins. Quantitative results for “free” rHcy turned out to be around 0.18–0.22 μM in serum samples from healthy individuals that could be directly analyzed without sample preconcentration.

Description: A click reaction is reported here for the first time as a useful technique to control the conformational stability of natural organic matter (NOM) suprastructures. Click conjugates were successfully formed between a previously butynylated NOM hydrophobic fraction and a hydrophilic polyethylene glycol (PEG)-amino chain. The click products were shown by size exclusion chromatography (HPSEC) hyphenated with Orbitrap mass spectrometry (MS) in electrospray ionization (ESI) (+), while precursors were visible in ESI (−). Despite their increase in molecular weight, HPSEC elution of click conjugates occurred after that of precursors, thus showing their departure from the NOM supramolecular association. This indicates that the click-conjugated NOM molecules were varied in their hydrophilic and cationic character and lost the capacity to accommodate in the original hydrophobic suprastructures. The most abundant product had the C 16 H 30 O 5 N 4 formula, a click conjugate of butanoic acid, while other products were short-chained (C 4 –C 8 ) linear unsaturated and hydroxylated carboxylic acids. Tandem MS revealed formation of triazole rings in clicked conjugates and their two fragmentations at the ester and the C-N alkyl–aryl bonds. The behavior of NOM molecules modified by click chemistry confirms that hydrophobicity and ionic charge of humic molecules play a pivotal role in stabilizing intermolecular forces in NOM. Moreover, the versatility of the click reaction may become useful to decorate NOM molecules with a variety of substrates, in order to alter NOM conformational and chemical properties and diversify its applications in the environment.