Surface Modification of Kraft Lignin by Mechanochemical Processing with Sodium Percarbonate.

In this article, we present a novel one-pot mechanochemical reaction for the surface activation of lignin. The process involves environmentally friendly oxidation with hydrogen peroxide, depolymerization of fractions with high molecular mass, and introduction of new carbonyl functions into the lignin backbone. Kraft lignin was ground with sodium percarbonate and sodium hydroxide in a ball mill at different time intervals. Analyses by infrared spectroscopy (IR), nuclear magnetic resonance spectroscopy (NMR), size exclusion chromatography (SEC), dynamic vapor sorption (DVS), and small-angle X-ray scattering (SAXS) showed significant improvements. After only 5 min of reaction, there was a 47% reduction in mass-average molecular weight and an increase in carboxyl functionalities. Chemical activation resulted in an approximately 2.8-fold increase in water adsorption. Principal component analysis (PCA) provided further insight into the correlations between IR spectra and SAXS parameters.

Simultaneous characterization of poly(acrylic acid) and polysaccharide polymers and copolymers.

Copolymer products that result from grafting acrylic acid and other hydrophilic monomers onto polysaccharides have recently gained significant interest in research and industry. Originating from renewable sources, these biodegradable, low toxicity, and polar copolymer products exhibit potential to replace polymers from fossil sources in several applications and industries. The methods usually employed to characterize these copolymers are, however, quite limited, especially for the measurement of bulk properties. With more sophisticated applications, for example, in pharmaceutics requiring a more detailed analysis of the chemical structure, we describe a new approach for this kind of complex polymers. Our approach utilizes chromatography in combination with several detection methods to separate and characterize reaction products of the copolymerization of acrylic acid and chemically hydrolyzed starch. These samples consisted of a mixture of homopolymer poly (acrylic acid), homopolymer hydrolyzed starch, and - in a lower amount - the formed copolymers. Several chromatographic methods exist that are capable of characterizing either poly (acrylic acid) or hydrolyzed starch. In contrast, our approach offers simultaneous characterization of both polymers. The combination of LC and UV/RI offered insight into the composition and copolymer content of the samples. Size exclusion chromatography experiments revealed the molar mass distribution of homopolymers and copolymers. FTIR investigations confirmed the formation of copolymers while ESI-MS gave more details on the end groups of hydrolyzed starches and poly (acrylic acids). Evidence of copolymer structures was obtained through NMR measurements. Finally, two-dimensional chromatography led to the separation of the copolymers from both homopolymers as well as the additional separation of sodium clusters. The methods described in this work are a powerful toolset to characterize copolymerization products of hydrolyzed starch and poly(acrylic acid). Together, our approach successfully correlates the physicochemical properties of such complex mixtures with their actual composition.

Covalently Fluorophore-Functionalized ZIF-8 Colloidal Particles as a Sensing Platform for Endocrine-Disrupting Chemicals Such as Phthalates Plasticizers.

We present the optical sensing of phthalate esters (PAEs), a group of endocrine-disrupting chemicals. The sensing takes place as changes in the fluorescence emission intensity of aminopyrene covalently bound to the organic ligands of the metal-organic framework compound ZIF-8. In the presence of PAEs, a quenching of the fluorescence emission is observed. We evaluated strategies to engineer colloidal size distribution of the sensing particles to optimize the sensory response to PAEs. A thorough characterization of the modified ZIF-8 nanoparticles included powder X-ray diffractometry, transmission electron microscopy, high-performance liquid chromatography, and photophysical characterization. The presented capability of the fluorophore-functionalized ZIF-8 to sense PAEs complements established methods such as chromatography-based procedures, which cannot be used on-site and paves the way for future developments such as hand-held quick sensing devices.

Power of Ultra Performance Liquid Chromatography/Electrospray Ionization-MS Reconstructed Ion Chromatograms in the Characterization of Small Differences in Polymer Microstructure.

From simple homopolymers to functionalized, 3-dimensional structured copolymers, the complexity of polymeric materials has become more and more sophisticated. With new applications, for instance, in the semiconductor or pharmaceutical industry, the requirements for the characterization have risen with the complexity of the used polymers. For each additional distribution, an additional dimension in analysis is needed. Small, often isomeric heterogeneities in topology or microstructure can usually not be simply separated chromatographically or distinguished by any common detector but affect the properties of materials significantly. For a drug delivery system, for example, the degree of branching and branching distribution is crucial for the formation of micelles. Instead of a complicated, time-consuming, and/or expensive 2D-chromatography or ion mobility spectrometry (IMS) method, that also has its limitations, in this work, a simple approach using size exclusion chromatography (SEC) coupled with electrospray ionization (ESI) mass spectrometry is proposed. The online coupling allows the analysis of reconstructed ion chromatograms (RICs) of each degree of polymerization. While a complete separation often cannot be achieved, the derived retention times and peak widths lead to information on the existence and dispersity of heterogeneities. Although some microstructural heterogeneities like short chain branching can for large polymers be characterized with methods such as light scattering, for oligomers where the heterogeneities just start to form and their influence is at the maximum, they are inaccessible with these methods. It is also shown that with a proper calibration even quantitative information can be obtained. This method is suitable to detect small differences in, e.g., branching, 3D-structure, monomer sequence, or tacticity and could potentially be used in routine analysis to quickly determine deviations.

Comparison of different methods for MP detection: What can we learn from them, and why asking the right question before measurements matters?

In recent years, an increasing trend towards investigating and monitoring the contamination of the environment by microplastics (MP) (plastic pieces < 5 mm) has been observed worldwide. Nonetheless, a reliable methodology that would facilitate and automate the monitoring of MP is still lacking. With the goal of selecting practical and standardized methods, and considering the challenges in microplastics detection, we present here a critical evaluation of two vibrational spectroscopies, Raman and Fourier transform infrared (FTIR) spectroscopy, and two extraction methods: thermal extraction desorption gas chromatography mass spectrometry (TED-GC-MS) and liquid extraction with subsequent size exclusion chromatography (SEC) using a soil with known contents of PE, PP, PS and PET as reference material. The obtained results were compared in terms of measurement time, technique handling, detection limits and requirements for sample preparation. The results showed that in designing and selecting the right methodology, the scientific question that determines what needs to be understood is significant, and should be considered carefully prior to analysis. Depending on whether the object of interest is quantification of the MP particles in the sample, or merely a quick estimate of sample contamination with plastics, the appropriate method must be selected. To obtain overall information about MP in environmental samples, the combination of several parallel approaches should be considered.

Critical Conditions for Liquid Chromatography of Statistical Copolymers: Functionality Type and Composition Distribution Characterization by UP-LCCC/ESI-MS.

Statistical ethylene oxide (EO) and propylene oxide (PO) copolymers of different monomer compositions and different average molar masses additionally containing two kinds of end groups (FTD) were investigated by ultra high pressure liquid chromatography under critical conditions (UP-LCCC) combined with electrospray ionization time-of flight mass spectrometry (ESI-TOF-MS). Theoretical predictions of the existence of a critical adsorption point (CPA) for statistical copolymers with a given chemical and sequence distribution1 could be studied and confirmed. A fundamentally new approach to determine these critical conditions in a copolymer, alongside the inevitable chemical composition distribution (CCD), with mass spectrometric detection, is described. The shift of the critical eluent composition with the monomer composition of the polymers was determined. Due to the broad molar mass distribution (MMD) and the presumed existence of different end group functionalities as well as monomer sequence distribution (MSD), gradient separation only by CCD was not possible. Therefore, isocratic separation conditions at the CPA of definite CCD fractions were developed. Although the various present distributions partly superimposed the separation process, the goal of separation by end group functionality was still achieved on the basis of the additional dimension of ESI-TOF-MS. The existence of HO-H besides the desired allylO-H end group functionalities was confirmed and their amount estimated. Furthermore, indications for a MSD were found by UPLC/MS/MS measurements. This approach offers for the first time the possibility to obtain a fingerprint of a broad distributed statistical copolymer including MMD, FTD, CCD, and MSD.

Ellman's and Aldrithiol Assay as Versatile and Complementary Tools for the Quantification of Thiol Groups and Ligands on Nanomaterials.

Simple, fast, and versatile methods for the quantification of thiol groups are of considerable interest not only for protein analysis but also for the characterization of the surface chemistry of nanomaterials stabilized with thiol ligands or bearing thiol groups for the subsequent (bio-) functionalization via maleimide-thiol chemistry. Here, we compare two simple colorimetric assays, the widely used Ellman's assay performed at alkaline pH and the aldrithiol assay executed at acidic and neutral pH, with respect to their potential for the quantification of thiol groups and thiol ligands on different types of nanoparticles like polystyrene nanoparticles, semiconductor nanocrystals (SC NC), and noble metal particles, and we derive criteria for their use. In order to assess the underlying reaction mechanisms and to obtain stoichiometry factors mandatory for reliable thiol quantification, both methods were studied photometrically and with electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS), thereby demonstrating the influence of different thiols on the reaction mechanism. Our results underline the suitability of both methods for the quantification of directly accessible thiol groups or ligands on the surface of 2D- and 3D-supports, here exemplarily polystyrene nanoparticles. Moreover, we could derive strategies for the use of these simple assays for the determination of masked (i.e., not directly accessible) thiol groups like disulfides such as lipoic acid and thiol stabilizing ligands coordinatively bound to Cd and/or Hg surface atoms of II/VI and ternary SC NC and to gold and silver nanoparticles.

Matrix-Assisted Ionization-Ion Mobility Spectrometry-Mass Spectrometry: Selective Analysis of a Europium-PEG Complex in a Crude Mixture.

The analytical utility of a new and simple to use ionization method, matrix-assisted ionization (MAI), coupled with ion mobility spectrometry (IMS) and mass spectrometry (MS) is used to characterize a 2-armed europium(III)-containing poly(ethylene glycol) (Eu-PEG) complex directly from a crude sample. MAI was used with the matrix 1,2-dicyanobenzene, which affords low chemical background relative to matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI). MAI provides high ion abundance of desired products in comparison to ESI and MALDI. Inductively coupled plasma-MS measurements were used to estimate a maximum of 10% of the crude sample by mass was the 2-arm Eu-PEG complex, supporting evidence of selective ionization of Eu-PEG complexes using the new MAI matrix, 1,2-dicyanobenzene. Multiply charged ions formed in MAI enhance the IMS gas-phase separation, especially relative to the singly charged ions observed with MALDI. Individual components are cleanly separated and readily identified, allowing characterization of the 2-arm Eu-PEG conjugate from a mixture of the 1-arm Eu-PEG complex and unreacted starting materials. Size-exclusion chromatography, liquid chromatography at critical conditions, MALDI-MS, ESI-MS, and ESI-IMS-MS had difficulties with this analysis, or failed. Graphical Abstract ᅟ.

Quantification of PEG-maleimide ligands and coupling efficiencies on nanoparticles with Ellman's reagent.

The surface modification of nanometer- and micrometer-sized particles and planar substrates with polyethylene glycol (PEG) ligands of varying length is a very common strategy to tune the hydrophilicity and biocompatibility of such materials, minimize unspecific interactions, improve biofunctionalization efficiencies, and enhance blood circulation times. Nevertheless, simple methods for the quantification of PEG ligands are comparatively rare. Here, we present a new concept for the quantification of PEG ligands for maleimide-functionalized PEG molecules and the determination of PEG coupling efficiencies, exploiting the quantitative reaction of maleimide with l-cysteine, and the subsequent determination of the unreacted thiol with the photometric Ellman's test. This is shown for heterobifunctional PEG spacers of varying length and amino-functionalized polystyrene nanoparticles (PS NP) without and with differently charged encoding dyes. The reaction of l-cysteine with the Ellman's reagent was monitored photometrically and with electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS) to derive the reaction mechanism and to obtain the stoichiometry factor for l-cysteine quantification. Mass balances and quantification of l-cysteine via its sulfur concentration using elemental analysis and inductively coupled plasma mass spectrometry (ICP-MS) confirmed the accuracy and reliability of this approach that can be extended to other surface groups and ligands.

Vibrational density of states of triphenylene based discotic liquid crystals: dependence on the length of the alkyl chain.

The vibrational density of states of a series of homologous triphenylene-based discotic liquid crystals HATn (n = 5, 6, 8, 10, 12) depending on the length of the aliphatic side chain is investigated by means of inelastic neutron scattering. All studied materials have a plastic crystalline phase at low temperatures, followed by a hexagonally ordered liquid crystalline phase at higher temperatures and a quasi isotropic phase at the highest temperatures. The X-ray scattering pattern for the plastic crystalline phase of all materials shows a sharp Bragg reflection corresponding to the intercolumnar distance in the lower q-range and a peak at circa 17 nm(-1) related to intracolumnar distances between the cores perpendicular to the columns as well as a broad amorphous halo related to the disordered structure of the methylene groups in the side chains in the higher q-range. The intercolumnar distance increases linearly with increasing chain length for the hexagonal columnar ordered liquid crystalline phase. A similar behaviour is assumed for the plastic crystalline phase. Besides n = 8 all materials under study exhibit a Boson peak. With increasing chain length, the frequency of the Boson peak decreases and its intensity increases. This can be explained by a self-organized confinement model. The peaks for n = 10, 12 are much narrower than for n = 5, 6 which might imply the transformation from a rigid system to a softer one with increasing chain length. Moreover the results can also be discussed in the framework of a transition from an uncorrelated to a correlated disorder with increasing n where n = 8 might be speculatively considered as a transitional state.

Elucidation of the structure of poly(γ-benzyl-l-glutamate) nanofibers and gel networks in a helicogenic solvent.

The synthesis, characterization, self-assembly, and gel formation of poly(γ-benzyl-l-glutamate) (PBLG) in a molecular weight range from ca. 7,000-100,000 g/mol and with narrow molecular weight distribution are described. The PBLG is synthesized by the nickel-mediated ring-opening polymerization and is characterized by size-exclusion chromatography coupled with multiple-angle laser light scattering, NMR, and Fourier transform infrared spectroscopy. The self-assembly and thermoreversible gel formation in the helicogenic solvent toluene is investigated by transmission electron microscopy, atomic force microscopy, small-angle X-ray scattering, and synchrotron powder X-ray diffraction. At concentrations significantly below the minimum gelation concentration, spherical aggregates are observed. At higher concentrations, gels are formed, which show a 3D network structure composed of nanofibers. The proposed self-assembly mechanism is based on a distorted hexagonal packing of PBLG helices parallel to the axis of the nanofiber. The gel network forms due to branching and rejoining of bundles of PBLG nanofibers. The network exhibits uniform domains with a length of 200 ± 42 nm composed of densely packed PBLG helices.

LC-MALDI-TOF imaging MS: a new approach in combining chromatography and mass spectrometry of copolymers.

A new approach that utilizes MALDI-TOF imaging mass spectrometry as a new detector for polymer chromatography is presented. For the first time, the individual retention behavior of single structural units of polyethylene oxide (PEO)/polypropylene oxide (PPO) copolymers and changes of the copolymer composition could be monitored. Composition specific calibration curves could be easily obtained by displaying the copolymer ion intensity data. This approach provides completely new insights in the chromatographic principle of copolymer separation and could be used to easily modify and adapt conditions for separation. In combination with electrospray deposition, homogeneous sample/matrix traces of surprisingly high spatial resolution could be obtained.

Cetirizine as pH-dependent cross-reactant in a carbamazepine-specific immunoassay.

High performance liquid chromatography (HPLC) was hyphenated with a previously reported carbamazepine-specific enzyme-linked immunosorbent assay (ELISA) as a screening approach to water analysis in order to identify possible interferences from transformation products. Treated wastewater was analysed and three substances were recognized by the antibody besides carbamazepine: the metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 2-hydroxycarbamazepine plus the structurally not obviously related antihistamine cetirizine. The molar cross-reactivity against cetirizine was found to be pH-dependent and assessed to be 400% at pH 4.5 and 22% at pH 10.5. Performing the ELISA at pH 10.5 greatly improved the accuracy when carbamazepine was determined in surface and wastewater samples.

Controlled folding of synthetic polymer chains through the formation of positionable covalent bridges.

Covalent bridges play a crucial role in the folding process of sequence-defined biopolymers. This feature, however, has not been recreated in synthetic polymers because, apart from some simple regular arrangements (such as block co-polymers), these macromolecules generally do not exhibit a controlled primary structure--that is, it is difficult to predetermine precisely the sequence of their monomers. Herein, we introduce a versatile strategy for preparing foldable linear polymer chains. Well-defined polymers were synthesized by the atom transfer radical polymerization of styrene. The controlled addition of discrete amounts of protected maleimide at precise times during the synthesis enabled the formation of polystyrene chains that contained positionable reactive alkyne functions. Intramolecular reactions between these functions subsequently led to the formation of different types of covalently folded polymer chains. For example, tadpole (P-shaped), pseudocyclic (Q-shaped), bicyclic (8-shaped) and knotted (α-shaped) macromolecular origamis were prepared in a relatively straightforward manner.

Structure and end-group analysis of complex hexanediol-neopentylglycol-adipic acid copolyesters by matrix-assisted laser desorption/ionization collision-induced dissociation tandem mass spectrometry.

Sequences and end groups of complex copolyesters were determined by fragmentation analysis by means of matrix-assisted laser desorption/ionization collision-induced dissociation tandem mass spectrometry (MALDI CID MS/MS). The complexity of the crude copolyester mixture was reduced by a chromatographic separation followed by a MALDI time-of-flight (TOF) investigation of fractions. Due to overlapping compositional and end-group information a clear assignment of end groups was very difficult. However, the fragmentation of suitable precursor ions resulted in typical fragment ion patterns and, therefore, enabled a fast and unambiguous determination of the end groups and composition of this important class of polymers.

Imaging mass spectrometry for examining localization of polymeric composition in matrix-assisted laser desorption/ionization samples.

The localization of polymeric composition in samples prepared for matrix-assisted laser desorption/ionization (MALDI) analysis has been investigated by imaging mass spectrometry. Various matrices and solvents were used for sample spot preparation of a polybutyleneglycol (PBG 1000). It was shown that in visibly homogeneous spots, prepared using the 'dried droplet' method, separation between matrix and polymer takes place. Moreover, using alpha-cyano-4-hydroxycinnamic acid (CCA) as matrix and methanol as solvent molecular mass separation of the polymer homologues in the spots was detectable. In contrast to manually spotted samples, dry spray deposition results in homogeneous layers showing no separation effects.

Determination of critical conditions of adsorption for chromatography of polymers.

Liquid chromatography (LC) at critical conditions of adsorption was used to separate various poly(ethylene oxides), poly(propylene oxides) and their copolymers. For the first time, the determination of the critical conditions by means of Ultra Performance Liquid Chromatography (UPLC) coupled to Electrospray Ionization Time-of-flight Mass Spectrometry (ESI-TOF MS) is reported. In contrast to established, mostly laborious routines to find suitable chromatographic separation conditions, this coupling enables a very fast adjustment of parameters. Similar to LC Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (LC/MALDI MS) coupling, a two-dimensional analysis of homo- and copolymers regarding its functionality type and molecular weight distribution, as well as copolymer composition, can be performed simultaneously. Furthermore, there is no need for using polymer standards for the determination of critical conditions or Size Exclusion Chromatography calibration.

Fractionation and solvent-free MALDI-MS analysis of polymers using liquid adsorption chromatography at critical conditions in combination with a multisample on-target homogenization/transfer sample preparation method.

A solvent-free homogenization/transfer matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) method is described for the preparation and precise transfer of up to 100 samples simultaneously on a single MALDI plate. This method is demonstrated using a poly(ethylene oxide) (PEO) mixture consisting of different molecular weights (500-6000) and end groups (PEO, dimethoxy-PEO, monomethoxy monomethacrylate-PEO, and dimethacrylate-PEO) that was fractionated using liquid adsorption chromatography at critical conditions. Off-line fractionation is performed prior to the on-target homogenization/transfer solvent-free sample preparation and MALDI mass analysis. The miniaturization of the solvent-free MALDI approach allowed analysis of less than 2 microg per PEO component per fraction corresponding to approximately 200 pmol for PEO 6000. The amounts of polymer sample used for LC separation and the quality of the MS results are equivalent to the "dry spray" method; however, three times more fractions were collected and analyzed with the newly developed hyphenated approach. The off-line method eliminates optimization of, for example, spray conditions or spreading of organic solvents on the MALDI plate that occurs with droplet deposition methods. The widespread applications of MALDI make this solvent-free, multisample method particularly important as it expands the capabilities for obtaining mass measurements with great efficiencies in areas with increased sample numbers. In addition, the solvent-free method is well suited for automated MALDI analysis as it virtually eliminates the "dead-spot" phenomenon.

A novel software tool for copolymer characterization by coupling of liquid chromatography with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

The results of copolymer characterization by coupling of chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) techniques and subsequent calculation of copolymer composition using a novel software tool 'MassChrom2D' are presented. For high-resolution mass analysis copolymer samples were fractionated by means of liquid adsorption chromatography (LAC). These fractions were investigated off-line by MALDI-TOF MS. Various mono-n-butyl ethers of polyethylene oxide-polypropylene oxide copolymers (PEO-co-PPO) were investigated. As well as the copolymer composition presented in two-dimensional plots, the applied approach can give additional hints on specific structure-dependent separation conditions in chromatography.

Principle of two-dimensional characterization of copolymers.

Two-dimensional polymer characterization is used for a simultaneous analysis of molar masses and chemical heterogeneities (e.g., end groups, copolymer composition, etc.). This principle is based on coupling of two different chromatographic modes. Liquid adsorption chromatography at critical conditions (LACCC) is applied for a separation according to the chemical heterogeneity, whereas in the second-dimension fractions are analyzed with regard to their molar mass distribution by means of size exclusion chromatography (SEC). Because appropriate standards for a calibration of the SEC are seldom available, matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) was used to substitute the SEC. The LACCC-MALDI MS coupling enables acquiring additional structural information on copolymer composition, which can considerably enhance the performance of this coupled method.