Examining functional group-dependent effects on the ionization of lignin monomers using supercritical fluid chromatography/electrospray ionization mass spectrometry.

The chemical and biological conversion of biomass-derived lignin is a promising pathway for producing valuable low molecular weight aromatic chemicals, such as vanillin or guaiacol, known as lignin monomers (LMs). Various methods employing chromatography and electrospray ionization-mass spectrometry (ESI-MS) have been developed for LM analysis, but the impact of LM chemical properties on analytical performance remains unclear. This study systematically optimized ESI efficiency for 24 selected LMs, categorized by functionality. Fractional factorial designs were employed for each LM to assess ESI parameter effects on ionization efficiency using ultra-high-performance supercritical fluid chromatography/ESI-MS (UHPSFC/ESI-MS). Molecular descriptors were also investigated to explain variations in ESI parameter responses and chromatographic retention among the LMs. Structural differences among LMs led to complex optimal ESI settings. Notably, LMs with two methoxy groups benefited from higher gas and sheath gas temperatures, likely due to their lower log P and higher desolvation energy requirements. Similarly, vinyl acids and ketones showed advantages at elevated gas temperatures. The retention in UHPSFC using a diol stationary phase was correlated with the number of hydrogen bond donors. In summary, this study elucidates structural features influencing chromatographic retention and ESI efficiency in LMs. The findings can aid in developing analytical methods for specific technical lignins. However, the absence of an adequate number of LM standards limits the prediction of LM structures solely based on ESI performance data.

Investigating Lignin-Derived Monomers and Oligomers in Low-Molecular-Weight Fractions Separated from Depolymerized Black Liquor Retentate by Membrane Filtration.

Base-catalyzed depolymerization of black liquor retentate (BLR) from the kraft pulping process, followed by ultrafiltration, has been suggested as a means of obtaining low-molecular-weight (LMW) compounds. The chemical complexity of BLR, which consists of a mixture of softwood and hardwood lignin that has undergone several kinds of treatment, leads to a complex mixture of LMW compounds, making the separation of components for the formation of value-added chemicals more difficult. Identifying the phenolic compounds in the LMW fractions obtained under different depolymerization conditions is essential for the upgrading process. In this study, a state-of-the-art nontargeted analysis method using ultra-high-performance supercritical fluid chromatography coupled to high-resolution multiple-stage tandem mass spectrometry (UHPSFC/HRMSn) combined with a Kendrick mass defect-based classification model was applied to analyze the monomers and oligomers in the LMW fractions separated from BLR samples depolymerized at 170-210 °C. The most common phenolic compound types were dimers, followed by monomers. A second round of depolymerization yielded low amounts of monomers and dimers, while a high number of trimers were formed, thought to be the result of repolymerization.

Identification of Phlorotannins in the Brown Algae, Saccharina latissima and Ascophyllum nodosum by Ultra-High-Performance Liquid Chromatography Coupled to High-Resolution Tandem Mass Spectrometry.

Phlorotannins are bioactive polyphenols in brown macroalgae that make these algae interesting as healthy food. Specific phlorotannins are, however, seldom identified, and extracts from different species are often only analysed for total phenolic content (TPC). In this study, our focus was to identify phlorotannin molecules from Saccharina latissima and Ascophyllum nodosum (a species rich in these compounds) using ultra-high-performance liquid chromatography coupled to high-resolution tandem mass spectrometry (UHPLC-HRMS2). Water and ethanol (30 and 80% v/v) were used at solid:liquid ratios, extraction times and temperatures, proposed to result in high TPC in extracts from other species. The S. latissima extracts, however, did not allow phlorotannin detection by either UHPLC-UV/Vis or UHPLC-HRMS2, despite a TPC response by the Folin-Ciocalteu assay, pinpointing a problem with interference by non-phenolic compounds. Purification by solid phase extraction (SPE) led to purer, more concentrated fractions and identification of four phlorotannin species in A. nodosum and one in S. latissima by UHPLC-HRMS2, using extracts in ethanol 80% v/v at a solid:liquid ratio of 1:10 for 20 h at 25 °C with an added 10 h at 65 °C incubation of remaining solids. The phlorotannin with the formula C12H10O7 (corresponding to bifuhalol) is the first identified in S. latissima.

Identification of lignin oligomers in Kraft lignin using ultra-high-performance liquid chromatography/high-resolution multiple-stage tandem mass spectrometry (UHPLC/HRMSn).

Kraft lignin is the main source of technically produced lignin. For the development of valuable products based on Kraft lignin, its molecular structure is important. However, the chemical composition of Kraft lignin is still not well known. So far, the analysis of Kraft lignin by mass spectrometry (MS) has been mainly focused on monomeric compounds. Previous MS studies on lignin oligomers (LOs) considered only synthesised LO standards and/or lignins produced by processes other than the Kraft process. Furthermore, published MS methods suffer from using high resolution only in the MS1 stage in multiple-stage tandem MS methods. A high resolution in all MSn stages would provide more detailed information about LO fragmentation pathways. Since lignin samples are complex mixtures of a large number of similar phenolic compounds, the selection of tentative LOs in the MS data is challenging. In this study, we present a method for non-targeted analysis of LOs in Kraft lignin using ultra-high-performance liquid chromatography/high-resolution multiple-stage tandem mass spectrometry (UHPLC/HRMSn). A pre-selection strategy for LOs has been established based on a data-dependent neutral loss MS3 method in combination with a principal component analysis-quadratic discriminant analysis classification model (PCA-QDA). The method was optimised using a design of experiments (DOE) approach. The developed approach improved the pre-selection of tentative LOs in complex mixtures. From 587 detected peaks, 36 peaks were identified as LOs. Graphical abstract ᅟ.

A sediment extraction and cleanup method for wide-scope multitarget screening by liquid chromatography-high-resolution mass spectrometry.

Previous studies on organic sediment contaminants focused mainly on a limited number of highly hydrophobic micropollutants accessible to gas chromatography using nonpolar, aprotic extraction solvents. The development of liquid chromatography-high-resolution mass spectrometry (LC-HRMS) permits the spectrum of analysis to be expanded to a wider range of more polar and ionic compounds present in sediments and allows target, suspect, and nontarget screening to be conducted with high sensitivity and selectivity. In this study, we propose a comprehensive multitarget extraction and sample preparation method for characterization of sediment pollution covering a broad range of physicochemical properties that is suitable for LC-HRMS screening analysis. We optimized pressurized liquid extraction, cleanup, and sample dilution for a target list of 310 compounds. Finally, the method was tested on sediment samples from a small river and its tributaries. The results show that the combination of 100 °C for ethyl acetate-acetone (50:50, neutral extract) followed by 80 °C for acetone-formic acid (100:1, acidic extract) and methanol-10 mM sodium tetraborate in water (90:10, basic extract) offered the best extraction recoveries for 287 of 310 compounds. At a spiking level of 1 µg mL-1, we obtained satisfactory cleanup recoveries for the neutral extract-(93 ± 23)%-and for the combined acidic/basic extracts-(42 ± 16)%-after solvent exchange. Among the 69 compounds detected in environmental samples, we successfully quantified several pharmaceuticals and polar pesticides.

Ultra-high-performance supercritical fluid chromatography with quadrupole-time-of-flight mass spectrometry (UHPSFC/QTOF-MS) for analysis of lignin-derived monomeric compounds in processed lignin samples.

The conversion of lignin to potentially high-value low molecular weight compounds often results in complex mixtures of monomeric and oligomeric compounds. In this study, a method for the quantitative and qualitative analysis of 40 lignin-derived compounds using ultra-high-performance supercritical fluid chromatography coupled to quadrupole-time-of-flight mass spectrometry (UHPSFC/QTOF-MS) has been developed. Seven different columns were explored for maximum selectivity. Makeup solvent composition and ion source settings were optimised using a D-optimal design of experiment (DoE). Differently processed lignin samples were analysed and used for the method validation. The new UHPSFC/QTOF-MS method showed good separation of the 40 compounds within only 6-min retention time, and out of these, 36 showed high ionisation efficiency in negative electrospray ionisation mode. Graphical abstract A rapid and selective method for the quantitative and qualitative analysis of 40 lignin-derived compounds using ultra-high-performance supercritical fluid chromatography coupled to quadrupole-time-of-flight mass spectrometry (UHPSFC/QTOF-MS).

Native lignin extraction from soft- and hardwood by green and benign sub/supercritical fluid extraction methodologies.

Lignin constitutes an impressive resource of high-value low molecular weight compounds. However, robust methods for isolation of the extractable fraction from lignocellulose are yet to be established. In this study, supercritical fluid extraction (SFE) and CO2-expanded liquid extraction (CXLE) were employed to extract lignin from softwood and hardwood chips. Ethanol, acetone, and ethyl lactate were investigated as green organic co-solvents in the extractions. Additionally, the effects of temperature, CO2 percentage and the water content of the co-solvent were investigated using a design of experiment approach employing full factorial designs. Ethyl lactate and acetone provided the highest gravimetric yields. The water content in the extraction mixture had the main impact on the amount of extractable lignin monomers (LMs) and lignin oligomers (LOs) while the type of organic solvent was of minor importance. The most effective extraction was achieved by using a combination of liquid CO2/acetone/water (10/72/18, v/v/v) at 60 °C, 350 bar, 30 min and 2 mL min-1 flow rate. The optimized method provided detection of 13 LMs and 6 lignin dimers (LDs) from the hardwood chips. The results demonstrate the potential of supercritical fluids and green solvents in the field of mild and bening lignin extraction from wood.

Separation of monomeric and dimeric phenolic compounds in lignosulphonate lignin on different stationary phases using ultrahigh-performance supercritical fluid chromatography.

Lignin is a promising renewable resource and its valorization could help to reduce our dependency on fossil carbon resources. Especially the production of small molecular weight and economically valuable compounds, such as vanillin, are of interest. A good separation of the sample components is crucial for a confident identification of compounds in complex sample mixtures using for instance mass spectrometry. In this work, the resolving power and selectivity of five different stationary phases for ultrahigh-performance supercritical fluid chromatography were studied for the class separation of lignin monomers (LMs) and dimers (LDs). A separation of LMs and LDs will help to identify such compounds in complex technical lignin samples. It could be shown that stationary phases with both hydrogen-bonding acceptor and donator groups offer high overall resolving power, while π-π-interactions are advantageous for the separation of the two compound classes. An almost complete separation combined with an improved overall resolving power was achieved with the 1-aminoanthracene stationary phase, which offers both hydrogen-bonding interactions and π-π-interactions.

A study of the spatial distribution patterns of airborne polycyclic aromatic hydrocarbons in crowberry (Empetrum nigrum) in Ilulissat, Greenland.

Polycyclic aromatic hydrocarbons (PAHs) are produced by anthropogenic activities, such as traffic and domestic heating. Due to their adverse effects to humans and natural habitats, the presence of PAHs in the environment needs to be monitored. Plants are known as natural accumulators of persistent organic pollutants (POPs) and can therefore be used for the monitoring of PAHs emitted into the environment. Contamination by PAHs also occurs in the Arctic such as Greenland due to long-range transport through air. However, as anthropogenic activities in the Arctic are increasing, there is a need to investigate the distribution of PAHs due to local emission sources. In this study, we present a systematic sampling approach to identify the influence of PAH sources in an area next to the town of Ilulissat in Greenland. Composite crowberry samples have been collected north of Ilulissat, where the town itself, an incineration site and Ilulissat airport are possible emission sources for PAHs. Matrix solid-phase extraction was used for the extraction of PAHs and the chemical analysis was performed by gas chromatography with mass spectrometry detection (GC-MS). In total, 18 out of 19 investigated PAHs could be detected in Empetrum nigrum in a concentration range of 0.69 to 93.01 µg/kgdry weight. Higher concentrations for most of the targeted PAHs were found close to the suspected emission sources and also along the road connecting them. For pyrene, the correlation between the concentration and the distance from the emission sources could be modelled and visualized using a two-dimensional exponential variogram and ordinary kriging. The range in which the samples were spatially correlated was approximately 500 m. Our results show that local emission sources contribute to the spatial distribution patterns of PAHs. Monitoring of pollution by airborne PAHs is therefore needed even in areas far from major pollution sources such as Ilulissat, Greenland. E. nigrum showed to be a feasible species for biomonitoring of PAHs due to its large abundance in the sampling area and its widespread availability in the Artic region.

Nontargeted Analysis Strategy for the Identification of Phenolic Compounds in Complex Technical Lignin Samples.

Lignin is the second most abundant biopolymer in nature and a promising renewable resource for aromatic chemicals. For the understanding of different lignin isolation and conversion processes, the identification of phenolic compounds is of importance. However, given the vast number of possible chemical transformations, the prediction of produced phenolic structures is challenging and a nontargeted analysis method is therefore needed. In this study, a nontargeted analysis method has been developed for the identification of phenolic compounds by using an ultrahigh-performance supercritical fluid chromatography-high-resolution multiple stage tandem mass spectrometry method, combined with a Kendrick mass defect-based classification model. The method is applied to a Lignoboost Kraft lignin (LKL), a sodium lignosulfonate lignin (SLS), and a depolymerized Kraft lignin (DKL) sample. In total, 260 tentative phenolic compounds are identified in the LKL sample, 50 in the SLS sample, and 77 in the DKL sample.

Biological valorization of low molecular weight lignin.

Lignin is a major component of lignocellulosic biomass and as such, it is processed in enormous amounts in the pulp and paper industry worldwide. In such industry it mainly serves the purpose of a fuel to provide process steam and electricity, and to a minor extent to provide low grade heat for external purposes. Also from other biorefinery concepts, including 2nd generation ethanol, increasing amounts of lignin will be generated. Other uses for lignin - apart from fuel production - are of increasing interest not least in these new biorefinery concepts. These new uses can broadly be divided into application of the polymer as such, native or modified, or the use of lignin as a feedstock for the production of chemicals. The present review focuses on the latter and in particular the advances in the biological routes for chemicals production from lignin. Such a biological route will likely involve an initial depolymerization, which is followed by biological conversion of the obtained smaller lignin fragments. The conversion can be either a short catalytic conversion into desired chemicals, or a longer metabolic conversion. In this review, we give a brief summary of sources of lignin, methods of depolymerization, biological pathways for conversion of the lignin monomers and the analytical tools necessary for characterizing and evaluating key lignin attributes.