Attenuated Total Reflectance Infrared Spectra Research Articles

Etalon-Assisted Study of the Strong CO Ligand Vibrations of the fac-[Re(CO)3(bpy)(CH3CN)]+ Octahedral Complex.

The strong CO ligand vibrations of an octahedral complex, fac-[Re (CO)3(bpy)(CH3CN)]+, in acetonitrile are observed at 2040 and 1932 cm-1. Facial rhenium tricarbonyl systems offer very strong and isolated CO vibrations with the potential for interactions between these vibrations. This work first identifies the dominant ion-pair species using attenuated total reflection infrared (ATR-IR) absorption spectra on a dilution series and then determines the strength of these CO ligand vibrations (as isolated vibrations) with a combination of ATR-IR and etalon-based measurements that determine the absolute complex index of refraction of the solution. Finally, the etalon experiments are modeled to study the interaction between vibrations, which is a property not embedded in the solution's complex index of refraction. The ATR-IR spectra are accomplished on a dilution series as well as a larger set of spectra as these solutions evaporated. The A'(1) CO ligand band at 2040 cm-1 is fit with a sum of three Lorentzian functions characterizing the distribution of free, solvent-separated, and contact ion pairs of this octahedral complex vs concentration. The other CO ligand band at 1932 cm-1 is broader and complicated by the dynamics of vibrational interactions, the unresolved splitting of the A'(2) and A″ CO vibrations, and ion-pair speciation. The etalon transmission measurements vs angle were on a 0.029 M solution, and Rabi splittings of 19 and 38 cm-1 were observed for the A'(1) CO vibration and the unresolved A'(2) + A″ CO vibrations, respectively. The great strength of the CO ligand vibrations is evident despite the use of a dilute solution. Integrated band intensities are reported in comparison to hybrid density functional calculations for isolated vibrations. Then, the observed Rabi splittings are modeled to obtain the coupling strength of the CO ligand vibration with etalon cavity modes and with each other. In summary, this work develops a method to determine the concentration of these solutions from the ATR-IR spectrum, characterizes the ion-pairing, shows that the index of refraction is not constant in the IR spectral region of interest, and develops an interaction Hamiltonian that characterizes cavity-vibration and vibration-vibration coupling.

Superoxide intermediate in the oxygen reduction on a zinc hydroxide model corrosion product.

The inhibition of the electrochemical oxygen reduction reaction (ORR) by zinc corrosion products plays an important role in the corrosion protection of galvanized steel. Hence, the electrocatalytic mechanism of the ORR on electrodeposited zinc hydroxide-based model corrosion products was investigated by in situ and operando attenuated total reflection infrared (ATR-IR) spectroscopy, supplemented by density functional theory (DFT) calculations. Model corrosion products containing flake-like crystalline Zn5(NO3)2(OH)8 were cathodically electrodeposited on germanium(100) electrodes from a zinc nitrate precursor electrolyte. Substantial amounts of the films are non-crystalline, and their surfaces predominantly consist of zinc oxide and hydroxide species, as evidenced by x-ray photoelectron spectroscopy. ATR-IR spectra show a peak at 1180 cm-1 during cathodic currents in O2-saturated NaClO4 solution. This peak is assigned to a surface-bound superoxide, the only ORR intermediate detected. Absorbance from the intermediate increases with increasing cathodic current, indicating an increase in surface concentration of superoxide intermediates at larger ORR current densities. The zinc hydroxide ages in the experiments, most likely by a transformation into zinc oxide, consistent with the observed decrease in absorbance over time of the OH bending mode of zinc hydroxide at 1380 cm-1. This aging is a time-dependent chemical process, implying that pure chemical aging is important in actual corrosion products as well. DFT calculations of adsorbed superoxide yield a Zn-O bond length similar to the bond length in Zn-O, thus enhancing superoxide interaction with undercoordinated tetrahedral Zn2+ sites on the surface. Thus, such active sites catalyze the first reduction step in the ORR.

Comparison of Vibrational Spectroscopic Techniques for Quantification of Water in Natural Deep Eutectic Solvents.

Vibrational spectroscopic techniques, i.e., attenuated total reflectance infrared (ATR-IR), near infrared spectroscopy (NIRS) and Raman spectroscopy (RS), coupled with Partial Least Squares Regression (PLSR), were evaluated as cost-effective label-free and reagent-free tools to monitor water content in Levulinic Acid/L-Proline (LALP) (2:1, mol/mol) Natural Deep Eutectic Solvent (NADES). ATR-IR delivered the best outcome of Root Mean Squared Error (RMSE) of Cross-Validation (CV) = 0.27% added water concentration, RMSE of Prediction (P) = 0.27% added water concentration and mean % relative error = 2.59%. Two NIRS instruments (benchtop and handheld) were also compared during the study, respectively yielding RMSECV = 0.35% added water concentration, RMSEP = 0.56% added water concentration and mean % relative error = 5.13% added water concentration, and RMECV = 0.36% added water concentration, RMSEP = 0.68% added water concentration and mean % relative error = 6.23%. RS analysis performed in quartz cuvettes enabled accurate water quantification with RMECV = 0.43% added water concentration, RMSEP = 0.67% added water concentration and mean % relative error = 6.75%. While the vibrational spectroscopic techniques studied have shown high performance in relation to reliable determination of water concentration, their accuracy is most likely related to their sensitivity to detect the LALP compounds in the NADES. For instance, whereas ATR-IR spectra display strong features from water, Levulinic Acid and L-Proline that contribute to the PLSR predictive models constructed, NIRS and RS spectra are respectively dominated by either water or LALP compounds, representing partial molecular information and moderate accuracy compared to ATR-IR. However, while ATR-IR instruments are common in chemistry and physics laboratories, making the technique readily transferable to water quantification in NADES, Raman spectroscopy offers promising potential for future development for in situ, sample withdrawal-free analysis for high throughput and online monitoring.

Highly Ordered Supramolecular Assembled Networks Tailored by Bioinspired H-Bonding Confinement for Recyclable Ion-Transport Materials

Highly Ordered Supramolecular Assembled Networks Tailored by Bioinspired H-Bonding Confinement for Recyclable Ion-Transport Materials

Monitoring Molecular Weight Changes during Technical Lignin Depolymerization by Operando Attenuated Total Reflectance Infrared Spectroscopy and Chemometrics.

Technical lignins are increasingly available at industrial scale, offering opportunities for valorization, such as by (partial) depolymerization. Any downstream lignin application requires careful tailoring of structural properties, such as molecular weight or functional group density, properties that are difficult to control or predict given the structure variability and recalcitrance of technical lignins. Online insight into changes in molecular weight (M w), to gauge the extent of lignin depolymerization and repolymerization, would be highly desired to improve such control, but cannot be readily provided by the standard ex‐situ techniques, such as size exclusion chromatography (SEC). Herein, operando attenuated total reflectance infrared (ATR‐IR) spectroscopy combined with chemometrics provided temporal changes in M w during lignin depolymerization with high resolution. More specifically, ex‐situ SEC‐derived M w and polydispersity data of kraft lignin subjected to aqueous phase reforming conditions could be well correlated with ATR‐IR spectra of the reaction mixture as a function of time. The developed method showed excellent regression results and relative error, comparable to the standard SEC method. The method developed has the potential to be translated to other lignin depolymerization processes.

Spectral and Texture Properties of Hydrophobic Aerogel Powders Obtained from Room Temperature Drying.

Attenuated Total Reflectance Infrared (ATR-IR) spectroscopy and texture measurements based on nitrogen adsorption-desorption isotherms are combined to characterize silica aerogel granules with different degrees of hydrophobicity. The aerogels were prepared from tetraethoxysilane via a room temperature hydrolysis-gelation process, solvent exchange, hydrophobization, and drying at subcritical conditions. The dependencies between the texture properties, pore architectures, surface fractal dimensions, and degree of hydrophobicity of the samples are extracted from the ATR-IR spectra and the adsorption-desorption isotherms. The IR absorption in the region of the Si-O-Si and Si-OH vibrations is used for a description of the structural and chemical changes in aerogel powders connected with their surface hydrophobization. The Frenkel–Halsey–Hill (FHH) theory is applied to determine the surface fractal dimension of the powder species.

Effect of elevated temperature and UV radiation on molecular structure of linoleic acid by ATR-IR and two-dimensional correlation spectroscopy

The effect of elevated temperature (44 °C) and ultraviolet (UV) radiation on molecular structure of linoleic acid (LA) was studied by Attenuated Total Reflection Infrared (ATR-IR) spectroscopy. To obtain more detailed information on molecular mechanism of these changes we applied moving-window analysis and two-dimensional correlation spectroscopy (2DCOS). Analysis of the time-dependent ATR-IR spectra of LA before and after UV irradiation revealed the structural changes in molecules of LA. The extent of these changes was significantly higher after an application of UV radiation. During 24 h experiment temperature was constant, therefore the spectral changes result from relatively slow processes (and requiring more energy), e.g. cis/trans isomerization, disruption of the C=C double bonds and partial breaking of hydrogen bonds in the cyclic dimers. As a side effect of these structural changes one can observe variations in the orientation of the chains. It is of note that the methyl and methylene groups reveal slightly different behaviour.

Enhanced corrosion inhibition of aniline derivatives electropolymerized coatings on copper: Preparation, characterization and mechanism modeling

Nanostructured polyaniline (PANi), poly(N-ethylaniline) (PNEA) and their molybdate-doped coatings (PANi-Mo and PNEA-Mo) were electropolymerized on passivated copper surface via voltammetric cycles in oxalic acid solution. The coating structures were featured by morphological observation, attenuated total reflectance infrared spectra (ATR-IR), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS); besides, anticorrosive performance of coatings was investigated for copper in 0.5 M H2SO4 solution through electrochemical and topographical measurements. ATR-IR and XRD results documented the occurrence of polymerization over copper surface, while the morphologies implied that the presence of ethyl and molybdate generated the extra increment in coating thickness. Owing to physical barrier of substituent and in-situ dissolving of molybdate, the corrosion of coated specimens was highly suppressed in H2SO4 solution. The protective effect of coatings followed the sequence of PNEA-Mo > PANi-Mo > PNEA > PANi with an inverse trend in porosity index. XPS analyses revealed that repeat units of polymer were in mixed state with predominantly oxidative type, which were inclined to parallelly deposit on copper surface with a strong anchoring strength. Molecular dynamics simulations well supported the configuration and forceful adhesion of polymers on Cu (1 1 1) plane, and indicated the favorable compatibility between polymer and dopant.

Structural FTIR analysis of cellulose functional groups isolated from Leucaena leucocephala pods using different bleaching agents.

Abstract In this study, extracted cellulose fibres were successfully isolated from Leucaena leucocephala pods through an efficient alkaline treatment and bleaching method using different bleaching agents such as 5 wt.% of sodium chlorite (NaClO 2 ), 5 wt.% of oxalic acid (C 2 H 2 O 4 ), 5 wt.% of potassium permanganate (KMnO 4 ) and 5 wt.% of sodium hypochlorite (NaClO), respectively. The functional group of the cellulose fibres were analysed by using attenuated total reflection infrared (ATR-IR) spectroscopy. The ATR-IR spectra reported that the Leucaena leucocephala fibres after being given chemical treatment with 5%w/w of NaClO lead the similar functional group with the commercial cellulose due to removal hemicellulose and lignin. The content cellulose of final product was 89.4% was measured by the chemical composition test. Meanwhile, the whiteness index of Leucaena leucocephala fibres treated with 5 wt.% NaClO was 78.5%, which was the highest. The results of ATR-IR, chemical composition test and whiteness index were indicated that the sample of Leucaena leucocephala pods presents the structure of cellulose.

Antitrypanosomal Effects of Zanthoxylum zanthoxyloides (Lam.) Zepern. & Timler Extracts on African Trypanosomes.

African trypanosomiasis is a disease caused by the parasitic protozoa of the Trypanosoma genus. Despite several efforts at chemotherapeutic interventions, the disease poses serious health and economic concerns to humans and livestock of many sub-Saharan African countries. Zanthoxylum zanthoxyloides (Lam.) Zepern. & Timler (Z. zanthoxyloides LZT) is a plant species of important phytochemical and pharmacological relevance in the subtropical zones of the African continent. However, the mechanisms of its antitrypanosomal effects in African trypanosomes remain to be elucidated. The aim of the study was to determine the in vitro effects and mechanisms of action of Z. zanthoxyloides LZT (root) fractions against Trypanosoma brucei. T. brucei (GUTat 3.1 strain), L. donovani (D10 strain), P. falciparum (3D 7 strain), Jurkat cells, and Chang liver cells were cultivated in vitro to the log phase in their respective media at 37°C. Crude extracts and fractions were prepared from air-dried pulverized plant material of Z. zanthoxyloides LZT (root) using the modified Kupchan method of solvent partitioning. Half-maximal inhibitory concentrations (IC50) were determined through the alamar blue cell viability assay. Effects of fractions on cell death and cell cycle of T. brucei were determined using flow cytometry. Fluorescence microscopy was used to investigate the effects of fractions on the morphology and distribution of T. brucei. Antitrypanosomal compounds of fractions were characterized using high-performance liquid chromatography (HPLC) and attenuated total reflectance infrared (ATR-IR) spectroscopy. Methanol, butanol, and dichloromethane fractions were selectively active against T. brucei with respective IC50 values of 3.89, 4.02, and 5.70 μg/ml. Moreover, methanol, butanol, and dichloromethane fractions significantly induced apoptosis-like cell death with remarkable alteration in the cell cycle of T. brucei. Furthermore, dichloromethane and methanol fractions altered the morphology, induced aggregation, and altered the ratio of nuclei to kinetoplasts in the parasite. The HPLC chromatograms and ATR-IR spectra of the active fractions suggested the presence of aromatic hydrocarbons with hydroxyl, carbonyl, amine, or amide functional groups. The results suggest that Z. zanthoxyloides LZT have potential chemotherapeutic effects on African trypanosomes with implications for novel therapeutic interventions in African trypanosomiasis.

Biosorption of cadmium by potential probiotic Pediococcus pentosaceus using in vitro digestion model.

An exponential increase of heavy metal toxicant in the human body is a growing health-related problem. In the present study, Pediococcus pentosaceus FB145 and FB181 strains were isolated from fermented seafood and served as highly Cd-resistant strains. The unchanged structural cells of P. pentosaceus strains after treatment with Cd cations were investigated using a scanning electron microscope energy dispersive X-ray analysis. Furthermore, both P. pentosaceus strains showed strong human gastrointestinal tract resistance properties. The Cd biosorption results fit the pseudo-second-order kinetics with capacities for P. pentosaceus FB145 and FB181 for Cd were 52.8 and 50.3 mg/g, respectively, whereas the maximum biosorption capacities were 52.65 and 50.35 mg/g, respectively. The equilibrium data were well fitted to the Freundlich isotherm model. The binding of Cd to bacterial cells may be caused by the presence of different functional groups such as carboxyl, amide, and phosphate on cell surface, which was confirmed by the Fourier transform infrared-attenuated total reflectance spectra. Moreover, these strains decreased the Cd bioaccessibility by 44.7-46.8% in the in vitro digestion model. These findings indicate that P. pentosaceus FB145 and FB181 are novel potent biosorbent for preventing cadmium toxicity and reducing its absorption into the human body.

Effect of surface-modified silica on the thermal and mechanical behaviors of poly(lactic acid) and chemically crosslinked poly(lactic acid) composites

This work studied the properties of poly(lactic acid) (PLA) and chemically crosslinked poly(lactic acid) (CrPLA) in the presence of different types of silica (SiO2): SiO2 from rice husk ash (RhSiO2) and commercially available SiO2 (CoSiO2). A mixture of triallyl isocyanate and dicumyl peroxide was used as a chemical crosslinking agent. The rice husk was burnt in a furnace to extract RhSiO2. The thermal and mechanical properties of the PLA and CrPLA composites were investigated as a function of the SiO2 content. The presence of C–H functional groups supports the compatibility of stearic acid and RhSiO2, thereby facilitating the esterification reaction observed in the Fourier transform infrared-attenuated total reflectance spectra. This played an important role in changing the properties of the PLA-based composites. The concave fracture surfaces were consistent with the tensile results, the thermal properties, and the degree of swelling, which suggested that there was a better reaction between stearic acid and RhSiO2. However, the heat deflection temperatures of both the PLA and the CrPLA composites were not significantly affected by blending with either CoSiO2 or RhSiO2.

The structural modification of pea protein concentrate with gum Arabic by controlled Maillard reaction enhances its functional properties and flavor attributes

The aim of this research was to investigate the impact of conjugation with gum Arabic (GA) on the functional properties and flavor profile of pea protein concentrate (PPC). The PPC-GA conjugates was prepared by incubating the mixture at a mass ratio of 1:4 at 60 °C and 79% relative humidity with variable times (0, 1, 3, and 5 days). The grafting of GA to PPC was confirmed using SDS-PAGE and Fourier transform infrared-attenuated total reflection spectra. The degree of conjugation between PPC and GA over the course of incubation was determined by non-specific Maillard reaction markers, the loss of free amino groups, and color development. The results suggested that the solubility and emulsification properties of PPC-GA conjugates was remarkably improved after 3 days incubation and further extending incubation time deteriorated the functionalities. Surface morphology modifications of the conjugates revealed by scanning electron microscope may explain the enhanced functionality. The corn oil-in-water emulsions stabilized by PPC-GA conjugates of 3 days incubation also showed great physical stabilities against pH (2–8), thermal processing (25, 37, and 72 °C), and ionic strength (0, 100, 300, and 500 mM), as compared to PPC alone or the mixture of PPC and GA. HS-SPME-GC-MS analysis and quantification of beany flavor markers in PPC-GA conjugates indicated that the beany flavors markers were remarkably diminished after 1 day incubation. The findings from this study indicated that controlled Maillard reaction could be an effective strategy to not only enhance the functionality of poor water soluble plant proteins, but also mitigate the overwhelming beany flavors, thus promoting the utilization of plant proteins as functional ingredients in food industry.

Effects of variable selection and processing of NIR and ATR-IR spectra on the prediction of extractive content in Eucalyptus bosistoana heartwood.

The use of quick attenuated total reflectance infrared (ATR-IR) spectroscopy and near infrared (NIR) spectroscopy to predict extractives content (EC) in heartwood of E. bosistoana with partial least squares regression (PLSR) models was studied. Different spectra pre-processing methods and variable selection were tested for calibration optimisation. While variable selection substantially improved the NIR-PLSR models, only small effects were observed for spectra pre-processing methods and ATR-IR-PLSR models. Both of the NIR-PLSR and ATR-IR-PLSR models yielded reliably EC results with high R2 and low root mean square error (RMSE). NIR based models performed better (RMSE 0.9%) than ATR-IR based models (RMSE 1.6%). Analysis showed that the models were based on IR signals assigned to chemical structures known from eucalyptus heartwood extracts. Combined with PLSR and variable selection, both, ATR-IR and the NIR spectroscopy, can be used to quickly predict EC in E. bosistoana, a measure needed in tree breeding and the quality control of for durable timber.

Infrared attenuated total reflection and 2D fluorescence spectroscopy for the discrimination of differently aggregated monoclonal antibodies.

Antibody aggregates may occur as undesirable by-products during the manufacturing process of biopharmaceutical proteins since parameters such as pH, temperature, ionic strength, protein concentration, oxygen, and shear forces can lead to aggregate formation. These aggregates have to be detected, quantified and removed cost extensively, since they may reduce the safety and efficacy of the product. Protein aggregates can range from small soluble dimers up to large visible agglomerates. Differently aggregated antibody samples were characterized for their soluble and insoluble aggregate concentration by size exclusion chromatography and fluorescence microscopy, respectively. The samples exhibited a high diversity of protein aggregates, which varied in amount, size and shape. For secondary structure characterization, infrared attenuated total reflection (IR-ATR) and two-dimensional fluorescence (2D-FL) spectroscopy were applied. Using direct spectroscopy, only marginal differences of various antibody aggregates were evident. However, using appropriate chemometric strategies, the evaluation of IR-ATR and 2D-FL spectra yielded the discrimination of differently aggregated antibody samples with yet unprecedented precision.

Insights into Self-Poisoning during Catalytic Hydrogenation on Platinum Surfaces Using ATR-IR Spectroelectrochemistry

Attenuated total reflection infrared (ATR-IR) spectroscopy has been combined with electrochemical methods to investigate molecular decomposition and self-poisoning processes on platinum surfaces under the conditions of catalytic hydrogenation. In aqueous 0.1 M H2SO4 the α-keto ester ethyl pyruvate (EP) is found to decompose on polycrystalline platinum electrodes to yield surface-adsorbed CO, but the observed behavior is highly dependent on the electrode potential, a parameter intimately linked to the surface-adsorbed hydrogen coverage. In the potential range −0.2 to −0.4 V (vs mercury/mercurous sulfate electrode) where the hydrogen coverage is negligible, CO is readily produced at the platinum surface along with other molecular fragments but the decomposition process becomes inhibited at high EP solution concentrations. At −0.5 V only very low coverages of CO are observed due to competing hydrogen adsorption at Pt(100) step sites which most favor EP decomposition. At more negative potentials, during the onset of catalytic EP hydrogenation, CO is generated rapidly but other intermediates or products are not observed in the ATR-IR spectra. Together these observations suggest two different mechanisms of EP decomposition, the first occurring directly upon EP adsorption and the second occurring after a single hydrogen atom transfer under hydrogen rich conditions. This ability to control substrate decomposition by tuning the surface hydrogen coverage may be used as a potential route to mitigating catalyst poisoning and deactivation during hydrogenation reactions.

Rapid intra-operative diagnosis of kidney cancer by attenuated total reflection infrared spectroscopy of tissue smears.

Herein, a technique to analyze air-dried kidney tissue impression smears by means of attenuated total reflection infrared (ATR-IR) spectroscopy is presented. Spectral tumor markers-absorption bands of glycogen-are identified in the ATR-IR spectra of the kidney tissue smear samples. Thin kidney tissue cryo-sections currently used for IR spectroscopic analysis lack such spectral markers as the sample preparation causes irreversible molecular changes in the tissue. In particular, freeze-thaw cycle results in degradation of the glycogen and reduction or complete dissolution of its content. Supervised spectral classification was applied to the recorded spectra of the smears and the test spectra were classified with a high accuracy of 92% for normal tissue and 94% for tumor tissue, respectively. For further development, we propose that combination of the method with optical fiber ATR probes could potentially be used for rapid real-time intra-operative tissue analysis without interfering with either the established protocols of pathological examination or the ordinary workflow of operating surgeon. Such approach could ensure easier transition of the method to clinical applications where it may complement the results of gold standard histopathology examination and aid in more precise resection of kidney tumors.

Centrifugal ultrafiltration of human serum for improving immunoglobulin A quantification using attenuated total reflectance infrared spectroscopy

Attenuated total reflectance infrared (ATR-IR) spectroscopy is a simple, rapid and cost-effective method for the analysis of serum. However, the complex nature of serum remains a limiting factor to the reliability of this method. We investigated the benefits of coupling the centrifugal ultrafiltration with ATR-IR spectroscopy for quantification of human serum IgA concentration. Human serum samples (n = 196) were analyzed for IgA using an immunoturbidimetric assay. ATR-IR spectra were acquired for whole serum samples and for the retentate (residue) reconstituted with saline following 300 kDa centrifugal ultrafiltration. IR-based analytical methods were developed for each of the two spectroscopic datasets, and the accuracy of each of the two methods compared. Analytical methods were based upon partial least squares regression (PLSR) calibration models - one with 5-PLS factors (for whole serum) and the second with 9-PLS factors (for the reconstituted retentate). Comparison of the two sets of IR-based analytical results to reference IgA values revealed improvements in the Pearson correlation coefficient (from 0.66 to 0.76), and the root mean squared error of prediction in IR-based IgA concentrations (from 102 to 79 mg/dL) for the ultrafiltration retentate-based method as compared to the method built upon whole serum spectra. Depleting human serum low molecular weight proteins using a 300 kDa centrifugal filter thus enhances the accuracy IgA quantification by ATR-IR spectroscopy. Further evaluation and optimization of this general approach may ultimately lead to routine analysis of a range of high molecular-weight analytical targets that are otherwise unsuitable for IR-based analysis.

Instrumental analyses of nanostructures and interactions with water molecules of biomass constituents of Japanese cypress

Nanostructures consisting of the biomass constituents of the denatured Japanese cypress (Chamaecyparis obtusa) were examined by instrumental analyses at multiple hierarchical levels. Delignification with NaClO2 solution smoothly proceeded to reveal a distorted cell by scanning electron microscopy; however, a trace amount of lignin still remained in the delignified sample according to attenuated total reflection infrared spectra (ATR-IR). Although hemicellulose could be removed by a treatment with NaOH solution, thermogravimetric analysis and 13C cross-polarization/magic angle spinning (CP-MAS) NMR showed a certain amount of hemicellulose remaining. Reaction of the delignified sample with NaOH solution produced a shrunken cell wall that consisted of cellulose with small amounts of lignin and hemicellulose, which were detected by ATR-IR and 13C CP-MAS NMR, respectively. These samples from which lignin and/or hemicellulose had been removed easily released water molecules, producing a decrease in the 1H signal intensity and longer 1H spin–lattice relaxation time (T1H) values in variable temperature 1H MAS NMR. The T1H values provided information about nano-scale molecular interaction difficult to obtain by other instrumental analyses and they greatly changed depending on the water content and ratio of the biomass constituents. The spin–lattice relaxation of all samples occurred via water molecules under humid conditions that provided sufficient water. Under heat-dried conditions, the spin–lattice relaxation mainly occurred via lignin for the samples with lignin remaining while it occurred via cellulose/hemicellulose for the samples without lignin. The variable temperature T1H analysis indicated that predominant spin–lattice relaxation route via lignin was caused by higher molecular mobility of lignin-containing samples compared with lignin-free samples.

Application of laboratory and portable attenuated total reflectance infrared spectroscopic approaches for rapid quantification of alpaca serum immunoglobulin G.

The objective of this study was to develop and compare the performance of laboratory grade and portable attenuated total reflectance infrared (ATR-IR) spectroscopic approaches in combination with partial least squares regression (PLSR) for the rapid quantification of alpaca serum IgG concentration, and the identification of low IgG (<1000 mg/dL), which is consistent with the diagnosis of failure of transfer of passive immunity (FTPI) in neonates. Serum samples (n = 175) collected from privately owned, healthy alpacas were tested by the reference method of radial immunodiffusion (RID) assay, and laboratory grade and portable ATR-IR spectrometers. Various pre-processing strategies were applied to the ATR-IR spectra that were linked to corresponding RID-IgG concentrations, and then randomly split into two sets: calibration (training) and test sets. PLSR was applied to the calibration set and calibration models were developed, and the test set was used to assess the accuracy of the analytical method. For the test set, the Pearson correlation coefficients between the IgG measured by RID and predicted by both laboratory grade and portable ATR-IR spectrometers was 0.91. The average differences between reference serum IgG concentrations and the two IR-based methods were 120.5 mg/dL and 71 mg/dL for the laboratory and portable ATR-IR-based assays, respectively. Adopting an IgG concentration <1000 mg/dL as the cut-point for FTPI cases, the sensitivity, specificity, and accuracy for identifying serum samples below this cut point by laboratory ATR-IR assay were 86, 100 and 98%, respectively (within the entire data set). Corresponding values for the portable ATR-IR assay were 95, 99 and 99%, respectively. These results suggest that the two different ATR-IR assays performed similarly for rapid qualitative evaluation of alpaca serum IgG and for diagnosis of IgG <1000 mg/dL, the portable ATR-IR spectrometer performed slightly better, and provides more flexibility for potential application in the field.