Rapid determination of protein contents in microencapsulated fish oil supplements by ATR-FTIR spectroscopy and partial least square regression (PLSR) analysis

Following the recent success in quantitative analysis of essential fatty acid compositions in a commercial microencapsulated fish oil (?EFO) supplement, we extended the application of portable attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopic technique and partial least square regression (PLSR) analysis for rapid determination of total protein contents-the other major component in most commercial ?EFO powders. In contrast to the traditional chromatographic methodology used in a routine amino acid analysis (AAA), the ATR-FTIR spectra of the ?EFO powder can be acquired directly from its original powder form with no requirement of any sample preparation, making the technique exceptionally fast, noninvasive, and environmentally friendly as well as being cost effective and hence eminently suitable for routine use by industry. By optimizing the spectral region of interest and number of latent factors through the developed PLSR strategy, a good linear calibration model was produced as indicated by an excellent value of coefficient of determination R2 = 0.9975, using standard ?EFO powders with total protein contents in the range of 140-450 mg/g. The prediction of the protein contents acquired from an independent validation set through the optimized PLSR model was highly accurate as evidenced through (1) a good linear fitting (R2 = 0.9759) in the plot of predicted versus reference values, which were obtained from a standard AAA method, (2) lowest root mean square error of prediction (11.64 mg/g), and (3) high residual predictive deviation (6.83) ranked in very good level of predictive quality indicating high robustness and good predictive performance of the achieved PLSR calibration model. The study therefore demonstrated the potential application of the portable ATR-FTIR technique when used together with PLSR analysis for rapid online monitoring of the two major components (i.e., oil and protein contents) in finished ?EFO powders in the actual manufacturing setting.

Tribology of lubricated nitrocarburised and titanium carbonitride surfaces

In the current work, two different coatings, nitrocarburised (CN) and titanium carbonitride (TiCN) on M2 grade high speed tool steel, were prepared by commercial diffusion and physical vapour deposition (PVD) techniques, respectively. Properties of the coating were characterised using a variety of techniques such as Glow-Discharge Optical Emission Spectrometry (GD-OES) and Scanning Electron Microscopy (SEM). Three non-commercial, oil-based lubricants with simplified formulations were used for this study. A tribological test was developed in which two nominally geometrically-identical crossed cylinders slide over each other under selected test conditions. This test was used to evaluate the effectiveness of a pre-applied lubricant film and a surface coating for various conditions of sliding wear. Engineered surface coatings can significantly improve wear resistance of the tool surface but their sliding wear performances strongly depend on the type of coating and lubricant combination used. These coating-lubricant interactions can also have a very strong effect on the useful life of the lubricant in a tribological system. Better performance of lubricants during the sliding wear testing was achieved hen used with the nitrocarburised (CN) coating. To understand the nature of the interactions and their possible effects on the coating-lubricant system, several surface analysis techniques were used. The molecular level investigation of Fourier Transform Infrared Spectroscopy (FTIR) revealed that oxidative degradation occurred in all used oil-based lubricants during the sliding wear test but the degradation behaviour of oil-based lubricants varied with the coating-lubricant system and the wear conditions. The main differences in the carbonyl oxidation region of the FTIR spectra (1900-1600 cm-1) between different coating-lubricant systems may relate to the effective lifetime of the lubricant during the sliding wear test. Secondary Ion Mass Spectrometry (SIMS) depth profiling shows that the CN coating has the highest lubricant absorbability among the tested tool surfaces. Diffusion of chlorine (C1), hydrogen (H) and oxygen (O) into the surface of subsurface of the tool suggested that strong interactions occurred between lubricant and tool surface during the sliding wear test. The possible effects of the interactions on the performance of whole tribological system are also discussed. The study of Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) indicated that the envelope of hydrocarbons (CmHn) of oil lubricant in the positive TOF-SIMS spectra shifted to lower mass fragment after the sliding wear testing due to the breakage of long-chain hydrocarbons to short-chain ones during the degradation of lubricant. The shift of the mass fragment range of the hydrocarbon (CmHn) envelope caries with the type of both tool surface and lubricant, again confirming that variation in the performance of the tool-lubricant system relates to the changes in surface chemistry due to tribochemical interactions at the tool-lubricant interface under sliding wear conditions. The sliding wear conditions resulted in changes not only in topography of the tool surface due to mechanical interactions, as outlined in Chapter 5, but also in surface chemistry due to tribochemical interactions, as discussed in Chapters 6 and 7.

Enhancing the carbon yield of cellulose based carbon fibres with ionic liquid impregnates

Here we show that ionic liquids (ILs), protic or aprotic in nature containing a phosphate anion, can be used as effective impregnating compounds resulting in a 50% improvement of the carbon yield of cellulose based carbon fibres and a 70 °C reduction in the onset of the depolymerization temperature. Using 13C NMR and FTIR spectra, we characterize the carbonized fibres with and without IL impregnates. The oxidative step in the formation of carbon fibres from cellulose precursors is very important in determining the final material properties, as such we examine this stage and show that the IL reduces the onset of the cellulose depolymerization temperature while improving the oxidative stability. This study highlights the ability of ILs to act as novel impregnates which can successfully reduce the formation of tar and volatile substances during carbonization of cellulose based carbon fibres resulting in an improved carbon yield and significant cost savings due to reduced maintenance and wear of equipment. This journal is

Simultaneous crystallization and decomposition of PVA/MMT composites during non-isothermal process

Decomposition of poly(vinyl alcohol)/montmorillonite clay (PVA/MMT) composites during melting-crystallization was experimentally confirmed by morphology and molecular structure changes. In particular, FTIR spectra show the shift of O-H stretching band as well as enhanced intensities of C-O stretching and CH2 rocking vibrational modes. Furthermore, Raman deconvolution indicates that C-H wagging, CH2-CH wagging, CH-CO bending and CH2 wagging modes in amorphous domains were all decreased greatly. Moreover, this decomposition leads to decreased melting enthalpy, melting point, crystallization enthalpy and crystallization temperature. Crystallization analysis shows that the MMT incorporated slows down the crystallization process in the PVA matrix regardless of the nucleation capability of MMT. Despite the severe decomposition, the crystallization kinetics still corroborated well with common classical models. As a result, molecular structure changes and crystallization retardation observed in this study clearly indicate the strong effects of the thermal degradation on the non-isothermal crystallization of PVA/MMT composites.

Understanding physicochemical changes in pretreated and enzyme hydrolysed hemp (Cannabis sativa) biomass for biorefinery development

The physicochemical properties of hemp biomass structure to pretreatment and enzymatic hydrolysis were investigated to improve upon reducing sugar production for biofuel development. Sodium hydroxide pretreated biomass (SHPB) yielded maximum conversion of holocellulose into reducing sugar (72 %). Scanning electron microscopy (SEM) revealed that enzymatic hydrolysis generated regular micropores in the fragmented biomass structure. The thermogravimetric analysis (TGA) curve suggested the degradation of hemicellulose and cellulose, which conformed well to the subsequent nuclear magnetic resonance (NMR) studies indicating the presence of α- and β-glucose (28.4 %) and α- and β-xylose (10.7 %), the major carbohydrate components commonly found in hydrolysis products of hemicellulose and cellulose. Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectra showed stretching modes of the lignin acetyl group, suggesting the loosening of the polymer matrix and thus the exposure of the cellulose polymorphs. X-ray diffraction pattern indicated that enzymatic hydrolysis caused a higher crystallinity index (36.71), due to the fragmentation of amorphous cellulose leading to the reducing sugar production suitable for biofuel development.

FTIR and thermal analysis of cashmere and other animal fibres

In this paper, sereval varieties of animal fibres including cashmere and Australian fine Merino wool have been analysed by Fourier transform infrared microscope and differential scanning calorimeter. The results showed that both Chinese and Australian cashmere fibres started absorbing heat at a relatively higher temperature but were thermally degraded quicker than other animal fibres tested. However, the mass changes (within the temperature range of 200oC to 350oC) and associated onset temperatures varied among fibre varieties. From the attenuated total reflectance spectra, the Chinese cashmere was clearly different from Australian cashmere and wool in a peak near 1040 cm-1 wavelength for S-O stretching of cysteic acid residues. The Chinese cashmere presented a stronger absorption at 1019 cm-1 wavelength, while Australian cashmere and wool peaked at 1079 cm-1 wavelength and had a weaker absorption. Combined with thermal analysis, the normalised R-SO3 - content of cysteic acid residues to the amide II peak of the protein backbone may have potential use in identifying Australian fine Merino wool from Chinese cashmere.

FTIR microspectroscopy for rapid screening and monitoring of polyunsaturated fatty acid production in commercially valuable marine yeasts and protists

The increase in polyunsaturated fatty acid (PUFA) consumption has prompted research into alternative resources other than fish oil. In this study, a new approach based on focal-plane-array Fourier transform infrared (FPA-FTIR) microspectroscopy and multivariate data analysis was developed for the characterisation of some marine microorganisms. Cell and lipid compositions in lipid-rich marine yeasts collected from the Australian coast were characterised in comparison to a commercially available PUFA-producing marine fungoid protist, thraustochytrid. Multivariate classification methods provided good discriminative accuracy evidenced from (i) separation of the yeasts from thraustochytrids and distinct spectral clusters among the yeasts that conformed well to their biological identities, and (ii) correct classification of yeasts from a totally independent set using cross-validation testing. The findings further indicated additional capability of the developed FPA-FTIR methodology, when combined with partial least squares regression (PLSR) analysis, for rapid monitoring of lipid production in one of the yeasts during the growth period, which was achieved at a high accuracy compared to the results obtained from the traditional lipid analysis based on gas chromatography. The developed FTIR-based approach when coupled to programmable withdrawal devices and a cytocentrifugation module would have strong potential as a novel online monitoring technology suited for bioprocessing applications and large-scale production.

Synchrotron-FTIR microspectroscopy enables the distinction of lipid accumulation in thraustochytrid strains through analysis of individual live cells

The superior characteristics of high photon flux and diffraction-limited spatial resolution achieved by synchrotron-FTIR microspectroscopy allowed molecular characterization of individual live thraustochytrids. Principal component analysis revealed distinct separation of the single live cell spectra into their corresponding strains, comprised of new Australasian thraustochytrids (AMCQS5-5 and S7) and standard cultures (AH-2 and S31). Unsupervised hierarchical cluster analysis (UHCA) indicated close similarities between S7 and AH-7 strains, with AMCQS5-5 being distinctly different. UHCA correlation conformed well to the fatty acid profiles, indicating the type of fatty acids as a critical factor in chemotaxonomic discrimination of these thraustochytrids and also revealing the distinctively high polyunsaturated fatty acid content as key identity of AMCQS5-5. Partial least squares discriminant analysis using cross-validation approach between two replicate datasets was demonstrated to be a powerful classification method leading to models of high robustness and 100% predictive accuracy for strain identification. The results emphasized the exceptional S-FTIR capability to perform real-time in vivo measurement of single live cells directly within their original medium, providing unique information on cell variability among the population of each isolate and evidence of spontaneous lipid peroxidation that could lead to deeper understanding of lipid production and oxidation in thraustochytrids for single-cell oil development.