Darryl M. Small

Development of a switchable multidimensional/comprehensive two-dimensional gas chromatographic analytical system.

In this study, a new system for analysis using a dual comprehensive two-dimensional gas chromatography/targeted multidimensional gas chromatography (switchable GC x GC/targeted MDGC) analysis was developed. The configuration of this system not only permits the independent operation of GC, GC x GC and targeted MDGC analyses in separate analyses, but also allows the mode to be switched from GC x GC to targeted MDGC any number of times through a single analysis. By incorporating a Deans switch microfluidics transfer module prior to a cryotrapping device, the flow stream from the first dimension column can be directed to either one of two second dimension columns in a classical heart-cutting operation. Both second columns pass through the cryotrap to allow solute bands to be focused and then rapidly remobilized to the respective second columns. A short second column enables GC x GC operation, whilst a longer column is used for targeted MDGC. Validation of the system was performed using a standard mixture of compounds relevant to essential oil analysis, and then using compounds present at different abundances in lavender essential oil. Reproducibility of retention times and peak area responses demonstrated that there was negligible variation in the system over the course of multiple heart-cuts, and proved the reliable operation of the system. An application of the system to lavender oil, as a more complex sample, was carried out to affirm system feasibility, and demonstrate the ability of the system to target multiple components in the oil. The system was proposed to be useful for study of aroma-impact compounds where GC x GC can be incorporated with MDGC to permit precise identification of aroma-active compounds, where heart-cut multidimensional GC-olfactometry detection (MDGC-O) is a more appropriate technology for odour assessment.

Investigation of folic acid stability in fortified instant noodles by use of capillary electrophoresis and reversed-phase high performance liquid chromatography

A single enzyme treatment with alpha-amylase, prior to the quantification of added folic acid (FA) in fortified instant fried Asian noodles with analysis performed by capillary zone electrophoresis (CZE) and reversed-phase high performance liquid chromatography (RP-HPLC) with UV detection, is described. The method was validated and optimized for capillary electrophoresis (CE) with separation achieved using a 8 mM phosphate-12 mM borate run buffer with 5% MeOH at pH 9.5. FA was well separated from matrix components with nicotinic acid (NA) employed as an internal standard. In a comparative study, separation of FA was performed using HPLC with a mobile phase consisting of 27% MeOH (v/v) in aqueous potassium phosphate buffer (3.5 mM KH(2)PO(4) and 3.2 mM K(2)HPO(4)), pH 8.5, and containing 5 mM tetrabutylammonium dihydrogen phosphate as an ion-pairing agent. For both methods, excellent results were obtained for various analytical parameters including linearity, accuracy and precision. The limit of detection was calculated to be 2.2 mg/L for CE without sample stacking and 0.10 mg/L with high performance liquid chromatography (HPLC). Sample extraction involved homogenization and enzymatic extraction with alpha-amylase. Results indicated that FA was stable during four main stages of instant fried noodle manufacturing (dough crumbs, cut sheets, steaming and frying).

Lactulose production from milk concentration permeate using calcium carbonate-based catalysts

Milk concentration permeate (MCP), a low-value by-product of ultrafiltration plants and calcium carbonate-based catalysts were used for lactulose production. The results obtained show the effectiveness of oyster shell powder and limestone for lactose isomerisation as a replacement for egg shell powder. With the reaction conditions of 12mg/ml catalyst loading, reflux time of 120min at 96°C, a maximum yield of 18-21% lactulose was achievable at a cost of <50% of original lactose degradation (measured by HPLC). De-proteination of MCP by acidification prior to isomerisation helped lactulose formation in the earlier stages, but did not significantly increase the yield. The resulting lactulose MCP (40°B) incorporated at the rate of 3-4% was effective in enhancing the growth rate and acid production of Lactobacillus acidophilus (LA-5) in probiotic products.

The behaviour of whey protein isolate in protecting Lactobacillus plantarum

There is increasing evidence that whey protein isolates (WPI), can be utilised to encapsulate and protect bioactive substances, including lactic acid bacteria, due to their physicochemical properties. However, little is known about what happens in the immediate vicinity of the cells. This study examined the protective behaviour of WPI for two strains of Lactobacillus plantarum, A17 and B21, during spray drying. B21 was found to be more hydrophobic than A17 and required 50% of the amount of WPI to provide comparably high survival (∼ 90%). We hypothesise that WPI protects the hydrophobic bacteria by initial attachment to the unfolded whey protein due to hydrophobic interactions followed by adhesion to the proteins, resulting in cells being embedded within the walls of the capsules. The encapsulated strains had a moisture content of approximately 5.5% and during storage trials at 20 °C retained viability for at least eight weeks.

Reduction of toxic gliadin content of wholegrain bread by the enzyme caricain

Increasingly the number of individuals being diagnosed with some form of sensitivity to the proteins in wheat grains represents a cause for concern. Currently, the treatment is dietary withdrawal of gluten, but commercial gluten-free bread presents some undesirable properties. The objective of this study has been to assess the ability of the enzyme caricain (from papaya latex) to detoxify gliadin in whole wheat flour and develop bread suitable for coeliacs and gluten intolerant individuals. Ion exchange chromatography was used to enrich the caricain in papaya latex and an enzyme-linked immunosorbent assay test kit was used for the analysis of gliadin residues in the baked bread. The partially purified enzyme was found to be more effective in reducing gliadin content than the crude papain and the resultant loaves had acceptable crumb and crust characteristics. Caricain appears to be capable of detoxifying gliadin and has the potential to mitigate the problems confronting coeliacs.

The effect of fermentation and addition of vegetable oil on resistant starch formation in wholegrain breads.

Resistant starch has potential health benefits but the factors affecting its formation in bread and baked products are not well studied. Here, the formation of resistant starch in wholemeal bread products was evaluated in relation to the processing conditions including fermentation time, temperature and the inclusion of palm oil as a vitamin source. The effects of each the factor were assessed using a full factorial design. The impact on final starch content of traditional sourdough fermentation of wholemeal rye bread, as well as the bulk fermentation process of wheat and wheat/oat blends of wholemeal bread, was also assessed by enzyme assay. Palm oil content was found to have a significant effect on the formation of resistant starch in all of the breads while fermentation time and temperature had no significant impact. Sourdough fermentation of rye bread was found to have a greater impact on resistant starch formation than bulk fermentation of wheat and wheat blend breads, most likely due the increased organic acid content of the sourdough process.

Controlled release of thiamin in a glassy κ-carrageenan/glucose syrup matrix.

The work dealt with the diffusional mobility of thiamin embedded in a high-solid matrix of κ-carrageenan with glucose syrup. It utilized thermomechanical analysis in the form of modulated differential scanning calorimetry and small-deformation dynamic oscillation in shear, Fourier transform infrared spectroscopy, wide angle X-ray diffraction, scanning electron microscopy and UV-vis spectrophotometry. The structural properties of the matrix were assessed in a temperature induced rubber-to-glass transformation. A thiamin-dye binding assay was employed to monitor the diffusion process of the vitamin from the high-solid preparation to ethylene glycol. The relationship between mechanical properties of the carbohydrate matrix and vitamin mobility was assessed via the application of the combined framework of the free volume theory and the predictions of the reaction rate theory. Results argue that the transport of the micronutrient is governed by the structural relaxation of the high-solid matrix. These were further treated with the concept of Fickian diffusion coefficient to provide the rate of the bioactive compound mobility within the present experimental settings.

Effect of a glassy gellan/polydextrose matrix on the activity of α-d-glucosidase

An investigation of the ability of the enzyme α-D-glucosidase to act on the substrate 4-nitrophenyl α-D-glucopyranoside (pNPG) while embedded in glassy carbohydrate matrices (deacylated gellan with polydextrose and polydextrose alone) is presented. Physicochemical characterisation of the matrices was achieved using the techniques of modulated differential scanning calorimetry, small deformation dynamic oscillation on shear, Fourier transform infra-red spectroscopy, wide angle X-ray diffraction and scanning electron microscopy. A UV-vis spectrophotometric procedure was adapted for the analysis of the activity of α-D-glucosidase in hydrolysing pNPG in the condensed carbohydrate systems. In order to derive a relationship between the structural properties of the matrix and the enzymatic activity, mechanical spectra were recorded using the combined framework of the Williams, Landel and Ferry equation with the time-temperature superposition principle. Theoretical modelling and experimental observations strongly argue for a pronounced effect of the gelling polysaccharide/co-solute mixture on enzymatic activity near the mechanical Tg of the matrix.

Assessment of arsenic in Australian grown and imported rice varieties on sale in Australia and potential links with irrigation practises and soil geochemistry.

Chronic dietary exposure to arsenic, particularly the inorganic forms (defined as elemental arsenic, predominantly As(3+) and As(5+), and all its inorganic compounds except arsine), is a matter of concern for human health. Ingestion of arsenic usually occurs via contaminated water but recent studies show there is also a risk of exposure from food, particularly Asian rice (Oryza sativa). Australia is a rice growing country, contributing around 2% of the world rice trade, and a large proportion of the population consumes rice regularly. In the present study we investigated concentrations of arsenic in both Australian grown and imported rice on sale in Australia and examined the potential links with irrigation practises and soil geochemistry. The results indicated a wide spread of arsenic levels of 0.09-0.33 mg kg(-1), with Australian grown Arborio and sushi varieties of O. sativa containing the highest mean value of ∼0.22 mg kg(-1). Arsenic levels in all samples were below the 1 mg kg(-1) limit set by Food Standards Australia New Zealand.

Impact of α-Amylases on Quality Characteristics of Asian White Salted Noodles Made From Australian White Wheat Flour

ABSTRACT The consumer acceptance of white salted Asian noodles depends on starch characteristics, and the purpose of this study has been to investigate the potential of exogenous α-amylases to enhance textural characteristics of this product. Noodles were prepared from commercial flours with low α-amylase activity, and the endogenous enzyme remained relatively stable during various processing and storage treatments. α-Amylase preparations of bacterial origin and from barley malt were incorporated, and the products were assessed by texture analysis and electron microscopy, as well as for color characteristics. On addition of the amylase preparations, noodles were softer when texture was assessed using either a flat cylinder probe or an axial blade. Some discoloration occurred in treated noodle sheets, although this was minimal in final products that had been cooked immediately after preparation or following drying. Scanning electron microscopy confirmed that the α-amylase of bacterial origin had greater im...