Eur Food Res Technol (2010) 231:441453 DOI 10.

1007/s00217-010-1297-4

ORIGINAL PAPER

Fundamental rheological and textural properties of doughs and breads produced from milled pearled barley Xour
Paul Sullivan John OFlaherty Nigel Brunton Elke Arendt Eimear Gallagher

Received: 11 February 2010 / Revised: 11 May 2010 / Accepted: 15 May 2010 / Published online: 5 June 2010 Springer-Verlag 2010

Abstract Barley has long been known as a good source of Wbre and in particular beta-glucan, but increased consumer awareness has meant that this is only now being exploited by food scientists and the food industry. In this study, doughs and breads were produced using pearled barley Xour (PBF) in diVerent ratios (30, 50, 70 and 100%) to wheat Xour. A 100% wheat Xour formulation was used as a control. The Xour formulations were evaluated for protein content; the dough rheological properties of the formulations were evaluated using fundamental oscillatory tests and uniaxial extension tests. The baked breads were evaluated for volume, texture, moisture, dietary Wbre and betaglucan. Digital image analysis of the crumb grain was also carried out. Protein analysis revealed that increasing the amount of PBF in the formulation leads to a signiWcant (p < 0.001) decrease in the protein content of the formulation. Starch pasting properties were also aVected by an increase in PBF concentration, with an increase in PBF signiWcantly increasing the peak viscosity, breakdown, setback and Wnal viscosity of the starch slurries of the formulations. Extensional rheology revealed that increasing the PBF in the doughs leads to the doughs rupturing at lower extensions and with a lower force. Bread volume between treatments was found to be signiWcantly (p 0.001) diVerent, with an increase in PBF inclusion leading to smaller loaves. Texture proWle analysis of the bread slices showed that breads containing higher ratios of

PBF had a signiWcantly harder (p 0.001) and less cohesive (p 0.001) crumb. Low levels of PBF (50%) did not signiWcantly eVect loaf volume or crumb texture, however, and the total dietary Wbre and beta-glucan levels of the loaves were signiWcantly increased (p 0.001) with increasing levels of PBF in the formulation, even at low levels of inclusion. Keywords Pearling Barley Milling Bread Beta-glucan Dietary Wbre

Introduction Barley has been used as a source of both human and animal feed for thousands of years, but with the increased cultivation of wheat and rice, it has become less commonly used for human consumption. However, due to its high soluble Wbre content and nutritional signiWcance, it has once again become a desirable food source. In 2007, barley ranked fourth behind maize (792 million metric tonnes), rice (660 million metric tonnes) and wheat (606 million metric tonnes) in quantity produced (133 million metric tonnes) and in area cultivation of cereal crops worldwide [13]. In recent years, the development of new functional foods has become a priority for both the food scientist and the food industry, primarily as a response to increased consumer awareness and demand. The utilisation of barley in breads has been reported to improve its nutritional value and thus its beneWts to human health [17]. The health-related importance of soluble Wbres such as beta-glucan has long been recognised; beta-glucans in particular have attracted much attention due to their functional properties [6, 30]. Beta-glucan has been shown to have a positive eVect on glycaemic insulin responsea 2006

P. Sullivan (&) J. OFlaherty N. Brunton E. Gallagher Ashtown Food Research Centre, Teagasc, Ashtown, Dublin 15, Ireland e-mail: paul.sullivan@teagasc.ie P. Sullivan J. OFlaherty E. Arendt University College Cork, Cork, Ireland

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study by Nilsson et al. [22], showed that barley could regulate blood sugar levels for up to 10 h after consumption. Also, their acceptance as bioactive ingredients is recognised worldwide [10]. Beta-glucan is a non-starch polysaccharide composed of linear chains of glucose with -(1 ! 3) and -(1 ! 4) linkages in the endosperm cell walls of cereals, particularly in oats and barley. -(1 ! 3) and -(1 ! 4)-glucan have been deWned by AACC International [2] as the edible parts of plants or analogous carbohydrates that are resistant to digestion and absorption in the human small intestine with complete or partial fermentation in the large intestine and can be either a soluble or insoluble dietary Wbre. Dry milling of grains provides milled fractions that contain both soluble and insoluble material. Such fractions are believed to oVer the combined beneWcial physiological eVects of soluble and insoluble dietary Wbre when incorporated into food systems. Fractions rich in beta-glucan have been obtained from cereal grains by dry milling (pin or roller), sieving and air classiWcation processes [18, 20]. These approaches resulted in fractions containing approximately 830% beta-glucans. Pearling is one of the oldest practices of processing barley. This is a process by which abrasive scouring is used to gradually remove the hull, pericarp, seedcoat, aleurone and subaleurone layers, as well as the embryo. Depending on the degree of pearling, the process can generate de-hulled barley, pot barley or pearled barley. De-hulling removes only the husk, whilst removal of the next kernel layer, the pericarp, produces pot barley. Further abrasion removes the seedcoat, aleurone and subaleurone layers, producing pearled barley. The pearled barley usually represents 4050% of the grain, but the degree of abrasion may be adjusted depending on what the Wnal product is to be used for [19]. This paper focuses on barley Xour produced from milled peeled and pearled barley. The work presented assessed some of the compositional characteristics of Xour from milled pearled barley (PB). The suitability of these Xours for bread making (when used on their own or as composites with wheat Xour) was also analysed.

Materials and methods Wheat Xour A standard commercial wheat Xour suitable for breadmaking (Odlum Group Ltd., Dublin, Ireland) was used in this study. Barley Xour Peeled and pearled barley Peeling and pearling of the barley was carried out by Bhler in Switzerland (Bhler AG, Switzerland). Peeling involved the removal of the outer layers of the barley grain (6.2% w/w) and was followed by pearling, where the peeled grain was polished, thus removing further layers of the grain (4.1% w/w). The peeled and pearled grain was packed into 30 kg bags and sent to Ashtown Food Research Centre for milling. Barley milling Milling of the pearled barley was carried out in a roller mill (Bhler AG, Switzerland) at Ashtown Food Research Centre, Dublin, and was milled to a milling degree of 70% following the AACC Bhler method [1]. The milling process produced two fractions of Xour (break Xour and reduction Xour) a bran fraction and a middlings fraction. The Xour fractions were combined, bagged and stored in a cool, dry room until use. Studies involving the use of the bran and middlings fractions will be reported on in future papers. Flour combinations and bread formulations After a number of preliminary baking tests using a wide range of barley Xour levels, a number of Xour combinations were selected for further analysis (Table 1). The baking recipe was Xour, 2 g salt 100 g1 Xour, 3 g bread fat 100 g1 Xour and 2.5 g fresh yeast 100 g1 Xour as well as water (as per the results of the farinograph measurements).

Table 1 Properties of pearled barley and wheat Xour combinations

Combination code A B C

Pearled barley Xour trial 100% Wheat Xour 70% Wheat Xour, 30% pearled barley Xour 50% Wheat Xour, 50% pearled barley Xour 30% Wheat Xour, 70% pearled barley Xour 100% Pearled barley Xour

Water absorption (%) 63.7 (0.3) 63.0 (0.5) 62.8 (0.3) 62.5 (0.0) 62.0 (0.5)

Protein (%)a

10.67 (0.27) 10.55 (0.22) 9.16 (0.18) 7.90 (0.17) 5.59 (0.10)

N 5.7 for wheat Xour and 6.25 for barley combinations and barley Xour

D E

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Flour analysis Mixing properties The mixing properties of the diVerent wheat Xour and barley combinations were examined using a Brabender Farinograph (Brabender OHG, Duisberg, Germany) following the farinograph British Standard Method No. 431920:1999 [7]. Each analysis was performed in triplicate, and mean values were calculated. Protein Protein content The wheat Xour and barley combinations were analysed for protein content using a Leco protein analyser (Leco FP-428, Nitrogen Analyzer, Leco Corporation, St. Joseph, MI, USA). Each analysis was performed in triplicate. Protein proWling The protein proWle of each Xour combination was analysed using Lab-on-a-chip capillary electrophoresis (Agilent Technologies, Palo Alto, CA). Total proteins were extracted from each Xour combination sample with 2 M Urea, 15% glycerol, 0.1 M Tris/HCL, pH 8.8 and 0.1 M DTT solution. The solution (0.4 ml) was added to each Xour combination (40 mg), and the samples were placed in a water bath at 36 C for 5 min before they were centrifuged at 14,000 rpm for 15 min. Aliquots of 4 l of the extracts were treated using the Agilent protein 230 assay reagents, following the assay protocol. The samples (6 l) were then applied to the sample wells of an Agilent protein 230 lab chip for analysis in the Agilent 2100 Bioanalyzer. The method allowed for analysis of 10 samples (in addition to a ladder included in the Agilent protein 230 assay reagents). Duplicate measurements of each sample (AE) were carried out, and the results were analysed using the Agilent 2100 Expert software (Agilent Technologies 20032007). Starch pasting properties The pasting properties of the Xours were determined with a Rapid ViscoAnalyser (RVA-4D, Newport ScientiWc, Sydney, NSW, Australia). The 13 min test proWle has a starting temperature of 50 C, which is held for 1 min, raised to 95 C in 3.7 min, held for 2.5 min, cooled to 50 C in 3.8 min and held for 2 min. Stirring speed is 960 rpm for 10 s (to ensure dispersion of the grist) and 160 rpm for the remainder of the test period. Results obtained from the test were as follows: peak viscosity (highest viscosity during heating); time to reach peak viscosity; trough (lowest viscosity after cooling); breakdown (peak viscosity minus trough); Wnal viscosity (maximum viscosity after the tem-

perature had returned to 50 C); and setback (Wnal viscosity minus trough) [31, 32]. All RVA analyses were performed in triplicate. Dough evaluation Uniaxial extension Uniaxial extension using the KieVer dough and gluten extensibility rig (Stable Micro Systems, Surrey, UK) was carried out on all the samples (AE) using a similar method to that described by Dunnewind et al. [12]. A texture analyzer (TA-XT2i, Stable Micro Systems, Surrey, UK) Wtted with the KieVer extensibility rig and equipped with a 5 kg load cell was used. The samples were prepared following the method described in the literature accompanying the KieVer extensibility rig. A piece of dough (25 g) was placed to rest in a proofer (Koma SDCC-1P/W, Koma Koeltechnische Industrie B.V., The Netherlands) with controlled temperature (30 C) and relative humidity (80%) for 20 min. The sample was then rolled by hand into a cylindrical shape, placed in the lubricated TeXon mould and compressed with the lubricated top TeXon plate. The sample was allowed to rest for another 45 min in the proofer at 30 C and 85% relative humidity. Before the start of the test, the sample was clamped between the plates of the KieVer rig. During the test, the maximum force in tension and the displacement were recorded. Three dough samples were used for each formulation, and the results were averaged. According to Dunnewind et al. [12], clamping of the sample results in a certain degree of sagging of the dough and therefore, the actual extension of the dough piece starts somewhat above the surface of the lower plate. Corrections were applied to account for this by measuring the distance the hook travelled from a predetermined starting point to the point where it touched the dough. The diVerence between this distance and the known distance from the starting point to the top of the bottom plate gave the height of sagging. Adding this to the point of the top surface of the bottom plate, the point of when the actual extension started was determined. The point when the hook touched the dough was determined as the point which the measured force deviated from zero, as described by Ktenioudaki et al. [21]. Fundamental rheology Rheological measurements were performed on a controlled stress rheometer (Anton Paar Gmbh, Graz, Austria) Wtted with parallel plates consisting of a 25-mm serrated probe and 25-mm serrated base plate. Dough samples (based on 300 g of Xour) were prepared as previously described in Flour combinations and bread formulations but without

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yeast. The samples were placed onto the base plate, and the upper plate was brought to a gap of 1.025 mm where excess was carefully trimmed. The plate was then lowered to 1 mm and testing began. The whole system was covered using a peltier hood, with a temperature setting of 30 C. The dough was allowed to rest for 5 min to allow relaxation of residual stresses. A frequency sweep from 0.1 to 10 Hz was performed with a target strain of 103 (0.1%). Preliminary tests (amplitude sweeps) indicated that the strain was within the linear viscoelastic region. Ten measuring points were recorded. The temperature was kept constant at 30 C. Each result is the average of four measurements. Bread evaluation Baking trials Loaves (300 g dough pieces in a 454 g tin) were prepared using the formulations described in Flour combinations and bread formulations following a straight dough baking procedure. The doughs were mixed in a Hobart mixer for the optimal mixing time, as measured by the farinograph plus 1 min. The doughs were moulded by hand and placed in the tins. The tins were transferred to a proofer (Koma SDCC 1P/W, Koma Koeltechnische Industrie B.V., The Netherlands) at 35 C and 80% relative humidity for 65 min. The loaves were baked at 220 C for 22 min in a deck oven (Tom Chandley Ltd., Manchester, UK). Six loaves were produced per batch. The loaves were allowed to cool for 2 h, and their loaf volume was tested using a volume measurer (BVML370, TexVol Instruments, Sweden). The loaves were then stored in polyethylene bags (2 loaves per bag) at room temperature. After 24 h, the Wrst bag was opened, and the two loaves were tested for crumb structure using image analysis with the C-Cell Bread Imaging System (Calibre Control International Ltd., UK) and for crumb texture using a Texture ProWle Analysis (TPA) programme using a texture analyzer (TA-XT2i, Stable Micro Systems, Surrey, UK). Two slices per loaf (4 slices in total) were used for image analysis and for crumb TPA, and the results were averaged. Finally, crumb and crust colour measurements took place using a colorimeter (Minolta CR-331, Konica Minolta Holdings Inc., Japan). L*, a*, and b* values for crust and crumb were recorded. L*, a* and b* coordinates indicate the colour of a sample on a three-dimensional scale. L* indicates the lightness of a sample (L = 0 refers to black, while L = 100 refers to white), a* indicates the colours position between magenta and green (negative a* values indicate the sample is more green than magenta, while positive a* values indicate the sample is more magenta than green) and Wnally, the b* values indicate the colours position between blue and

yellow (negative b* values indicate the colour is more blue than yellow, while positive b* values indicate the colour is more yellow than blue. The loaf volume results were the average of 6 loaves per replicate; for the image analysis, TPA and colour measurements, the results were the average of 4 measurements per replicate (2 slices from each loaf). This process was then repeated on the second bag after 72 h and the third bag after 120 h. Fibre Fibre analysis was carried out according to AOAC Method 991.43 [4], adapted for use with Foss Fibertec Equipment. Duplicate samples (pre-dried, de-sugared and defatted as required) were enzymatically digested to remove protein and starch. After precipitation of soluble dietary Wbre with alcohol, the total residue was dried and weighed. One duplicate was analysed for protein and the other for ash. Total dietary Wbre was calculated as the weight of residue minus the weight of protein plus ash in the sample. Beta-glucan All bread samples (approximately 50 g) were freeze dried and ground to pass a 0.5-mm screen using a Fritsch pulverisette 14 (Fritsch GmbH Idar-Oberstein, Germany). The samples (80120 mg) were added to glass centrifuge tubes (16 120 mm; 17 ml capacity). A volume of 0.2 ml of aqueous ethanol (50% v/v) was added to the tubes in order to aid dispersion of the samples. Following the addition of ethanol, sodium phosphate buVer (4.0 ml, 20 mM, pH 6.5) was added and the tubes vortexed. On mixing, the tubes were placed in a boiling water bath and incubated for 60 s. The samples were vigorously stirred on a vortex mixer and incubated at 100 C for a further 2 min before being stirred again. The tubes were incubated at 50 C and allowed to equilibrate for 5 min. Lichenase was added (0.2 ml, 10 U) to the samples and stirred before being sealed with paraWlm and incubated for 1 h at 50 C, being stirred regularly on a vortex mixer. After 1 h, sodium acetate buVer (5.0 ml, 200 mM, pH 4.0) was added to the tubes which were vortexed. The tubes were then allowed to equilibrate to room temperature for 5 min and centrifuged (1,000g, 10 min). From the samples, 0.1 ml aliquots were dispensed into three test tubes (12 ml capacity). Beta-glucosidase (0.1 ml, 0.2 U) in 50 mM sodium acetate buVer (pH 4.0) was added to two of these tubes (the reaction). To the third (the reaction blank), 50 mM acetate buVer (0.1 ml, pH 4.0) was added. The tubes were incubated at 50 C for 10 min. After incubation, 3.0 ml of GOPOD Reagent (Megazyme International Ireland Ltd., Ireland) was added to each tube and incubated at 50 C for a further 20 min. The tubes were

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removed from the water bath, and their absorbances were measured at 510 nm within 1 h. The experiment was run in triplicate, and the results were calculated as a mean of these triplicates. Taste panel Sensory analysis was conducted on the Wve bread samples at 24 h post baking by 20 tasters. Panellists were asked to assess the breads for acceptability, and to mark a 6 cm line (0 = unacceptable, 6 = very acceptable) in accordance with their opinion. Results were analysed (ANOVA) as 5 samples 20 trained tasters. Statistical analysis ANOVA one-way statistical analysis was carried out using Minitab (Minitab version 15.1.1.0., Minitab Ltd., UK) to determine signiWcant diVerences measured in the properties of the Xours, doughs and breads. Pearsons correlation analysis between the various parameters was also carried out using Minitab (Minitab version 15.1.1.0., Minitab Ltd., UK).

concentration decreased the water absorption of the formulation. The dough development time, stability and degree of softening (data not shown) were not found to diVer signiWcantly between Xour combinations and showed no apparent pattern as to the eVect of barley addition on these parameters. Protein Protein content There were signiWcant (p < 0.01) diVerences between the protein contents of the diVerent Xour formulations (Table 1). Increasing the pearled barley Xour level in the formulation leads to a signiWcant decrease in protein content. Andersson et al. [3] reported on the protein content of a number of varieties of milled barley fractions and found their protein content of the diVerent fractions ranged from 7.7 to 9%, which is signiWcantly higher than the result found in this study, where pearled barley Xour had a protein content of 5.6%, suggesting that the cultivar of barley used in this study, Sebastien (Spring 2007) has a low protein content. This diVerence may be due to genomic, environmental or agronomic factors. A low protein content in Xour usually indicates that a Xour will have a poor baking quality when compared to a Xour which has a high protein content, this would therefore suggest that increasing the amount of barley Xour in the bread formulation may produce a lower quality bread. Protein proWling Protein proWle analysis of the Xour formulations produced from the milling of pearled barley combined with standard bakers wheat Xour are shown in Fig. 1. Electropherograms give an indication of the abundance of proteins found in a sample. From the results, it is apparent that increasing the pearled barley Xour concentration of

Results and discussion Flour analysis Mixing properties Mean farinograph results for the various Xour combinations are shown in Table 1. From the results, it is apparent that the addition of barley to a wheat Xour formulation had a slight eVect on the resulting dough. Increasing the barley

Fig. 1 Electropherograms of A: 100% wheat Xour; B: 70% wheat, 30% PBF; C: 50% wheat, 50% PBF; D: 30% wheat, 70% PBF and E: 100% PBF

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the combinations leads to an overall decrease in the abundance of protein in the combinations, and this is particularly apparent from the peaks found at the 35 s retention time (4663 Kda), where their intensity falls from approximately 450 Xuorescent units in 100% wheat Xour to less than 100 Xuorescent units in 100% pearled barley Xour. There was, however, a small increase in the abundance of proteins at a retention time of 30 s, which relates to the 15 28 Kda range. Salcedo et al. [24] found the 1528 Kda range to contain A-hordeins, suggesting the substitution of barley Xour for wheat Xour increases the concentration of A-hordeins in the Xour. This would agree with the results of the protein content analysis described previously. Starch pasting properties The pasting properties of starch pastes from pearled barley Xour and wheat Xour composites are shown in Table 2. The substitution of pearled barley Xour for wheat Xour had a signiWcant eVect on the starch pasting properties of the starch slurries of the resulting composite Xours (p < 0.01). The breakdown in viscosity during the test increased with the increased inclusion of pearled barley in the composite Xours. High breakdown values are correlated with high peak viscosities. This also relates to the degree of swelling of the starch granules during heating. More starch granules with a high swelling capacity result in a higher peak viscosity [23]. The highest breakdown was measured for 100% pearled barley Xour. A 2006 study by Ragaee and Abdel-Aal [23] found that the substitution of 15% barley into wheat formulae increased the peak viscosity of the pastes as well as the breakdown and setback of the pastes when compared to the wheat Xour. The peak time was found to be similar in barley Xours to wheat Xours and also similar to the barley substituted wheat Xour, which is in agreement with the results of this study. Increasing the level of pearled barley Xour increased the setback (starch retrogradation) of the paste. Therefore, breads containing higher levels of pearled barley Xour could stale more rapidly than those containing low pearled barley levels. Bhatty and Rossnagel [5] suggested pearled

barley varieties which exhibited high peak viscosity and setback do so due to higher beta-glucan contents in their starch slurries. These results would suggest that higher levels of pearled barley Xour in a bread formulation may aVect both the shelf life and textural qualities of the resulting breads, leading to a lower quality Wnal product. Dough analysis Fundamental rheology The storage modulus of a material refers to the deformation energy stored in the material after oscillation is removed. It is a measurement of a materials elastic properties. The higher the value is, the more elastic the material will be. The loss modulus of a material refers to the energy lost from the sample during oscillation. If energy is lost, the sample cannot go back to its original shape, which is an indication of viscous behaviour. In this study, all doughs had a higher storage modulus (G) than loss modulus (G), indicating that the doughs had a solid, elastic-like behaviour which is to be expected (Fig. 2). SigniWcant diVerences (p < 0.01) were observed in the rheological properties of the composite doughs (i.e. the doughs containing both wheat and barley Xour) as the level of barley Xour addition increased. Skendi et al. [27] found that the inclusion of beta-glucan increased the G values of Xours with good breadmaking quality Xour doughs, whereas decreased the G of poor quality wheat cultivar, which would agree with these results as the wheat Xour used in the present study was a Bakers Xour of good quality. The authors suggested that the changes in the dough rheology may be due to beta-glucans having the highest aYnity for water uptake in the dough followed by gluten and starch. Consequently, the distribution of water in the dough structure would be altered by the addition of betaglucans. The pure doughs (i.e. the 100% wheat and the 100% barley Xour doughs) had the lowest values of loss and storage modulus. The composite doughs had signiWcantly higher storage and loss moduli than the pure Xour doughs

Table 2 Rapid visco analysis of pearled barley (PBF) Xour and wheat combinations Formulation code A B C D E Combination Peak viscosity (cp) 1,825 (19) 3,722 (22) 3,779 (23) 4,648 (28) 5,839 (35) Breakdown (cp) 929 (7) 1,488 (9) 1,495 (9) 1,735 (11) 2,380 (14) Final viscosity (cp) 1,891 (85) 3,641 (22) 3,754 (22) 4,588 (28) 5,326 (32) Setback (cp) 996 (36) 1,408 (8) 1,470 (19) 1,676 (10) 1,867 (11) Peak time (min) 5.76 (0.03) 5.87 (0.04) 5.93 (0.02) 6.00 (0.05) 5.80 (0.05)

100% Wheat 70% Wheat, 30% PBF 50% Wheat, 50% PBF 30% Wheat, 70% PBF 100% PBF

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Eur Food Res Technol (2010) 231:441453 Fig. 2 Frequency sweep of pearled barley dough combinations, WF wheat Xour, PBF pearled barley Xour
|G'| |G''| [Pa]
10000000

447

1000000

100000

0.1

10

Frequency (Hz)
G' 100% PBF G' 50% WF 50% PBF G' 100% WF G'' 100% PBF G'' 50% WF 50% PBF G'' 100% WF G' 30% WF 70% PBF G' 70% WF 30% PBF G'' 30% WF 70% PBF G'' 70% WF30% PBF

(p < 0.01). The diVerence between the pure doughs and the composite doughs could be attributed to a diVerence in the interaction of starchgluten in composite doughs. A 2004 study by Sivaramakrishnan et al. [26], on the rheological properties of rice Xour doughs also found that composite doughs diVered rheologically when compared to pure doughs. The complex moduli of the dough combinations are shown in Fig. 3. It is apparent from the results that the complex moduli diVer signiWcantly between Xour combinations (p < 0.05). As with the results for the storage and loss moduli, the doughs can be divided into two categories, composite combinations and pure combinations. The composite combinations have a higher complex modulus than the pure combinations, with the higher barley
Fig. 3 Complex modulus of pearled barley dough combinations from 0.1 to 10 Hz. A: 100% wheat Xour; B: 70% wheat Xour, 30% PBF; C: 50% wheat Xour, 50% PBF; D: 30% wheat Xour, 70% PBF; E: 100% PBF

content in the composite doughs measuring the highest values of complex modulus. This indicates that the addition of barley to the composite doughs increases resistance to deformation, producing a more Wrm dough. Between the two pure Xour doughs (i.e. 100% wheat and 100% barley), the wheat dough has a signiWcantly higher complex modulus than the barley dough, suggesting the wheat dough was more resistant to deformation. This is most likely due to the gluten matrix of the wheat dough providing the dough with increased structural strength. The phase angle is an important measurement when studying the rheology of doughs as it indicates how a viscoelastic sample behaves. The phase angles of all the samples during the frequency sweeps were within the 1020 region (from 18.20 to 16.55 at 0.1 Hz and from 15.75 to
10000000

Complex Modulus |G*| [Pa]

1000000

100000 0.1 1 10

Frequency
|G*| A |G*| B |G*| C |G*| D |G*| E

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13.15 at 10 Hzdata not shown). This indicates that the samples had more elastic than viscous attributes, as expected in a dough. Such a narrow phase angle range would suggest that there is little diVerence in the viscoelastic properties of doughs produced from diVerent barley Xour inclusion levels. Uniaxial extension Uniaxial extension can be measured using a KieVer dough and gluten extensibility rig, which extends a clamped piece of dough until rupture. Doughs that are characterised by high extensibility (i.e. an increased distance before rupture) and high maximum resistance to extension (i.e. an increased force required to rupture the dough) are likely to have good bread making qualities; therefore, this method can be useful to determine the eVect that the substitution of barley Xour for wheat will have on potential Wnal products. An increase in barley concentration in the dough signiWcantly decreased both the length the dough could be extended before rupture, and the force required to bring about the rupture (Fig. 4). This would suggest that the elasticity of the dough was decreased by the addition of the barley Xour. Skendi et al. [27] studied how the inclusion of betaglucan into a dough system aVected a doughs strength. The authors found that the increased inclusion of beta-glucan weakened the gluten network of the dough and suggested that this was due to the beta-glucan disrupting intermolecular associations of gluten proteins. As a result, doughs containing higher levels of beta-glucan will require less force to disrupt and will also be less elastic.

Bread analysis Loaf volume Bread formulations with high concentration of pearled barley had signiWcantly (p 0.001) decreased loaf volume in comparison with the wheat control (Fig. 5). Kkrbic et al. [28] and Cleary et al. [9] also found the addition of barley Xour to a wheat Xour formulation leads to a decrease in bread volume. This is to be expected, as the protein/gluten concentration of the Xour formulations was decreased with increasing levels of lower protein barley Xours. This gluten dilution, coupled with less retention of CO2 gas, would lead to a decrease in baking quality [25]. In the present study, it was found that inclusions of up to 50% barley Xour did not signiWcantly aVect loaf volumes. Crust and crumb colour The levels of barley incorporation signiWcantly aVected the colour properties of the loaves (Fig. 6). Higher levels of barley inclusions signiWcantly increased the L* values of the bread crust, i.e. the crust was signiWcantly lighter (p 0.001). With L* values ranging from 47.5 for 100% wheat Xour to L* values of 74.5 for 100% barley Xour breads. This diVerence was clearly visible to the naked eye, with the increased barley inclusion leading to a paler crust. Gill et al. [15] also found that there was a gradual lightening of crust colour with an increased incorporation of barley Xour into wheat bread formulations. Texture proWle analysis Crumb textural properties of the slices using TPA software on a TAXT2i Texture Analyser at 24, 72 and 120 h after

25
A

900 800 700

20 15
C B

a b

Volume (cm3)
50 60 70

Force (N)

600 500 400 300 200 100

10 5 0 0 10
E

20

30

40

Displacement (mm)
100% WF 70% WF 30% PBF 50% WF 50% PBF 30% WF 70% PBF 100% PBF

Formulation

Fig. 4 Uniaxial extension of doughs made from wheat Xour and pearled barley Xour composites, WF wheat Xour, PBF pearled barley Xour. A: 100% wheat Xour; B: 70% wheat Xour, 30% PBF; C: 50% wheat Xour, 50% PBF; D: 30% wheat Xour, 70% PBF; E: 100% PBF

Fig. 5 Loaf volumes of breads produced from diVerent whole barley Xour with wheat and pearled barley Xour with wheat composites. A: 100% wheat Xour; B: 70% wheat Xour, 30% PBF; C: 50% wheat Xour, 50% PBF; D: 30% wheat Xour, 70% PBF; E: 100% PBF. * Columns with diVerent superscripts are statistically signiWcant (p 0.05)

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80.00 70.00 60.00 50.00
a a,b b,c c d

449

40.00 30.00 20.00 10.00 0.00 A B C D E

loaves containing 70 and 100% barley Xour had the lowest cohesiveness values after 120 h storage. This agrees with the study of Gujral et al. [16] who found that the addition of barley Xour to a wheat bread formulation leads to a decrease in the cohesiveness of the bread slices, with a 100% wheat Xour bread slice having a cohesiveness of 0.51 after 1 day compared to 0.31 in an 80% wheat, 20% barley containing bread. Hardness Crumb hardness for pearled barley Xour bread slices is shown in Fig. 8. It can be seen that the control (100% wheat) resulted in breads with the softest crumb, and increasing the levels of barley in the formulations leads to a signiWcant increase (p 0.001) in the crumb hardness. Hardness was particularly increased above 50% barley Xour inclusion. This would suggest that to produce a satisfactory bread product, the level of pearled barley Xour substitution should not exceed 50% if a bread of a satisfactory quality is to be produced. Crumb structure (digital image analysis) Crumb structure is an important characteristic when evaluating a Xour for bread making suitability. A good bread making Xour should produce a crumb that consists of a large number of small sized, thin walled cells. Two slices of 1 cm thickness from the centre of each loaf were scanned, and the images analysed using the C-Cell software (Calibre, UK). Typical raw images of the slices from the bread formulations can be seen in Fig. 9. From the Wgure, it is apparent that increasing the barley levels in the bread formulations produced loaves with a decreased height and volume. The results of Kkrbic et al. [28] described barley containing
5000

L*

Formulation

Fig. 6 Crust colour (L*) of the loaves made from diVerent barley inclusion levels. A: 100% wheat Xour; B: 70% wheat Xour, 30% PBF; C: 50% wheat Xour, 50% PBF; D: 30% wheat Xour, 70% PBF; E: 100% PBF. * Columns with diVerent superscripts are statistically signiWcant (p 0.05)

baking. Two way statistical analysis (ANOVA) revealed signiWcant diVerences (p 0.001) among the formulations for resilience, chewiness, springiness and adhesiveness, but in particular the cohesiveness and hardness of the crumb. Cohesiveness Crumb cohesion is a desirable property in breads; by measuring crumb cohesiveness, one can understand the force required to break apart the crumb. Figure 7 shows the results for crumb cohesiveness properties. The control (100% wheat) breads had the highest crumb cohesiveness properties. An increased barley inclusion in the bread formulations leads to a signiWcant decrease (p 0.001) in the crumb cohesiveness. Crumb cohesiveness of the control was 0.52 and decreased to 0.43 after 120 h storage. Crumb cohesiveness of all samples decreased over the storage period (p 0.001). In particular,

0.600 0.500

4500 4000

Hardness (g)

3500 3000 2500 2000 1500 1000 500 0 A B C D E

Cohesiveness

0.400 0.300 0.200 0.100 0.000 A B C D E

Formulation
Day 1 Day 3 Day 5

Formulation
Day 1 Day 3 Day 5

Fig. 7 InXuence of pearled barley Xour inclusion on the cohesiveness of bread slices. A: 100% wheat Xour; B: 70% wheat Xour, 30% PBF; C: 50% wheat Xour, 50% PBF; D: 30% wheat Xour, 70% PBF; E: 100% PBF. F-test: treatment (T) (p 0.001; sed 0.008), time (t) (p 0.001; sed 0.003), interaction (T * t) (p 0.05; sed 0.008)

Fig. 8 Crumb hardness of the baked pearled barley Xour samples tested as measured with texture analyser during TPA. A: 100% wheat Xour; B: 70% wheat Xour, 30% PBF; C: 50% wheat Xour, 50% PBF; D: 30% wheat Xour, 70% PBF; E: 100% PBF. F-test: treatment (T) (p 0.001; sed 134.3), time (t) (p 0.001; sed 67.7), interaction (T * t) (p 0.001; sed 182.5)

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450 Fig. 9 Digital images of bread slices of barley Xour inclusion levels. (LR) A: 100% wheat Xour; B: 70% wheat Xour, 30% PBF; C: 50% wheat Xour, 50% PBF; D: 30% wheat Xour, 70% PBF; E: 100% PBF

Eur Food Res Technol (2010) 231:441453

bread samples as having a Xat shape due to the lower levels of gluten in the breads and therefore, reduced oven spring, and this is evident from the Wgure. Slice area and slice height Slice area and slice height are important properties when assessing the quality of a bread loaf. Increased slice area and slice heights would suggest increased levels of expansion during prooWng, leading to a more desirable end product. The level of barley level inclusion had a signiWcant impact on both the slice area and the height of the bread slices (p 0.001). Increasing the barley Xour inclusion leads to a decrease in the slice area (Fig. 10). The volume of the loaves were shown to have been signiWcantly reduced on inclusion of barley Xour into the formulation, and so this result would also be expected. Increasing the ratio of barley Xour in the bread formulation leads to a decrease in the slice area of the resulting loaf. This would be expected, as both the volume of the loaves from which the slices were taken and the slice area were also found to decrease signiWcantly (p 0.001) on the increased addition of barley Xour. Although the slice area did decrease on addition of barley Xour, the decrease was not signiWcant up to a barley inclusion level of 50%, again suggesting that if a quality bread product is to be produced, it cannot exceed the 50% level. Number of cells The area of cells in a bread slice gives an indication of the size of the CO2 bubbles captured during
6000 90
b b c c

prooWng. For a bread slice to be considered to be of good quality, usually a large number of small sized cells are desirable. The number of cells was signiWcantly (p < 0.01) aVected by the increased inclusion of barley Xour. The number of cells decreased when the level of barley inclusion was increased. This was particularly evident in the 100% barley Xour formulation, which had an average of 2,431 cells compared to 3,226 in the control. This can be partly explained by the lower loaf volumes associated with barley containing breads but is also an indication of a more coarse structure above 50% barley inclusion when compared to the control. Area of holes The area of holes was signiWcantly increased by the increased inclusion of barley Xour, particularly above 50% barley Xour, with the 70% barley Xour formulation having an average hole area of 3.2 mm3, compared to 0.25 mm3 in the control. This would again suggest that increasing barley above a 50% inclusion level can have detrimental eVects on the Wnal product. From the above results, it can be seen that the inclusion of barley Xour increased the area of holes in the bread while decreasing both slice height and area and the number of cells in the bread. A dough requires a strong gluten matrix to produce a good quality Wnal bread product. By substituting barley Xour for wheat Xour, the gluten concentration of the dough will be decreased and therefore, the doughs ability to form a strong gluten matrix may be aVected. With a weak gluten matrix, the dough is less likely to be able to trap CO2 bubbles during prooWng, and so these will escape leading to a decreased slice area, slice height and also a decrease in the number of cells, which are formed by the CO 2 bubbles trapped in the dough. The increase in the number of holes may also be explained by a weak gluten matrix, where the matrix has ruptured allowing CO2 bubbles to aggregate producing holes. The gluten matrix may be weakened further by the increased beta-glucan levels associated with barley inclusion. A previous study by the authors found pearled barley Xour to contain higher levels of beta-glucan than wheat Xour [29]. Beta-gluten has been shown to have a higher aYnity to water than gluten. Therefore, during mixing, the presence of beta-gluten may lead to a further decrease in gluten matrix strength by disrupting the glutens ability to form.

a,b a

Slice area (mm2 )

5000 4000 3000 2000 1000 0 A B

Slice area (mm 2 )

80 70 60 50 40 30 20 10 0

Formulation
Height (mm)

Fig. 10 Slice area and slice height of breads containing diVering levels of barley Xour. A: 100% wheat Xour; B: 70% wheat Xour, 30% PBF; C: 50% wheat Xour, 50% PBF; D: 30% wheat Xour, 70% PBF; E: 100% PBF

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Height (mm)

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451
6.00

Fibre and beta-glucan analysis The Wbre content of the breads increased signiWcantly (p < 0.01) with increasing levels of barley in the formulation (Fig. 11). A previous study by the authors on the Wbre content of barley Xour found barley Xour to have a higher Wbre content than wheat [29] consequently this result is to be expected. Increasing the barley levels in the formulations leads to a signiWcant increase in the beta-glucan levels of the breads. Cavallero et al. [8] and Dhingra and Jood [11] also found that the substitution of barley Xour into a wheat Xour formulation produced bread with a signiWcantly higher betaglucan concentration than the wheat control. From a nutritional perspective, a high level of barley inclusion would be recommended to increase the beta-glucan levels and therefore, improve the nutritional value of the bread product. According to the recommendation of the United States FDA [14], food that contains barley must provide at least 0.75 g of soluble beta-glucan per serving to qualify for the health claims stating that the product reduces the risk of coronary heart disease. Given that the 50% barley Xour breads contained approximately 0.6% beta-glucan, it was calculated that a serving size of 125 g would be required to meet the claim. This equates to 6 slices of bread from a standard 800 g loaf, which would suggest that although the beta-glucan content of the bread has been increased signiWcantly by the addition of 50% pearled barley Xour, further studies to increase the beta-glucan content of the breads are required to produce a quality bread product that would qualify for this health claim. Taste panel Breads containing barley inclusion levels of up to 50% did not diVer signiWcantly from the control (Fig. 12), however,
5 4.5 4 3.5 3 b a d b,c b,c c

Sensory Score (cm)

5.00 a 4.00 3.00 2.00 1.00 0.00 A B C D E a a b c

Formulation

Fig. 12 Taste panel scores of breads containing diVering levels of barley. A: 100% wheat Xour; B: 70% wheat Xour, 30% PBF; C: 50% wheat Xour, 50% PBF; D: 30% wheat Xour, 70% PBF; E: 100% PBF. * Columns with diVerent superscripts are statistically signiWcant (p < 0.05)

above the 50% level, signiWcant diVerences were found. This would suggest that a bread containing 50% barley has the potential to be commercialised as it would be an acceptable alternative to pure wheat bread. Data correlation No correlations between the rheological properties of the doughs and the bread textural characteristics were found when both the pure Xour doughs and the composite doughs were considered. However, when correlating the composite doughs with their respective breads, a number of correlations could be found. There were strong correlations between the phase angle of the composite Xours and a number of bread structural characteristics, in particular, the phase angle at 10 Hz was found to strongly correlate with the area of holes in the crumb (R2 = 0.818) as well as crumb cohesiveness (R2 = 0.795). Therefore, there is potential for rheological tests to be used to predict bread quality in future studies. Correlations were found between the starch pasting properties of the Xour formulations and the uniaxial extensions of the resulting doughs. Peak viscosity correlated very strongly with the fracture force of the dough (R2 = 0.979) while setback correlated with the distance to fracture of the dough (R2 = 0.960). The protein content of the Xour formulations was also found to correlate strongly with a number of dough and bread characteristics. Protein content and the distance to dough fracture were highly correlated (R2 = 0.817), as was protein content with starch breakdown (R2 = 0.860). Starch pasting properties correlated very strongly with the beta-glucan content of the breads, which suggests that it has the potential to be used as a predictor for beta-glucan content of wheat-barley Xour formulations. Setback

2.5 2 1.5 1 0.5 0 A a

b B

b,c

Formulation
% Beta Glucan % Total Dietary Fibre

Fig. 11 Beta-glucan and total dietary Wbre contents of breads containing diVering levels of barley Xour. A: 100% wheat Xour; B: 70% wheat Xour, 30% PBF; C: 50% wheat Xour, 50% PBF; D: 30% wheat Xour, 70% PBF; E: 100% PBF

123

452

Eur Food Res Technol (2010) 231:441453

(R2 = 0.969), Wnal viscosity (R2 = 0.9545) breakdown (R2 = 0.946) and peak viscosity (R2 = 0.964) all showed potential as beta-glucan content indicators.

Conclusions In this study, pearled barley Xour was milled and substituted into a standard wheat bread formula. The resulting breads were compared with a wheat bread control. Comparing the Xours of the diVerent formulations, it was apparent that increasing the pearled barley Xour content of the formulations leads to a signiWcantly lower protein content when compared to the wheat Xour. Proteolytic studies of the Xour conWrmed this decrease. A low protein content usually signiWes a reduced baking quality, so this result would suggest that the inclusion of barley Xour into the bread formulation would decrease the quality of the resulting breads. The starch pasting properties of the Xour combinations showed that increasing the barley Xour content of the Xour formulation leads to an increased Wnal viscosity of the paste and an increased starch retrogradation. These increases would suggest that the bread produced from formulations with a high barley Xour concentration would be more likely to stale rapidly when compared to wheat breads. A number of rheological tests were undertaken on the doughs of the formulations. The pure doughs (100% wheat Xour and 100% pearled barley Xour) had lower storage and loss moduli than the composite doughs (doughs containing both wheat and barley Xour); therefore, the composite Xour formulations produced Wrmer doughs. This Wrmness may be a consequence of the barley Xour particles disrupting the gluten matrix created by the wheat Xour. Increasing the barley Xour content in the formulation decreased the force required to rupture the doughs. The distance by which the doughs could be extended before rupturing was also reduced by the increased inclusion of barley Xour. A decreased force and distance of extension would suggest that the increased barley concentration decreased the strength of the dough, and so the breads produced from these doughs was likely to be of a lower quality than breads containing little or no barley Xour. Analysis of the breads conWrmed the results of the Xour and dough analysis. The volumes of the breads made from diVering barley Xour levels were found to diVer signiWcantly. Increasing the pearled barley Xour concentration signiWcantly decreased the volume of the breads. Texture proWle analysis of the breads also yielded signiWcant diVerences, in particular to the cohesiveness, chewiness and hardness of the breads. Hardness was found to increase signiWcantly with an increased inclusion of barley Xour and was also found to signiWcantly increase the rate of staling of

the breads, as was predicted from the starch retrogradation results of the Xours. Digital image analysis revealed that the slice area and height of the bread decreased with an increase in the level of barley Xour in the formulation. The Wbre contents of the breads increased signiWcantly with the increase in pearled barley Xour to the formulation. Beta-glucan levels were found to signiWcantly increase with the increase in pearled barley Xour in the Xour formulation. Taste panel results indicate that the addition of barley Xour to a wheat Xour formulation does not have a signiWcant eVect on bread acceptability up to 50% barley Xour addition. The results would indicate that there is potential for a bread product containing up to 50% barley Xour.
Acknowledgments This project is funded by the Department of Agriculture, Fisheries and Food under the Food Institutional Research Measure.

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