Eur Food Res Technol (2010) 230:437445 DOI 10.

1007/s00217-009-1184-z

ORIGINAL PAPER

Baking properties and microstructure of pseudocereal Xours in gluten-free bread formulations

Laura Alvarez-Jubete Mark Auty Elke K. Arendt Eimear Gallagher

Received: 30 June 2009 / Revised: 22 September 2009 / Accepted: 3 November 2009 / Published online: 25 November 2009 Springer-Verlag 2009

Abstract In the present study, the baking properties of the pseudocereals amaranth, quinoa and buckwheat as potential healthy and high-quality ingredients in gluten-free breads were investigated. Scanning electron micrographs were taken of each of the Xours. The pasting properties of these Xours were assessed using a rapid visco analyser. Standard baking tests and texture pro Wle analysis were performed on the gluten-free control and pseudocerealcontaining gluten-free breads. Confocal laser scanning microscopy (CLSM) images were also obtained from the baked breads and digital image analysis was conducted on the bread slices. Bread volumes were found to signiWcantly increase for the buckwheat and quinoa breads in comparison with the control. In addition, the pseudocereal-containing breads were characterised by a signiWcantly softer crumb texture eVect that was attributed to the presence of natural emulsiWers in the pseudocereal Xours and conWrmed by the confocal images. No signiWcant diVerences were obtained in the acceptability of the pseudocereal-containing gluten-free breads in comparison with the control. Keywords Pseudocereals Gluten free Bread Microscopy Baking properties Pasting properties
L. Alvarez-Jubete E. Gallagher (&) Ashtown Food Research Centre, Teagasc, Ashtown, Dublin 15, Ireland e-mail: Eimear.Gallagher@teagasc.ie L. Alvarez-Jubete E. K. Arendt Department of Food and Nutritional Sciences, National University of Ireland, Cork, Ireland M. Auty National Food Imaging Centre, Moorepark Food Research Centre, Teagasc, Moorepark, Fermoy, Co. Cork, Ireland

Introduction To date, the only treatment available for celiac disease is a strict lifelong adhesion to a gluten-free diet [1]. Therefore, celiac patients must avoid the consumption of gluten-containing foods. However, this may prove a diYcult and overwhelming task for the celiac patients as the majority of the cereal-based foods available in the market (such as pasta, baked products, snacks and breakfast cereals) are prepared with gluten-containing grains, such as wheat [2]. Although gluten-free alternatives are readily available, Wnding goodquality gluten-free products has been reported as a major issue for celiac patients who are trying to adhere to a gluten-free diet [3, 4]. Despite recent advances in the formulation of highquality gluten-free products, the replacement of gluten in cereal-based products, such as bread, biscuit, cake and pasta, still represents a signiWcant technological challenge [5]. The formulation of gluten-free breads possibly represents the greatest challenge, due to the fundamental role of gluten in breadmaking [6]. Gluten is an essential structure-building protein that provides viscoelasticity to the dough, good gas-holding ability and good crumb structure of the resulting baked product [5]. Some of the most important approaches developed to date to mimic the properties of gluten in gluten-free bakery products involve the use of gums, hydrocolloids and protein-based ingredients [6]. Considerably, fewer studies have been dedicated to improving the nutritional quality of gluten-free products. Gluten-free cereal foods are frequently made using reWned gluten-free Xour or starch and are generally not enriched or fortiWed [7]. As a result, many gluten-free cereal foods do not contain the same levels of B-vitamins, iron and Wbre as their gluten-containing counterparts [7, 8]. A need to

123

438

Eur Food Res Technol (2010) 230:437445 Table 1 Bread formulations Ingredient (% Xour/ starch base) Rice Xour Potato starch Amaranth Xour Quinoa Xour Buckwheat Xour Yeast Sugar Salt Xanthan gum SunXower oil Water Gluten-free control (GFC) 50 50 3 3 2 0.5 6 87 Amaranth (A) Quinoa (Q) Buckwheat (B)

improve their nutritional quality has been raised by many medical and nutritional experts [2, 9]. Several gluten-free grains exist, such as the pseudocereals amaranth, quinoa and buckwheat. These seeds are also characterised by an excellent nutrient proWle. Besides being important energy sources due to their starch content, amaranth, quinoa and buckwheat provide good-quality protein, dietary Wbre and lipids rich in unsaturated fats [10]. Moreover, they contain adequate levels of important micronutrients, such as minerals and vitamins and signiWcant amounts of other bioactive components, such as saponins, phytosterols, squalene, fagopyritols and polyphenols [1114]. A recent trend by researchers has focused on their use in the formulation of high-quality healthy gluten-free products, such as bread and pasta. In a series of recent studies, the nutritional properties and baking characteristics of amaranth, quinoa and buckwheat have been assessed [10, 12, 15]. The authors found that the replacement of potato starch with a pseudocereal Xour resulted in gluten-free breads with an increased content of important nutrients, such as protein, Wbre, calcium, iron and vitamin E. The resulting breads also had a signiWcantly higher content of polyphenol compounds and their in vitro antioxidant activity was increased. In the present study, technological aspects (i.e. batter/ dough and baking properties) related to the application of the pseudocereals as ingredients in the production of gluten-free breads were evaluated.

50 50 3 3 2 0.5 6 87

50 50 3 3 2 0.5 6 87

50 50 3 3 2 0.5 6 87

Materials and methods Bread ingredients

were mixed together for 1 min using an A120 Hobart mixer (Hobart Food Equipment, Sydney, Australia) at speed 1, yeast was dissolved in the water and added to the dry ingredients together with the oil and the batter formed was mixed for a further minute. After scraping the base of the bowl, the batter was further mixed for 2 min at speed 2. The batter was then scaled into baking tins (400 g) and placed in a proofer (Koma, Roermond, The Netherlands) for 30 min at 35 C and 80% relative humidity. The loaves were baked in a deck oven (Tom Chandley Ovens, Manchester, UK) at 220225 C for 25 min. They were then cooled to room temperature and stored in polyethylene bags. Six loaves were produced per bake and the preparation of the breads was done in triplicate (i.e. 3 bakes per each type of bread). Flour pasting properties

Amaranth Xour (Ziegler & Co., Wunsiedel, Germany), quinoa Xour (Ziegler & Co., Wunsiedel, Germany), rice Xour (S&B Herba, Orpington, Kent, UK), potato starch (Healy Chemicals Ltd, Dublin, Ireland), wheat Xour (Odlum Group, Dublin, Ireland), sunXower oil (Flora, Liverpool, UK), xanthan gum (All In All Ingredients, Dublin, Ireland), fresh yeast (Yeast Product, Dublin, Ireland), salt (Imeos Enterprises, Runcorn, Cheshire, UK) and caster cane sugar (Tate & Lyle, London, UK) were the materials used in the study. Preparation of breads The diVerent bread formulations are presented in Table 1. The amount of water used in the control and in each of the pseudocereal-containing breads was kept the same; the only diVerence in the formulation of the breads was the type of Xour used as a composite with rice Xour. The gluten-free batter was prepared as follows: dry ingredients

The pasting properties of the Xours and starches were evaluated using a Rapid Visco Analyser (RVA, Newport ScientiWc Pty. Ltd, Warriewood, Australia). The method used was the RVA General Pasting Method (Newport ScientiWc Pty. Ltd, 1998). Bread evaluation Loaf volume was measured using a volume meter (TexVol BVM-L370, Sweden). Loaf weight was recorded and loaf speciWc volume (ml/g) calculated. Bake loss deWned as the amount of water and organic material (sugars fermented and released as CO2) lost during baking was also calculated ([weight of the loaf before baking weight of the loaf after baking and cooling]/[weight of the loaf before baking] 100). Moisture was measured following a procedure based on the ICC method 110.1 [16] using a Brabender moisture oven (Brabender, Duisberg, Germany).

123

Eur Food Res Technol (2010) 230:437445

439

Crust and crumb colour were measured using a Minolta Chromameter (Minolta CR-100, Osaka, Japan) and results were expressed using the L*, a*, b* colour scale. Crumb structure of the loaves was evaluated using the C-Cell Bread Imaging System (Calibre Control International Ltd., UK). The procedure followed in this study consists of the standardised procedure described by the C-Cell Bread Imaging System manufacturer (Calibre Control International Ltd., UK). Crumb texture was assessed by conducting a texture proWle analysis (TPA) using a texture analyser (TA-XT2i, Stable Micro Systems, Surrey, UK) equipped with a 25 Kg load cell and a 36 mm aluminium cylindrical probe. Pre-test, test and post-test speed were 2, 1 and 5 mm/ s, respectively, and compression was set at 40%. All bread evaluation analysis were conducted 24 h after baking (day 1) and moisture and TPA analysis were repeated 72 and 120 h after baking (days 3 and 5, respectively). Scanning electron microscopy (SEM) Flour samples were sprinkled onto double-sided carbon tape Wxed to an aluminium specimen stub and examined under high vacuum in a Zeiss Aupra 40VP Weld emission scanning electron microscope (Carl Zeiss SMT, Cambridge, UK). Secondary electron images were acquired at an accelerating voltage of 1 kV. Confocal laser scanning microscopy (CLSM)

Massachusetts, US). Data were analysed using analysis of variance (ANOVA) and the mean were separated by the TukeyKramer test. DiVerences of p < 0.05 were considered signiWcant.

Results Scanning electron microscopy of the Xours SigniWcant diVerences can be observed in the scanning electron micrographs of the pseudocereal Xours, rice Xour, wheat Xour and potato starch (Fig. 1). In particular, the size of the Xour particles seems to diVer considerably among the Xours under study. Smallest particle size can be observed in potato starch and wheat Xour, followed by rice, buckwheat, amaranth and quinoa Xours. Also, considerable diVerences can be observed in the size and shape of the starch granules. The size of the starch granules in amaranth and quinoa Xours is signiWcantly smaller (<2 m) than in the rest of the Xours, and their shape is polygonal. These Wndings are in agreement with previously published studies [11, 13, 17]. Buckwheat starch granules are also polygonal in shape, while rice starch granules are irregular in shape and wheat granules show an oval shape. Potato starch granules are signiWcantly larger than in the rest of the Xours, and their shape is oval. Pasting properties of the Xours

Bread samples approximately 5 5 3 mm thick were cut with a razor blade, placed on a microscope slide and 50 ml of aqueous Nile Blue (0.1% w/w) added to the surface. A coverslip was placed on top and the samples were imaged in a Lecia SP5 confocal scanning laser microscope. Dual channel images were acquired with a 63 (1.4 NA) objective, using 488 nm argon ion laser excitation to image fat (pseuodocoloured green) and 633 nm helium neon laser excitation to reveal protein (pseudocoloured bright red) and gelatinised starch (pseudocoloured dull red). Images, 512 512 pixels, 8 bit depth were acquired. Sensory analysis Sensory analysis was conducted on all the breads tested by a panel consisting in 17 non-celiac consumers. 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. Statistical analysis Results were analysed using the statistics toolbox of the software Matlab 7.6 R2008a (Mathworks, Natick,

The results obtained for the pasting properties of rice Xour, potato starch and the pseudocereal Xours amaranth, quinoa and buckwheat are summarised in Table 2, and representative graphs of each Xour are presented in Fig. 2. SigniWcant diVerences were observed in the pasting proWles of the samples under study. The pasting proWle of potato starch exhibited high peak viscosity and breakdown and small setback. These are typical pasting characteristics of root starches. Peak viscosity diVered signiWcantly among the rice and pseudocereal Xours, and decreased in the order rice > buckwheat > quinoa > amaranth. Breakdown was signiWcantly lower for the pseudocereal Xours compared with rice Xour, which suggests an increased ability of the pseudocereals to withstand heating and shear stress. Highest Wnal viscosity was observed for the rice and buckwheat Xours, followed by amaranth and quinoa (p < 0.05). Similarly, setback was highest for rice Xour followed by buckwheat, amaranth and quinoa Xours (p < 0.05). Final viscosity is an important indicator of the strength of the gel formed upon cooling, and represents an important quality parameter. These Wndings are in agreement with previously published studies [1820]. The pasting proWles obtained in the present study for the pseudocereal Xours are consistent with previously reported

123

440

Eur Food Res Technol (2010) 230:437445

Fig. 1 Scanning electron micrographs of amaranth, quinoa, buckwheat and rice Xour, potato starch, and wheat Xour. Scale bars row a 100 m; b 20 m; c 2 m Table 2 Pasting properties of amaranth, quinoa, buckwheat and rice Xour, and potato starch PV Amaranth Quinoa Buckwheat Rice Xour Potato starch 273.0 2.3 a 288.0 1.9 b 341.4 10.6 c 429.6 0.1 d 479.2 1.8 e TV 225.5 1.7 a 293.4 1.2 b 321.0 6.9 c 303.1 0.6 d 174.9 0.2 e BD 47.5 0.6 a 5.4 0.7 b 19.9 3.2 c 126.5 0.4 d 304.2 1.5 e FV 321.6 2.5 a 191.8 1.8 b 606.5 5.7 c 599.0 4.6 c 201.7 1.2 d SB 96.0 0.8 a 101.6 0.5 b 285.1 1.6 c 298 2.1 d 26.7 1.0 e Ptime (min) 5.7 0.0 a 7.0 0.0 b 6.4 0.1 c 5.5 0.0 d 3.4 0.0 e

PV peak viscosity, TV trough viscosity, BD breakdown (PV TV), FV Wnal viscosity, SB setback (FV TV), Ptime (min), peak time

values ranging from 3.5 to 19.6% for quinoa starch [18, 19, 22]. The amylose content in amaranth seeds has been reported to be lower than 8% [19, 23]. Contrarily, the content of buckwheat has been reported to be as high as 47% [20], although similar values to those found in other common cereals (2526%) have also been reported [24]. Bread evaluation Loaf volume, bake loss and crust/crumb colour The results for the loaf volume, bake loss (%) and colour of the baked breads are presented in Table 3. The replacement of potato starch by each of the pseudocereal Xours had a variable eVect on loaf volume. Loaf volumes were increased (p < 0.05) for buckwheat (1.63 ml/g) and quinoa (1.4 ml/g) breads in comparison with the control (1.3 ml/g). However, no diVerence in volume was found between the control breads and those containing amaranth. Bake loss diVered slightly between the gluten-free control and the pseudocereal-containing gluten-free breads; however, the diVerences were not signiWcant. In relation to the crust colour of the baked breads, the pseudocereal-containing gluten-free breads were signiWcantly darker (lower L* values) compared with the glutenfree control. The darkening of the crust colour brought

Fig. 2 Pasting proWle of amaranth, quinoa, buckwheat and rice Xour, and potato starch

levels of amylose in these Xours, and with the small size of their starch granules (see previous section). The amylose content of amaranth and quinoa starch is much lower than that found in other cereals [21]. In the case of quinoa starch, considerable variability exists within the literature, with

123

Eur Food Res Technol (2010) 230:437445 Table 3 SpeciWc volume (ml/g), bake loss (%), crust L* and crumb L*/b* of the baked breads Bread Amaranth Quinoa Buckwheat Gluten-free C SpeciWc volume (ml/g) 1.31 0.03 a 1.40 0.02 b 1.63 0.05 c 1.29 0.03 a Bake loss (%) 9.0 0.5 a 9.1 0.4 a 9.0 0.3 a 9.4 0.4 a Crust L* 56.0 1.4 a 52.7 2.7 a 51.4 0.9 a 69.7 1.4 b

441

Crumb L*/b* 3.9 0.1 a 4.1 0.1 a 5.6 0.3 b 6.4 0.4 c

Mean value of three replicates SD. Mean values followed by the same letter are not statistically diVerent (p < 0.05)

about by the replacement of potato starch by a pseudocereal Xour is desirable as gluten-free breads tend to have a lighter crust colour than white wheat breads which sometimes appear artiWcial [25]. Crumb colour (L*/b*) (white/yellow ratio) was also inXuenced and the pseudocereal-containing gluten-free breads were characterised by a signiWcantly darker crumb colour in comparison with the control. Digital image analysis The results for the crumb grain analysis of the baked breads are summarised in Table 4, and the images obtained are presented in Fig. 3. Crumb structure diVered signiWcantly in terms of number of cells, cell volume and wall thickness. Crumb grain represents an important attribute when deWning bread quality [26]. In the present study, largest number of cells was

for buckwheat bread followed by quinoa, GFC and amaranth breads. Smallest cell volume was found in gluten-free control bread, followed by quinoa, amaranth and buckwheat breads. Cell wall was thinnest in quinoa bread and increased subsequently in the order quinoa < gluten-free control < buckwheat < amaranth. Texture proWle analysis (TPA) of bread crumb The results for the TPA analysis of the baked breads, as well as their moisture content, are presented in Fig. 4. All the pseudocereal-containing gluten-free breads had a softer crumb in comparison with the gluten-free control, with amaranth bread having the softest crumb (p < 0.05) over the entire testing period. Overall, a similar trend was found in crumb cohesiveness, where all of the pseudocereal-containing gluten-free breads had a more cohesive crumb in comparison with the control (p < 0.05). Again, most cohesive crumb was detected in amaranth bread (p < 0.05). Also, all the pseudocereal-containing glutenfree breads had signiWcantly higher crumb springiness in comparison with the control (p < 0.05). Despite the diVerences observed in crumb texture, no signiWcant diVerences were recorded in the moisture content of the bread samples. No studies could be found in the literature to with compare the TPA results obtained in the present study. Previously published studies on the baking properties of the pseudocereals focused on their impact on bread volume and sensory analysis when added at diVerent levels as composites with

Table 4 Crumb structure (digital image analysis) of amaranth, quinoa, buckwheat and gluten-free control breads Bread Amaranth Quinoa Buckwheat Gluten-free control Number of cells 2,589 145 a 3,176 334 b, c 3,340 87 b 2,992 128 c Wall thickness 0.46 0.01 a 0.42 0.02 b 0.44 0.00 b 0.43 0.01 b Cell volume 21.2 1.5 a 18.7 2.7 a, c 22.9 0.7 b 17.9 2.0 c

Fig. 3 Raw (a) and cell (b) images of amaranth, quinoa, buckwheat and gluten-free control breads

123

442

Eur Food Res Technol (2010) 230:437445

a
Crumb hardness (g)

6000 5000 4000

24 h

b
Crumb cohesiveness

0.45 0.4 0.35 0.3 0.25 0.2 0.15 0.1 0.05 0 Amaranth Quinoa Buckwheat GFC
24 h 72 h 120 h

3000 2000 1000 0 Amaranth Quinoa Buckwheat GFC

72 h 120 h

c
Crumb springiness

1 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 Amaranth Quinoa Buckwheat GFC

d
Moisture (%)

48.5 48 47.5 47 46.5 46 45.5 45 Amaranth Quinoa Buckwheat GFC

24 h 72 h 120 h

24 h 72 h 120 h

Fig. 4 Moisture and TPA proWle of the breads after 24, 72 and 120 h post-baking: (a) crumb hardness; (b) crumb cohesiveness; (c) crumb springiness; (d) moisture (%). Mean value of three replicates SD

wheat Xour, and did not measure their impact on the texture proWle of the resulting breads [2730]. The eVect of storage time (120 h) on crumb structure was also investigated (Fig. 4). Overall, crumb hardness increased with storage time. In the case of gluten-free control, amaranth and quinoa breads, the increase in crumb hardness with storage time was not signiWcant. For buckwheat bread, the increase in crumb hardness was only signiWcant after 120 h. In addition, crumb cohesiveness was found to decrease with storage time for all bread samples apart from those containing amaranth. A similar trend was identiWed for crumb springiness and, although values for this parameter were found to decrease with storage time, these diVerences were only signiWcant in the gluten-free control and quinoa breads. Also no signiWcant diVerences were recorded in the moisture content of the pseudocerealcontaining gluten-free breads during the storage period, with the exception of buckwheat bread, the moisture content of which decreased signiWcantly after 120 h. These results suggest that the pseudocereal-containing gluten-free products may be used in the production of gluten-free breads with a softer crumb structure. This is a desirable characteristic as gluten-free breads are often characterised by a hard texture [6]. Also the increased cohesiveness and springiness found in the pseudocereal-containing glutenfree breads can be considered beneWcial, as gluten-free breads are often characterised by a crumbly, brittle texture [31].

Confocal laser scanning microscopy (CLSM) The images obtained by confocal laser scanning microscopy of the bread samples are presented in Fig. 5. As expected, signiWcant variation was observed between the diVerent breads. Starch gelatinisation appears to have occurred to a greater degree in the gluten-free control bread compared with the pseudocereal-containing gluten-free breads, with starch granules fusing together and losing their original structure. Partial gelatinisation seems to have occurred in the pseudocereal-containing gluten-free breads, and as a result, a greater number of starch granules have retained their integrity. A more homogenous structure is apparent for the pseudocereal-containing gluten-free breads, with less gas voids and a more even distribution of fat, protein and starch. Also, the images reveal the importance of the fat globules in forming complexes with starch granules and/or stabilising gas cells. This eVect appears particularly more predominant in the pseudocereal-containing gluten-free breads in comparison with the gluten-free control. Sensory analysis The acceptability scores of the baked breads as determined by the taste panellists are displayed in Fig. 6. No signiWcant diVerences were observed in the acceptability of the baked breads, showing that pseudocereal Xours may be introduced

123

Eur Food Res Technol (2010) 230:437445 Fig. 5 Confocal laser scanning micrographs of gluten-free control, amaranth, quinoa and buckwheat breads. Scale bars row a 0250 m; b 0250 m; c 050 m

443

6 5 4 3 2 1 0 Amaranth Quinoa Buckwheat GFC

Fig. 6 Acceptability scores of baked breads

into a gluten-free bread formulation to enhance crumb softness and cohesiveness and without adversely aVecting the sensory properties of the loaves.

Discussion Some of the more important properties when assessing the quality of baked breads are loaf volume and crumb texture. This study showed how bread volume can be signiWcantly increased following the incorporation of pseudocereal Xours, such as quinoa and buckwheat. The volume of bread

depends on a number of factors, such as viscosity of the batters, amylose/amylopectin ratio, the presence of surface-active components and/or the occurrence of protein aggregation upon heating [32]. In gluten-free breads, the viscosity of the batters prior to starch gelatinisation is crucial to prevent the Xour particles from settling and gas cells from rising and thus, maintain a homogenous system during prooWng and baking until starch gelatinisation [32]. Therefore, factors, such as peak viscosity of the batters seem to have implications in relation to the Wnal quality of the resultant baked bread. In the present study, bread volume of the pseudocereal-containing gluten-free breads was found to increase accordingly with peak viscosity of the pseudocereal Xour as measured by the rapid visco analyser. Also, the amylose/amylopectin ratio is a crucial factor determining bread volume and crumb structure as amylose contributes signiWcantly to crumb setting due to its quick retrogradation rate [32]. As discussed in the previous section, the amylose content in amaranth and quinoa Xours is lower than in wheat, whereas buckwheat has higher levels of amylose than those found in cereals. These Wndings are consistent with the obtained results in this study: bread volumes and crumb structure were found to improve according to the previously reported amylose contents of the pseudocereal Xours. The lipid content and composition in amaranth, quinoa and buckwheat seeds may also have implications in relation with functionality during bread making. It has been shown that polar lipids naturally present in cereals may contribute

123

444

Eur Food Res Technol (2010) 230:437445

towards the stabilisation of gas bubbles during bread making [32, 33]. The confocal laser scanning micrographs of the baked breads obtained in the present study show that lipids in amaranth, quinoa and buckwheat may act as surface-active agents and thus contribute to gas cell stabilisation prior to starch gelatinisation. This eVect was particularly relevant in quinoa breads. However, no such eVect was observed in the gluten-free control bread. Lipid content in amaranth and quinoa seeds has been reported as being 23 times higher than that in buckwheat or in common cereals, such as wheat [10]. The content of polar lipids in quinoa seeds is very high and represents approximately 25% of total lipids [34]. The polar lipid content in amaranth seeds has been reported to be approximately 10% of total lipids [35], whereas in buckwheat it ranges between 13.5 and 15.5% total lipids [36]. Thus, the high level of polar lipids in pseudocereal seeds, and in particular in quinoa seeds, may have functionality as gas cell stabilising agents during bread making. Also, the high levels of fats present in amaranth and quinoa Xours may have implications in relation with both crumb structure and crumb texture. The use of emulsiWers in baking has been shown to have a softening eVect on bread crumb [37]. Fatty acids from lipid, such as monoglycerides can form complexes with amylose, thus limiting starch swelling during baking and leaching of amylose into solution [37, 38]. As a result, fewer entanglements between starch granules and amylose in solution take place, leading to breads with a softer crumb structure [32]. Monoglycerides in amaranth Xour have been reported to be in the range 3.03.8% total lipid content [35]. In quinoa seeds monoglyceride levels are lower (approximately 2%) but the levels of free fatty acids are high (19% total lipid content) [34]. As previously seen, the confocal laser scanning micrographs of the pseudocereal-containing baked breads showed fat molecules surrounding starch compounds and possibly, forming complexes with starch molecules. This eVect thus, supports the hypothesis that the emulsiWers naturally present in the pseudocereal Xours may have a positive eVect, resulting in breads with a softer crumb. However, as previously discussed, amylose is necessary for good crumb structure, and the presence of emulsiWers may thus have a weakening eVect on crumb structure. The low levels of amylose in amaranth and quinoa Xour, as well as the high fat content characteristic of these Xours may be responsible for their soft texture, but also for their relatively weak crumb structure when processed into breads, in comparison with buckwheat Xour. Other aspects in relation to the functionality of these Xours in gluten-free systems remain to be investigated, such as the application of hydrocolloids other than xanthan gum. For example, hydroxypropylmethylcellulose (HPMC) is a hydrocolloid with surface-active properties, thus its use in

the formulation of pseudocereal-containing gluten-free breads may result in breads with improved crumb structure and volume. Also, the pseudocereal-containing breads may beneWt from higher water levels, especially in the case of buckwheat and quinoa breads, due to the higher water binding capacity upon heating of buckwheat and quinoa Xours, as seen in their respective pasting proWles. In addition, amaranth and quinoa breads may beneWt from the presence as composites of gluten-free Xours with high amylose content, such as buckwheat Xour, to compensate for their intrinsic low amylose content. Finally, the role of natural emulsiWers present in the pseudocereal Xours may have potential in the production of gluten-free breads characterised by an improved crumb texture. The production of high-quality gluten-free breads containing pseudocereal Xours would represent a signiWcant step towards ensuring an adequate intake of nutrients in people with celiac disease.

Conclusions The gluten-free breads containing buckwheat or quinoa Xour had a signiWcantly higher volume in comparison with the gluten-free control. All the pseudocereal-containing gluten-free breads were characterised by a signiWcantly softer crumb. This eVect was attributed to the presence of natural emulsiWers in the pseudocereal Xours and was conWrmed by confocal laser scanning microscopy. Results from the sensory panel showed that pseudocereal Xours may be introduced into a gluten-free bread formulation without adversely aVecting the sensory properties of the loaves. The pseudocereal Xours represent feasible ingredients in the manufacture of good-quality, healthy gluten-free breads.
Acknowledgment Enterprise Ireland. The present study is Wnancially supported by

References
1. Catassi C, Fasano A (2008) In: Arendt EK, Dal Bello F (eds) Gluten-free cereal products and beverages. Academic Press, London 2. Kupper C (2005) Dietary guidelines and implementation for celiac disease. Gastroenterol 128:S121S127 3. Case S (2005) The gluten-free diet: how to provide eVective education and resources. Gastroenterol 128:S128S134 4. Pietzak M (2005) Follow-up of patients with celiac disease: achieving compliance with treatment. Gastroenterol 128:S135 S141 5. Gallagher E, Gormley TR, Arendt EK (2004) Recent advances in the formulation of gluten-free cereal-based products. Trends Food Sci Technol 15:143152 6. Arendt EK, Morrisey A, Moore MM, Dal Bello F (2008) In: Arendt EK, Dal Bello F (eds) Gluten-free cereal products and beverages. Academic Press, London

123

Eur Food Res Technol (2010) 230:437445 7. Thompson T (1999) Thiamin, riboXavin, and niacin contents of the gluten free diet: is there cause for concern? J Am Diet Assoc 99:858862 8. Thompson T (2000) Folate, Iron, and dietary Wber contents of the gluten-free diet. J Am Diet Assoc 100:13891395 9. Thompson T, Dennis M, Higgins LA, Lee AR, Sharrett MK (2005) Gluten-free diet survey: are Americans with coeliac disease consuming recommended amounts of Wbre, iron calcium and grain foods? J Hum Nutr Diet 18:163169 10. Alvarez-Jubete L, Arendt EK, Gallagher E (2009) Nutritive value and chemical composition of pseudocereals as gluten-free ingredients. Int J Food Sci Nutr 60(suppl 4):240257 11. Berghofer E, Schoenlechner R (2002) In: Belton PS, Taylor JRN (eds) Pseudocereals and less common cereals: grain properties and utilization potential. Springer, Berlin 12. Alvarez-Jubete L, Wijngaard HH, Arendt EK, Gallagher E (2010) Polyphenol composition and in vitro antioxidant activity of amaranth, quinoa and buckwheat as aVected by sprouting and bread baking. Food Chem 119:770778 13. Taylor JRN, Parker ML (2002) In: Belton PS, Taylor JRN (eds) Pseudocereals and less common cereals: grain properties and utilization. Springer, Berlin 14. Wijngaard HH, Arendt EK (2006) Buckwheat. Cereal Chem 83:391401 15. Alvarez-Jubete L, Holse M, Hansen A, Arendt EK, Gallagher E (2009) Impact of baking on the vitamin E content of the pseudocereals amaranth, quinoa and buckwheat. Cereal Chem 86(5):511515 16. ICC (1976) Standard methods. International Association for Cereal Chemistry, Vienna 17. Wijngaard HH, Renzetti S, Arendt EK (2007) Microstructure of buckwheat and barley during malting observed by confocal laser scanning microscopy and scanning electron microscopy. J Inst Brew 113:3441 18. Lindeboom N, Chang PR, Falk KC, Tyler RT (2005) Characteristics of starch from eight quinoa lines. Cereal Chem 82:216222 19. Qian J, Kuhn M (1999) Characterization of Amaranthus cruentus and Chenopodium quinoa starch. Starch Strke 51:116120 20. Qian J, Rayas-Duarte P, Grant L (1998) Partial characterization of buckwheat (Fagopyrum esculentum). Starch Cereal Chem 75:365373 21. Schoenlechner R, Siebenhandl S, Berghofer E (2008) In: Arendt EK, Dal Bello F (eds) Gluten-free cereal products and beverages. Academic Press, London

445 22. Wright KH, Huber KC, Fairbanks DJ, Huber CS (2002) Isolation and characterization of Atriplex hortensis and sweet Chenopodium quinoa starches. Cereal Chem 79:715719 23. Hunjai C, Wansoo K, Malshick S (2004) Properties of Korean Amaranth starch compared to waxy millet and waxy sorghum starches. Starch Starke 56:469477 24. Yoshimoto Y, Egashira T, Hanashiro I, Ohinata H, Takase Y, Takeda Y (2004) Molecular structure and some physicochemical properties of buckwheat starches. Cereal Chem 81:515520 25. Gallagher E, Gormley TR, Arendt EK (2003) Crust and crumb characteristics of gluten-free breads. J Food Eng 56:153161 26. Scanlon MG, Zghal MC (2001) Bread properties and crumb structure. Food Res Int 34:841864 27. Adekunle Ayo J (2001) The eVect of amaranth grain Xour on the quality of bread. Int J Food Prop 4:341351 28. Chauhan GS, Zillman RR, Eskin MA (1992) Dough mixing and breadmaking properties of quinoa-wheat Xour blends. Int J Food Sci Technol 27:701705 29. Park SH, Morita N (2005) Dough and breadmaking properties of wheat Xour substituted by 10% with germinated quinoa Xour. Food Sci Technol Int 11:471476 30. Samiyi M, Ashraf HL (1993) Iranian breads supplemented with amaranth Xour. Int J Food Sci Technol 28:625628 31. Moore MM, Schober TJ, Dockery P, Arendt EK (2004) Textural comparisons of gluten-free and wheat-based doughs, batters, and breads. Cereal Chem 81:567575 32. Schober T (2009) In: Gallagher E (ed) Gluten-free food science and technology. Wiley-Blackwell, Oxford 33. Gan Z, Ellis PR, SchoWeld JD (1995) Gas cell stabilisation and gas retention in wheat bread dough. J Cereal Sci 21:215230 34. Przybylski R, Chauhan GS, Eskin NAM (1994) Characterization of quinoa (Chenopodium quinoa) lipids. Food Chem 51:187192 35. Gamel TH, Mesallam AS, Damir AA, Shekib LA, Linssen JP (2007) Characterization of amaranth seed oils. J Food Lipids 14:323334 36. Mazza G (1988) Lipid content and fatty acid composition of buckwheat seed. Cereal Chem 65:122126 37. Hoseney RC (1998) Principles of cereal science and technology. AACC International, St. Paul 38. Belitz HD, Grosch W, Schieberle P (2004) Food chemistry. Springer, Berlin

123