Eur Food Res Technol (2010) 231:2735 DOI 10.

1007/s00217-010-1247-1

ORIGINAL PAPER

The inXuence of germination conditions on beta-glucan, dietary Wbre and phytate during the germination of oats and barley
Florian Hbner Tonya ONeil Kevin D. Cashman Elke K. Arendt

Received: 20 October 2009 / Revised: 20 January 2010 / Accepted: 13 February 2010 / Published online: 9 March 2010 Springer-Verlag 2010

Abstract This study aimed to quantify the changes caused by varying germination conditions on the contents of some bioactive compounds in barley and oats. Samples of the two grains were germinated at temperatures between 10 and 20 C for a period of 26 days, using a two-dimensional central composite design. The germination temperature had only minor eVect in comparison with the germination time. Slight changes in the mineral content of the malts were observed, mainly caused by steeping. Phytate has been seen as an anti-nutritional compound, as it complexes minerals and lowers their bioavailability. The phytate content in barley malts was considerably lower than in the native kernels. Variations in the germination conditions did not have a signiWcant eVect on phytate content. In oats, degradation of phytate was signiWcantly enhanced by prolonging the germination period. It was possible to retain the amounts of soluble dietary Wbre, when short germination periods were applied. However, long germination periods caused an extensive breakdown of soluble dietary Wbre, especially beta-glucan. The content of insoluble Wbre, however, was increased by applying long germination periods for oat malts. Keywords Barley Beta-glucan Dietary Wbre Malting Oats Phytate

Introduction Cereals have been a staple food in many cultures for millennia. Today, growth area and production Wgures see cereals as the most produced food variety worldwide [1]. In many cases, cereals are the main energy source in the human diet with starch as the main carbohydrate source. However, cereal grains are also a major source of proteins, dietary Wbre and minerals and some micronutrients. Germination has long been used as a way to modify taste, appearance and technological properties of grains and has been widely used as a measure to produce malts for the use in brewing and distilling industry as well as food ingredient [1, 2]. Sprouting of cereals has also been found to increase the nutritive value of grains [3, 4], as the contents of some vitamins and antioxidants can be increased and some anti-nutritional factors decreased. Amongst the main compounds associated with healthpromoting eVects in cereals is dietary Wbre (DF). A number of macromolecules contribute to DF in oats and barley. Some of them are insoluble in water, e.g., celluloses, hemicelluloses and lignin, while others are soluble in water, e.g., beta-glucan and arabinoxylans [5]. Health beneWts and technological properties of dietary Wbre in cereals have been extensively investigated and reviewed in recent years [5, 6]. The main fraction of water soluble dietary Wbre in barley and oats are beta-glucans. Beta-glucans are part of the endosperm cell wall [7] and are degraded during the germination process and result in very low contents of beta-glucan in barley malts as well as oat malts [1, 8]. A number of health beneWts associated with beta-glucan intake have been reported and reviewed [9]. Consequently, the use of beta-glucan as functional ingredient for beverages has been investigated [10] and possibilities for the use as ingredient

F. Hbner T. ONeil K. D. Cashman E. K. Arendt (&) Department of Food and Nutritional Sciences, University College Cork, College Road, Cork, Co. Cork, Ireland e-mail: e.arendt@ucc.ie F. Hbner Bio Transfer Unit, University College Cork, Cork, Ireland

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for functional food have been discussed [11]. Wilhelmson [12] and coworkers developed a germination process for oats with a high retention of beta glucan. In addition to dietary Wbre, other compounds found in cereals are interesting from a nutritional point of view, amongst them certain minerals. Minerals play an important role in a wide range of physiological functions of the body. Cereals, and especially oats, are potentially a rich source of minerals and trace elements like Mg, Zn and Cu [1, 13]. However, a signiWcant proportion of the mineral content present may be of poor bioavailability, due to the fact that the ions form complexes with inhibitory factors such as tannins, some dietary Wbre and especially phytate [14]. The latter compound is a myo-inositol molecule esteriWed with phosphate group on all six hydroxyl groups. Due to its strongly negative charge, phytate complexes some metal ions [15] and thereby decreases their bioavailability [16]. The mineral-phytate complexes in the grains can only be poorly utilised by human beings, as these lack the necessary phytases to break down the complexes [17]. Germination has been discovered to increase the phytate activity of cereals [18, 19] and results in a breakdown of phytate. A decrease of the phytate content has been seen to improve the absorption of iron and zinc from whole grain oat products [20, 21]. However, in recent time, phytate has been connected with antioxidant activity and the prevention of certain types of cancer, probably caused by complexing of pro-oxidant minerals [22]. This paper aims to gather information about the inXuence of malting and especially the eVect of germination time and temperature on the contents of minerals, phytate, dietary Wbre and beta glucan. Previous studies were aimed at Wnding ways to inXuence the respective factor and resulted in optimal germination and processing conditions which were very diVerent to the ones applied in commercial malting and therefore might pose problems. The present study, however, attempted to identify germination conditions for the retention of beta glucan or the breakdown of phytate under conditions close to the ones commonly applied. Mathematical modelling was used as a tool to investigate the chosen range of germination conditions.

Buckwheat had a germination capacity of 96% (H2O2) and oats of 93%. Malting and sample preparation The grains were malted in a micro malting machine (Joe White Malting Systems, Perth, Australia). The grains were steeped to a water content of 45% in varying wet and dry phases of 3-h duration (7 cycles each). The steeping water used was tab water, pre-treated with a Filtromat Wlter (Elga Process Water, Celbrigde, Ireland). Water quality was assessed using methods 1.1.10.2 and 1.1.10.3 of Mitteleuropische Brautechnische Analysenkommission. The water had a total hardness of 3.2dH and a carbon hardness of 1.68dH. Germination conditions were varied as explained in the following paragraphs. Kilning conditions were 45 C for 8 h and subsequently 55 C for 15 h. The malts were vacuum-packed and stored at 20 C. Rootlets were removed manually before milling with a Bhler laboratory-scale disc mill (Bhler GmbH, Braunschweig, Germany), set at a gap width of 0.2 mm. Experimental design RSM [23] was applied in order to investigate the impact of germination conditions on the nitrogenous compounds, proteolytic activity and the molecular weight of the proteins. A circumscribed, two-dimensional central composite design was developed featuring variations in germination time between 48 and 144 h and germination temperature between 10 and 20 C. Nine combinations of time and temperature were used and the centre point done in duplicate. The measured responses were analysed by Wtting quadratic models to the data with least square regression in order to identify signiWcant (p < 0.05) eVects of the variations in the germination conditions on the responses. For the analysis of the data, the calculation of the models and the interpretation of the results, the program Statistica 7.1. (StatSoft Inc., Tulsa, USA) was used. In order to validate the models, barley and oats were soaked, germinated for 115 h at 13 C and kilned as described earlier. The contents measured in these malts were compared with the ones achieved by calculation from the models. Minerals

Materials and methods Raw materials Barley, variety Sebastien, was harvested in Ireland in 2005 and obtained from Cork Malting Company (Cork, Ireland). Oats of unknown variety was harvested in Finland in 2005 and obtained from Raisio Inc. (Raisio, Finland). The germination capacity of both grains was measured using the method of the European Brewing Convention (EBC) 3.5.2.

Minerals (calcium, magnesium, iron, zinc and copper) were analysed by atomic absorption spectrophotometry (Varian SpectraAA-600, Varian International Australia Ltd., Melbourne, Australia). Grain and malt samples were preashed over a Bunsen Xame and subsequently ashed at

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Table 1 InXuence of germination conditions on the contents of Calcium (Ca), Magnesium (Mg), Copper (Cu), Iron (Fe) and Zinc (Zn) in oats and barley malts in comparison with the contents in unmalted kernels Time (h) Temp Oats Barley (C) Ca Mg Cu Fe Zn Ca Mg Cu Fe Zn (mg/100 g) (mg/100 g) (mg/100 g) (mg/100 g) (mg/100 g) (mg/100 g) (mg/100 g) (mg/100 g) (mg/100 g) (mg/100 g) Unmalted 48 48 62 96 96 96 96 130 144 144 10 20 15 12 15 15 18 15 10 20 20.8 32.5 31.1 32.0 32.4 31.9 31.0 30.8 31.5 31.7 31.1 114 122 122 121 126 123 130 126 128 126 130 0.2 1.3 1.1 0.9 1.2 1.3 1.5 0.9 1.1 1.1 0.5 4.0 3.7 3.7 3.6 3.7 3.6 3.8 3.7 3.8 4.0 3.9 2.7 3.6 3.6 3.4 4.0 3.7 3.9 3.5 3.6 3.5 3.5 17.1 23.1 22.2 23.7 22.0 23.4 22.3 22.9 23.7 22.6 22.1 106 103 105 106 106 106 111 113 112 115 112 0.2 1.5 1.4 1.0 1.5 1.7 1.9 1.4 1.8 1.5 1.1 3.7 2.8 2.8 2.8 2.7 2.7 2.9 2.9 2.9 2.9 2.7 2.0 2.4 2.4 2.3 2.4 2.6 2.4 2.5 2.8 2.8 2.5

All values given are mg per 100 g dry matter

550 C for 4 h in a muZe furnace. The ashed samples were dissolved in 1 N HNO3 and Wltered prior to the spectrophotometry. In the cases of calcium and magnesium, LaCl3 (BDH Ltd, Poole, UK) was added. Skim milk powder (Community Bureau of Reference Materials, Brussels, Belgium) was used as standard reference material. Measurements were performed in triplicate. Phytate Phytate content was measured after acid extraction of inositol phosphates following the procedure used in the commercially available assay kit (K-PHYT 05/07, Megazyme Ltd., Bray, Ireland). The concentration of free phosphates in the extracts was subtracted from the concentration after subsequent breakdown of myo-inositol phosphates by phytases and alkaline phosphatase. The content of bound phytate was then divided by 0.282 in order to calculate the amount of phytate. All measurements were performed in duplicate. Analysis for dietary Wbre, beta-glucan and beta-glucanase activity The raw material and malt samples were analysed for insoluble and soluble dietary Wbre according to Association of Analytical Communities (AOAC) method 991.43 Total, Soluble and Insoluble Dietary Fiber in Foods. The enzymes used in the procedures were obtained from Megazyme as part of their Total Dietary Fibre Assay Procedure kit (Catalogue number K-TDFR 12/05). Beta glucan content in grains and malts was determined according to the McCleary method EBC method 4.16.1

with enzymes and chemicals from the Megazyme kit (Catalogue Nr K-BGLU 04/06). Beta glucanase activity was measured using the relevant Megazyme kit (K-MBGL). Analysis for beta glucan and dietary Wbre were performed in duplicate and for glucanase activity in triplicate.

Results and discussion Minerals Oats and to a lesser degree barley are seen as important source of minerals in the human nutrition. Due to the high amounts of cereal-based products consumed, they can make a signiWcant contribution to the mean daily intake of a number of minerals essential for the metabolic function of the human body. The results of our mineral analysis, covering Mg, Ca, Fe, Zn and Cu are shown in Tables 1 and 2. Slightly higher amounts of minerals were found in oats than barley, which is consistent with the literature [1]. Some diVerences between the malted and the unmalted samples were seen in the contents of the measured minerals. Ca and Zn increased as a consequence of the steeping, while Fe contents were found in similar (oats) or lower amounts (barley) in the malted samples. Mg on the other hand was found in lower amounts in unmalted oats than in the corresponding malts, while the Mg content in native barley was at the lower end of the range found in barley malts. Cu content in both unmalted oats and barley was very low (0.2 mg/100 g), while oat malts contained between 0.5 and 0.3 mg/100 g and barely malts even higher

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Table 2 p-Values for the inXuence of germination time and temperature as well as model Wts (R2) for the mathematical models for the contents of Calcium (Ca), Magnesium (Mg), Copper (Cu), Iron (Fe) and Zinc (Zn) in oat and barley malts InXuencing factor Oats Ca Time (linear) Time (quadratic) Temperature (linear) Temperature (quadratic) Interactive Model Wt R2 >0.5 >0.5 0.046 >0.5 >0.5 0.719 Mg 0.049 >0.5 >0.5 >0.5 >0.5 0.702 Cu >0.5 >0.5 >0.5 >0.5 >0.5 0.627 Fe 0.017 >0.5 >0.5 >0.5 >0.5 0.850 Zn >0.5 >0.5 >0.5 >0.5 >0.5 0.460 Barley Ca >0.5 >0.5 >0.5 >0.5 >0.5 0.630 Mg 0.026 >0.5 >0.5 >0.5 >0.5 0.763 Fe >0.5 >0.5 >0.5 >0.5 >0.5 0.226 Cu >0.5 >0.5 >0.5 >0.5 >0.5 0.321 Zn >0.5 >0.5 >0.5 >0.5 >0.5 0.713

SigniWcant inXuencing factors in bold print

values, between 1.1 and 1.9 g/100 g. Lintschinger and coworkers [24] showed that mineral contents of germinating cereals can be inXuenced by the mineral concentration in the steeping water. During the steeping, consequently a change in the mineral content due to leeching or uptake from the steeping liquid can be expected. This explains the diVerences seen between the native and malted samples in our trials. The biggest diVerence was seen for Cu, the increase was likely caused by corrosion of the copper pipes used in the heat exchanger unit of the malting machine. Although some diVerences in the mineral contents of the malt samples were seen, there were only few cases in which the variations in the germination had a signiWcant impact on the content. Magnesium content in malts of both oats and barley was increased as a consequence of prolonged germination time, as was the content of Fe in barley malts, while high temperatures resulted in lower contents of Ca. Model Wts were very low, likely due to comparatively small variations in the samples; therefore, it is not helpful to use the models for predictive purposes or to validate the model Wts. During germination, minerals are transported into the forming rootlets, which, in the case of barley, were reported to have and ash content of about 8.7% [25]. This results in a loss of minerals from the kernel. On the other hand, there is also the loss of organic material for rootlet formation as well as losses due to respiration. While the two eVects seem to be fairly balanced, so that little overall changes occur, higher malting losses are expected for longer germination periods as well as higher temperatures and this might result in slightly increased contents of minerals. Phytate Phytate has been shown to considerably decrease the bioavailability of some minerals, due to the formation of complexes [16] and therefore has traditionally been seen as anti-nutritional factor. This has been identiWed as a reason for poor uptake of minerals from unprocessed whole grain,

especially oats [14, 26]. Germination was proposed and investigated to increase the comparably low activity of phytate degrading enzymes [27, 28] in oats. Results of phytate measurements in ungerminated barley and oats as well as in the corresponding malts are shown in Table 3. Unmalted oats contained 0.683% phytate, while unmalted barley contained lower values (0.568%). Larsson and Sandberg reported 17.3 mol phytate per g oats, which corresponds to 1.12% phytate [27]. Belitz et al. reported 1.13% for oats and 1.19% for barley [1], while Centeno and collaborators found 0.18 mg phytate phosphorus per g barley [18], which is equivalent to about 0.64% phytate. It appears that the contents measured in our study are slightly lower than the reported ones in the case of barley but much lower than the ones expected in oats. Table 3 shows the inXuence of the germination conditions on the phytate content in the malts of oats and barley, respectively. No signiWcant eVects (Table 4) of the varied germination conditions were seen in the case of barley malt, the values ranging between 419 and 485 mg/100 g. Apparently, the main loss of phytate in barley happened during steeping and the Wrst 2 days of germination. Formation of phytate-degrading enzymes as well as the breakdown of phytate during the germination of barley has been followed in detail [18]. In contrast to our study, they found only a minor breakdown of myo-inositol-hexa-phosphates during the Wrst day of germination at 22 C. The contribution of lower myo-inositol phosphate esters to the total was about 10%, most of it myo-inositol-penta-phosphates. No accumulation of myo-inositol penta-, tetra-, or tri-phosphates was observed during the germination period. This means that, although the calculation in our study assumes that 100% of the bound phosphate was released from myo-inositol-hexa-phosphates, the calculated values are likely close to the real ones. Centeno et al. also reported that 60% of the original phyate was broken down after 5 days of germination [18], while Sandberg and Larsson found a loss of 50% of phytate after germination for 30 40 h at 15 C [27]. This is a stronger degradation than the

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Table 3 InXuence of germination conditions on the contents of phytate, insoluble (IDF), soluble (SDF) and total dietary Wbre (TDF), beta-glucan and beta-glucanase activity (GA) in oat and barley malts in comparison with ungerminated kernels Time (h) Temp Oats (C) Phytate IDF SDF TDF Beta-glucan (% dm) (% dm) (% dm) (% dm) (% dm) 0.683 10 20 15 12 15 15 18 15 10 20 13 0.645 0.560 0.650 0.559 0.603 0.588 0.541 0.530 0.467 0.517 0.551 0.533 22.3 24.7 26.7 24.5 27.7 26.6 27.1 27.1 28.2 28.6 31.2 27.4 28.0 3.4 3.2 3.6 2.1 1.7 1.6 1.7 1.2 1.4 1.5 1.4 1.3 1.6 25.7 27.9 30.3 26.5 29.0 28.2 29.3 28.4 29.6 30.2 31.6 28.8 29.6 4.2 2.6 2.2 1.3 1.3 0.3 0.2 0.3 0.2 0.2 0.3 0.26 0.3 Barley GA Phytate (U/g dm) (% dm) n.m. 41 129 60 102 73 102 68 90 91 75 88 80 0.568 0.451 0.430 0.469 0.485 0.474 0.441 0.419 0.467 0.412 0.437 0.460 0.449 IDF SDF TDF Beta-glucan (% dm) (% dm) (% dm) (% dm) 14.1 14.9 14.9 15.5 14.5 15.7 15.2 16.8 15.9 16.5 16.3 15.7 15.5 3.1 4.1 4.5 3.7 2.6 2.3 2.3 2.3 1.3 1.8 1.4 1.9 2.1 17.2 17.0 17.3 17.2 16.0 16.9 16.5 18.0 17.2 18.2 17.7 17.0 17.6 3.8 2.4 1.8 1.6 1.7 0.3 0.3 0.3 0.3 0.3 0.4 0.48 0.4 GA (U/g dm) n.m. 338 440 405 538 584 542 585 583 514 474 580 562

Unmalted 48 48 62 96 96 96 96 130 144 144 Predicted 115

Predicted values using the models for a germination period of 115 h at 13 C and the measured values in these samples (n.m. = not measured)

Table 4 p-Values for the inXuence of germination time and temperature as well as model Wts (R2) for the mathematical models for the contents of phytate, insoluble (IDF), soluble (SDF) and total dietary Wbre (TDF), beta-glucan and beta-glucanase activity (GA) in oats and barley InXuencing factor Oats Phytate IDF Time (linear) Time (quadratic) Temperature (linear) Temperature (quadratic) Interactive Model Wt R2 0.002 >0.05 >0.05 0.002 >0.05 >0.05 SDF TDF Beta-glucan GA 0.004 >0.05 >0.05 >0.05 >0.05 0.923 >0.05 >0.05 >0.05 >0.05 >0.05 0.607 Barley Phytate IDF >0.05 >0.05 >0.05 >0.05 >0.05 0.537 >0.05 >0.05 >0.05 >0.05 >0.05 0.483 SDF TDF Beta-glucan GA 0.018 >0.05 >0.05 >0.05 >0.05 0.830 0.008 0.030 >0.05 >0.05 >0.05 0.928

0.004 >0.05 >0.05 >0.05 >0.05 >0.05 0.924 >0.05 >0.05 >0.05 >0.05 0.793

<0.001 >0.05 >0.05 >0.05 >0.05 >0.05 0.986 >0.05 >0.05 >0.05 >0.05 0.625

0.045 >0.05 0.023 >0.05 0.952 0.940

SigniWcant inXuencing factors in bold print

one found in our study. A possible explanation for the diVerence is the fact that steeping and germination procedures were used which are vastly diVerent from the ones used in commercial malting or the micro malting used in our study. The comparatively high temperatures in the study by Centeno and collaborators [18] might have contributed to the diVerence. In contrast to these Wndings, Alminger found a phytate reduction by 34% after 4 days of germination [29] and Briggs reported a net decrease of phytate between 10 and 30% during malting [2], which corresponds well with the values measured in the present study. In oat malts the phytate concentration, ranging from 0.645 to 0.467%, was signiWcantly decreased as a consequence of prolonged germination times. In addition, a signiWcant eVect of the quadratic temperature term and the interactive term were seen. A graph illustrating these eVects

is shown in Fig. 1. To our knowledge, no recent publication investigated the breakdown of phytate under malting/germination conditions close to the ones used in commercial malting. Two studies by Larsson and Sandberg [28, 29] showed the impact of germination on phytate in oats. After 3040 h of germination at 15 C, they found a decrease in phytate content by 16% in oats, which was also reported [29]. As already seen for barley, this reported breakdown is stronger than the one found in our study, a diVerence which likely is mostly caused by the germination procedure. However, in contrast to barley, there was a signiWcant eVect of the germination conditions. This is likely caused by the activity of phytases. Larsson and Sandberg found that the optimal temperature for breakdown of phytate by indigenous phytases in oat is at temperatures between 37 and 40 C [28], which is lower than for other cereals. Temperatures in the

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0.68 0.66 0.64 0.62 0.60 0.58 0.56 0.54 0.52 0.50 0.48 0.46 0.44 0.42 40 60 80 100 120

140 160

Germ

in

tem ation

ure perat

/ C

Fig. 1 Model for the inXuence of germination time and temperature on the phytate content in oats; model Wt R2 = 0.952

drying grain during kilning in our trials (air temperatures of 45 C) might be lower initially, due to energy used for evaporation of water, and therefore favour the breakdown of phytate at these temperatures. At the stage where temperatures are high enough to favour the phytases in barley, water content in the grains might be low enough to prevent high enzymatic activity in spite of favourable temperatures. Dietary Wbre Dietary Wbre (DF) has been identiWed as an important reason for the positive health eVect caused by the uptake of whole grain cereals [30]. In this study, the contents of soluble and insoluble dietary Wbre were measured and from these two fractions the content of total dietary Wbre was calculated. The relevant results are shown in Table 3 and key values for the mathematical models in Table 4, respectively. Higher values of total DF (TDF) were measured in oats, native kernels and malts, than in barley and barley malts. The reason for this is clearly the high content of husk in oats, which is reported to contain high amounts of dietary Wbre [25]. Barley contains high amounts of cellulose, arabinoxylan (AX) and beta-glucan [31]. While beta-glucans and AX are partly soluble, celluloses are not. Oats is known to contain high levels of beta-glucan but only small amounts of AX [7, 8]. DiVerences in the conformation of AX and crosslinks with other macromolecules aVect the properties, amongst them the solubility. The main fraction of the total dietary Wbre in oats as well as in barley is insoluble dietary Wbre (IDF). Native oats and barley kernels contained 22.3 and 14.1% IDF, respectively. IDF in oat malts increased as

a consequence of prolonged germination times. Germination periods of 48 h resulted in IDF contents of 24.7 and 26.7%, respectively, for germination temperatures of 10 and 20 C. In contrast, malts germinated for 144 h had contents of 28.6 and 31.2% for the respective germination temperatures. Although some variations in the content of DF were seen as a consequence of the germination temperatures, these were not statistically signiWcant (p = 0.0522). In barley, variations in IDF between 14.9 and 16.8% were measured in the malts, with slight trend towards higher contents in malts germinated for longer periods. However, neither this trend nor any eVect of the germination temperature was signiWcant. The model Wt for this parameter was very poor, likely caused by a comparatively high value measured for the samples germinated for 96 h at 18 C. This might have been an eVect of poor sampling or a slight variation in the raw material used in the malting trials. A repetition of the measurement with freshly prepared sample resulted in the same values. Therefore, the measurement was fully taken into consideration. The increase in IDF can partly be explained by the loss of compounds such as starch due to respiration. Some material is also used for the build-up of the rootlets, which were removed before analysing for the Wbre content. During germination, a degradation and solubilisation of some dietary Wbre components takes place. Amongst the aVected compounds are beta-glucans and arabinoxylans, which are, as partly soluble Wbre, discussed in more detail in the following section. Finally, a build-up of compounds such as celluloses and hemi-celluloses as structural elements of the growing acrospires can be expected, as seen in germinating rice [32]. Further research will have to show to what intent the increase in insoluble dietary Wbre is caused by the loss of other nutrients. While increased values of dietary Wbre might be positive from a nutritional point of view, an increased loss of starch and proteins is not desirable from an economical perspective. Higher losses of starch and other organic materials reduce the amount of malt produced from a Wxed amount of raw material. Soluble dietary Wbre Many of the health beneWts associated to the intake of dietary Wbre, e.g., lowering of blood glucose level, eVects on the immune system in the small intestine are mainly caused by soluble dietary Wbre (SDF). SDF increase the viscosities of solutions and thereby slow intestinal transit, delay gastric emptying and reduce glucose and sterol absorption. There is also evidence for decreases of serum cholesterol, postprandial blood glucose and insulin contents in the human body [5]. The main compounds contributing to the SDF in barley are arabinoxylans and beta-glucans [2, 33].

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Eur Food Res Technol (2010) 231:2735 Fig. 2 RSM models for the inXuence of germination time and temperature on the content of soluble dietary Wbre in a oat malts, model Wt R2 = 0.924, and b barley malts, R2 = 0.968

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(a)
SDF / % dm

4.0 3.5 3.0 2.5 2.0 1.5 1.0 40

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SDF / % dm

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18 16 14 12 10 8

Both of them are components of the cell wall and are mainly concentrated in the endosperm. Unmalted oats contained 3.6% SDF and unmalted barley 3.1%. During the beginning phases of the germination, a solubilisation of DF took place. While not visible in oats (samples germinated for 48 h contained similar levels than the unmalted ones) were seen, there was an increase in barley from 3.1% in the native kernel to contents of 4.5 and 4.7, respectively, for barley malts germinated for 48 h at 10 and 20 C. The reason for this increase is a solubilisation of the relevant macromolecules. Cleavage of intermolecular bonds and breakdown of protein structures can thereby increase the amount of SDF. In both barley and oats, a signiWcant decrease in the content of SDF during the course of germination was observed (Tables 3, 4), while the germination temperature in the applied range had no signiWcant impact in either case. The model Wts are very high for both models, graphs of which are shown in Fig. 2a and b. Comparing with the values for beta-glucan (see below), it appears that degradation of betaglucans was largely responsible for the loss of SDF. Khler and co-workers found no signiWcant increase or decrease in the content of SDF within the 96 h of germination in wheat and a signiWcant increase for germination periods longer 96 h at temperatures of 15 and 25 C [34]. This is in accordance with the low beta-glucan content in wheat, which might be broken down, but is compensated by the solubilisation of other compounds, such as AX. Beta-glucan and -glucanase activity The health eVects of beta-glucan have been intensely studied and the relevant health claims have been approved [9]. Barley and oats can be seen as the most important sources of beta-glucans in the human diet. During germination, beta-glucan is quickly degraded in both cereals. Unmalted

barley contained 3.8% beta-glucan and unmalted oats 4.2%. Even short germination times (i.e., 48 h) led to a considerable breakdown of beta-glucan in both grains. Oats, after soaking and germination for 48 h and kilning, contained 2.6% beta-glucan and barley 2.4%, a decrease of 37 and 68%, respectively. In samples germinated for longer than 96 h, only small amounts of residual beta-glucan were found, 0.2% in oats and slightly higher 0.3% in barley. The RSM models for both oats and barley are shown in Fig. 3. The numbers found for barley are consistent with the ones typically found in literature [2]. Peterson [8] and Wilhelmson et al. [12] reported similar levels of beta-glucan in a wide range of oat malts which were germinated at 15 C. In contrast to our study, only a small loss of beta-glucan within the Wrst 2 days of germination was seen. However, the named samples were not kilned but freeze dried, cutting out a period of time in which beta-glucans can also be broken down. No signiWcant impact of the germination temperature on the breakdown of beta-glucan was found. The measured data show slightly higher values after 48 h of germination at 10 C than at 20 C for barley malts as well as oat malts. However, the diVerences are comparatively small and were not observed in samples germinated for longer periods. Wilhelmson and coworkers found beta-glucans in oat degraded at approximately the same rate at 25 C as at 15 C, but less quickly at 5 C [12]. The temperature range investigated in the present study was not large enough to replicate this. However, applying very low temperatures might require the use of additional equipment and energy and a prolonged germination process in order to reach the required changes in the functional and nutritional properties. Therefore, this might be unsuitable for commercial malting. The breakdown of beta-glucans during germination is caused by beta-glucanases. In barley malts, the activity

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34 Fig. 3 RSM models for the inXuence of germination time and temperature on the content of beta glucan in a oat malts, model Wt R2 = 0.923, and b barley malts, R2 = 0.830

Eur Food Res Technol (2010) 231:2735

(a)
% dm Beta-glucan /

3.0 2.5 2.0 1.5 1.0 0.5

40

(b)
% dm Beta-glucan /
22

3.0 2.5 2.0 1.5 1.0 0.5

0 Ge 6 80 rm 0 ina 10 20 tio n 1 40 tim 1 60 e /h 1 40

22 20 18

Ge

60

rm

80

ina

tio

0 10

nt

0 12

im

e/

0 14

0 16

Ge

rm

in

on ati

tem

tu era

re

C /

20 18 16 14 12 10 8

Ge

rm

in

on ati

tem

tur era

e/

16 14 12 10 8

increased as a consequence of prolonged germination periods until a maximum was reached and then slightly decreased. In oats, however, no clear trend was seen. Wilhelmsson et al. reported that in their studies there was a signiWcant impact of the germination temperatures, i.e., 5, 15 and 25 C [12]. However, this did not apply for the temperature range between 10 and 20 C. It might be that the assay procedure used is not favourable for the determination of oat beta-glucanase activity.

Acknowledgments The help of J. C. Oliveira, Department of Food and Process Engineering, University College Cork with developing the experimental design, is gratefully acknowledged. Funding for this research was provided under the Irish National Development Plan, through the Food Institutional Research Measure, administered by the Department of Agriculture, Fisheries & Food, Ireland.

References
1. Belitz H-D, Grosch W, Schieberle P (2001) Lehrbuch der Lebensmittelchemie, 5th edn. Springer, Berlin 2. Briggs DE (1998) Malts and malting. Blackie Academic and Professional, London 3. Aufhammer W (2003) RohstoV Getreide. Eugen Ulmer GmbH & Co, Stuttgart 4. Kaukavirto-Norja A, Wilhelmson A, Poutanen K (2004) Agric Food Sci 13:100112 5. Charalampopoulos D, Wang R, Pandiella SS, Webb C (2002) Int J Food Microbiol 79:131141 6. Voragen AGJ (1998) Trends Food Sci Tech 9:328335 7. Cui SW, Wang Q (2009) Struct Chem 20:291297 8. Peterson DM (1998) Cereal Chem 75(2):230234 9. Woods PJ (2007) J Cereal Sci 46:230238 10. Temelli F, Bansema C, Stobbe K (2004) Cereal Chem 81:499503 11. Brennan CS, Cleary LJ (2005) J Cereal Sci 42:113 12. Wilhelson A, Oksman-Caldentey K-M, Laitila A, KaukovirtaNorja A, Poutanen K (2001) Cereal Chem 78(6):715720 13. Frlich W, Nyman M (1988) J Cereal Sci 7:7382 14. Sandstrm B, Almgren A, Kvist B, Cederblad (1987) J Nutr 117:18981902 15. Harland BF, Morris ER (1995) Nutr Res 15(5):733754 16. Zhou JR, Erdman JW (1995) Crit Rev Food Sci Nutr 35:495508 17. Reddy NR, Sathe SK, Sahlunke DK (1982) Adv Food Res 28:192 18. Centeno C, Viveros A, Brenes A, Canales R, Lozano A, de la Cuadra C (2001) J Agric Food Chem 49:32083215 19. Bartnik M, Szafranszka I (1987) J Cereal Sci 5(1):2328 20. RossanderHulten L, Gleerup A, Hallbergh L (1990) Eur J Clinic Nutr 44:783791 21. Sandstrm B, Cederblad A, Stenquist B, Andersson H (1990) Eur J Clinic Nutr 44(10):705708 22. Graf E, Eaton JW (1993) Nutr Cancer 19:1119 23. Box GEP, Hunter W, Hunter J (1978) Statistics for experimentiers. Wiley Inc, New York

Conclusions Malting of oats and barley can be used as a tool to inXuence the nutritional properties of the grains. The breakdown of phytate, regarded as anti-nutritional factor, in oats is favoured by long germination periods, while this was not seen in barley. While ungerminated barley had higher contents of phytate then all the corresponding malts, variation in the germination conditions was not a way to inXuence the phytate contents. Mineral appeared to be mainly inXuenced by the steeping, or else the loss of organic material due to respiration of the embryo. Short germination periods can result in increased contents of SDF in comparison with the native kernels. Longer germination periods, however, considerably decreased the content of SDF as well as betaglucans, while the content of ISF in oats was increased. Variation in the germination temperature between 10 and 20 C had little overall eVect on Wbre and beta-glucan. While the majorities of malts is used in the brewing and distilling industries, malts might be interesting ingredients for functional food and beverages. For these applications, it is important to choose a raw material and malting conditions which will enhance the desired compound, e.g., short germination periods for high Wbre, but long germination times for degradation of phytate in oats.

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Eur Food Res Technol (2010) 231:2735 24. Lintschinger J, Fuchs N, Moser J, Kuehnelt D, Goessler W (2000) J Agric Food Chem 48:53625368 25. Salama A-RA, El-Sahn MA, Mesallam AS, El-Tabey S (1997) J Sci Food Agric 75:5056 26. Ekholm P, Virkki L, Ylinen M, Johansson L (2003) Food Chem 80:165170 27. Larsson M, Sandberg A-S (1992) J Food Sci 57(4):994997 28. Larsson M, Sandberg A-S (1995) J Cereal Sci 21:8795 29. Alminger M (2006) Sveriges Utsaedesfoerenings Tidskrift 116(1/2):313

35 30. Slavin JL, Jacobs D, Marquart L (2001) Critic Rev Biotechnol 21(1):4966 31. Holtekjlen AK, Uhlen AK, Brthen E, Sahlstrm S, Knutsen SH (2006) Food Chem 94:348358 32. Lee YR, Kim JY, Woo KS, Hwang IG, Kim KH, Kim KJ, Kim JH, Jeong HS (2007) Food Sci Biotechnol 16(6):10061010 33. Sungurtas J, Swanston JS, Davies HV, McDougall GJ (2004) J Cereal Sci 39(2):273281 34. Khler P, Hartmann G, Wieser H, Rychlik M (2007) J Agric Food Chem 55:46784683

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