Research Article Received: 25 April 2011 Revised: 16 July 2011 Accepted: 24 July 2011 Published online in Wiley Online Library: 14 September 2011 (wileyonlinelibrary.com) DOI 10.1002/jsfa.4628 Effects of germination on nutritional composition of waxy wheat Pham Van Hung,a∗ Tomoko Maeda,b Syota Yamamotoc and Naofumi Moritac,d Abstract BACKGROUND: Germination is considered to improve the nutritive value, antioxidant capacity and functional properties of grains In this study, changes in the chemical composition, nutritive value and antioxidant capacity of waxy wheat during germination were determined RESULTS: Over a 48 h period of germination the protein and free lipid contents of germinated waxy wheat were not significantly different from those of the control, whereas the bound lipid content decreased significantly An increase in levels of ash and dietary fibre was clearly observed for the 48 h-germinated wheat The total free amino acid content of the 48 h-germinated wheat was 7881 mg kg−1 flour (dry basis (d.b.)), significantly higher than that of the ungerminated wheat (2207 mg kg−1 flour, d.b.) In particular, γ -aminobutyric acid increased from 84 mg kg−1 flour (d.b.) in the control to 155 mg kg−1 flour (d.b.) in the 48 h-germinated wheat Germination did not significantly affect the fatty acid composition of both free and bound lipids of waxy wheat, whereas free phenolic compounds increased during germination, resulting in an increase in antioxidant capacity of germinated wheat CONCLUSION: Germinated waxy wheat had a better nutritional composition, such as higher dietary fibre, free amino acid and total phenolic compound contents, than ungerminated waxy wheat Therefore germinated waxy wheat should be used to improve the nutritional quality of cereal-based products c 2011 Society of Chemical Industry Keywords: waxy wheat; germination; nutritional composition; antioxidant INTRODUCTION J Sci Food Agric 2012; 92: 667–672 flavour mainly through the formation of Maillard compounds during baking.8 In particular, γ -aminobutyric acid (GABA), a free amino acid widely distributed in nature, has several physiological functions such as neurotransmission and induction of hypotensive, diuretic and tranquillising effects.9 Free lipids, the major lipids in wheat, contain large amounts of essential fatty acids (linoleic and linolenic acids), which play an important role in the human body, where they cannot be synthesised because humans lack the enzymes required for their production The remaining bound lipids also have technological importance in reducing stickiness, improving freeze/thaw stability and retarding retrogradation of ∗ Correspondence to: Pham Van Hung, School of Biotechnology, International University, Vietnam National University, Quarter 6, Linh Trung Ward, Thu Duc District, HoChiMinh City, Vietnam E-mail: hungphambk@yahoo.com a School of Biotechnology, International University, Vietnam National University, Quarter 6, Linh Trung Ward, Thu Duc District, HoChiMinh City, Vietnam b Department of Life and Health Sciences, Hyogo University of Teacher Education, 942-1, Shimokume, Yashiro, Hyogo 673-1494, Japan c Laboratory of Food Chemistry, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1, Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan d Department of Food Packaging Technology, Toyo College of Food Technology, 4-3-2, Minami-Hanayashiki, Kawanishi, Hyogo 666-0026, Japan www.soci.org c 2011 Society of Chemical Industry 667 Waxy (amylose-free) wheat (Triticum aestivum L.) produced by new physicochemical and biological techniques is considered to have superior functional properties for breadmaking, resulting in formation of a new texture of breads with soft, viscous and glutinous breadcrumbs and retardation of bread staling.1 Several recent studies have focused on the application of waxy wheat for making whole-grain breads because of the high amounts of dietary fibre, resistant starch, vitamins, minerals and microconstituents located in the bran and germ of waxy wheat grains.2,3 The consumption of whole-grain foods is considered to have many physiological benefits related to life-threatening diseases such as coronary heart disease, colon cancer and diabetes.4 The dietary fibre located in the germ and bran of grains is usually removed by refining However, the consumption of dietary fibre can reduce the risk of heart disease and prevent colorectal cancer and metabolic and inflammatory bowel diseases such as diabetes and diverticulitis For example, soluble dietary fibre consumption was found to significantly lower blood cholesterol and help stabilise blood glucose levels,5,6 while insoluble dietary fibre consumption helped to protect against colon cancer and other bowel disorders of the intestinal tract.7 Free amino acids, free and bound lipids and free and bound phenolic compounds in whole wheat grains also contribute to the nutritive value of whole-wheat foods and have significant health benefits The qualitative and quantitative compositions of free amino acids of fermented doughs affect bread www.soci.org starch.10 Phenolic compounds in whole wheat exist mainly in the bound form and are found in the bran associated with cell wall materials.11 These phenolic compounds play an important role in combating oxidative stress in the human body by maintaining a balance between oxidants and antioxidants.12 Therefore the consumption of whole-grain foods has become popular, although they can be unattractive to consumers, because the large amounts of bran and germ in whole-grain flour tend to deteriorate the quality and sensory value of the end-use products Recently, several researchers have reported that germination improves not only enzyme activities but also nutritional composition, such as free amino acids, dietary fibre, vitamins, minerals and phenolic compounds.13 – 19 Koehler et al.15 found that gluten proteins of wheat were substantially degraded whereas soluble dietary fibre and total folate increased during germination Tkachuk16 reported that the free amino acid content after 122 h of germination at 10, 16.5 and 25 ◦ C was respectively 4×, 10× and 7× that of ungerminated wheat Increased levels of vitamins, minerals and phytoestrogens in soybean, wheat and alfalfa after germination were found by Plaza et al.,17 while a significant reduction in levels of tannins and phytic acid and an increase in digestible protein of sorghum cultivars during germination were reported by Mohamed Nour et al.18 Increased reducing sugars, total sugars and damaged starch with respect to the germination time of amaranth grains were also detected by Colmenares de Ruiz et al.14 During germination, ferulic, vanillic and syringic acid levels were found to rise as the germination time increased.13,19 Hung et al.13 also reported an increase in total phenolic compounds and antioxidant capacity of germinated wheat compared with ungerminated wheat Therefore germination helps to improve the chemical composition, nutritive value and acceptability characteristics of food products With the hope that waxy wheat grains will prove popular for enhancing whole-grain products, the objective of this study was to investigate changes in the chemical composition, nutritive value and antioxidant capacity of waxy wheat grains during germination MATERIALS AND METHODS Materials Waxy wheat grains (∼30 g kg−1 amylose) named Urara-mochi (Nohrin-mochi 163) were harvested in Nagano Prefecture in Japan in 2004 For the control sample, whole waxy wheat grains without germination were milled into whole-grain flour using a Retsch ZM1 milling apparatus (Haan, Germany) with a 0.5 mm mesh 1,1-Diphenyl-1-picrylhydrazyl (DPPH), Folin–Ciocalteu phenol reagent and other chemicals were purchased from Wako Chemical Co (Osaka, Japan) 668 Germination conditions Waxy wheat grains (∼50 g) were first rinsed with distilled water four times The grains were then soaked in excess distilled water for 30 at room temperature After steeping, the kernels were drained of water, placed separately on sterilised pre-soaked blotting paper and incubated in the dark in a cabinet at 30 ◦ C and 85% relative humidity for 0, 6, 12, 24, 36 and 48 h After incubation the samples were washed carefully with distilled water, frozen at −84 ◦ C and then freeze-dried The freeze-dried samples were ground in a Retsch ZM1 milling apparatus with a 0.5 mm mesh wileyonlinelibrary.com/jsfa PV Hung et al Determination of proximate composition The proximate composition of waxy wheats was determined by the following standard methods: AACC20 methods 44-15A, 08-01 and 30-10 for moisture, ash and lipid contents respectively; AACC20 method 46-10 for protein content using a Kjeltec Auto Sampler System 1035 analyser (Tecator Ltd, Tokyo, Japan); AOAC21 method 985.29 for dietary fibre content using a TDF-100A total dietary fibre assay kit (Sigma-Aldrich Co Ltd St Louis, MO, USA) Free and bound lipids were extracted using a Soxhlet system according to CEC standard procedures.22 Free lipid content was determined after h of Soxhlet extraction of a g sample with 110 mL of hot diethyl ether The residual solvent was removed on a rotary evaporator under reduced pressure at 35 ◦ C and the lipid extract was dried to constant weight at 105 ◦ C Bound lipid was extracted from the free lipid-removed residue Prior to ether extraction the residue was subjected to acid hydrolysis for h in 100 mL of hot 1.5 mol L−1 HCl The hydrolysed residue was then washed with at least 800 mL of distilled water and dried overnight at 70–75 ◦ C Finally, it was extracted with diethyl ether as described above Total lipid was calculated as the sum of free and bound lipids Free amino acid determination Free amino acids of germinated waxy wheat were determined according to the method of Saikusa et al.23 Wheat flour (1.6 g) was homogenised with mL of 80 mL L−1 trichloroacetic acid solution in a test tube (2 cm × 16 cm) using a homogeniser for and then shaken (100 strokes min−1 , cm amplitude) at 30 ◦ C for h The suspension was centrifuged at 14 000 × g for 15 at ◦ C The supernatant was filtered through a 0.45 µm membrane filter (Advantec Co., Ltd, Tokyo, Japan) and then analysed for free amino acids using an LC-11 amino acid analyser (Yanaco Co., Kyoto, Japan) Fatty acid determination The free fatty acid composition of free and bound lipids of germinated waxy wheat was determined by gas/liquid chromatography Free and bound lipids were extracted with nhexane (as described above) and used to prepare methylated fatty acids with 140 g L−1 boron trifluoride in methanol according to the method of Christie.24 The fatty acid composition was analysed using a Yanaco G 3800 gas/liquid chromatograph (Osaka, Japan) Determination of total phenolic compounds and antioxidant capacity of wheat extracts Total phenolic compounds and antioxidant capacity of waxy wheat extracts were investigated by the methods of Hung and Morita.25 Free phenolic compounds were extracted from g of waxy wheat flour by shaking with 10 mL of chilled 800 mL L−1 ethanol in water for 20 The extraction was repeated three times and the combined supernatants were evaporated at 45 ◦ C and reconstituted with methanol to a final volume of 10 mL The extract was then stored at −40 ◦ C until use Bound phenolic compounds were extracted six times with diethyl ether/ethyl acetate (1 : v/v) after alkaline hydrolysis of the residue from free phenolic compound extraction The combined diethyl ether/ethyl acetate extracts were evaporated to dryness and bound phenolic compounds were reconstituted in 10 mL of methanol and then stored at −40 ◦ C until use Total phenolic compounds and antioxidant capacity of free and bound phenolic extracts were determined using the Folin–Ciocalteu and DPPH radical-scavenging methods respectively as previously described by Hung and Morita.25 c 2011 Society of Chemical Industry J Sci Food Agric 2012; 92: 667–672 Nutritional composition of germinated waxy wheat www.soci.org Statistical analysis All tests were performed at least in duplicate Analysis of variance was carried out together with Duncan’s multiple range test to compare treatment means at P < 0.05 using SPSS Version 16 (SPSS Inc., Chicago, IL, USA) RESULTS AND DISCUSSION Proximate composition of waxy wheat during germination Changes in nutritional composition of waxy wheat grains during germination are shown in Table The ash content of waxy wheat (18.0 ± 0.2 g kg−1 , dry basis (d.b.)) did not change significantly during the first 36 h of germination However, the ash content of the 48 h-germinated wheat (19.4 ± 0.1 g kg−1 , d.b.) was significantly higher than that of the control During 48 h of germination the protein and free lipid contents of germinated waxy wheat were not significantly different from those of the control (154 ± and 18.3 ± 1.1 g kg−1 , d.b., respectively), whereas the amount of bound lipid decreased significantly An increase in total dietary fibre was clearly observed for the 48 h-germinated wheat, whereas the amount of starch decreased significantly after 12 h of germination Several previous studies have also found changes in nutritive value of sprouted wheat Lemar and Swanson26 reported changes in composition of wheat germinated for 1–3 days to sprout lengths of 0.25 and inch The most notable differences in proximate composition of sprouted and control flours were the significantly higher crude fat and crude protein concentrations determined for all four sprouted flours, whereas the crude fibre and ash contents did not change significantly.26 Later, Ranhotra et al.27 studied changes in nutritive value of sprouted hard red winter wheat during germination for 3–5 days and reported that the protein and lipid contents of sprouted wheat increased appreciably with the duration of sprouting The increase in crude fibre was most pronounced by the fifth day, whereas the ash content of the sprouted wheat changed little.27 These results not conflict with the results of our study, because the protein and lipid contents reported increased over a longer period of germination (>48 h) The loss of starch due to germination probably meant that the weight of grain decreased, while ash and fibre remained the same but increased as a proportion of total grain weight Changes in free amino acids during germination Changes in free amino acid composition of waxy wheat during germination are shown in Table The amounts of free amino acids in waxy wheat increased significantly with increasing germination time The essential amino acids isoleucine, leucine, phenylalanine Table Germination time (h) Amino acid Essential amino acids Ile 64a 88b Leu 58a 120b Lys 43a 74d Met 29a 51b Phe 53a 85b Thr 73a 176e Val 119a 175b Semi-essential amino acids Arg 118a 170c Gly 77b 94c His 76a 83b Tyr 91a 100b Non-essential amino acids Ala 281a 308b Asn 476b 561c Asp 151b 167c Cys 16a 16a GABA 84a 107b Glu 183a 311b Orn 7b 6b Pro 68a 65a Ser 138a 151bc Total 2207 2906 12 24 36 48 83b 117b 63c 60c 85b 163d 173b 153c 230c 70d 71d 152c 228f 769c 334d 392e 50b 71d 249d 111b 1990e 337d 277d 37a 136e 239d 146c 1546d 139b 59a 145d 110c 120a 72b 236f 163d 181d 114d 108c 229e 183d 74b 171e 260f 303b 579d 198d 14a 101b 382c 5b 73b 158c 3011 523c 659e 230e 19b 102b 794d 3a 370c 230e 5193 790e 446a 132a 24c 105b 892e 9c 667d 147b 7040 622d 775f 247f 108d 155c 1150f 43d 1178e 197d 7881 Values are mean of duplicate measurements Means with the same letter in the same row are not significantly different (P < 0.05) and valine reached maximum levels of 334, 392, 249 and 1990 mg kg−1 flour (d.b.) respectively after 36 h of germination, compared with levels of 64, 58, 53 and 119 mg kg−1 flour (d.b.) in the control Of the other essential amino acids, threonine and methionine were highest after 24 and 48 h of germination respectively, whereas lysine peaked after h of germination and then decreased dramatically The amounts of the semi-essential amino acids histidine and glycine were highest after 24 and 36 h of germination respectively, whereas arginine and tyrosine were highest after 48 h of germination Almost all non-essential amino acids increased significantly with increasing germination time and reached their highest levels after 48 h In particular, GABA, Changes in proximate composition (g kg−1 , d.b.) of waxy wheat grains during germination Germination time (h) 12 24 36 48 Table Changes in free amino acid composition (mg kg−1 flour, d.b.) of waxy wheat grains during germination Ash Protein Free lipid Bound lipid Total dietary fibre Starch 180 ± 0.2b 17.3 ± 0.1a 17.6 ± 0.1a 17.6 ± 0.1a 18.6 ± 0.2c 19.4 ± 0.1d 154 ± 1a 155 ± 1a 156 ± 2a 157 ± 1a 157 ± 1a 157 ± 2a 18.3 ± 1.1b 17.9 ± 0.3a 18.1 ± 1.1ab 18.1 ± 0.3ab 18.2 ± 0.0ab 18.2 ± 0.5ab 8.6 ± 0.2e 8.0 ± 0.3d 7.5 ± 0.1c 7.1 ± 0.2b 6.9 ± 0.2ab 6.7 ± 0.1a 177 ± 2a 175 ± 11a 184 ± 1b 181 ± 1b 180 ± 3b 190 ± 2c 624 ± 5c 627 ± 3c 617 ± 4b 619 ± 3b 619 ± 4b 609 ± 4a J Sci Food Agric 2012; 92: 667–672 c 2011 Society of Chemical Industry wileyonlinelibrary.com/jsfa 669 Values are mean ± standard deviation of duplicate (lipid contents only) or triplicate measurements Means with the same letter in the same column are not significantly different (P < 0.05) www.soci.org Table PV Hung et al Changes in fatty acid composition (% of total fatty acids) of waxy wheat grains during germination Fatty acid C14 : Myristic C16 : Palmitic C16 : Palmitoleic C18 : Stearic C18 : Oleic C18 : Linoleic C18 : Linolenic C20 : Arachidic C20 : Eicosenoic C20 : Eicosatrienoic C20 : Eicosapentaenoic Saturated Monounsaturated Polyunsaturated Total Free lipid Bound lipid Germination time (h) Germination time (h) 12 24 36 48 12 24 36 48 0.1 19.0 0.1 1.0 14.4 59.9 4.0 0.2 0.9 0.0 0.1 20.3 15.5 64.0 99.8 0.1 18.9 0.2 1.0 14.5 59.5 3.9 0.2 1.0 0.0 0.1 20.3 15.8 63.6 99.6 0.1 19.0 0.2 1.0 14.1 59.6 4.0 0.2 1.0 0.0 0.1 20.5 15.4 63.7 99.6 0.0 19.1 0.1 1.0 13.8 60.4 4.2 0.2 1.0 0.0 0.0 20.3 15.0 64.5 99.8 0.1 18.8 0.2 1.0 14.3 59.7 4.1 0.2 0.0 0.0 0.1 20.1 14.6 64.0 98.6 0.0 19.0 0.3 1.1 14.1 59.9 4.2 0.2 0.9 0.0 0.0 20.4 15.3 64.1 99.7 0.0 19.2 0.0 0.8 10.3 59.6 3.8 0.0 6.0 0.4 0.0 20.0 16.3 63.7 100 0.0 19.5 0.0 1.0 10.6 61.4 3.9 0.4 3.1 0.0 0.0 20.9 13.7 65.3 100 0.0 19.4 0.0 1.2 10.4 62.0 4.0 0.5 2.2 0.0 0.0 21.4 12.6 66.0 100 0.0 18.1 0.0 1.4 11.4 56.1 3.8 0.0 8.4 0.5 0.0 19.5 19.8 60.4 99.7 0.1 18.4 0.1 1.2 9.7 57.3 4.9 0.5 6.3 0.5 0.2 20.3 16.1 62.9 99.3 0.0 19.1 0.2 1.1 9.9 59.5 4.3 0.4 4.5 0.3 0.2 20.6 14.6 64.2 99.4 Values are mean of duplicate measurements a functional ingredient with health benefits, increased during germination Total free amino acids increased significantly from 2207 mg kg−1 flour (d.b.) for the control to 7040 mg kg−1 flour (d.b.) after 36 h of germination and to 7881 mg kg−1 flour (d.b.) for the 48 h-germinated waxy wheat The largest contributors to the increased amounts of total amino acids were proline (from 68 to 1178 mg kg−1 ), valine (from 119 to 1990 mg kg−1 ), leucine (from 58 to 392 mg kg−1 ) and glutamic acid (from 183 to 1150 mg kg−1 ) Increases in free amino acid composition during germination have also been investigated in common wheat16,28 and buckwheat.29 670 Changes in fatty acids during germination Changes in fatty acid composition of waxy wheat during germination are shown in Table Eleven fatty acids, representing 98.6–100% of total fatty acids, were present in the control and germinated waxy wheat Linoleic (C18 : 2), palmitic (C16 : 0), oleic (C18 : 1), linolenic (C18 : 3) and stearic (C18 : 0) acids were the major components of both free and bound lipids of waxy wheat, and their percentages in free and bound lipids were not significantly different (56.1–62.0, 18.1–19.5, 9.7–14.5, 3.8–4.9 and 0.8–1.4% respectively) Myristic (C14 : 0), palmitoleic (C16 : 1), arachidonic (C20 : 0), eicosatrienoic (C20 : 3) and eicosapentaenoic (C20 : 5) acids were trace components in both free and bound lipids, whereas eicosaenoic (20 : 1) acid was a trace component in free lipid but a major component in bound lipid of waxy wheat The results also indicate that germination did not significantly affect the fatty acid composition of both free and bound lipids of waxy wheat, where the major components were polyunsaturated fatty acids (60.4–66.0% of total fatty acids), followed by saturated (19.5–21.4%) and monounsaturated (12.6–19.8%) fatty acids Linoleic and linolenic acids are essential fatty acids that play an important role in the human body, because these fatty acids cannot be synthesised as humans lack the enzymes required for their production In this study, these essential fatty acids in waxy wheat did not change significantly during 48 h of germination However, the amount of linoleic acid was found to decrease in wileyonlinelibrary.com/jsfa quinoa seeds after 72 h of germination.30 Thus the germination time should not be longer than 48 h in order to retain the essential fatty acids in germinated grains Changes in total phenolic content of waxy wheat extracts during germination Figure shows changes in total phenolic content (TPC) of waxy wheat during germination determined by the Folin–Ciocalteu method and expressed as mg ferulic acid equivalent (FAE) kg−1 flour The TPC of free phenolic extracts did not change significantly during the first 24 h of germination However, the amounts of free phenolic compounds increased significantly after germination for 36 and 48 h The TPC of bound phenolic extracts first decreased after 12 and 24 h of germination but then increased significantly after 36 and 48 h of germination The increase in free phenolic content is due to the increase in syringic acid as reported by Hung et al.13 However, bound phenolic compounds were first lost by hydrolysis of polyphenolic compounds bound to cell walls as reported by Yang et al.19 and then ferulic acid accumulation due to phenolic biosynthesis contributed to the increase in bound phenolic content after 24 h of germination Hung et al.13 also reported that sprouted Canadian Western Red Spring (CWRS) and Canadian Western Amber Durum (CWAD) wheats exhibited significantly higher free TPC than their corresponding controls, and the bound TPC extracted with alkali from steeped, then incubated subsamples in both CWRS and CWAD classes was also found to be significantly greater than that from their corresponding steeped subsamples Changes in antioxidant capacity of waxy wheat extracts during germination The antioxidant capacity of waxy wheat extracts determined by the DPPH radical-scavenging method is shown in Fig Free phenolic extracts of waxy wheat germinated for 36 and 48 h exhibited significantly higher DPPH radical scavenging than those of the c 2011 Society of Chemical Industry J Sci Food Agric 2012; 92: 667–672 Nutritional composition of germinated waxy wheat www.soci.org Total phenolics (mg FAE kg-1 flour, db) 2500 Free phenolics b b Bound phenolics a a b 2000 ab 1500 c b 1000 a a a a 500 0 12 24 Germination time (h) 36 48 Figure Changes in total phenolic content of waxy wheat grains during germination Values are mean and standard deviation of triplicate measurements Means with the same letter in the same series are not significantly different (P < 0.05) 45 DPPH scavenging (%) 40 bc 35 a ab ab bc c 30 c 25 b 20 15 a a a a 10 Free phenolics Bound phenolics 0 12 24 Germination time (h) 36 48 Figure DPPH radical-scavenging capacity of free and bound phenolic extracts from germinated waxy wheat grains (DPPH concentration 0.075 mmol L−1 ) Values are mean and standard deviation of triplicate measurements Means with the same letter in the same line are not significantly different (P < 0.05) control and the and 12 h-germinated wheat The DPPH radical scavenging of bound phenolic extracts first decreased after h of germination and then increased again with longer germination time These results are due to the changes in TPC of both free and bound phenolic extracts of germinated waxy wheat In addition, other antioxidant compounds such as vitamin C and tocopherols also increased with the length of germination, which might also increase the antioxidant activity of germinated waxy wheat.19 CONCLUSION J Sci Food Agric 2012; 92: 667–672 REFERENCES Hung PV, Maeda T and Morita N, Waxy and high-amylose wheat starches and flours – characteristics, functionality and application Trends Food Sci Technol 17:448–456 (2006) Hung PV, Maeda T and Morita N, Dough and bread qualities of flours with whole waxy wheat flour substitution Food Res Int 40:273–279 (2007) Hung PV, Maeda T, Fujita M and Morita N, Dough properties and breadmaking qualities of whole waxy wheat flours and effects of additional enzymes J Sci Food Agric 87:2538–2543 (2007) Marquart L, Jacobs Jr DR, McIntosh GH, Poutanen K and Reicks M (eds), Whole Grains and Health Blackwell, Ames, IA (2007) Anderson JW, Deakins DA, Floore TL, Smith BM and Whitis SE, Dietary fiber and coronary heart disease Crit Rev Food Sci Nutr 29:95–147 (1990) Brown L, Rosner B, Willett W and Sacks F, Cholesterol-lowering 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DISCUSSION Proximate composition of waxy wheat during germination Changes in nutritional composition of waxy wheat grains during germination are shown in Table The ash content of waxy wheat (18.0 ±... acid composition during germination have also been investigated in common wheat1 6,28 and buckwheat.29 670 Changes in fatty acids during germination Changes in fatty acid composition of waxy wheat. .. in nutritional composition of waxy wheat during 48 h of germination were investigated in this study The free amino acid composition was found to increase significantly during germination of waxy