Determination of the activity of acidic phytate-degrading enzymes in cereal seeds

Five different methods were compared to elucidate the total activity of the acidic phytate-degrading enzymes present in the seeds of rye, wheat, and barley. Phytate-degrading activity was studied at pH 5.0 by quantifying the liberated phosphate. Rye showed the highest acid phytate-degrading activity among the cereals studied. Using an aqueous extract, only 30-50% of the activity was found (rye, 3443 mU g(-1) of grain; wheat, 1026 mU g(-1) of grain; barley, 1032 mU g(-1) of grain) compared to that found by the direct incubation of the dry-milled cereal grains in a buffered phytate-containing solution (rye, 6752 mU g(-1) of grain; wheat, 2931 mU g(-1) of grain; barley, 2093 mU g(-1) of grain). Extending the extraction time resulted in an increase in extractable phytate-degrading activity by, at maximum, 10-15%. Extraction of phytate-degrading activity is strongly enhanced in the presence of Triton X-100 and the protease inhibitor phenylmethylsulfonyl fluoride (rye, 6536 mU g(-1) of grain; wheat, 2873 mU g(-1) of grain; barley, 2023 mU g(-1) of grain), suggesting at least a partial association with membrane structures and a degradation by proteolytic activity during extraction. In addition, it was shown that determining phytate-degrading activity by quantification of the liberated inorganic phosphate is more robust and precise than determining phytate-degrading activity by quantification of the residual phytate.     

Moderate decrease of pH by sourdough fermentation is sufficient to reduce phytate content of whole wheat flour through endogenous phytase activity

Whole wheat bread is an important source of minerals but also contains considerable amounts of phytic acid, which is known to impair their absorption. An in vitro trial was performed to assess the effect of a moderate drop of the dough pH (around 5.5) by way of sourdough fermentation or by exogenous organic acid addition on phytate hydrolysis. It was shown that a slight acidification of the dough (pH 5.5) with either sourdough or lactic acid addition allowed a significant phytate breakdown (70% of the initial flour content compared to 40% without any leavening agent or acidification). This result highlights the predominance of wheat phytase activity over sourdough microflora phytase activity during moderate sourdough fermentation and shows that a slight drop of the pH (pH value around 5.5) is sufficient to reduce significantly the phytate content of a wholemeal flour. Mg "bioaccessibility"of whole wheat dough was improved by direct solubilization of the cation and by phytate hydrolysis.     

Phytase and acid phosphatase activities in plant feedstuffs

A total of 183 samples representing 24 feedstuffs were analyzed for total phosphorus, phytate phosphorus content, phytase (Phy), and acid phosphatase (AcPh) activities with the objective to predict the capacity to hydrolyze phytic acid and to contribute to formulating environmentally adequate diets for monogastric animals. Of the cereals and cereal byproducts analyzed, only rye (5147 U kg(-)(1); 21 955 U g(-)(1)), wheat (1637 U kg(-)(1); 10 252 U g(-)(1)), rye bran (7339 U kg(-)(1); 56 722 U g(-)(1)), and wheat bran (4624 U kg(-)(1); 14 106 U g(-)(1)) were rich in Phy and AcPh activities. Legume seeds and oilseeds contained negligible Phy activity and a moderate amount of AcPh activity, except for kidney bean (33 433 U g(-)(1)) and full-fat linseed meal (13 263 U g(-)(1)). On the other hand, a significant linear regression between phytate phosphorus (y) and total phosphorus (x) was observed in cereal byproducts (R(2) = 0. 95; y = 0.8458x - 0.0367; P < 0.001) and oil seeds (R(2) = 0.95; y = 0.945x - 0.20; P < 0.001). Phy and AcPh were positively correlated with respect to phytate phosphorus in cereals, cereal byproducts, and other byproducts and negatively correlated in legume seeds and oilseeds. Except for cereals, the highest correlation between enzyme activities and phytate phosphorus was found for phytase. It is not possible to predict Phy and AcPh activities from phytate phosphorus content by linear and quadratic regressions. Finally, only highly significant and positive correlation was found between Phy and AcPh activities for cereals, cereal byproducts, and oilseeds.     

Synthesis of angiotensin I-converting enzyme (ACE)-inhibitory peptides and gamma-aminobutyric acid (GABA) during sourdough fermentation by selected lactic acid bacteria

This article aimed at investigating the synthesis of angiotensin I-converting enzyme (ACE)-inhibitory peptides and gamma-aminobutyric acid (GABA) during sourdough fermentation of white wheat, wholemeal wheat, and rye flours. Sourdough lactic acid bacteria, selected previously for proteinase and peptidase activities toward wheat proteins or for the capacity of synthesizing GABA, were used. The highest ACE-inhibitory activity was found by fermenting flour under semiliquid conditions (dough yield 330) and, especially, by using wholemeal wheat flour. Fourteen peptides, not previously reported as ACE-inhibitory, were identified from the water/salt-soluble extract of wholemeal wheat sourdough (IC 50 0.19-0.54 mg/mL). The major part of the identified peptides contained the well-known antihypertensive epitope VAP. The synthesis of GABA increased when the dough yield was decreased to 160. The highest synthesis of GABA (258.71 mg/kg) was found in wholemeal wheat sourdough.     

Methodological aspects of measuring phytase activity and phytate phosphorus content in selected cereal grains and digesta and feces of pigs

This study was to examine the time course of sample-specific linearity of intrinsic phytase hydrolysis in major cereal grains and in ileal digesta and fecal samples and to determine the time course of the microbial phytase-catalyzed hydrolysis of various sources of phytate for estimating phytate phosphorus (P) content. The intrinsic phytase activity in barley, corn, oat, and wheat samples was measured over multiple time points from 0 to 120 min at 1.5 mmol.L(-1) of sodium phytate at pH 5.5 and 37 degrees C. Time courses of hydrolysis of purified phytate and phytate associated with the cereal grain samples and the pig digesta and fecal samples were examined with the Natuphos microbial phytase over multiple time points from 0 to 48 h of incubation. The intrinsic phytase hydrolysis was linear (P < 0.05) for up to 120 min for the barley, corn, and wheat samples, whereas in the oat sample the hydrolysis was linear (P < 0.05) for only up to 30 min of incubation. The intrinsic phytase activities (phytase unit: mumol.kg(-1) of dry matter.min(-1)) for the barley, corn, and wheat samples were estimated to be 693, 86, and 1189 by linear regression analysis. Intrinsic phytase activity (412 phytase units) for the oat sample based on a 30-min incubation was considerably higher than the value (103 phytase units) determined from the 120-min incubation for the same oat sample. There were quadratic with plateau relationships (P < 0.05) between the hydrolytic release of inorganic P from various sources of phytate and the incubation time. The minimal incubation times required for the complete hydrolysis of phytate were estimated to be 4, 3, and 11 h for the purified phytate, the cereal grain samples, and the pig digesta and feces, respectively. It was concluded that multiple time point experiments need to be conducted to determine valid intrinsic phytase activity and phytate P content in samples through intrinsic and microbial phytase hydrolysis incubations.     

Changes in Phytate Content in Whole Meal Wheat Dough and Bread Fermented with Phytase‐Active Yeasts

The degradation of inositol hexakisphosphate (IP6) was evaluated in whole meal wheat dough fermented with baker's yeast without phytase activity, different strains of Saccharomyces cerevisiae (L1.12 or L6.06), or Pichia kudriavzevii with extracellular phytase activity to see if the degradation of IP6 in whole meal dough and the corresponding bread could be increased by fermentation with phytase‐active yeasts. The IP6 degradation was measured after the dough was mixed for 19 min, after the completion of fermentation, and in bread after baking. Around 60–70% of the initial value of IP6 in the flour (10.02 mg/g) was reduced in the dough already after mixing, and additionally 10–20% was reduced after fermentation. The highest degradation of IP6 was seen in dough fermented with the phytase‐active yeast strains S. cerevisiae L1.12 and P. kudriavzevii L3.04. Activity of wheat phytase in whole meal wheat dough seems to be the primary source of phytate degradation, and the degradation is considerably higher in this study with a mixing time of 19 min compared with earlier studies. The additional degradation of IP6 by phytase‐active yeasts was not related to their extracellular phytase activities, suggesting that phytases from the yeasts are inhibited differently. Therefore, the highest degradation of IP6 and expected highest mineral bioavailability in whole meal wheat bread can be achieved by use of a phytase‐active yeast strain with less inhibition. The strain S. cerevisiae L1.12 is suitable for this because it was the most effective yeast strain in reducing the amount of IP6 in dough during a short fermentation time.     

Improved bioavailability of dietary phenolic acids in whole grain barley and oat groat following fermentation with probiotic Lactobacillus acidophilus , Lactobacillus johnsonii , and Lactobacillus reuteri

The aim of this study was to improve the bioavailability of the dietary phenolic acids in flours from whole grain barley and oat groat following fermentation with lactic acid bacteria (LAB) exhibiting high feruloyl esterase activity (FAE). The highest increase of free phenolic acids was observed after fermentation with three probiotic strains, Lactobacillus johnsonii LA1, Lactobacillus reuteri SD2112, and Lactobacillus acidophilus LA-5, with maximum increases from 2.55 to 69.91 μg g(-1) DM and from 4.13 to 109.42 μg g(-1) DM in whole grain barley and oat groat, respectively. Interestingly, higher amounts of bound phenolic acids were detected after both water treatment and LAB fermentation in whole grain barley, indicating higher bioaccessibility, whereas some decrease was detected in oat groat. To conclude, cereal fermentation with specific probiotic strains can lead to significant increase of free phenolic acids, thereby improving their bioavailability.     

Use of fungal phytase to improve breadmaking performance of whole wheat bread.

The possible use of phytase as a breadmaking improver has been tested in whole wheat breads by adding different amounts of fungal phytase. The effect of phytase addition on the fermentation stage and the final bread quality was analyzed. The phytase addition shortened the fermentation period, without affecting the bread dough pH. Regarding the whole wheat bread, a considerable increase of the specific bread volume, an improvement of the crumb texture, and the width/height ratio of the bread slice were obtained. An in vitro assay revealed that the improving effect of phytase on breadmaking might be associated with the activation of alpha-amylase, due to the release of calcium ions from calcium-phytate complexes promoted by phytase activity. As a conclusion, phytase offers excellent possibilities as a breadmaking improver, with two main advantages: first, the nutritional improvement produced by decreasing phytate content, and second, all the benefits produced by alpha-amylase addition can be obtained by adding phytase, which promotes the activation of endogenous alpha-amylase.     

Effects of Oat Grain Hydrothermal Treatments on Wheat-Oat Flour Dough Properties and Breadbaking Quality

Hydrothermal treatments, which are routine in oat processing, have profound effects on oat flour dough rheological properties. The influence of roasting and steam treatments of oat grain on dough mixing and breadbaking properties was investigated when hydrothermally treated oat flour was blended with wheat flour. Roasting of oat grain (105°C, 2 hr) resulted in oat flours that were highly detrimental to wheat flour dough mixing properties and breadbaking quality. Steaming (105°C, 20 min) or a combination of roasting and steaming of oat grain significantly improved the breadbaking potential of the oat flours. The addition of oat flours increased water absorption and mixing requirements of the wheat flour dough and also decreased bread loaf volume. However, at the 10% substitution level, steamed oat flours exhibited only a gluten dilution effect on bread loaf volume when wheat starch was used as a reference. Oat flour in the breadbaking system decreased the retrogradation rate of bread crumb starch. The results indicate that adequate hydrothermal treatments of oat grain are necessary for oat flour breadbaking applications. Steamed oat flours used at a 10% level retarded bread staling without adversely affecting the loaf volume.     

Prolonged fermentation of whole wheat sourdough reduces phytate level and increases soluble magnesium

This work was designed to compare the effects of different leavens (yeast, sourdough, and a mixture of both) on phytic acid (PA) degradation and to assess the repercussions of PA breakdown on phosphorus and magnesium solubility during bread-making. Sourdough fermentation was more efficient than yeast fermentation in reducing the phytate content in whole wheat bread (-62 and -38%, respectively). Furthermore, lactic acid bacteria present in sourdough enhanced acidification, leading to increased magnesium and phosphorus solubility. To intensify phytate breakdown, bran was incubated with microorganisms (yeast or sourdough) before bread-making. Using this new method, the percentage of phytate breakdown was near 90%, whereas 40% of phytate remained in traditional French bread. In conclusion, a prolonged fermentation with sourdough still leads to improved Mg and P solubility by decreasing phytate content and through acidification.     

Development of Predictive Models for Optimization of Phytate Degradation in Wheat and Rye During Hydrothermal Processing

Whole kernels of wheat (Triticum aestivum ‘Kosack’), rye (Secale cereale ‘Motto’) and rice (Oryza sativa) were hydrothermally treated to reduce the phytate content. The hydrothermal processes used in this study included alternating wet and dry steeping periods. Two hydrothermal processes were used for wheat and rye with two and three wet and dry steeping steps, respectively. The process using two wet and dry steeping steps (process 1) was optimized for phytate reduction with respect to temperature during the process and lactic acid concentration used in the wet steeping steps. Optimal conditions for phytate breakdown in wheat and rye in process 1 were 55°C during the whole process and 1.3–1.5% lactic acid solution as the soaking agent in the wet steeping periods for wheat and 1.3% for rye. The hydrothermal process with three wet and dry steeping periods for wheat and rye (process 2) showed that the temperature used in the third wet and dry steeps did not significantly influence (P > 0.05) the phytate reduction in wheat and rye during these steps. The maximal phytate reduction was 94.4–95.6% in wheat and 99.0–99.5% in rye during hydrothermal processing. In rice, hydrothermal processing resulted, at best, in a 99.8% phytate reduction.     

Strains of lactic acid bacteria isolated from sour doughs degrade phytic acid and improve calcium and magnesium solubility from whole wheat flour

Five strains of lactic bacteria have been isolated from sour doughs and examined for their ability to degrade phytic acid. In white flour medium in which phytic acid was the only source of phosphorus, the disappearance of phytate and an elevation of inorganic phosphate were observed after only 2 h of incubation in all strains tested (-30 and +60%, respectively). Both phenomena correspond to phytate breakdown. No difference was observed in the levels of phytic acid hydrolysis among strains, suggesting that phytase enzymes are similar among these bacteria. Using whole wheat flour medium naturally rich in phytic acid in the presence of Leuconostoc mesenteroides strain 38, a 9 h fermentation established that the degradation of PA and the production of lactic acid lead to greater Ca and Mg solubility than in control medium.     

Effect of Cooking on Lignans Content in Whole‐Grain Pasta Made with Different Cereals and Other Seeds

Lignans are of increasing interest because of their potential anticarcinogenic, antioxidant, estrogenic, and antiestrogenic activities. In this work, mixed‐cereal pastas manufactured by adding 60% whole‐grain flours of different cereals (wheat, oat, rye, barley, and rice) to durum wheat semolina, a multigrain pasta with different grains (cereals, legumes, and flaxseed), and a traditional industrial durum wheat semolina were analyzed for their lignans content both in the raw and in the cooked state, ready for consumption. For raw mixed‐cereal pastas, total lignans were within the range 94.91–485.62 μg/100 g d.w. After cooking, total lignans losses of about 35.5, 18.31, and 5.46% were observed respectively in oat‐, rye‐, and rice‐added pastas, whereas increases of 5.74 and 13.62% were observed in barley‐added and whole durum wheat pastas. Interesting results were obtained for the multigrain pasta: the raw product exhibited a total lignans content of 9,686.17 ± 287.03 μg/100 g d.w., and the major contribution was given by secoisolariciresinol. This highest total lignans value resulted from its rich and varied composition in seeds of different origin, legumes, and flaxseed in particular. Our findings showed that mixed‐cereal and multigrain pastas can be considered a good source of lignans. The effect of cooking was not the same for each product, and it depended on the different lignans profile of each grain, on the different chemical structure of each lignan, and on the nature of the food matrix.     

Calcium-binding capacities of different brans under simulated gastrointestinal pH conditions. In vitro study with (45)Ca

The present study was performed to investigate calcium-binding characteristics of different brans under simulated gastrointestinal pH conditions and to explore the significance of dietary fiber, oxalate, and phytate for calcium binding. Different brans (rice, rye, soy, fine wheat, coarse wheat, and oat) and CaCl(2) solution containing (45)Ca were incubated at 37 degrees C at gastric pH (2.2) followed by buffering steps of 1 degree from pH 3.0 to pH 8.0. Total calcium binding and calcium-binding capacity of the pH 2.2 soluble bran fraction were determined. Additionally, oxalate and phytate contents of brans and solubility profiles of phytic acid were investigated. Calcium-binding capacities of brans showed a clear pH dependence. At gastric pH calcium binding was low in all brans, ranging from 0.022 to 0.040 mmol of calcium/g of bran. Soy bran, nearly phytate-free, showed higher binding values up to pH 4.0 and lower values between pH 5.0 and 8.0. In all other brans, binding values increased strongly with increasing pH in the quantitative order rice bran > coarse wheat bran > fine wheat bran > rye bran > oat bran. The solubility profiles indicate that in the cases of rye, wheat, and rice bran phytate accounts for 70-82% of their total calcium-binding capacities. The results suggest that dietary fiber makes no important contribution to calcium binding, except for soy and oat brans. Oxalate plays only a minor role in calcium binding by brans.     

Important aroma compounds in freshly ground wholemeal and white wheat flour-identification and quantitative changes during sourdough fermentation

An investigation on the odor-active compounds of wholemeal (WWF) and white wheat flour (WF 550) by aroma extract dilution analysis (AEDA) and by quantitative studies using stable isotope dilution assays (SIDA) revealed a significant number of odor-active compounds, such as (E)-2-nonenal, (E,Z)- and (E,E)-2,4-decadienal, (E)-4,5-epoxy-(E)-2-decenal, 3-hydroxy-4,5-dimethyl-2(5H)-furanone, and vanillin, with high odor activities in both wheat flours. The amounts and, consequently, the aroma potencies of vanillin, (E,E)-2,4-decadienal, and 3-(methylthio)propanal were much higher in the WWF than in the WF 550 samples. Fermentation of suspensions of both flours with lactic acid bacteria did not generate new odorants; however, many compounds, such as acetic acid or 3-methylbutanal, were increased, whereas aldehydes (formed from the degradation of unsaturated fatty acids) were decreased. Comparing the odorant concentrations present before and after fermentation gave evidence that the main influence of the microorganisms on sourdough aroma is to either enhance or decrease specific volatiles already present in the flour. A comparison with literature data indicated that most of these odorants are also important for the bread crumb aroma present after baking of the dough.     

Studies on rye (Secale cereale L.) lines exhibiting a range of extract viscosities. 2. Rheological and baking characteristics of rye and rye/wheat blends and feeding value for chicks of wholemeals and breads

Five rye lines exhibiting a wide range of extract viscosities were evaluated for the rheological and baking properties of their flours, individually and in blends with hard red spring wheat flour. Commercial cultivars of rye and triticale were included in the study as controls. Extract viscosities of rye flours were higher than those of corresponding wholemeals, indicating shifting of water-extractable arabinoxylan into flour during roller milling. Falling numbers of the rye flours correlated positively with their extract viscosities in the presence (r = 0.73, p < 0.05) or absence (r = 0.65, p < 0.05) of an enzyme inhibitor. Farinograms revealed the weakness of rye and triticale flours compared to wheat flour. Extract viscosities of rye flours were negatively correlated (r = -0.65, p < 0.05) with mixing tolerance index and positively correlated (r = 0.64, p < 0.05) with dough stability, suggesting a positive impact of extract viscosity on dough strength. Extract viscosity was negatively correlated (r = -0.74, p < 0.05) with loaf volume and specific volume (r = -0.73, p < 0.05) and positively correlated (r = 0.73, p < 0.05) with loaf weight of rye/wheat bread. Overall, the results indicated that 30% of flour from high or low extract viscosity rye could be incorporated into rye/wheat breads without seriously compromising bread quality. Inclusion of rye, particularly high extract viscosity rye, in chick diets seriously impeded growth performance and feed efficiency. Part of the arabinoxylan survived bread-making and exerted an effect on chicks, although substantially lower digesta viscosities were observed in chicks fed rye bread diets than in those fed rye wholemeals.     

Effect of several germination conditions on total P, phytate P, phytase, and acid phosphatase activities and inositol phosphate esters in rye and barley.

Two assays were conducted to study the evolution of rye and barley phosphatases (phytase and acid phosphatase) and the degradation of its substrates (inositol phosphate esters) during seed germination. In this manner we could obtain a low-phytate, endogenous phosphatase rich ingredient to be used in animal nutrition. In the first assay, the seeds were soaked for 1 and 14 h and germinated for 3 and 5 days with and without the addition of gibberellic acid (GA3). In the second assay, the seeds were soaked for 1 h and germinated for 1, 3, and 5 days with GA3. Phytase (up to 5739 and 3151 U x kg(-1)) and acid phosphatase (up to 18288 and 3151 U x g(-1)) activities, and IP6 (6.09 and 6.01 mg x g(-1)), IP5 (0.48 and 0.48 mg x g(-1)), and IP4 (0.13 and 0.06 mg x g(-1)) were detected in ungerminated rye and barley, respectively. The germination process caused a significant increase of Phy and AcPh activities in rye (up to 112 and 213%) and barley (up to 212 and 634%) and a reduction in the phytate phosphorus content (up to 84 and 58%, respectively). Phytate phosphorus content was affected only by soaking time in the case of rye. Finally, during the course of germination, IP6 and IP5 were rapidly degraded in rye (88 and 79%) and barley (67 and 52%), and IP4 was only a short-living intermediate, which was increased during hydrolysis and degraded to IP3. In conclusion, a marked increase of Phy and AcPh activities in rye and barley with a concomitant decrease in phytate phosphorus content and an increase in the content of lower inositol phosphates were observed during the rye and barley germination.     

Phytate degradation by lactic acid bacteria and yeasts during the wholemeal dough fermentation: a 31P NMR study

myo-Inositol hexaphosphate (IP6) is the main source of phosphorus in cereal grains, and therefore, in bakery products. Different microorganisms such as yeasts and lactic acid bacteria have phytase enzymes able to hydrolyze IP6 during the wholemeal breadmaking. In this paper, the phytase activity of Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus curvatus, and Saccharomyces cerevisiae strains, isolated from southern Italian sourdoughs, is assayed using the (31)P NMR technique. The sourdough technology based on the use of lactic acid bacteria in the breadmaking is finally suggested.     

Quantitative analysis of phytate globoids isolated from wheat bran and characterization of their sequential dephosphorylation by wheat phytase

Wheat phytase was purified to investigate the action of the enzyme toward its pure substrate (phytic acid - myo-inositol hexakisphosphate) and its naturally occurring substrate (phytate globoids). Phytate globoids were purified to homogeneity from wheat bran, and their nutritionally relevant parameters were quantified by ICP-MS. The main components of the globoids were phytic acid (40% w/w), protein (46% w/w), and several minerals, in particular, K > Mg > Ca > Fe (in concentration order). Investigation of enzyme kinetics revealed that K(m) and V(max) decreased by 29 and 37%, respectively, when pure phytic acid was replaced with phytate globoids as substrate. Time course degradation of phytic acid or phytate globoids using purified wheat phytase was followed by HPIC identification of inositol phosphates appearing and disappearing as products. In both cases, enzymatic degradation initiated at both the 3- and 6-positions of phytic acid and end products were inositol and phosphate.     

Contents of phenolic acids, alkyl- and alkenylresorcinols, and avenanthramides in commercial grain products

The contents of free and total phenolic acids and alk(en)ylresorcinols were analyzed in commercial products of eight grains: oat (Avena sativa), wheat (Triticum spp.), rye (Secale cerale), barley (Hordeum vulgare), buckwheat (Fagopyrum esculentum), millet (Panicum miliaceum), rice (Oryza sativa), and corn (Zea mays). Avenanthramides were determined in three oat products. Free phenolic acids, alk(en)ylresorcinols, and avenanthramides were extracted with methanolic acetic acid, 100% methanol, and 80% methanol, respectively, and quantified by HPLC. The contents of total phenolic acids were quantified by HPLC analysis after alkaline and acid hydrolyses. The highest contents of total phenolic acids were in brans of wheat (4527 mg/kg) and rye (4190 mg/kg) and in whole-grain flours of these grains (1342 and 1366 mg/kg, respectively). In other products, the contents varied from 111 mg/kg (white wheat bread) to 765 mg/kg (whole-grain rye bread). Common phenolic acids found in the grain products were ferulic acid (most abundant), ferulic acid dehydrodimers, sinapic acid, and p-coumaric acid. The grain products were found to contain either none or only low amounts of free phenolic acids. The content of avenanthramides in oat flakes (26-27 mg/kg) was about double that found in oat bran (13 mg/kg). The highest contents of alk(en)ylresorcinols were observed in brans of rye (4108 mg/kg) and wheat (3225 mg/kg). In addition, whole-grain rye products (rye bread, rye flour, and whole-wheat flour) contained considerable levels of alk(en)ylresorcinols (524, 927, and 759 mg/kg, respectively).