Betaine absorbed more rapidly in pigs fed a supplement versus when present in cereal dietary fiber

The net absorption of betaine and choline was determined for 4 h after the first meal of the day in three experiments with porto-arterial catheterized pigs in which betaine was added as a supplement to a low-betaine diet (n = 4 pigs) and compared to the net absorption of betaine and choline from high-fiber breads differing in amount and source of dietary fiber (two experiments, n = 6 pigs each). Plasma betaine peaked after 30 min when betaine was fed as a supplement, whereas it peaked after 120-180 min when high-fiber breads were fed. Plasma betaine showed no diet x time interaction after feeding with high-fiber breads, indicating that the absorption kinetic did not differ between fiber sources. The net absorption of choline was not affected by the experimental diets. In conclusion, betaine in cereal sources has to be liberated from the matrix prior to absorption, causing delayed absorption.

Hedemann, M.S., et al., Distinct Difference in Absorption Pattern in Pigs of Betaine Provided as a Supplement or Present Naturally in Cereal Dietary Fiber. J Agric Food Chem, 2015

Postprandial plasma betaine and other methyl donor-related responses after consumption of minimally processed wheat bran or aleurone

The bran and particularly the aleurone fraction of wheat are high in betaine and other physiological methyl donors, which may exert beneficial physiological effects. We conducted two randomised, controlled, cross-over postprandial studies to assess and compare plasma betaine and other methyl donor-related responses following the consumption of minimally processed bran and aleurone fractions (study A) and aleurone bread (study B). For both studies, standard pharmacokinetic parameters were derived for betaine, choline, folate, dimethylglycine (DMG), total homocysteine and methionine from plasma samples taken at 0, 0.5, 1, 2 and 3 h. In study A (n 14), plasma betaine concentrations were significantly and substantially elevated from 0.5 to 3 h following the consumption of both bran and aleurone compared with the control; however, aleurone gave significantly higher responses than bran. Small, but significant, increases were also observed in DMG measures; however, no significant responses were observed in other analytes. In study B (n 13), plasma betaine concentrations were significantly and substantially higher following consumption of the aleurone bread compared with the control bread; small, but significant, increases were also observed in DMG and folate measures in response to consumption of the aleurone bread; however, no significant responses were observed in other analytes. Peak plasma betaine concentrations, which were 1.7-1.8 times the baseline levels, were attained earlier following the consumption of minimally processed aleurone compared with the aleurone bread (time taken to reach peak concentration 1.2 v. 2.1 h). These results showed that the consumption of minimally processed wheat bran, and particularly the aleurone fraction, yielded substantial postprandial increases in plasma betaine concentrations. Furthermore, these effects appear to be maintained when aleurone was incorporated into bread.

Keaveney, E.M., et al., Postprandial plasma betaine and other methyl donor-related responses after consumption of minimally processed wheat bran or wheat aleurone, or wheat aleurone incorporated into bread. Br J Nutr, 2015: p. 1-9

Betaine may be a key contributor to reduction of disease risk associated with greater intake of whole grain

Dietary fibre alone does not fully explain the frequent association between greater intake of whole grains and reduced risk of disease in observational studies, and other phytochemicals or food structure may also play an important role. For all the observational evidence for the benefits of a whole-grain-rich diet, we have only limited knowledge of the mechanisms behind this reduction in disease risk, aside from the action of specific cereal fibres on reduction of blood cholesterol and the post-prandial glucose peak. Nutritional metabolomics, the global measurement and interpretation of metabolic profiles, assesses the interaction of food with the endogenous gene-protein cascade and the gut microbiome. This approach allows the generation of new hypotheses which account for systemic effects, rather than just focusing on one or two mechanisms or metabolic pathways. To date, animal and human trials using metabolomics to investigate mechanistic changes to metabolism on eating whole grains and cereal fractions have led to new hypotheses around mechanistic effects of whole grains. These include the role of cereals as a major source of dietary glycine betaine, a possible effect on phospholipid synthesis or metabolism, the role of branched-chain amino acids and improvements in insulin sensitivity, and the possibility that whole grains may have an effect on protein metabolism. These hypotheses help explain some of the observed effects of whole grains, although mechanistic studies using stable isotopes and fully quantitative measures are required to confirm these potential mechanisms.

Ross, A.B., Whole grains beyond fibre: what can metabolomics tell us about mechanisms? Proc Nutr Soc, 2014: p. 1-8

Cereal foods are the major source of betaine in the Western diet

Betaine and its precursor choline are important components of one-carbon metabolism, remethylating homocysteine into methionine and providing methyl groups for DNA methylation. Cereals are the main source of betaine in the diet, though there is little literature available on the content of betaine in cereal products, nor on betaine intake from cereals. Betaine and free-choline concentrations were measured by liquid-chromatography with tandem mass spectrometry in a wide range of commercially available cereal foods and cereal fractions. Whole grain wheat and related fractions were the best overall common source of betaine, while the pseudocereal quinoa had the highest amount of betaine measured (3900mug/g). Based on estimates of dietary intake data cereal foods provide approximately 60-67% of betaine in Western diets, and 20-40% of betaine in South-East Asian diets. Average intake of betaine was 131mg/d, well below those used in intervention studies using betaine to lower blood homocysteine.

Ross, A.B., A. Zangger, and S.P. Guiraud, Cereal foods are the major source of betaine in the Western diet - Analysis of betaine and free choline in cereal foods and updated assessments of betaine intake. Food Chem, 2014. 145C: p. 859-865

Metabolomics reveals the metabolic shifts following an intervention with rye bread in postmenopausal women- a randomized control trial

BACKGROUND: Epidemiological studies have consistently shown that whole grain (WG) cereals can protect against the development of chronic diseases, but the underlying mechanism is not fully understood. Among WG products, WG rye is considered even more potent because of its unique discrepancy in postprandial insulin and glucose responses known as the rye factor. In this study, an NMR-based metabolomics approach was applied to study the metabolic effects of WG rye as a tool to determine the beneficial effects of WG rye on human health. METHODS: Thirty-three postmenopausal Finnish women with elevated serum total cholesterol (5.0-8.5 mmol/L) and BMI of 20-33 kg/m2 consumed a minimum of 20% of their daily energy intake as high fiber WG rye bread (RB) or refined wheat bread (WB) in a randomized, controlled, crossover design with two 8-wk intervention periods separated by an 8-wk washout period. At the end of each intervention period, fasting serum was collected for NMR-based metabolomics and the analysis of cholesterol fractions. Multilevel partial least squares discriminant analysis was used for paired comparisons of multivariate data. RESULTS: The metabolomics analysis of serum showed lower leucine and isoleucine and higher betaine and N,N-dimethylglycine levels after RB than WB intake. To further investigate the metabolic effects of RB, the serum cholesterol fractions were measured. Total- and LDL-cholesterol levels were higher after RB intake than after WB. CONCLUSIONS: This study revealed favorable shifts in branched amino acid and single carbon metabolism and an unfavorable shift in serum cholesterol levels after RB intake in postmenopausal women, which should be considered for evaluating health beneficial effects of rye products.

Moazzami, A.A., et al., Metabolomics reveals the metabolic shifts following an intervention with rye bread in postmenopausal women- a randomized control trial. Nutr J, 2012. 11: p. 88

Thermal and refining processes, not fermentation, tend to reduce lipotropic capacity of plant-based foods

Plant-based foods (PBF) are relevant and diversified sources of lipotropes, which are compounds preventing excess hepatic fat deposits. In a first study, we defined the lipotropic capacity (LC, %) of raw PBF as the means of 8 lipotrope densities (LD, mg/100 kcal), each expressed relative to that of a reference food ranking the highest considering its mean 8 LD ranks (LCraw asparagus = 100%) (A. Fardet, J.-F. Martin and J. M. Chardigny, J. Food Comp. Anal., 2011, DOI: 10.1016/j.jfca.2011.1003.1013). We showed that vegetables appeared as the best source of lipotropes on a 100 kcal-basis compared to legumes, cereals, fruits and nuts. The main objective of this second study was to quantify the effect of processing on LD and LC of raw PBF based on lipotrope contents collected in a USDA (United State Department of Agriculture) database and the literature, i.e. betaine, choline, myo-inositol, methionine, magnesium, niacin, pantothenic acid and folate contents. Choline and betaine densities were not significantly affected by processing while methionine and lipotropic micronutrient densities were significantly decreased, especially for magnesium, pantothenate and folates. Myo-inositol density decreases were insignificant due to lower product number resulting from limited literature data. Lipotropic micronutrient densities were more affected by processing than other densities. Fermentations increased betaine (median change of +32%) and choline (+34%) densities. Canning and boiling vegetables increased choline densities (+26%). Globally, processing significantly reduced LC by 20%, fermentations being less drastic (median change of −5%) than refining (−33%) and thermal treatments (−16%). More specifically, canning increased LC of beetroot (536 vs 390%) and common bean (40 vs 36%) as fermentation towards LC grape (14 vs 7% for wine). Results were then mainly discussed based on percentages of lipotrope content changes on a dry-weight basis. Results of this study also showed that the LC is quite a relevant index to estimate effect of processing on lipotropic potential of PBF.

Fardet, A., J.F. Martin, and J.M. Chardigny, Thermal and refining processes, not fermentation, tend to reduce lipotropic capacity of plant-based foods. Food Funct, 2011. 2(8): p. 483-504.

Metabolomics reveals the metabolic shifts following an intervention with rye bread in postmenopausal women- a randomized control trial

Prostate cancer (PC) is the most common cancer in the Western world and the second most important cancer causing male deaths, after lung cancer, in the United States and Britain. Lifestyle and dietary changes are recommended for men diagnosed with early-stage PC. It has been shown that a diet rich in whole grain (WG) rye reduces the progression of early-stage PC, but the underlying mechanism is not clear. This study sought to identify changes in the metabolic signature of plasma in patients with early-stage PC following intervention with a diet rich in WG rye and rye bran product (RP) compared with refined white wheat product (WP) as a tool for mechanistic investigation of the beneficial health effects of RP on PC progression. Seventeen PC patients received 485 g RP or WP in a randomized, controlled, crossover design during a period of 6 wk with a 2-wk washout period. At the end of each intervention period, fasting plasma was collected and used for (1)H NMR-based metabolomics. Multilevel partial least squares discriminant analysis was used for paired comparisons of multivariate data. A metabolomics analysis of plasma showed an increase in 5 metabolites, including 3-hydroxybutyric acid, acetone, betaine, N,N-dimethylglycine, and dimethyl sulfone, after RP. To understand these metabolic changes, fasting plasma homocysteine, leptin, adiponectin, and glucagon were measured separately. The plasma homocysteine concentration was lower (P = 0.07) and that of leptin tended to be lower (P = 0.07) after RP intake compared to WP intake. The increase in plasma 3-hydroxybutyric acid and acetone after RP suggests a shift in energy metabolism from anabolic to catabolic status, which could explain some of the beneficial health effects of WG rye, i.e., reduction in prostate-specific antigen and reduced 24-h insulin secretion. In addition, the increase in betaine and N,N-dimethylglycine and the decrease in homocysteine show a favorable shift in homocysteine metabolism after RP intake.

Moazzami, A.A., et al., Metabolomics reveals the metabolic shifts following an intervention with rye bread in postmenopausal women- a randomized control trial. Nutr J, 2012. 11: p. 88

Consumption of wheat aleurone-rich foods increases fasting plasma betaine and modestly decreases fasting homocysteine and LDL-cholesterol

There is strong evidence that whole-grain foods protect against heart disease. Although underlying mechanisms and components are unclear, betaine, found at high levels in wheat aleurone, may play a role. We evaluated the effects of a diet high in wheat aleurone on plasma betaine and related measures.

In a parallel, single-blinded intervention study, 79 healthy participants (aged 45–65 y, BMI ≥ 25 kg/m2) incorporated either aleurone-rich cereal products (27 g/d aleurone) or control products balanced for fiber and macronutrients into their habitual diets for 4 wk. Fasting blood samples were taken at baseline and postintervention (4 wk) from participants. Compared with the control, the aleurone products provided an additional 279 mg/d betaine and resulted in higher plasma betaine (P < 0.001; intervention effect size: 5.2 μmol/L) and lower plasma total homocysteine (tHcy) (P = 0.010; −0.7 μmol/L). Plasma dimethylglycine and methionine, which are products of betaine-mediated homocysteine remethylation, were also higher (P < 0.001; P = 0.027) relative to control. There were no significant effects on plasma choline or B vitamins (folate, riboflavin, and vitamin B-6). However, LDL cholesterol was lower than in the control group (P = 0.037).

We conclude that incorporating aleurone-rich products into the habitual diet for 4 wk significantly increases plasma betaine concentrations and lowers tHcy, which is attributable to enhanced betaine-homocysteine methyltransferase-mediated remethylation of homocysteine. Although this supports a role for betaine in the protective effects of whole grains, concomitant decreases in LDL suggest more than one component or mechanism may be responsible.

Price et al (2010). "Consumption of Wheat Aleurone-Rich Foods Increases Fasting Plasma Betaine and Modestly Decreases Fasting Homocysteine and LDL-Cholesterol in Adults." The Journal of Nutrition 140(12): 2153-2157.

A whole-grain diet increases plasma betaine, and decreases total and LDL-cholesterol compared with a refined-grain diet

Epidemiological studies have repeatedly found that whole-grain (WG) cereal foods reduce the risk of several lifestyle-related diseases, though consistent clinical outcomes and mechanisms are elusive.

To compare the effects of a WG-rich diet with a matched refined-grain (RG) diet on plasma biomarkers and bowel health parameters, seventeen healthy subjects (eleven females and six males) completed an exploratory cross-over study with a 2-week intervention diet based on either WG- or RG-based foods, separated by a washout of at least 5 weeks. Both diets were the same except for the use of WG (150 g/d) or RG foods. Subjects undertook a 4 h postprandial challenge on day 8 of each intervention diet. After 2 weeks, the WG diet tended to decrease plasma total and LDL-cholesterol (both P = 0.09), but did not change plasma HDL-cholesterol, fasting glucose, C-reactive protein or homocysteine compared with the RG diet. Plasma betaine and alkylresorcinol concentrations were elevated after 1 week of the WG diet (P = 0.01 and P < 0.0001, respectively). Clostridium leptum populations in faeces were increased after the WG diet, along with a trend for decreased faecal water pH (P = 0.096) and increased stool frequency (P < 0.0001) compared with the RG diet. A short controlled intervention trial with a variety of commercially available WG-based products tended to improve biomarkers of CVD compared with a RG diet.

Changes in faecal microbiota related to increased fibre fermentation and increased plasma betaine concentrations point to both fibre and phytochemical components of WG being important in mediating any potential health effects.

Ross et al (2011). "A whole-grain cereal-rich diet increases plasma betaine, and tends to decrease total and LDL-cholesterol compared with a refined-grain diet in healthy subjects." Br J Nutr: 1-12.

Betaine is an important component of whole grain health benefits

Epidemiological studies have clearly shown that whole-grain cereals can protect against obesity, diabetes, CVD and cancers. The specific effects of food structure (increased satiety, reduced transit time and glycaemic response), fibre (improved faecal bulking and satiety, viscosity and SCFA production, and/or reduced glycaemic response) and Mg (better glycaemic homeostasis through increased insulin secretion), together with the antioxidant and anti-carcinogenic properties of numerous bioactive compounds, especially those in the bran and germ (minerals, trace elements, vitamins, carotenoids, polyphenols and alkylresorcinols), are today well-recognised mechanisms in this protection.

Recent findings, the exhaustive listing of bioactive compounds found in whole-grain wheat, their content in whole-grain, bran and germ fractions and their estimated bioavailability, have led to new hypotheses. The involvement of polyphenols in cell signalling and gene regulation, and of sulfur compounds, lignin and phytic acid should be considered in antioxidant protection. Whole-grain wheat is also a rich source of methyl donors and lipotropes (methionine, betaine, choline, inositol and folates) that may be involved in cardiovascular and/or hepatic protection, lipid metabolism and DNA methylation. Potential protective effects of bound phenolic acids within the colon, of the B-complex vitamins on the nervous system and mental health, of oligosaccharides as prebiotics, of compounds associated with skeleton health, and of other compounds such as α-linolenic acid, policosanol, melatonin, phytosterols and para-aminobenzoic acid also deserve to be studied in more depth. Finally, benefits of nutrigenomics to study complex physiological effects of the ‘whole-grain package’, and the most promising ways for improving the nutritional quality of cereal products are discussed.

Fardet A. New hypotheses for the health-protective mechanisms of whole-grain cereals: what is beyond fibre? Nutrition Research Reviews. 2010;23(01):65-134.

Betaine content of cereal products

LC-MS/MS analysis was used to analyze 47 plasma samples, 32 cereal flours and cereal fractions, and 51 cereal products.

Whole-grain wheat and rye flours, and products based on these were the best whole cereal sources of betaine (747-1508 mug/g) and to a lesser extent choline (76-159 mug/g), while the bran fraction contained the highest concentrations of betaine and free-choline (2350-2899 mug/g and 366-384 mug/g respectively). Refined wheat flour and products contained lower concentrations, while rice and maize contained only very low and no detectable amounts of betaine respectively (0-10 mug/g), and low amounts of free-choline (<31 mug/g).

These results were mirrored in cereal products analyzed, with whole-grain wheat or rye-based cereal products having the highest concentrations of the two metabolites. Plasma concentrations for betaine and free-choline in a group of 47 subjects ranged from 15.2-66.3 and 9.8-18.5 mumol/L respectively, within the range of previous reports. This LC-MS/MS method can be used to rapidly and sensitively quantify betaine and free-choline in plasma and cereal products.

Whole-grain cereal products and products containing cereal bran appear to be excellent dietary sources of betaine and free-choline.

Bruce et al (2010). "Quantitative Measurement of Betaine and Free Choline in Plasma, Cereals and Cereal Products by Isotope Dilution LC-MS/MS." J Agric Food Chem. Epub Jan 26

Plasma betaine increases upon intake of high-fiber rye buns

An NMR-based metabonomic study explored the biochemical effects of a rye based fiber-rich diet in hypercholesterolemic pigs.

The pigs were fed high-fat, high-cholesterol rye- (n = 9) or wheat- (n = 8) based buns with similar levels of dietary fiber for 9-10 wk. Fasting plasma samples were collected 2 days before and after 8 and 12 days on the experimental diets, while postprandial samples taken after 58-67 days, and 1H NMR spectra were acquired on these.

The NMR spectra demonstrated a high intensity for the spectral region at 3.29 ppm in the rye diet. The 3.29 ppm signal is ascribed to N(CH3)3 protons in betaine, which may be an important contributor to the health promoting effects of rye.

Bertram et al (2009). "NMR-based metabonomics reveals that plasma betaine increases upon intake of high-fiber rye buns in hypercholesterolemic pigs." Mol Nutr Food Res. epub 14 July 2009

Characterization of industrial rye and wheat brans

Six different rye brans from Sweden, Denmark and Finland were analysed and compared with two wheat brans regarding colour, particle size distribution, microscopic structures and chemical composition including proximal components, vitamins, minerals and bioactive compounds.

There were many variations within rye brans, and differences from wheat bran. Rye bran had less betaine (194-278 mg/100 g) than wheat brans (431-441 mg/100 g).

Kamal-Eldin et al (2009). "Physical, microscopic and chemical characterisation of industrial rye and wheat brans from the Nordic countries." Food Nutr Res 53 DOI: 10.3402/fnr.v53i0.1912.

Betaine and choline content of wheat fractions

This present study suggests that the wheat aleurone layer contains the greatest concentration of both betaine and choline (1553 and 210 mg/100 g of sample, respectively). The bran fraction contained 867 and 102 mg/100 g of sample of betaine and choline, respectively, while the flour fraction contained 23 mg/100 g of sample (betaine) and 28 mg/100 g of sample (choline). The betaine content for the bran was lower, and the choline content was higher compared to previous studies, although it is known that there is large variation in betaine and choline contents between wheat cultivars. The study further emphasizes the superior phytonutrient composition of the aleurone layer.

Graham et al (2009). "Analysis of Betaine and Choline Contents of Aleurone, Bran, and Flour Fractions of Wheat (Triticum aestivum L.) Using 1H Nuclear Magnetic Resonance (NMR) Spectroscopy." J Ag Food Chem 57(5): 1948-1951.

Dietary patterns, food groups, and nutrients as predictors of plasma choline and betaine

Higher plasma betaine was predicted by consumption of high-fiber bread, complex carbohydrates, fiber, folate, thiamine and total energy. Lower plasma betaine was correlated with high-fat dairy products and western dietary pattern (with a high loading for meat, pizza, sugar, and fat). Higher plasma choline was predicted by egg consumption and cholesterol intake.

Konstantinova et al (2008). "Dietary patterns, food groups, and nutrients as predictors of plasma choline and betaine in middle-aged and elderly men and women." Am J Clin Nutr 88(6): 1663-1669.

Antioxidants, Cereals and Betaine

Fardet (2008) "Fardet et al (2008). "Is the in vitro antioxidant potential of whole-grain cereals and cereal products well reflected in vivo?" Journal of Cereal Science 48(2): 258-76."

This paper suggests that betaine has long been neglected as a source of antioxidant from cereals.

Papers 2004-7:

Fardet et al (2007). "Whole-Grain and Refined Wheat Flours Show Distinct Metabolic Profiles in Rats as Assessed by a 1H NMR-Based Metabonomic Approach." J Nutr 137(4): 923-29.

Likes et al (2007). "The betaine and choline content of a whole wheat flour compared to other mill streams." J. Cereal Sci. 46(1): 93-5.

Price et al (2007). "Plasma uptake of methyl donors from wheat fractions." Proc Nutr Soc 66(S1): 114A.