Betaine in Cereal Grains: History
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Betaine is a non-essential nutrient which performs several important physiological functions in organisms. Abundant data exist to suggest that betaine has a potential for prevention of chronic diseases and that its dietary intake may contribute to overall health enhancement. 

  • betaine
  • cereals
  • pseudocereals
  • gluten-free
  • stability
  • cooking
  • baking
  • extrusion

1. Introduction

Betaine (N,N,N-trimethylglycine, glycine betaine) is an organic nitrogenous compound, found for the first time in sugar beet juice (Beta vulgaris).
Betaine is a zwitterion of quaternary ammonium which is still named trimethylglycine and glycine betaine (Figure 1). It is a methyl derivative of the amino acid glycine ((CH3)3N+CH2COO and molecular weight 117.2). It is characterized as methylamine due to its three free methyl groups [1].
Figure 1. Betaine chemical structure.
Various analogues of glycine betaine exist in plants: proline betaine (stachydrine), trigonelline, arsenobetaine, betonicine, butirobetaine, ergothionine, propionobetaine, and sulfur analogues. The sulfur analogues are several in type: β-alaninebetaine, dimethylsulfonioacetate, and dimethylsulfoniopropionate (DMSP). The food survey study by de Zwart et al. [2] showed that only some betaine analogues were present in food at appreciable levels (>10 µg/g)—glycine betaine, proline betaine, trigonelline, and DMSP. Slow et al. [3] indicated glycine betaine as dominant in grain products, proline betaine in citruses, and trigonelline in coffee. Most recently, some rare forms of betaine were identified in the grains of most common cereals: pipercolic acid betaine in rye flour and valine betaine and glutamine betaine in flours of barley, rye, oat, durum, and winter wheat [4]. The content of betaine analogues was found to be vastly variable in grains; higher betaine levels seem to be induced by plant growth under stress conditions (drought, salt stress, cold, freezing, hypoxia, etc.) [2][3]. Since the potential health effects of betaine analogues, particularly trigonelline and proline, have not yet been fully resolved, currently only glycine betaine has dietary relevance.
Betaine represents a bioactive compound that has significant physiological functions in the human organism as an osmolite and donor of methyl groups for many biochemical processes. As such, it is indispensable to preserve the health of kidneys, liver, and heart [5]. This compound has an important role in preventing and treating many chronic diseases, among which lowering of plasma homocysteine levels has gained the most attention [5][6][7]. High serum homocysteine levels have been associated with increased risk for cardiovascular diseases (stroke, heart attack, atherosclerosis), cancer, peripheral neuropathy, etc. Moreover, betaine has been shown to improve athletic performance by enhancing muscle endurance [7][8].

2. Cereal Grains as a Source of Betaine

Data on the distribution of betaine in various cereals and pseudocereals are scarce and there is definitely a lack of detailed study. Most data come from various studies that were focused on estimation of betaine dietary intake. Nevertheless, available studies report on wide variations in betaine content in cereals. Different types of cereals may have different amounts of betaine [9]. The following ranges were found by de Zwart et al. [2]: 270–1110 µg/g (dry solids) in wheat flour, and 200–1000 µg/g in oats. More detailed overview of betaine levels in various cereals and pseudocereals from different studies is displayed in Table 1. The displayed data showed that betaine content spanned in wide ranges within the studied grains. According to Corol et al. [9], betaine content in cereals varies depending on multiple factors including genotype and environmental differences such as geographical and/or year-to-year variations and their interactions with genotype. This study revealed a three-fold difference in glycine betaine content within bread wheat genotypes and a 3.8-fold difference across six environments. The highest glycine betaine levels were found in Hungarian wheat grains whereas the lowest in those grown in the UK [9]. Slow et al. [3] and de Zwart et al. [2] indicated that the level of betaine depends on the level of stress under which the crop grows. This is due to osmoprotectant and cryoprotectant function of betaine. For example, growth under drought can cause higher levels of betaine compared to well-watered crops.
Table 1. Betaine content in different samples of cereals and pseudocereals.

Cereals and Pseudocereals

Betaine

References

(µg/g Dry Weight)

Wheat (Triticum aestivum)

   

raw grain

1150–1320

[10]

 

490–574

[11]

bran

5047–5383

[11]

 

2717

[12]

 

2300–7200

[3]

aleurone

4538–6242

[13]

germ

3414

[13]

wholegrain flour

792

[13]

 

730 *

[14]

 

604

[15]

 

540

[11]

refined flour

718 *

[16]

 

700 *

[14]

 

415–593

[12][11]

 

398

[13]

 

180 *

[4]

 

141.2

[15]

flour (not specified by origin)

270–1110

[2]

Wheat Emmer (T. dicoccum)

   

raw grain

830–940

[10]

refined flour

195 *

[4]

Wheat Einkorn (T. monococcum)

   

refined flour

367.3 *

[4]

Durum wheat (T. durum)

   

semolina

1227

[11]

 

483

[12]

 

683

[13]

refined flour

253–303

[11]

 

310

[12]

wholegrain flour

713

[13]

 

245 *

[4]

Spelt wheat (T. aestivum ssp. spelta)

   

raw grain

973–2723

[11]

 

565–714

[12]

wholegrain flour

1296–1442

[11]

 

1370–1430

[10]

refined flour

978

[13]

 

522–593

[11]

410

[12]

Kamut wheat, Khorasan (T. turgidum ssp. turanicum)

   

raw grains

1100

[14]

Triticale (xTriticosecale)

   

raw grain

986–1030

[11]

Rye

   

raw grain

2213

[11]

 

1530–1760

[10]

 

444

[12]

bran

1651

[15]

refined flour

310 *

[4]

wholegrain flour

1500 *

[14]

 

1182

[11]

 

986

[12]

Barley

   

raw grain

460

[10]

raw grain from naked var.

980

[10]

wholegrain flour

776–1023

[11]

 

779

[12]

refined flour

250 *

[4]

flour from naked var

424

[12]

 

574

[11]

pearled grain

274

[12]

Oats

   

raw grain

280

[10]

 

388

[12]

raw grain from naked var.

440

[10]

wholegrain flour

310 *

[14]

flour

404–688

[11]

 

53 *

[4]

bran

200 *

[14]

 

190

[13]

Maize

   

raw grain

107–304

[11]

 

175

[12]

wholegrain meal

120 *

[14]

degermed meal

4 *

[14]

semolina

3–22

[13]

refined corn grits

37

[13]

flour, enriched

20 *

[14]

refined flour

2.1 *

[4]

bran

184

[12]

 

104

[11]

 

46 *

[14]

flakes

103–120

[11]

 

7–9

[13]

 

n.d.

[12]

starch

n.d.

[12]

popped

19

[13]

 

n.d.

[12]

Rice

   

grain

1–5

[13]

 

n.d.

[12]

refined flour

8.4 *

[4]

expanded

n.d.

[12]

starch

n.d.

[12]

Amaranth (Amaranthus cruentus)

   

raw grain

7420

[11]

 

680 *

[14]

 

646

[13]

expanded grain

669

[11]

 

607

[12]

flour

895–1225

[11]

 

871

[12]

Proso millet

   

sample type not specified

95–112

[13]

dehulled grain

281

[11]

refined flour

1320 *

[4]

Buckwheat

   

wholegrain flour

108

[11]

 

7–20

[13]

refined flour

n.d.

[12]

groats, roasted

10 *

[4]

 

26 *

[14]

Sorghum

   

refined flour

425 *

[4]

Quinoa

   

grains

6300 *

[14]

3042–4428

[13]

610.8 *

[4]

n.d. not detected; * result expressed on wet weight.

3. Betaine Content in Cereal-Based Products

The betaine content in cereal products depends on the processing method. Two to four times lower betaine content were found in refined grain products compared to equivalent whole grain products [13]. Betaine content is notably dependent on the loss of bran fraction during processing. The higher the abrasion of aleurone layer, the lower the betaine content in the product. Outstanding betaine levels were determined in wheat bran, up to 7200 µg/g (Table 1). Likes et al. [16] analyzed the betaine contents in different milling streams and reported the lowest betaine level in the cleanest milling fractions. In the study of de Zwart et al. [2], a wide range of different foods was analyzed for betaine content and flour was denoted as an item high in betaine (730 µg/g), however it was not specified the type of flour, except that it was available from retail markets. Betaine ranges in bread, pasta, breakfast cereals and snacks are given in Table 2. As it can be seen, the variation within each product category is high due to versatility of ingredients in product formulations. In each product category, the highest betaine content was reported for wholegrain products or products containing bran or germ. Among breads, rye, spelt, and wholemeal breads were abundant in betaine. Moderate to high betaine contents were reported for pasta products, but it must be noted that mainly uncooked samples were analyzed (Table 2). Breakfast cereals are a mixture of cereal and non-cereal ingredients and the betaine content will depend on the contribution of each ingredient. In the study of Filipčev et al. [11], two samples of commercially available breakfast cereals were analyzed, one of which contained no detectable levels of betaine whereas the other had 471 µg/g (on dry solids). A similar concluded was made by Ross et al. [13] for muesli and muesli bars which were found to contain only low-to-moderate betaine levels. These products were mainly based on oats and contained other low-betaine ingredients such as dried fruits. In contrast to Ross et al. [13], the USDA data [14] report on much wider span of betaine in breakfast cereals, from 7 µg/greaching to as much as 3600 µg/g (on wet weight) betaine.
Table 2. Betaine content in various grain-based products.

Product

Betaine Content

References

(µg/g Dry Weight)

Bread

   

rye bread

855–1377

[13]

wholegrain spelt

913

[13]

wholemeal

670–790

[3]

wholegrain

499–781

[13]

 

560–620

[3]

multigrain

247–678

[13]

white (refined)

360–520

[3]

 

174–287

[13]

various (white, sourdough)

310–590 *

[14]

 

380 *

[14]

 

579

[15]

wheat tortilla

311

[13]

Pasta

   

wholegrain wheat pasta

710–1286

[13]

 

375

[15]

pasta, not specified

480–1350

[2]

refined wheat pasta

628–706

[13]

refined wheat (T. aestivum) pasta, uncooked

253

[12]

durum wheat pasta, uncooked

188

[12]

one–egg spelt pasta

243–516

[13]

barley pasta

211

[13]

noodles with egg, enriched, uncooked

1300 *

[14]

noodles with egg, enriched, cooked

190 *

[14]

refined couscous

691

[13]

bulghur

1311

[13]

cooked bulghur

830 *

[14]

Breakfast cereals

   

ready-to-eat wheat germ, toasted, plain

4100 *

[14]

ready-to-eat wheat bran, toasted

3200 *

[14]

wholegrain rye flakes

1640

[13]

wholegrain wheat-based cereals

732–915

[13]

wholegrain oat and wheat-based muesli

310

[13]

wholegrain oat-based muesli

117–226

[13]

breakfast cereals, not specified

180–300

[12]

muesli bar

171

[13]

wholegrain porridge oats

128–167

[13]

extruded whole grain oat cereals

73–91

[13]

cereal bar

74–75

[13]

various ready-to-eat cereals

7–3600 *

[14]

Snacks, cookies, crackers, crispbread, cakes, pastry

   

wholegrain rye crispbread

1428–1527

[13]

frozen, read-to-eat pancakes

690–720 *

[14]

wholegrain wheat crackers

293–649

[13]

crackers, classic, saltines, cheese

340–580 *

[14]

wholegrain wheat rusks

556–564

[13]

wholegrain wheat muffin

437–501

[13]

various commercial cakes

190–480 *

[14]

wholegrain wheat biscuit

425

[12]

Graham cookies

390 *

[14]

doughnuts

270–380 *

[14]

English muffins

220–360 *

[14]

extruded spelt

308

[12]

refined wheat crackers

258–332

[13]

digestive biscuit

271–309

[13]

apple pie, commercial

160 *

[14]

biscuit

4–144

[13]

Danish pastry, fruit enriched

140 *

[14]

plain Danish pastry

81 *

[14]

* Result expressed on wet weight.

4. Betaine Content in Gluten-Free Cereal Products

Gluten-free products have been generally recognized to be low in betaine content [13][15]. In the majority of commercially available gluten-free products, a very low level of betaine (<50 μg/g) was observed [13]. Table 3 lists the betaine levels reported for commercial gluten-free products from several studies. In the bread and biscuits category, betaine levels ranged from non-detectable to 107 µg/g. Similar findings were reported by Kojić et al. [12], who also found that gluten-free samples (starch, corn extrudates, pasta, cornflakes, and rice) contained no detectable levels of betaine. Gluten-free cereals contained much lower amounts of betaine in comparison to glutenous cereals: corn had 107–304 µg/g betaine [11]; teff and millet between 50–150 µg/g [13], proso millet 280 µg/g [11]. Buckwheat is a frequent ingredient in gluten-free products. According to Ross et al. [13], buckwheat was among those ingredients low in betaine (<20 µg/g) although as high as 390 µg/g betaine was found in buckwheat uncooked pasta (Table 3).
Table 3. Betaine content in gluten-free products.

Product

Betaine Content

(µg/g Dry Weight)

References

Bread and biscuits

   

gluten-free crispbread

9–107

[13]

savory biscuits

n.d.–104

[11]

wholegrain gluten-free bread

12–68

[13]

oatmeal biscuits

3

[13]

gluten-free flour enriched with fibers

1

[13]

sweet biscuits

n.d.

[12]

flour mixture for gluten-free bread

n.d.

[12]

gluten-free cookies with almonds, crackers, salty sticks

n.d.

[12]

expanded maize

n.d.

[12]

Pasta

   

buckwheat pasta, uncooked

390

[11]

 

382

[13]

 

175

[12]

maize-based pasta

2–20

[13]

maize and rice-based pasta, uncooked

n.d.

[12]

rice-based pasta, uncooked

n.d.

[12]

Breakfast cereals and related products

   

soy bran

182

[12]

unseasoned popcorn

19

[13]

cornflakes

14

[13]

buckwheat flakes

10

[13]

rice-based breakfast cereals

4–5

[13]

expanded rice

n.d.

[12]

n.d. not detected.

5. Stability of Betaine in Grain-Based Products

Betaine is known to be a thermostable compound that survives the severe treatment during sugar beet processing (extracting with water, treatment with CaOH2 and CO2, concentration, crystallization) and almost quantitatively accumulates in molasses [17]. Pure anhydrous betaine decomposes at > 245 °C. Since food processing practices do not employ such high temperatures, betaine losses caused by food thermal treatments were initially not expected [18]. However, some data suggest that certain cooking and baking losses of betaine may exist in spite of its thermostability in the pure form. Being a water-soluble compound with a small molecule, it is not unlikely that some betaine losses will occur, depending on the type of food processing and cooking. Available data suggest that losses are very high if processing involves water removal after cooking or boiling due to its solubility in water. Very high losses were observed during the baking of betaine-enriched bread, implying that fermentation by baker’s yeast may be one of the causes but future research is needed to understand the possible mechanisms.

This entry is adapted from the peer-reviewed paper 10.3390/foods7040049

References

  1. Yancey, P.H.; Clark, M.E.; Hand, S.C.; Bowlus, R.D.; Somero, G.N. Living with stress: Evolution of osmolyte systems. Science 1982, 217, 1214–1222.
  2. De Zwart, F.J.; Slow, S.; Payne, R.J.; Lever, M.; George, P.M.; Gerrard, J.A.; Chambers, S.T. Glycine betaine and glycine betaine analogues in common foods. Food Chem. 2003, 83, 197–204.
  3. Slow, S.; Donaggio, M.; Cressey, P.J.; Lever, M.; George, P.M.; Chambers, S.T. The betaine content of New Zealand foods and estimated intake in the New Zealand diet. J. Food Compos. Anal. 2005, 18, 473–485.
  4. Servillo, L.; D’Onofrio, N.; Giovane, A.; Casale, R.; Cautela, D.; Ferrari, G.; Castaldo, D.; Balestrieri, M.L. The betaine profile of cereal flours unveils new and uncommon betaines. Food Chem. 2018, 239, 234–241.
  5. Craig, S.A. Betaine in human nutrition. Am. J. Clin. Nutr. 2004, 80, 539–548.
  6. Olthof, M.R.; Van Vliet, T.; Boelsma, E.; Verhoef, P. Low dose betaine supplementation leads to immediate and long term lowering of plasma homocysteine in health men and women. J. Nutr. 2003, 133, 4135–4138.
  7. Steenge, G.R.; Verhoef, P.; Katan, M.B. Betaine supplementation lowers plasma homocysteine in healthy men and women. J. Nutr. 2003, 133, 1291–1295.
  8. Hoffman, J.R.; Ratamess, N.A.; Kang, J.; Rashti, S.L.; Faigenbaum, A.D. Effect of betaine supplementation on power performance and fatigue. J. Int. Soc. Sports Nutr. 2009, 6, 7–17.
  9. Corol, D.I.; Ravel, C.; Raksegi, M.; Bedo, Z.; Charmet, G.; Beale, M.H.; Ward, J.L. Effects of genotype and environment on the contents of betaine, choline, and trigonelline in cereal grains. J. Agric. Food Chem. 2012, 60, 5471–5481.
  10. Hefni, E.M.; Schaller, F.; Witthöft, M.C. Betaine, choline and folate content in different cereal genotypes. J. Cereal Sci. 2018, 80, 72–79.
  11. Filipčev, B.V.; Brkljača, J.S.; Krulj, J.A.; Bodroža-Solarov, M.I. The betaine content in common cereal-based and gluten-free food from local origin. Food Feed Res. 2015, 42, 129–137.
  12. Kojić, J.; Krulj, J.; Ilić, N.; Lončar, E.; Pezo, L.; Mandić, A.; Bodroža-Solarov, M. Analysis of betaine levels in cereals, pseudocereals and their products. J. Funct. Foods 2017, 37, 157–163.
  13. Ross, A.B.; Zangger, A.; Guiraud, S.P. 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, 145, 859–865.
  14. Patterson, K.Y.; Bhagwat, S.A.; Williams, J.R.; Howe, J.C.; Holden, J.M. USDA Database for the Choline Content of Common Foods—Release 2. Available online: (accessed on 5 January 2018).
  15. Bruce, S.J.; Guy, P.A.; Rezzi, S.; Ross, A.B. Quantitative measurement of betaine and free choline in plasma, cereals and cereal products by isotope dilution LC-MS/MS. J. Agric. Food Chem. 2010, 58, 2055–2061.
  16. Likes, R.; Madl, R.L.; Zeisel, S.H.; Craig, S.A.S. The betaine and choline content of a whole wheat flour compared to other mill streams. J. Cereal Sci. 2007, 46, 93–95.
  17. Šušić, S.; Sinobad, V. Ispitivanja u cilju unapređenja industrije šećeraJugoslavije. Hem. Ind. 1989, 43 (Suppl. 1–2), 10–21.
  18. The Scientific Panel on Dietetic Products, Nutrition and Alergies. Opinion on the scientific panel on dietetic products, nutrition and allergies on a request from the Commission related to an application concerning the use of betaine as a novel food in the EU. EFSA J. 2005, 191, 1–17.
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