Assignment

VITAMIN B1 AND B2

Submitted By: Nazir Ullah 53794

Ahsan Ullah 53781
Fida Hussain 537
BS 4th

Submitted To: Mam Anum Aslam

Assistant Professor
Department of Biochemistry
Hazara University Mansehra

DEPARTMENT OF CHEMISTRY
HAZARA UNIVERSITY MANSEHRA
2021
 Table of Contents
Thiamine B1...................................................................................................................3
Introduction................................................................................................................3
History........................................................................................................................4
Thiamine deficiency...................................................................................................6
Prenatal supplementation............................................................................................6
Adverse effects...........................................................................................................7
Chemistry...................................................................................................................7
Biosynthesis................................................................................................................8
Foods..........................................................................................................................8
Benefits.......................................................................................................................9
Deficiency symptoms...............................................................................................10
Health risks...............................................................................................................11
Sugar and salt intake.............................................................................................11
Calorie intake........................................................................................................12
May reduce risk of certain cancers.......................................................................12
Maintaining blood sugar levels.............................................................................12
May aid weight loss..............................................................................................13
Nutrition...................................................................................................................13
Occurrence in foods..............................................................................................13
Dietary recommendations.....................................................................................13
Antagonists...........................................................................................................14
Food fortification......................................................................................................14
Vitamin B2 (Riboflavin)..............................................................................................15
History......................................................................................................................15
Riboflavin foods.......................................................................................................16
Benefits.....................................................................................................................17
Deficiency and dosage..............................................................................................17
Causes.......................................................................................................................18
Diagnosis..................................................................................................................19
Treatment..................................................................................................................20
Side effects...............................................................................................................20

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Function....................................................................................................................21
Medical uses.............................................................................................................22
Biosynthesis..............................................................................................................22
Chemistry.................................................................................................................22
Industrial uses...........................................................................................................22
Fluorescent spectra of riboflavin..........................................................................23
Industrial synthesis...............................................................................................23
References................................................................................................................24

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Thiamine B1

Introduction

Thiamine, also known as thiamin or vitamin B1, is a vitamin found in food and

manufactured as a dietary supplement and medication. Food sources of thiamine

include whole grains, legumes, and some meats and fish. Grain processing removes

much of the thiamine content, so in many countries cereals and flours are enriched

with thiamine. Supplements and medications are available to treat and prevent

thiamine deficiency and disorders that result from it, including beriberi and Wernicke

encephalopathy. Other uses include the treatment of maple syrup urine disease and

Leigh syndrome. They are typically taken by mouth, but may also be given by

intravenous or intramuscular injection.

Thiamine supplements are generally well tolerated. Allergic reactions, including

anaphylaxis, may occur when repeated doses are given by injection. Thiamine is in

the B complex family. It is an essential micronutrient, which cannot be made in the

body. Thiamine is required for metabolism including that of glucose, amino acids, and

lipids. Thiamine was discovered in 1897, was the first B vitamin to be isolated in

1926, and was first made in 1936. It is on the World Health Organization's List of

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Essential Medicines. Thiamine is available as a generic medication, and as an over-

the-counter drug.

History

In 1884, Takaki Kanehiro (1849–1920), a surgeon general in the Japanese navy,

rejected the previous germ theory for beriberi and hypothesized that the disease was

due to insufficiencies in the diet instead. Switching diets on a navy ship, he

discovered that replacing a diet of white rice only with one also containing barley,

meat, milk, bread, and vegetables, nearly eliminated beriberi on a nine-month sea

voyage. However, Takaki had added many foods to the successful diet and he

incorrectly attributed the benefit to increased nitrogen intake, as vitamins were

unknown substances at the time. The Navy was not convinced of the need for so

expensive a program of dietary improvement, and many men continued to die of

beriberi, even during the Russo-Japanese war of 1904–5. Not until 1905, after the

anti-beriberi factor had been discovered in rice bran (removed by polishing into white

rice) and in barley bran, was Takaki's experiment rewarded by making him a baron in

the Japanese peerage system, after which he was affectionately called "Barley Baron".

The specific connection to grain was made in 1897 by Christiaan Eijkman (1858–

1930), a military doctor in the Dutch Indies, who discovered that fowl fed on a diet of

cooked, polished rice developed paralysis, which could be reversed by discontinuing

rice polishing. He attributed beriberi to the high levels of starch in rice being toxic. He

believed that the toxicity was countered in a compound present in the rice polishings.

An associate, Gerrit Grijns (1865–1944), correctly interpreted the connection between

excessive consumption of polished rice and beriberi in 1901: He concluded that rice

contains an essential nutrient in the outer layers of the grain that is removed by

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polishing. Eijkman was eventually awarded the Nobel Prize in Physiology and

Medicine in 1929, because his observations led to the discovery of vitamins.

In 1910 a Japanese scientist Umetaro Suzuki first isolated the compound which he

described as aberic acid. In translation from the Japanese paper in which it was

claimed to be a new finding this claim was omitted. In 1911 a Polish biochemist

Casimir Funk isolated the antineuritic substance from rice bran (the modern thiamine)

that he called a "vitamine" (on account of its containing an amino group). However,

Funk did not completely characterize its chemical structure. Dutch chemists, Barend

Coenraad Petrus Jansen (1884–1962) and his closest collaborator Willem Frederik

Donath (1889–1957), went on to isolate and crystallize the active agent in 1926,

whose structure was determined by Robert Runnels Williams (1886–1965), a US

chemist, in 1934. Thiamine was named by the Williams team as "thio" or "sulfur-

containing vitamin", with the term "vitamin" coming indirectly, by way of Funk, from

the amine group of thiamine itself (by this time in 1936, vitamins were known to not

always be amines, for example, vitamin C). Thiamine was synthesized in 1936 by the

Williams group.

Thiamine was first named "aneurin" (for anti-neuritic vitamin). Sir Rudolph Peters, in

Oxford, introduced thiamine-deprived pigeons as a model for understanding how

thiamine deficiency can lead to the pathological-physiological symptoms of beriberi.

Indeed, feeding the pigeons upon polished rice leads to an easily recognizable

behavior of head retraction, a condition called opisthotonos. If not treated, the animals

died after a few days. Administration of thiamine at the stage of opisthotonos led to a

complete cure within 30 minutes. As no morphological modifications were observed

in the brain of the pigeons before and after treatment with thiamine, Peters introduced

the concept of a biochemical lesion.

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When Lohman and Schuster (1937) showed that the diphosphorylated thiamine

derivative (thiamine diphosphate, ThDP) was a cofactor required for the oxydative

decarboxylation of pyruvate, a reaction now known to be catalyzed by pyruvate

dehydrogenase, the mechanism of action of thiamine in the cellular metabolism

seemed to be elucidated. At present, this view seems to be oversimplified: pyruvate

dehydrogenase is only one of several enzymes requiring thiamine diphosphate as a

cofactor; moreover, other thiamine phosphate derivatives have been discovered since

then, and they may also contribute to the symptoms observed during thiamine

deficiency. Lastly, the mechanism by which the thiamine moiety of ThDP exerts its

coenzyme function by proton substitution on position 2 of the thiazole ring was

elucidated by Ronald Breslow in 1958.

Thiamine deficiency

Thiamine is used to treat thiamine deficiency which when severe can prove fatal. In

less severe cases, non-specific signs include malaise, weight loss, irritability and

confusion. Well-known disorders caused by thiamine deficiency include beriberi,

Wernicke–Korsakoff syndrome, optic neuropathy, Leigh's disease, African Seasonal

Ataxia, and central pontine myelinolysis. In Western countries, thiamine deficiency is

seen mainly in chronic alcoholism. Thiamine deficiency is often present in alcohol

misuse disorder. Also at risk are older adults, persons with HIV/AIDS or diabetes, and

persons who have had bariatric surgery. Varying degrees of thiamine deficiency have

been associated with the long-term use of high doses of diuretics, particularly

furosemide in the treatment of heart failure.

Prenatal supplementation

Women who are pregnant or lactating require more thiamine. For pregnant and

lactating women, the consequences of thiamine deficiency are the same as those of the

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general population but the risk is greater due to their temporarily increased need for

this nutrient. In pregnancy, this is likely due to thiamine being preferentially sent to

the fetus and placenta, especially during the third trimester. For lactating women,

thiamine is delivered in breast milk even if it results in thiamine deficiency in the

mother. Pregnant women with hyperemesis gravidarum are also at an increased risk

for thiamine deficiency due to losses when vomiting. Thiamine is an important aspect

for not only mitochondrial membrane development, but also synaptosomal membrane

function. It has also been suggested that thiamine deficiency plays a role in the poor

development of the infant brain that can lead to sudden infant death syndrome (SIDS).

Adverse effects

Thiamine is generally well tolerated and non-toxic when administered orally. Rarely,

adverse side effects have been reported when thiamine is given intravenously

including allergic reactions, nausea, lethargy, and impaired coordination.

Chemistry

Thiamine is a colorless organosulfur compound with a chemical formula

C12H17N4OS. Its structure consists of an aminopyrimidine and a thiazolium ring

linked by a methylene bridge. The thiazole is substituted with methyl and

hydroxyethyl side chains. Thiamine is soluble in water, methanol, and glycerol and

practically insoluble in less polar organic solvents. It is stable at acidic pH, but is

unstable in alkaline solutions. Thiamine, which is a persistent carbene, is used by

enzymes to catalyze benzoin condensations in vivo. Thiamine is unstable to heat, but

stable during frozen storage. It is unstable when exposed to ultraviolet light and

gamma irradiation. Thiamine reacts strongly in Maillard-type reactions.

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Biosynthesis

Complex thiamine biosynthesis occurs in bacteria, some protozoans, plants, and

fungi.The thiazole and pyrimidine moieties are biosynthesized separately and then

combined to form thiamine monophosphate (ThMP) by the action of thiamine-

phosphate synthase (EC2.5.1.3). The biosynthetic pathways may differ among

organisms. In E. coli and other enterobacteriaceae, ThMP may be phosphorylated to

the cofactor thiamine diphospate (ThDP) by a thiamine-phosphate kinase (ThMP +

ATP → ThDP + ADP, EC 2.7.4.16). In most bacteria and in eukaryotes, ThMP is

hydrolyzed to thiamine, which may then be pyrophosphorylated to ThDP by thiamine

diphosphokinase (thiamine + ATP → ThDP + AMP, EC 2.7.6.2).

The biosynthetic pathways are regulated by riboswitches. If there is sufficient

thiamine present in the cell then the thiamine binds to the mRNAs for the enzymes

that are required in the pathway and prevents their translation. If there is no thiamine

present then there is no inhibition, and the enzymes required for the biosynthesis are

produced. The specific riboswitch, the TPP riboswitch (or ThDP), is the only

riboswitch identified in both eukaryotic and prokaryotic organisms.

Foods

 Meat, fish, and grains are a good source of Vitamin B1

 There are high concentrations of Vitamin B1 in the outer layers and germ of

cereals, as well as in yeast, beef, pork, nuts, whole grains, and pulses.

 Fruit and vegetables that contain it include cauliflower, liver, oranges, eggs,

potatoes, asparagus, and kale.

 Other sources include brewer’s yeast and blackstrap molasses.

 Breakfast cereals and products made with white flour or white rice may be

enriched with vitamin B.

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  In the United States, people consume around half of their vitamin B1 intake in

foods that naturally contain thiamin, while the rest comes from foods that are

fortified with the vitamin.

 Heating, cooking, and processing foods, and boiling them in water, destroy

thiamin. As vitamin B1 is water-soluble, it dissolves into cooking water.

White rice that is not enriched will contain only one tenth of the thiamin

available in brown rice.

 The National Institutes of Health (NIH) Office of Dietary Supplements (ODS)

note that one serving of fortified breakfast cereal provides 1.5 milligrams (mg)

of thiamin, which is more than 100 percent of the daily recommended amount.

 One slice of whole wheat bread contains 0.1 mg, or 7 percent of the daily

requirement. Cheese, chicken, and apples contain no thiamin.

 Humans need a continuous supply of vitamin B1, because it is not stored in

the body. It should be part of the daily diet.

Benefits

 Vitamin B1, or thiamin, helps prevent complications in the nervous system,

brain, muscles, heart, stomach, and intestines. It is also involved in the flow of

electrolytes into and out of muscle and nerve cells.

 It helps prevent diseases such as beriberi, which involves disorders of the

heart, nerves, and digestive system.

 Uses in medicine

 Patients who may receive thiamin to treat low levels of vitamin B1 include

those with peripheral neuritis, which is an inflammation of the nerves outside

the brain, or pellagra.

 Some athletes take thiamin supplements to boost their performance.

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  People with ulcerative colitis, persistent diarrhea, and poor appetite may also

receive thiamin. Those who are in a coma may be given thiamin injections.

 Some athletes use thiamin to help improve their performance. It is not a

prohibited substances for athletes in the U.S. Other conditions in which

thiamin supplements may help include:

 AIDS

 canker sores

 cataracts

 glaucoma and other vision problems

 cerebellar syndrome, a type of brain damage

 cervical cancer

 diabetic pain

 stress

 heart disease

 kidney disease in patients with diabetes type 2

 motion sickness

 a weakened immune system.

 Not all of these uses have been definitively confirmed by research.

Deficiency symptoms

 A deficiency of vitamin B1 commonly leads to beriberi, a condition that

features problems with the peripheral nerves and wasting.

 Weight loss and anorexia can develop.

 There may be mental problems, including confusion and short-term memory

loss.

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  Muscles may become weak, and cardiovascular symptoms can occur, for

example, an enlarged heart.

 In the U.S., the recommended daily allowance (RDA) of thiamin taken by

mouth is 1.2 mg for males and 1.1 mg for females over the age of 18 years.

Pregnant or breastfeeding women of any age should consume 1.4 mg each

day.

 People with poor diet, cancer, “morning sickness” during pregnancy, bariatric

surgery, and hemodialysis are at risk of thiamin deficiency.

 People who regularly drink alcohol to excess may have a deficiency, as they

may not absorb thiamin from their food.

 Wernicke-Korsakoff syndrome is a disorder that affects people with chronic

alcoholism. It is linked to a lack of thiamin, and it can be fatal if not treated.

 People with Wernicke-Korsakoff syndrome and those who are withdrawing

from alcohol may receive thiamin injections to help them recover.

 Other diseases, such as HIV, can reduce the absorption of nutrients, and this

can lead to a deficiency of vitamin B1.

Health risks

When eating peanut butter in moderation and as part of a balanced diet, there is little

risk to a person’s health. People aiming to follow a healthful diet may wish to eat pure

peanut butter, which contains no other ingredients.

Sugar and salt intake

Flavored peanut butter products can contain added sugar. Although foods with added

sugar can be part of a healthful diet, it is best to limit the intake of this substance as

much as possible for optimal health. Those who need to follow a low sodium diet

should choose unsalted peanut butter when possible.

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Calorie intake

Additionally, a 2-tablespoon serving of peanut butter has around 188 calories. For a

person consuming 2,000 calories per day, this much peanut butter would be 9.4% of

their daily calorie intake. Those who eat more calories than they burn are likely to

gain weight, which may lead to some people having overweight or obesity, potentially

causing other health issues. It is important for a person to be mindful of what they eat

and try to stick to a healthful, balanced diet.

May reduce risk of certain cancers

Research shows a link between consuming nuts and peanut butter and a lower risk of

certain cancers. For example, a 2017 study found that women who consumed more

nuts and peanut butter had a reduced risk of certain types of breast cancer.

A 2015 study found that people who had a high intake of nuts may have a lower risk

of cardiovascular disease mortality. The researchers also recommended peanuts as a

cost-effective way to boost heart health.

Maintaining blood sugar levels

Some research suggests that people consuming a meal with peanut butter had a lower

blood glucose level than when they had the same meal without it. These results

suggest that peanut butter may help improve blood sugar control, which is important

for optimal health and disease prevention.

May aid weight loss

Peanut butter is full of fiber, fat, and protein, which can help make a person feel fuller

for longer. This feeling of fullness could reduce a person’s chances of snacking on

less healthful foods.

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Nutrition
Occurrence in foods
Thiamine is found in a wide variety of processed and whole foods. Whole grains,

legumes, pork, fruits, and yeast are rich sources. The salt thiamine mononitrate, rather

than thiamine hydrochloride, is used for food fortification, as the mononitrate is more

stable, and does not absorb water from natural humidity (is non-hygroscopic),

whereas thiamine hydrochloride is hygroscopic. When thiamine mononitrate dissolves

in water, it releases nitrate (about 19% of its weight) and is thereafter absorbed as the

thiamine cation.

Dietary recommendations
In the U.S. the Estimated Average Requirements (EARs) and Recommended Dietary

Allowances (RDAs) for thiamine were updated in 1998, by the Institute of Medicine

now known as the National Academy of Medicine (NAM). The European Food

Safety Authority (EFSA) refers to the collective set of information as Dietary

Reference Values, with Population Reference Intake (PRI) instead of RDA, and

Average Requirement instead of EAR. AI and UL defined the same as in United

States. For women (including those pregnant or lactating), men and children the PRI

is 0.1 mg thiamine per megajoule (MJ) of energy consumed. As the conversion is 1

MJ = 239 kcal, an adult consuming 2390 kilocalories should be consuming 1.0 mg

thiamine. This is slightly lower than the U.S. RDA.[33] The EFSA reviewed the same

safety question and also reached the conclusion that there was not sufficient evidence

to set a UL for thiamine

Antagonists
Thiamine in foods can be degraded in a variety of ways. Sulfites, which are added to

foods usually as a preservative, will attack thiamine at the methylene bridge in the

structure, cleaving the pyrimidine ring from the thiazole ring. The rate of this reaction

13
is increased under acidic conditions. Thiamine is degraded by thermolabile

thiaminases (present in raw fish and shellfish). Some thiaminases are produced by

bacteria. Bacterial thiaminases are cell surface enzymes that must dissociate from the

membrane before being activated; the dissociation can occur in ruminants under

acidotic conditions. Rumen bacteria also reduce sulfate to sulfite, therefore high

dietary intakes of sulfate can have thiamine-antagonistic activities.

Plant thiamine antagonists are heat-stable and occur as both the ortho- and para-

hydroxyphenols. Some examples of these antagonists are caffeic acid, chlorogenic

acid, and tannic acid. These compounds interact with the thiamine to oxidize the

thiazole ring, thus rendering it unable to be absorbed. Two flavonoids, quercetin and

rutin, have also been implicated as thiamine antagonists.

Food fortification
Refining grain removes its bran and germ, and thus subtracts its naturally occurring

vitamins and minerals. In the United States, B-vitamin deficiencies became common

in the first half of the 20th century due to white flour consumption. The American

Medical Association successfully lobbied for restoring these vitamins by enrichment

of grain, which began in the US in 1939. The UK followed in 1940 and Denmark in

1953. As of 2016, about 85 countries had passed legislation mandating fortification of

wheat flour with at least some nutrients, and 28% of industrially milled flour was

fortified, often with thiamine and other B vitamins.

Vitamin B2 (Riboflavin)
Riboflavin, also known as vitamin B2, is a vitamin found in food and used as a dietary

supplement. It is required by the body for cellular respiration. Food sources include

eggs, green vegetables, milk and other dairy product, meat, mushrooms, and almonds.

Some countries require its addition to grains.

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As a supplement it is used to prevent and treat riboflavin deficiency. At amounts far in

excess of what is needed to meet dietary needs as a nutrient, riboflavin may prevent

migraines. Riboflavin may be given by mouth or injection. It is nearly always well

tolerated. Normal doses are safe during pregnancy. Riboflavin was discovered in

1920, isolated in 1933, and first synthesized in 1935. It is on the World Health

Organization's List of Essential.

History
The name "riboflavin" comes from "ribose" (the sugar whose reduced form, ribitol,

forms part of its structure) and "flavin", the ring-moiety which imparts the yellow

color to the oxidized molecule (from Latin flavus, "yellow"). The reduced form,

which occurs in metabolism along with the oxidized form, is colorless.

Vitamin B" was originally considered to have two components, a heat-labile vitamin

B1 and a heat-stable vitamin B2. In the 1920s, vitamin B2 was initially thought to be

the factor necessary for preventing pellagra. In 1923, Paul Gyorgy in Heidelberg was

investigating egg-white injury in rats; the curative factor for this condition was called

vitamin H, which is now called biotin. Since both pellagra and vitamin H deficiency

were associated with dermatitis, Gyorgy decided to test the effect of vitamin B2 on

15
vitamin H deficiency in rats. He enlisted the service of Wagner-Jauregg in Kuhn's

laboratory. In 1933, Kuhn, Gyorgy, and Wagner found that thiamin-free extracts of

yeast, liver, or rice bran prevented the growth failure of rats fed a thiamin-

supplemented diet.

Further, the researchers noted that a yellow-green fluorescence in each extract

promoted rat growth, and that the intensity of fluorescence was proportional to the

effect on growth. This observation enabled them to develop a rapid chemical and

bioassay to isolate the factor from egg white in 1933. The same group then isolated

the same preparation (a growth-promoting compound with yellow-green fluorescence)

from whey using the same procedure (lactoflavin). In 1934, Kuhn's group identified

the structure of so-called flavin and synthesized vitamin B2, leading to evidence in

1939 that riboflavin was essential for human health.

Riboflavin foods

Vitamin B2 is a water-soluble vitamin that is flushed out of the body daily, so it must

be restored each day. The best way to get this vitamin is by eating foods that are rich

in riboflavin. Riboflavin is found in eggs, nuts, dairy products, meats, broccoli,

brewer's yeast, Brussel sprouts, wheat germ, wild rice, mushrooms, soybeans, green

leafy vegetables and whole grain and enriched cereals and bread, according to the

University of Maryland Medical Center.

Benefits
Riboflavin is a vitamin that is needed for growth and overall good health. It helps the

body break down carbohydrates, proteins and fats to produce energy, and it allows

oxygen to be used by the body. “Riboflavin is also used for the development and

function of the skin, lining of the digestive tract, blood cells and other vital organs,”

Dr. Sherry Ross, women’s health expert at Providence Saint John’s Health Center in

Santa Monica, California, told Live Science. Vitamin B2 is also important for eye
16
health. According to the University of Michigan, this vitamin is needed to protect

glutathione, which is an important antioxidant in the eye. The U.S. National Library

of Medicine (NLM) reports that eating a diet rich in riboflavin can lower the risk of

developing cataracts. Taking supplements containing riboflavin and niacin may also

be helpful in preventing cataracts. Levels of certain vitamins, chemicals and minerals

in the bloodstream seem to be dependent on healthy levels of B2, as well. For

example, riboflavin changes vitamin B6 and folate (vitamin B9) into forms that the

body can use. According to the American Journal of Clinical Nutrition, riboflavin is

important to how the body processes iron. Without it, research shows that the body is

more likely to develop anemia. Taking riboflavin can also reduce homocysteine

levels in the blood by 26 to 40 percent, according to the NLM.

B2 may be important to pregnancy health, as well. According to a study by the

University Women's Hospital, Heidelberg, Germany, riboflavin deficiency may be a

factor in causing preeclampsia, a condition that causes high blood pressure in late

pregnancy. Those suffering from migraines may find that taking doses of B2 may

help. A study by the department of neurology of Humboldt University of Berlin found

that those taking high doses of riboflavin had significantly fewer migraines.

Deficiency and dosage

Deficiency of riboflavin is rare in developed countries because it is a vitamin found in

many common foods. Some people are more prone to deficiency than others. “This is

more common in people on extreme diets who are underweight or those with

digestive problems such as celiac disease,” Dr. Kristine Arthur, internist at Orange

Coast Memorial Medical Center in Fountain Valley, California, told Live Science.

Teens, alcoholics and the elderly are also more susceptible to vitamin B2 deficiency

because of poor diet. Deficiency can cause anemia, sore throat, mouth or lip sores,

inflammation of the skin and swelling of soft tissue in the mouth. These symptoms
17
can show up after just a few days of deficiency, according to the American Journal of

Clinical Nutrition.

The normal recommended daily allowance (RDA) of riboflavin is dependent on age,

gender and reproductive status. “RDA is 1.3 milligrams daily for men and 1.1 mg for

women. A higher dose of 3 mg per day can help to prevent cataracts. Higher doses up

to 400 mg can be used to treat migraine headaches,” said Arthur. A cup of chopped

kale has 0.1 mg, while a hard-boiled egg has 0.3 mg and a glass of whole milk has 0.4

mg, according to the U.S. Department of Agriculture. One cup of whole almonds has

1.4 mg of riboflavin, or 85 percent of the RDA. As a supplement, riboflavin is usually

included in multivitamins and B-complex vitamins. It also is available separately in

doses of 25 mg, 50 mg and 100 mg. Relatively nontoxic, riboflavin is considered safe

at high doses because excess is disposed of through the urinary tract. There may be

some side effects from taking higher doses of B2, though. “Some people notice their

urine turning yellow-orange in color and having diarrhea when taken in higher doses,”

said Ross.

Causes
Riboflavin is continuously excreted in the urine of healthy individuals, making

deficiency relatively common when dietary intake is insufficient. Riboflavin

deficiency is usually found together with other nutrient deficiencies, particularly of

other water-soluble vitamins. A deficiency of riboflavin can be primary – poor

vitamin sources in one's daily diet – or secondary, which may be a result of conditions

that affect absorption in the intestine, the body not being able to use the vitamin, or an

increase in the excretion of the vitamin from the body. Subclinical deficiency has also

been observed in women taking oral contraceptives, in the elderly, in people with

eating disorders, chronic alcoholism and in diseases such as HIV, inflammatory bowel

disease, diabetes and chronic heart disease. The Celiac Disease Foundation points out
18
that a gluten-free diet may be low in riboflavin (and other nutrients) as enriched wheat

flour and wheat foods (bread, pasta, cereals, etc.) are a major dietary contribution to

total riboflavin intake.

Diagnosis
Overt clinical signs are rarely seen among inhabitants of the developed countries. The

assessment of riboflavin status is essential for confirming cases with unspecific

symptoms where deficiency is suspected. Glutathione reductase is a nicotinamide

adenine dinucleotide phosphate (NADPH) and FAD-dependent enzyme, and the

major flavoprotein in erythrocytes. The measurement of the activity coefficient of

erythrocyte glutathione reductase (EGR) is the preferred method for assessing

riboflavin status. It provides a measure of tissue saturation and long-term riboflavin

status. In vitro enzyme activity in terms of activity coefficients (AC) is determined

both with and without the addition of FAD to the medium. ACs represent a ratio of

the enzyme's activity with FAD to the enzyme's activity without FAD. An AC of 1.2

to 1.4, riboflavin status is considered low when FAD is added to stimulate enzyme

activity. An AC > 1.4 suggests riboflavin deficiency. On the other hand, if FAD is

added and AC is < 1.2, then riboflavin status is considered acceptable. Tillotson and

Bashor reported that a decrease in the intakes of riboflavin was associated with

increase in EGR AC. In the UK study of Norwich elderly, initial EGR AC values for

both males and females were significantly correlated with those measured 2 years

later, suggesting that EGR AC may be a reliable measure of long-term biochemical

riboflavin status of individuals. These findings are consistent with earlier studies.

Experimental balance studies indicate that urinary riboflavin excretion rates increase

slowly with increasing intakes, until intake level approach 1.0 mg/d, when tissue

saturation occurs. At higher intakes, the rate of excretion increases dramatically. Once

intakes of 2.5 mg/d are reached, excretion becomes approximately equal to the rate of
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absorption At such high intake a significant proportion of the riboflavin intake is not

absorbed. If urinary riboflavin excretion is <19 µg/g creatinine (without recent

riboflavin intake) or < 40 µg per day are indicative of deficiency.

Treatment
Multi-vitamin dietary supplements often contain 100% of the U.S. Daily Value (1.3

mg) for riboflavin, and can be used by persons concerned about an inadequate diet.

Over-the-counter dietary supplements are available in the United States with doses as

high as 100 mg, but there is no evidence that these high doses have any additional

benefit for healthy people.

Side effects
In humans, there is no evidence for riboflavin toxicity produced by excessive intakes,

in part because it has lower water solubility than other B vitamins, because absorption

becomes less efficient as doses increase, and because what exceeds the absorption is

excreted via the kidneys into urine. Even when 400 mg of riboflavin per day was

given orally to subjects in one study for three months to investigate the efficacy of

riboflavin in the prevention of migraine headache, no short-term side effects were

reported. Any excess at nutritionally relevant doses is excreted in the urine, imparting

a bright yellow color when in large quantities. The limited data available on

riboflavin's adverse effects do not mean, however, that high intakes have no adverse

effects, and the Food and Nutrition Board urges people to be cautious about

consuming excessive amounts of riboflavin.

Function
Flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) function as

cofactors for a variety of flavoprotein enzyme reactions:

 Flavoproteins of electron transport chain, including FMN in Complex I and

FAD in Complex II

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  FAD is required for the production of pyridoxic acid from pyridoxal (vitamin

B6) by pyridoxine 5'-phosphate oxidase

 The primary coenzyme form of vitamin B6 (pyridoxal phosphate) is FMN

dependent

 Oxidation of pyruvate, α-ketoglutarate, and branched-chain amino acids

requires FAD in the shared E3 portion of their respective dehydrogenase

complexes

 Fatty acyl CoA dehydrogenase requires FAD in fatty acid oxidation

 FAD is required to convert retinol (vitamin A) to retinoic acid via cytosolic

retinal dehydrogenase

 Synthesis of an active form of folate (5-methyltetrahydrofolate) from 5,10-

methylenetetrahydrofolate by methylenetetrahydrofolate reductase is FADH2

dependent

 FAD is required by Kynurenine 3-monooxygenase to convert tryptophan to

niacin (vitamin B3)

 Reduction of the oxidized form of glutathione (GSSG) to its reduced form

(GSH) by glutathione reductase is FAD dependent

Medical uses
Corneal ectasia is a progressive thinning of the cornea; the most common form of this

condition is keratoconus. Collagen cross-linking by applying riboflavin topically then

shining UV light is a method to slow progression of corneal ectasia by strengthening

corneal tissue. A 2017 review found that riboflavin taken daily in amounts roughly

200 to 400 times the Recommended Dietary Allowance (RDA) may be useful to

prevent migraines in adults, but found that clinical trials in adolescents and children

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had mixed outcomes. A hypothesis has been proposed that riboflavin improves

mitochondrial energy production.

Biosynthesis
The biosynthesis of one riboflavin molecule requires one molecule of GTP and two

molecules of ribulose 5-phosphate as substrates. The imidazole ring of GTP is

hydrolytically opened, yielding a 4, 5-diaminopyrimidine which is converted to 5-

amino-6-ribitylamino-2, 4 (1H,3H)-pyrimidinedione by a sequence of deamination,

side chain reduction and dephosphorylation. Condensation of 5-amino-6-ribitylamino-

2,4 (1H,3H)-pyrimidinedione with 3, 4-dihydroxy-2-butanone 4-phosphate obtained

from ribulose 5-phosphate affords 6,7-dimethyl-8-ribityllumazine. Dismutation of the

lumazine derivative yields riboflavin and 5-amino-6-ribitylamino-2,4(1H,3H)-

pyrimidinedione, which is recycled in the biosynthetic pathway. The structure of the

biosynthetic enzyme, 6,7-dimethyl-8-ribityllumazine synthase, has been studied in

considerable detail.

Chemistry
As a chemical compound, riboflavin is a yellow-orange solid substance with poor

solubility in water compared to other B vitamins. Visually, it imparts color to vitamin

supplements (and bright yellow color of urine in persons taking it).

Industrial uses
Fluorescent spectra of riboflavin
Because riboflavin is fluorescent under UV light, dilute solutions (0.015–0.025%

w/w) are often used to detect leaks or to demonstrate coverage in an industrial system

such a chemical blend tank or bioreactor.

Industrial synthesis
Large cultures of Micrococcus luteus growing on pyridine (left) and succinic acid

(right). The pyridine culture has turned yellow from produced riboflavin.

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The industrial scale production of riboflavin using diverse microorganisms, including

filamentous fungi such as Ashbya gossypii, Candida famata and Candida flaveri, as

well as the bacteria Corynebacterium ammoniagenes and Bacillus subtilis. The latter

organism, genetically modified to both increase the production of riboflavin and to

introduce an antibiotic (ampicillin) resistance marker, is employed at a commercial

scale to produce riboflavin for feed and food fortification. The chemical company

BASF has installed a plant in South Korea, which is specialized on riboflavin

production using Ashbya gossypii. The concentrations of riboflavin in their modified

strain are so high that the mycelium has a reddish/brownish color and accumulates

riboflavin crystals in the vacuoles, which will eventually burst the mycelium.

Riboflavin is sometimes overproduced, possibly as a protective mechanism, by some

bacteria in the presence of high concentrations of hydrocarbons or aromatic

compounds. One such organism is Micrococcus luteus (American Type Culture

Collection strain number ATCC 49442), which develops a yellow color due to

production of riboflavin while growing on pyridine, but not when grown on other

substrates, such as succinic acid.

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References
1. Zempleni J, Galloway JR, McCormick DB (January 1996). "Pharmacokinetics

of orally and intravenously administered riboflavin in healthy humans". The

American Journal of Clinical Nutrition. 63 (1): 54–66.

doi:10.1093/ajcn/63.1.54. PMID 8604671.

2. European Food Safety Authority (February 2006). "Tolerable Upper Intake

Levels for Vitamins and Minerals" (PDF). EFSA. Retrieved 18 June 2018.

3. "Nutrient reference values for Australia and New Zealand" (PDF). National

Health and Medical Research Council. 9 September 2005. Retrieved 19 June

2018.

4. Royer-Morrot MJ, Zhiri A, Paille F, Royer RJ (1992). "Plasma thiamine

concentrations after intramuscular and oral multiple dosage regimens in

healthy men". European Journal of Clinical Pharmacology. 42 (2): 219–22.

doi:10.1007/BF00278489. PMID 1618256. S2CID 19924442.

5. American Society of Health-System Pharmacists. "Thiamine Hydrochloride".

Drugsite Trust (Drugs.com). Retrieved 17 April 2018.

6. "Thiamine: MedlinePlus Drug Information". medlineplus.gov. Retrieved 30

April 2018.

7. Guidelines on food fortification with micronutrients (PDF). WHO and FAO.

2006. pp. 13–14. ISBN 92-4-159401-2. Retrieved 5 May 2018.

8. "Thiamine". www.drugbank.ca. Retrieved 30 April 2018.

9. Kliegman RM, Stanton B (2016). Nelson Textbook of Pediatrics. Elsevier

Health Sciences. p. 322. ISBN 9781455775668. There are no cases of adverse

effects of excess thiamine... A few isolated cases of puritis...

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