Introduction to brewing

0:06Skip to 0 minutes and 6 secondsAt the end of a hectic day, how relaxing to sit down with feet up and a
refreshing brew. Former British Prime Minister, William Gladstone, was clearly a fan when he said, ‘if you are
cold, tea will warm you. If you are too heated, it will cool you. If you are depressed, it will cheer you. If you are
excited, it will calm you.’ But it is not only
0:26Skip to 0 minutes and 26 secondsGladstone that enjoys a brew: Around 2 billion cups of coffee are
consumed worldwide every day and tea remains the most consumed drink in the world, after water. In the UK
alone, the tea market is worth around £700 million per year. Our tour of beverages concentrates on the
chemistry of beer, tea and coffee. We will start by looking at the beer-making process, from milling to maturing.
Of particular interest will be the key flavour components. We will see how a compound found in hops,
undergoes a rearrangement reaction during the brewing process, to form compounds called isohumulones -
these give a pint of bitter its name.
1:09Skip to 1 minute and 9 secondsThe bitterness of beer is measured according to the International Bitterness
Units scale, with one IBU corresponding to just one part-per-million of isohumulone. We will then look at
brewing a cup of tea, and what organic compounds are present, including polyphenol antioxidants. Tea is rich
in antioxidants including catechins, which generally give a bitter taste to tea. Hardly a week goes by without a
claim that drinking a cup of tea is good, or bad. So we will look at the underlying chemistry and the structures of
the organic compounds involved in these claims. Finally, we will explore coffee, which contains over 1000
aroma compounds. Again, the reported health effects of drinking coffee can be conflicting, including that for
caffeine.
1:54Skip to 1 minute and 54 secondsFor example, a large US study found that women who drank two or more
cups of coffee a day were less likely to get depressed. Caffeine is known to improve memory and the speed
with which our brains process information - it increases the production of neurotransmitters such as dopamine.
It also increases breathing and heart rate, resulting in a short burst of energy. The average daily intake of
adults is around 200 mg of caffeine - equivalent to two mugs or four cups of coffee. For those of you who prefer
coffee without caffeine, we will investigate how decaffeinated coffee is prepared - the use of supercritical
carbon dioxide is particularly effective, removing 97-99% of the caffeine from the beans.
2:38Skip to 2 minutes and 38 secondsThis section should certainly quench your thirst for knowledge!

Welcome back to the course and our third week of study, which explores brewing!

Brewing is both an art and a science. For a pleasing brew, whether it be a pint of bitter, or a cup of filter coffee,
there are various devices to aid the process.

For example, “a beer making kit” typically includes a fermenter, a valve and tubing, a funnel, thermometer,
hydrometer (that measures the specific gravity (relative density) of liquids), a spoon, bottles and caps. Those of
you who make drip or filter coffee will need filter paper, a kettle, a balance, a grinder and a spoon as part of
your essential kit.

So, the kitchens of connoisseurs of a good brew can sometimes resemble an organic chemistry laboratory
(indeed, there are some products that are a hybrid between laboratory glassware and modern drinkware). A
modern organic chemistry laboratory, like our professional standard teaching laboratories at York, will have
standard kit ranging from mechanical stirrers to balances, thermometers, and lots of glassware. This includes
Quickfit glassware, required, for example, when carrying out a distillation.

Then there are the more specialised pieces of equipment, such as a rotary evaporator (for evaporation of
solvents from samples), a Büchner funnel (used in filtration) and glassware, such as Kipp’s apparatus (for making
small volumes of gases) and a Kjeldahl flask (round-bottomed and long-necked to trap splashes from material
being boiled for analysis). Indeed, some pieces of laboratory glassware are so intricate and beautifully
designed that they may not be out of place in an art gallery! (At York, we are fortunate to have our own
chemistry glassblower.)
Scientific equipment is constantly being refined and nature is a constant source of inspiration, even for pipettes.
The sex organs of liverworts have inspired a design for a plastic pipette that can pick up and transfer precise
amounts of water - the shape of a ‘dipper’ is designed so that the water is gripped by surface tension!

So what is your favourite piece of chemistry-related equipment or glassware, and why? This may be
something in your home, perhaps a test tube spice rack or a mortar and pestle, or for those of you doing a
course or job involving practical chemistry, something from your laboratory. Alternatively, it may be something
you remember from your school days, perhaps the Bunsen burner, so popular it has its own celebration day!

https://youtu.be/YKve8mt1-vg

scientific_equipment.pdf
Chemistry of flavor

0:06Skip to 0 minutes and 6 secondsFlavour is the sensory impression of food or other substances, determined
mainly by taste and smell. Working together with our sense of smell, around 10 000 taste buds in our tongue
elicit various sensations. Flavour is a complicated subject and it is very rare for just one substance to be
responsible for the flavour of a particular food or drink. Normally a cocktail of low-boiling organic compounds
are present. For example, chocolate contains around 300 flavour components. One area of current interest is
understanding why certain flavours go together, for example, strawberries and cream. Perhaps both
components have many important aroma compounds that are the same, or similar?
0:51Skip to 0 minutes and 51 secondsIt is also interesting to understand why different people have different
reactions to the same food or drink - remember the Marmite marketing slogan ‘love it, or hate it’? The taste,
aroma and mouth feel all contribute to the overall
1:06Skip to 1 minute and 6 secondsflavour of a food or drink: two of which are chemical senses, the taste and
the aroma. The expectations of the taster, the look of the food or drink, and the order in which foods are
consumed all play a contributing role to the overall flavour and, when combined with chemical phenomena
occurring when you taste something, the science of flavour becomes an incredibly complicated system.
Located on the tongue, the taste buds are where a person’s sense of taste originates. Each taste bud contains
many taste cells; taste cells are long and thin and stretch from the opening of the taste bud in the mouth, to the
nerve cell.
1:46Skip to 1 minute and 46 secondsThere are several taste cells in every taste bud, which are able to recognise
five main tastes - sweet, salty, bitter, sour and umami. Umami has only recently been acknowledged as a
separate taste. It is the taste of savoury things, like cheese and meat, and the taste comes from the amino acid
L-glutamic acid. Each taste cell reacts to different taste-provoking molecules, or tastants. Sweet, bitter and
umami taste signals trigger a response without entering the taste cell. The tastants are small molecules that
bind to the receptor proteins that are on the surface of the taste receptor cell. The binding is very specific, as
the tastant has exactly the right shape for interaction with the receptor protein.
2:34Skip to 2 minutes and 34 secondsOnce bound, the receptor protein changes shape, which initiates a series
of chemical reactions that leads to the release of a neurotransmitter that activates nerves to the brain,
registering the taste. Salty and sour taste signals operate differently. Ions, such as Na+ and Cl- in sodium
chloride (or common salt), can bind to ion channels, which are proteins that assist the ions entering inside the
taste receptor cell. This leads to neurotransmitters being released that activate nerves to the brain, registering
it as a salty or sour taste. In acidic foods and drinks, the acids (such as citric acid) liberate hydrogen ions, or
protons, that bind to the ion channels.
3:16Skip to 3 minutes and 16 secondsWhile humans have only five primary tastes, there are a much larger
number of aroma receptors. When a food is placed into the mouth, food vapours reach the nasal lining and
interact with the aroma sensors, which are combined with the taste sensors to give a food or drink a specific
flavour. In taste sensors, the taste bud (a specialised sense organ) is attached to nerve cells. Whereas aroma
sensors are nerve cells themselves, which are directly connected to the brain. Without smell foods and drinks
tend to taste bland and have no flavour. Research is still being undertaken on aroma sensors and how they
bind to compounds, as the process is still not fully understood.
3:58Skip to 3 minutes and 58 secondsThe last contributing factor to flavour is the mouth feel of a food or drink. As
well as taste sensors (taste buds) in the mouth, there are also other sensors that measure the mouth feel;
reacting to pressure, temperature and irritating substances, picking up heat, cold, textures and the feel of
carbonated drinks on the tongue, among others. All of these contribute to the mouth feel and, hence, the
flavour of the final food or drink. Overall, there are three main contributors to the flavour of a substance; taste,
aroma and mouth feel. Throughout this week we will look at the main flavour components of beer, tea and
coffee and see how different organic compounds contribute to the final taste of a brew.

Favourite smells
A survey of 2000 UK adults in 2015 showed that freshly baked bread was their favourite smell, followed
by bacon, newly-mown grass then coffee. Interestingly, other than newly-mown grass, they are things
that can help make a home more warm and welcoming. Food and drink smells featured heavily in the
poll with orange, coconut, Christmas cake, lime and doughnuts proving popular. Cherry, marzipan,
popcorn and wine also made the top 50 as well as cheese and biscuits.

At the other end of the spectrum, bins were named the most disliked smell, along with drains, body odour,
sewage, vomit and rotten milk and food.

Researchers found that 60% of people believed there are certain smells that remind them of particular people
or places, while 70% associated a smell with a memory.

What is your favourite food or drink smell and does it remind you of anything?

For the thiol fans, you might like to know that according to the 2000 Guinness Book of World Records,
ethanethiol (CH3CH2SH) is the ‘smelliest substance in existence’.

Smells good?
Aside from foods and drinks, perhaps one or more of these everyday fragrances will interest you? (Organic
compounds that contribute to the aroma are shown in brackets.)

New car smell (toluene, ethylbenzene, styrene, xylenes)

Wet dog smell (benzaldehyde, phenylethanal, ethanal, phenol, 2-methylbutanal)

Halitosis or bad breath (methanethiol, hydrogen sulfide, dimethyl sulfide)

Rain (palmitic acid, stearic acid)

Old books (benzaldehyde, vanillin, ethylbenzene, toluene, 2-ethylhexanol)

Newly-mown grass ((Z)-3-hexenal, (E)-2-hexenal, (Z)-3-hexenyl ethanoate, methanol, ethanol)

Headline news – Love it or hate it

Researchers at York have discovered a potential link between eating Marmite and activity in the brain. It appears
that Marmite increases levels of a specific neurotransmitter – known as GABA (γ-aminobutyric acid,
H2NCH2CH2CH2CO2H) – in the brain. This is likely due to the high concentration of vitamin B12 in Marmite. So,
dietary intervention may affect GABA imbalances, which are associated with a variety of neurological disorders.

Tongues detect odours

Research indicates the tongue detects not just taste, but also odours. In this work, human taste cells responded
to fragrances, including a clove-scented compound called eugenol (C10H12O2), even though the concentration
of these substances was below the level necessary to trigger a taste response. It opens up the possibility of
using odours to trick us into healthier eating. For example, adding a very low concentration of an odour to a
food could make us think it is sweeter than it is – thereby reducing the need for sugar and helping tackle the
obesity crisis.
Serial extraction of coffee and tea

0:06Skip to 0 minutes and 6 secondsIn Britain it is estimated that a staggering 165 million cups of tea, and 70
million cups of coffee, are drunk each day. Back in 1946 George Orwell said that there are 11 rules for perfect
tea making, rules from which nobody should dare depart. To mark the 100th anniversary of Orwell’s birth, the
Royal Society of Chemistry (RSC) commissioned research, concluding that Orwell’s rules were quite wrong -
for example, it argued against adding cold milk to hot tea as this makes denaturation, or degradation, of the
proteins in the milk more likely. But what is the best way to make a perfect cup of tea?
0:46Skip to 0 minutes and 46 secondsThis is partly related to the steeping, or soaking, time - how long the hot
water is allowed to be in contact with the tealeaf. Leaving a tea bag in longer, in boiling water, will clearly make
a stronger cup of tea. Interestingly, the International Organisation for Standardisation has specified a period of
six minutes for brewing tea. It is not intended to produce a good cup of tea, but it is designed to produce a
consistent one for taste testing. So, adding hot water to the tea bag allows the flavoursome compounds to
become extracted into your pot, or cup, or tea. But, not all of the compounds are extracted at the same time, so
different flavours ‘escape’ the leaf at different times.
1:27Skip to 1 minute and 27 secondsThis activity is designed to investigate the different extraction rates of the
aroma and flavour compounds in tea and coffee. First set out four or five cups. For tea, take a tea bag in a
hand-held sieve and slowly pour hot water onto the bag and let water drip into the first cup, then move the bag
over the next cup and so on, to give four or five fractions. For coffee, use the drip-brewing filtered coffee
method - this is where hot water is poured over roasted ground coffee beans contained within a cone of filter
paper. Drip filter ground coffee into each cup in turn.
2:04Skip to 2 minutes and 4 secondsNow smell and taste each fraction in turn - for coffee, does the first fraction
taste acidic or sour? Do you notice a change in the aroma as you move from cup to cup? You might like to try
using hot water at different temperatures - use a thermometer, record the temperature, and see if changing the
temperature affects the extraction process, by altering the flavour and aroma of the different fractions. Before
finishing it is worth combining all four or five fractions and taste how they complement one another in the
combined brew. At each stage of the extraction, describe the aroma and flavour, and record your findings.
2:39Skip to 2 minutes and 39 secondsPost your results - don’t forget to identify the specific tea or coffee you
used, and let’s see what information we can extract!

The following ingredients will be required:

 Ground coffee
 Boiling water

The following items will be required:

 Measuring spoons
 Measuring jug
 Five cups
 Filter paper
 Timer

Ensure that you read through all of the instructions before beginning the experiment to ensure that you
understand exactly what will happen at each step.
Step one
Measure out two tablespoons of coffee into some filter paper and position the full filter paper into the bottom of
the first cup, as shown in the figure below. Then place the four other cups in a row alongside the first, for the
following fractions.

Step two
Boil the kettle and measure out 500 mL of boiling water into a measuring jug. You are going to need ~100 mL
of water for each cup that you have. Not all the water will pass through the filter paper in the time allowed for
the extractions, but it's useful to have extra water in case you start to run out.

Step three
Set the timer to 30 seconds. As you start to pour the boiling water, slowly into the first cup, begin the timer.
Stop pouring the water when the filter paper looks about half full – but keep topping it up, as it gets low. If you
fill the filter paper up too much, you risk spilling the water as you try to transfer it between cups, which will ruin
the experiment.

Step four
After the 30 seconds have passed, transfer the filter paper to the next cup, starting the timer again (for another
30 seconds) and ensuring to top up the water level in the filter paper when it looks like it is getting low.
Continue until you have poured 100 mL of water into each cup. Look out for colour differences between the
fractions.

Step five
Now you should taste each fraction of your coffee – remembering to leave enough to combine them at the end,
so you can taste a complete cup of coffee – and you should record what each fraction tastes like. We have
provided a table below of our results and a blank one in the downloads section for you to fill in. Make sure to
take note of what kind of coffee you used.

Type of coffee: LavAzza Qualita Rossa Filter Coffee

Fraction number Time of extraction Taste description

(seconds)
1 0 to 30 slightly acidic

very acidic, slightly

2 30 to 60
sour

strong flavour, very

3 60 to 90
heavy

4 90 to 120 bitter

very weak coffee

5 120 to 150
flavour

After completing the table, mix each of your fractions into one cup and taste them all together. What does the
resultant coffee mixture taste like?

For the tea drinkers among you, a further ‘tea extraction experiment’, blank results tables
and the risk assessment can be located in the additional PDFs found at the bottom of the
section. Remember to upload your results to our open Padlet (we have included some
examples from a previous course to help inspire you) and/or
on Twitter or Instagram hashtag #FLchemistry. Alternatively, you might like to explain your
results using the comments section below. We are eager to see what you have been able to
come up with!

We hope to get lots of interaction and that your extraction gives you satisfaction, and a
positive reaction! Why not give it a go?

Risk_assesment_for_coffee_extractions.pdf
Tables_for_Results.pdf

Tea_Extraction_Experiment.pdf
Tea and taste

Tea chemistry is complicated. Harvested from the bushy Camellia sinensis plant, tea leaves are mostly
comprised of water. When plucked, the leaves begin to lose water and wilt, which is called withering.
As they wither, their cell walls begin to break down and carbohydrates, proteins and fats break down into
simpler sugars, proteins and lipids. This is intensified by rolling, crushing or twisting the leaves, and
the thousands of compounds which were once inside the leaves come into contact with oxygen and
each another and react in oxidation reactions.

For making black tea, hot air is blown over the leaves in a process called firing, to degrade the enzymes so the
leaf does not mould or break down. During this process, green pigments in the tea leaf are oxidised to form
other pigments, which give the characteristic brown-black colour.

In contrast, green tea (which is made from the same plant leaves) involves the rapid steaming or pan firing of
the freshly harvested leaves to inactivate the enzymes. Typically, black tea and green tea contain around the
same amount of caffeine… but caffeine does not have much effect on the taste of the tea.

Tea producers control the natural tendency of tea leaves to wilt and oxidise in order to produce a finished tea
that has a desirable appearance, aroma, flavour, and taste, ready for packaging. Manufactured teas are sized,
graded and evaluated for flavour and infusion colour by professional tea tasters. For hundreds of years, tea
makers have produced drinkable teas using the principles of withering and oxidation with no knowledge of the
underlying chemistry.

Brewing and steeping

Brewing is the act of making tea, while steeping is the process involved in this. Brewing the perfect cup of tea
requires carefully steeping loose tea leaves or tea bags (round, square or pyramid-shaped) in fresh water
heated correctly. Various types of teas steep within different times, and understanding this may explain the
difference between drinking a bland cup of tea and enjoying a great cup of tea. Dr Stuart Farrimond, an expert
tea maker, argues that the best brew time is 5 minutes and that we should drink it out of a orange or red mug
(apparently, it makes the tea taste sweeter!). The age of the tea leaf or poor storage conditions can also have a
significant effect and make the tea tasteless and faded in colour.

When we steep tea leaves, our senses are awakened by thousands of low-boiling organic compounds,
collectively called the 'aroma complex'. These come from the tea liquor and the thousands of non-volatile
compounds, not all of which are water-soluble. The solubility of the compounds is related to the properties of
the water used for soaking the leaves, such as temperature and pH. Consequently, it is difficult to generalise
and say that a specific organic compound is responsible for a particular taste.

Around 98% of British people drink their tea with milk. When tea lovers add milk, the chemistry of tea can
change. There are conflicting opinions as to the effect adding milk to tea has on the organic compounds
contained therein, especially compounds called polyphenols, which have been shown to
have antioxidant properties (which can have health benefits). Studies have indicated that proteins in milk can
bind to polyphenols in black tea and inhibit their antioxidant properties. Other research suggests that this does
not actually affect the concentration of antioxidants in the blood after ingestion, because the binding can be
broken down during digestion. So, the jury is out. What we do know is the polyphenols present in tea leaves
are important for the colour and astringency of a cup of tea.

Milk first or last?

A much-debated question is whether to add milk first or last – historically, the order of steps was taken as an
indication of class. The 'milk-first' approach gradually brings the contents of the cup up from fridge-cool. In
contrast, the 'milk-last' approach rapidly heats the first drop of milk almost to the temperature of the teapot,
breaking down the proteins in milk to a greater degree and so developing a more 'boiled milk' flavour. Some
people say the boiled milk taste is less pleasant, but taste is subjective and others will argue otherwise.

If you do not add milk to tea, you may add lemon juice, which causes an immediate change to the colour.
Adding acidic lemon juice, which contains citric acid (C6H8O7), typically lightens the colour of a cup of black tea
from dark brown to an orangey-yellow colour – the coloured compounds are protonated, which causes them to
absorb light in a different way.

For example, the OH groups of polyphenols (abbreviated Ar–OH) can be deprotonated (to form Ar–O –) or
protonated (to form Ar–+OH2); each species absorbs light of different wavelength, and the concentration of
each species depends on the pH of the tea. The addition of citric acid lowers the pH of the tea, which increases
the concentration of the protonated species, hence the change in colour. [A dramatic colour change, due to a
change in pH, is seen using butterfly pea flower tea, which, due to anthocyanins (pigments which are found in
many flowers and fruits), is coloured blue - on protonation of the anthocyanins the colour changes to red.]

Various compounds found in tea have been organised into broad groups. Collectively we have some idea of
what happens to these groups during processing and for which flavours and aromas they are responsible for,
as we will see next.

Why do tea plants make polyphenols?

Polyphenols are described as secondary metabolites of plants, which means they are not directly involved in
the normal growth, development, or reproduction of an organism. They are generally involved in defence
against ultraviolet radiation or pathogens, parasites and plant predators.
Interestingly, plants make a greater array of secondary metabolites than animals because they do not have the
same physical mobility to escape from predators, so they have developed a chemical defence against such
predators (e.g. phenolic compounds can be unpalatable).

Too much of a good thing!

In 2013, a US woman lost all of her teeth, attributed to excessive tea drinking – she drank a pitcher of iced tea
(around 6.5 pints) every day for 17 years! The loss of teeth was linked to skeletal fluorosis, a bone disease
caused by consuming high levels of fluoride (F–). Fluoride is needed for healthy bones and teeth and it is
added in very low levels to drinking water (in England, water contains 1 mg per litre, or 1 ppm of fluoride). Tea
drinkers get an extra dose of fluoride as this in present, in variable amounts (from 103 to 839 milligrams (mg)
per kilogram (kg)), in tea leaves. Research shows economy teas may use the older leaves on the tea plant,
which may contain higher levels of fluoride – it concludes that people drinking 1 litre of cheap tea a day may be
consuming more fluoride than the daily recommended amount. (By the way, the LD50 of sodium fluoride on oral
administration in rats is 52 mg/kg.)

On the subject of fluoride, one continuing conspiracy theory is that adding fluoride to public water supplies to
reduce dental decay is harmful. But there is no convincing scientific evidence that water fluoridation at around
1 mg per litre, the UK standard, is harmful to health. Fluoride ions interact with tooth enamel, producing the
mineral fluorohydroxyapatite (HO– ions in hydroxyapatite are replaced by F– ions), which reduces the likelihood
of the tooth surface dissolving. A 2015 analysis showed fluoridation resulted in children having 26-35% fewer
decayed, missing or filled teeth.

Tea temperature
Some interesting research has been conducted on how the serving temperature affects our enjoyment of food
and drink. Steaming hot dishes may smell delicious, but research has shown that the perception of taste
decreases when the temperature goes above 35 °C. It may be that the burning session, when consuming really
hot food, masks the taste sensation to warn us of the danger of hurting ourselves. Consequently, the
temperature of what you drink will also affect the food you are eating. Typically, North American people like ice-
cold water at mealtimes, whereas Europeans are happy with not-far-below room temperature, and Asian
people often drink hot water or tea while eating. So, where do you stand on tea temperature – should it be
scalding hot, cool enough to glug, or somewhere in-between?

Also, in 2017, an Australian scientist sparked controversy with his claim that the best way to
make tea is by using a microwave. Where do you stand on using a kettle or a microwave? If
you are not sure, why not try a blind tasting test and let us know what you find.

Yorkshire tea
By the way, coming from York, we must extoll the virtues of Yorkshire Tea. A strong black tea, one of the top
selling brands in the U.K., produced by The Bettys and Taylors Group, based in Harrogate, Yorkshire. The
same company owns Bettys Café Tea Rooms, which are traditional tea rooms serving traditional meals -
afternoon tea in one of the York branches is a lovely treat! (York is regularly named one of the best places to
live and visit in the UK, and in 2022 it was named as the best night out in the UK.)
The organic flavour components of tea

0:06Skip to 0 minutes and 6 secondsAmazingly, for hundreds of years tea makers have produced drinkable teas,
from tealeaves, using principles of withering and oxidation, with no understanding of the underlying chemistry.
Today, we now know the most important organic compounds in fresh tealeaves, responsible for producing teas
with the most desirable appearance, aroma, flavour, and taste. These include polyphenols, largely responsible
for astringency, and amino acids, which give tea its brothiness. During tea processing the organic compounds
undergo changes to produce a ‘finished’ or ‘made’ tea - that is, one that has been processed and is ready for
packaging or steeping. In steeped tea, polyphenols are mainly responsible for astringency (the ‘drying’
sensation experienced in the mouth after drinking tea).
0:52Skip to 0 minutes and 52 secondsThese compounds are produced as a defence against insects and other
animals and are the most abundant compounds in tea comprising as much as 30-40% of both freshly plucked
tealeaves and solids in tea liquor. A tea drinker typically consumes 180 to 240 mg of polyphenols from a strong
cup of tea. There are an estimated 30,000 polyphenolic compounds in tea, including (-)-epicatechin, a member
of a class of polyphenolic compounds called flavanoids, which have similar structures, all containing 15-carbon
atoms. One for the key distinctions between black teas and green teas is the production of another type of
polyphenol, called theaflavins, which includes theaflavin and related compounds.
1:43Skip to 1 minute and 43 secondsTheaflavin gives a bright red-orange appearance to the tea, due to the
presence of a benzotropolone ring system, and is formed from two molecules of a flavonoid. Polyphenol
oxidase and peroxidase are the most important enzymes in tealeaves. They are responsible for the enzymatic
browning of tealeaves that occurs when the cell walls in the leaves are broken and the polyphenols exposed to
oxygen. These enzymes may be deactivated using heat so that browning cannot occur; this is one of the first
steps in green tea production and explains why finished green tea leaves remain green. Amino acids give tea
its savoury taste. Tealeaves contain many amino acids, the most abundant one being L-theanine.
2:30Skip to 2 minutes and 30 secondsL-Theanine is responsible for promoting alpha brain wave activity, which
can promote relaxation. Plant pigments give leaves their colour and are responsible for absorbing light for
photosynthesis. The two major groups of pigments in fresh tealeaves are chlorophylls and carotenoids. During
oxidation, the green colour of tea chlorophylls is converted to black pigments called pheophytins. This
conversion leads to the dark appearance of finished oxidized teas. Chlorophyll comprises two organic
compounds with similar structures; one is called chlorophyll a. When chlorophyll a loses the magnesium ion
from the centre of the ring, and it is replaced with two hydrogens, it forms an olive-brown coloured compound
called pheophytin a.
3:19Skip to 3 minutes and 19 secondsThe aroma of tea is made up of hundreds, perhaps thousands, of flavour
and aroma compounds that exist in trace amounts. Many of these aromatic compounds do not exist in fresh
tealeaves, but are formed during processing. The flavour and aroma of each tea depends on a wide variety of
combinations of these compounds, hence it is called aroma complex. So, how do we know that these organic
compounds are present in tea? Analytical techniques, particularly high performance liquid chromatography
(HPLC) has proved very useful in this respect. A pump is used to pass a pressurized liquid solvent through a
column filled with a solid called the stationary phase.
4:04Skip to 4 minutes and 4 secondsA sample is injected into the solvent stream and each component in the
sample interacts slightly differently with the stationary phase, causing different flow rates for the different
components and leading to the separation of the components as they flow out the column. The presence of an
organic compound can be detected using an ultraviolet absorbance detector, which produces a UV spectrum.
An HPLC-UV spectrum is a plot of UV absorbance versus time. The retention time can be used to identify the
organic compound by comparison with standards (comparing the retention times of known compounds injected
into the same type of column, operated under the same conditions) and the area under the peak can identify its
concentration.
4:43Skip to 4 minutes and 43 secondsSo, this method is extremely useful in separating, detecting and quantifying
the components in tea, including caffeine.
A Sweet Solution
Sugar (sucrose, C12H22O11) is a very popular additive, and understanding how it interacts with water and
other molecules is important when considering how it influences the flavours of food and drinks.
Research at York has shown that sugar has an important effect in reducing the bitterness of tea and
coffee, not just by masking it but by affecting its fundamental chemistry.

Caffeine (C8H10N4O2), as well as acting as a stimulant, is partly responsible for the bitter taste in tea and coffee.
In water, caffeine molecules tend to stick to each other, and this is further enhanced by the addition of sugar.
As sugar causes the caffeine molecules to clump together, it takes away the bitterness of tea and coffee – as
they clump together there is less surface area to arouse our tastebuds and so we find it harder to taste them.

For the first time, work at York suggests that the underlying cause of the clumping is the affinity between sugar
molecules and water, which in turn makes the caffeine molecules stick together (or aggregate) in order to avoid
the sugar. This work will help food scientists in their development of new tastes.

Sugar-Tax?
In the UK, a levy on sugar-sweetened drinks has been introduced to combat child obesity. It was argued the
NHS could save billions of pounds and also thousands of lives in a generation by weaning the public off its
sweet tooth.

Today, research has shown that children and teenagers are consuming three times the recommended level of
sugar. But, some people object to being taxed for something that causes no harm if eaten in moderation and
they don't want the 'nanny state' interfering in their choices.

What do you think?

The Aspartame Controversy

Instead of using sugar to sweeten tea, many people use the artificial sweetener aspartame (C 14H18N2O5). The
use of this sweetener has attracted controversy, with claims that it is linked to health problems, including
cancer. However, following numerous scientific studies (it is one of the most rigorously tested food ingredients),
aspartame hasn’t been linked conclusively to any specific health problems, other than for people with
phenylketonuria (a rare genetic disorder in which the body can’t break down the amino acid phenylalanine
(C9H11NO2), which is produced when aspartame is broken down in the body). Although there are no
substantive doubts over the toxicity of aspartame, there is an ongoing debate about the possible influences of
non-caloric sweeteners on body weight, and so scientific studies continue.

Those of you who are interested in practical aspects, and who would like to know how to separate a mixture
using High Performance Liquid Chromatography (HPLC) (as mentioned in the video above), might like to see
this video, made to help prepare our undergraduates before their practicals.

https://youtu.be/RcnyZbpMyUk
Taste test

Described as one of the best jobs in the world, but one that few know exists, tea tasting is an
art as esoteric as whisky blending or wine connoisseurship. It takes five years to train a
palate to identify, blindfolded, the origin and blend of each sample and an expert is
expected to be able to detect not only the country that produced a certain batch, but the
region. The flavour of tea is influenced by the soil, altitude and climate of where it is grown
and the timing of the leaf plucking – as there are 1,500 different varieties, from 36 countries,
which all need to graded, it is far from straightforward. Expert tasters sip, grade and blend
while moving along dozens of tea samples, slurping the liquid from a spoon to ensure it
hits all the taste buds at high speed, then expelling it into a wheeled spittoon. Each day, a
taster can sample between 200 and 1,000 teas, adjusting blends of different teas to ensure
that the company’s brand remains constant – a popular supermarket brand contains a
blend of up to 35 different teas, each batch remaining constant in quality, character and
flavour. Ensuring that favourite tea brands always taste the same is big business. In 2014,
one manufacturer insured its master blender’s tastebuds for £1 million!

The highly sensitive palates of professional tea tasters would no doubt have detected any traces of
poisons in tea, so well-liked by crime writers. Agatha Christie fans may remember morphine
poisoning in ‘Sad Cypress’, while Sherlock Holmes enthusiasts may recall the dramatic restaurant
scene in the film ‘A Game of Shadows’, when Irene Adler sips from a contaminated teacup. Now is
your turn to turn detective to analyse the components of a brew. The following are descriptions of
compounds A-F that have all found their way into a pot of tea.

A: An odourless naturally occurring acid with a sour taste that is used as a food additive and
preservative (it is an E number); at a dose of 417 mg/kg it can cause vomiting

B: A hydroxide that may cause severe burns, accelerates corrosion and rusting of many metals,
and may cause electrical failures

C: A common naturally occurring antioxidant acid that has been shown to have anticancer
properties; it is used to make dyes and inks and on heating to over 200 °C produces carbon
dioxide

D: Made by microorganisms in the rumen of sheep and cows, it helps keep the body’s nervous
system and blood cells healthy; the recommended adult daily allowance is 1.5-4 mgs a day

E: A natural fructofuranoside that can cause gum disease and is a primary cause of cardiovascular
disease; on heating it melts and changes colour from white to yellow, brown then black

F: Triggers a cooling sensation when inhaled, eaten, or applied to the skin, and is used in
decongestants, perfumes and some cigarettes; prices for pure crystals are typically in the £7-15/kg
range

Which of A-F is most unlikely to find its way into your favourite cup of tea?
Almost 500 years ago, chemist and Swiss physician Paracelsus specified the basic principle of toxicology: “All
things are poison and nothing is without poison; only the dose makes a thing not a poison.” This is often
shortened to: “The dose makes the poison.” So, a substance that contains toxic properties can cause harm
only if it occurs in a high enough concentration. In other words, any chemical (even water and oxygen) can be
toxic if too much is ingested or absorbed into the body. The toxicity of a specific substance depends on a
variety of factors, including how much of the substance a person is exposed to, how they are exposed, and for
how long.

Also, does anyone remember the ‘dihydrogen monoxide hoax’? This called for dihydrogen monoxide, or water,
to be banned, regulated, and labeled as hazardous. It illustrated how a lack of scientific literacy and an
exaggerated analysis can lead to misplaced fears!

Here are the names of the compounds:

A = Citric acid (in lemon juice)

B = Water
C = Gallic acid (in tea leaves)
D = Vitamin B12 (in milk)
E = Sucrose or table sugar
F = Menthol (in peppermint tea)

Did you pick the right one?

The stages of brewing beer
Beer is a fermented aqueous drink that is made from four main ingredients: water, hops, yeast and
barley malt. Barley malt is sometimes partly made from rice, corn or wheat, all of which are very starch-
rich. Each of the four components is added at different times during the beer making process.

We begin the beer making process with the mash, which is the name of the slurry formed from mixing barley
malt and water. Barley malt makes up the body of the beer, and typically 200 g of barley malt is needed to
make a litre of beer. The amount of barley malt added to the water determines the strength of the resultant
beer, and the type of malt (rice, corn or wheat) used controls the colour of the final product.

The mash is then heated to around 60 °C. At this temperature, the enzymes in the malt – called amylase and
protease – are able to catalyse the breakdown of starch and proteins in the mash. This produces a mixture of
sugars and peptides. To ensure that the amylase and protease enzymes are present in mash, harvested barley
has to go through a process called germination to turn it into the barley malt that is used in the mash.

To start germination, barley is left to soak up water in steeping tanks for a few days. The soaked barley is then
aerated at around 15 °C in a large room, where it is constantly turned to promote sprouting. The aeration takes
around five days and the resultant barley is called the green malt. The green malt is then dried at temperatures
over 48 °C, which stops sprout growth. Changing the drying temperature will change the type of malt produced.
This dried barley is the barley malt that goes into the mash; it is full of sugars, starch and the enzymes that are
needed in the mash to catalyse the protein and starch degradation.

After the enzymes have catalysed the starch and protein degradation, the mash is filtered. What remains is a
sugar solution that is called the 'wort'. The wort is put into a brewing kettle, then boiled, and hops are added, for
at least an hour. Only a few grams of hops are needed per litre of beer (much less than the barley malt), but it
is a very important ingredient in the beer making process. When the hops are added, they form insoluble
complexes by bonding with polypeptides and proteins, which is important in stabilising the beer foam and acts
as a steriliser.

When the wort has been cooled to room temperature, and the used hops are removed by filtration, the
resulting solution is called 'hopped wort'. This solution is then moved into a fermentation vessel and the yeast is
added. The yeast cells in the hopped wort convert sugars into ethanol and carbon dioxide; this process is
called anaerobic (without oxygen or air) fermentation.

Top versus bottom fermentation

There are two types of fermentation called top and bottom. In top fermentation the yeast is able to function at
room temperature and it does not get denatured until the concentration of ethanol reaches 12%. When the
activity ends, the yeast collects at the top of the fermenter with the consistency of a dense foam. Conversely,
in bottom fermentation the yeast is only active below 5 °C and when the ethanol concentration is less than 5%.
Once the yeast stops working, it settles on the bottom of the fermenter.

The fermentation process is completed in one week, and yields a mixture called 'young' or 'green' beer. This
young beer is not drinkable, as it contains bad-tasting and smelly compounds that are a result of the
fermentation process. Two compounds that occur in high concentrations are butane-2,3-dione and pentane-
2,3-dione (both contain two ketone functional groups, hence the name dione) and they have very buttery
tastes, which is highly undesirable in lager.
The final act – conditioning
To allow these unwanted, foul tasting compounds to decompose, the beer has to undergo conditioning. This
period can take anywhere from a few days (for ale) to several weeks (for lager). The young beer remains in
contact with the yeast to allow the development of a clearer appearance and a more mature flavour. During the
conditioning, enzymes within the yeast are able to reduce both butane-2,3-dione (C4H6O2) and pentane-2,3-
dione (C5H8O2), converting them into butane-2,3-diol (C4H10O2) and pentane-2,3-diol (C5H12O2) respectively,
until they fall below critical values (<100 ppb or 0.1 mg per litre of lager).

The reduction of butane-2,3-dione occurs sequentially; first to form 3-hydroxybutanone and then butane-2,3-
diol. The reduction of pentane-2,3-dione is also sequential; unlike butane-2,3-dione, pentane-2,3-dione is
unsymmetrical, therefore, on reduction, two structural isomers are formed depending on which ketone is reduced
first. Note that both hydroxy-ketone isomers form the same reduced product (pentane-2,3-diol).

Once conditioning is complete the yeast is removed by filtration through porous rock. Yeast that has been used
in the beer making process is then sold as baker's yeast (yeast used as a leavening agent for bread-making
and baking), as bedding for growing mushrooms, for use in ethanol fuel and as a feeding supplement for
livestock due to its high protein and vitamin B levels.

The beer is then packaged and sold. An important step in the packaging process is the addition of CO 2, under
pressure, resulting in the formation of beer foam. For the stability of the packaged beer, it is important to
ensure there is no oxygen in the beer containers, as we will see later.
Historically, beer has faded in and out of popularity. For example, it was a necessity in medieval society due to
the practicality of drinking beer over dirty water, while it is currently outlawed in most of the Middle East. But the
current trend in increasing production levels suggests the world’s fondness for beer isn't going to fade anytime
soon.

See in the downloads section below for a set of images that summarise the key stages of brewing beer (that
include the chemical structures above).

Finally, a toast
Started in 2016 in Yorkshire, Toast Ale is a company that brews beer from unsold bread and the discarded
heels of loaves from sandwich makers (apparently, in the U.K. up to 44% of bread is wasted - staggeringly
high!). All profits go to fighting global food waste.

3.7_Brewing_process.pdf
The organic flavour components of beer
0:06Skip to 0 minutes and 6 secondsBeer is one of the most popular and largely consumed alcoholic drinks in the
world. The total world’s beer production amounts to around 200 million kilolitres. It is a complex alcoholic
beverage, containing many flavour-active compounds, present in a wide range of concentrations, including
compounds called humulones. Beer flavour is a delicate balance of all these compounds, and for the brewers,
the challenge is to produce their products with consistent flavour, and to maintain the flavour balance for as
long as possible in the market place. Malting and brewing technology have remained very traditional, but the
efficiency of the process has improved through a better understanding of the technology and the underpinning
chemistry.
0:53Skip to 0 minutes and 53 secondsInnovation in the brewing industry is driven by cost reduction, for example,
by more efficient use of the raw materials and lower energy consumption, and the need for improved quality
and safety of the final product. There are hundreds of compounds present in the hops used to produce beer, of
these, the most interesting are the humulones, specifically the non-bitter tasting (-)-humulone. Humulones have
a carbon atom bonded to four different groups. The two ways of arranging the four substituents around the
chiral centre, shown as (-)-humulone and ( )-humulone, are mirror images of each other (and so they are
enantiomers); however, only (-)-humulone is present in hops.
1:36Skip to 1 minute and 36 secondsDuring the brewing process, (-)-humulone undergoes a rearrangement
reaction to form cis- and trans-isohumulones; the compounds responsible for the majority of the bitterness in
beer. In cis-isohumulone, the -OH and the alkene (Me2C=CHCH2) groups are arranged on the same side of
the planar five-membered carbon ring, whereas, in trans-isohumulone, the -OH and the alkene Me2C=CHCH2)
groups are arranged on opposite sides. The rearrangement reaction of (-)-humulone is known as an
isomerisation reaction. This is because cis- and trans-isohumulones are both structural isomers of (-)-
humulone; they all have the same structural formula (C21H30O5) but the atoms are arranged in different ways.
Isohumulone is the compound responsible for the main bitterness in beer and is formed from boiling hops
during the manufacturing process.
2:27Skip to 2 minutes and 27 secondsBitterness units (BU) are used to describe the extent of the bitterness of a
beer. A single bitterness unit is equal to one milligram of isohumulone per litre of beer. The bitterness units in
different beers ranges from around 10 bitterness units in a standard lager, to around 45 bitterness units in
certain dark, malty beers, where higher levels of bitterness are needed to counter the sweetness of lots of malt.
As well as the bitterness unit, the flavour components of beer can be measured in units called flavour units. A
flavour unit of a flavour-active compound is measured in multiples of the flavour threshold (the lowest
concentration of detection by aroma or taste) and is recorded as a concentration.
3:11Skip to 3 minutes and 11 secondsBeer has primary, secondary and tertiary flavour compounds. Primary
flavour compounds are found in beers at concentrations greater than two flavour units, these include
isohumulone (ranging from 2.0 flavour units in standard larger to 12 flavour units in dark, malty beers), ethanol
(found at about 2.9 flavour units in a standard beer), CO2 (at concentrations between 3.0 to 5.5 flavour units)
and various other related hop compounds (2.0 to 12 flavour units). Secondary flavour compounds have
concentrations ranging from 0.5 to 2.0 flavour units; it is the secondary compounds that create the distinct
flavour of a beer. These compounds include fruity-flavoured esters (such as, 3-methylbutyl ethanoate, or
isoamyl acetate, and ethyl hexanoate), alcohols with carbon-chain lengths greater than two, and polyphenols.
3:48Skip to 3 minutes and 48 secondsTertiary flavour compounds are found in the concentration range of 0.1 to
0.5 flavour units; an example being ethyl octanoate, which imparts an apple flavour to the beer. The removal of
a single tertiary flavour compound would not directly affect the resultant flavour of the beer, but a combination
of all the tertiary flavour compounds, working together, does influence the final taste. Overall, with a beer
containing hundreds of organic compounds and around 2000 ales, lagers and keg beers being produced in the
UK alone, it is important for breweries to ensure they are aware of flavour and bitterness compounds to
develop unique and desirable brews.

Different beer types

Originating in the Middle Ages or later, the majority of beers that are drunk today are considered to be
recent inventions. There are several-dozen beer styles currently in acceptance (though many new
brews still fight for a chance to be accepted), and we are going to look at the different families into
which beers are divided.
Flavour, strength, appearance of the head and colour of the beer are all relevant in determining the
characteristics of a beer style. These characteristics are influenced by the quantities and types of the
ingredients used in the brew, as well as the brewing processing conditions.

General classifications of beer style families are split by the town of manufacture and the colour of the resultant
brew, however chemistry-wise this is very unrevealing; hence, the following classifications attempt to
subjectively sort beers by their flavour characteristics.

The difference between lager and ale

To a drinker, the look, smell and taste determine the differences between a lager and an ale. Whereas ales
have a fruity 'estery' taste, lagers are described as more 'crisp'.

However, the main differences between the two styles of beer come down to the brewing process. In 'Stages
of Brewing Beer' we discussed the two different types of yeast that can be used in the beer making process:
top or bottom fermenting yeast.

When brewing ales, top fermenting yeasts are used and the barrels are kept at the cellar temperature (warm).
Top fermentation means that the ethanol concentration can reach up to 12%, a much higher concentration than
the bottom fermenting yeast would allow.

Conversely, lagers use bottom fermenting yeasts and ferment at cold temperatures, which would make the top
fermenting yeast stop working. After fermentation the beer is 'lagered'; this is the period of time after the yeast
has been added when the beer is left in the cold for several weeks in order to age. The slow process of
fermentation at cold temperatures is what results in the 'clean' taste.

Bitter ales: For bitter ales, a moderate colour is seen. The bitterness can stretch from moderately high to very
intense, whereas the maltiness is generally low for this style.

Bitter lagers: The malty flavour in many lagers is not very noticeable, as the dominating flavours in the brew
are bitter.

Malty ales: This family of beer styles incorporates ales that have the dominant flavour of maltiness; the
maltiness is a product of heat treatments (where the Maillard reaction occurs during browning, as we will see
later), which also results in most malty ales having a dark colour (due to compounds called melanoidins).

Phenolic ales: This class of beer is described as having the flavour of cloves (due to the presence of 2-
methoxy-4-vinylphenol), and is usually characteristic of wheat-based and barley-based ales. The phenolic
flavour is considered to be an off-flavour in other beer styles.

Speciality beers: The speciality beer family encompasses all beers with flavours not usually found in beers
such as spices, smoky flavours, and fruit beers among others.

Overall, there are many different types of ales and lagers, allowing beer drinkers to pick and choose the brews
that they find most pleasant tasting.

Does beer contain any nutritional value?

A popular question! Research has shown that in small amounts, beer can have slight nutritional and also health
benefits. Studies evaluating the relative benefits of wine, for which the health benefits are already well
understood, versus beer, suggest that moderate consumption is also associated with lower rates of
cardiovascular disease. From a nutritional standpoint, beer contains more protein and B vitamins than wine
and the antioxidant content is equivalent to that of wine, but the specific antioxidants are different (because
hops and grapes contain different antioxidants). However, these nutrients exist in small amounts, especially
compared with the large amount of carbohydrates in beer (a pint of beer, 568 mL, contains around 150-180
calories). A heavy intake of beer (typically >500 mL per day) is often associated with bigger waists, which has
been explained by the liver metabolising (or ‘burning’) the alcohol in beer, rather than our body fat (especially in
the belly). Although not everyone agrees! However, it is generally advised not to drink beer or any other alcoholic
beverage for health benefits, while over-consumption wipes out any positive health benefits and increases the
risk of liver cancer, cirrhosis, alcoholism, and obesity.

Different countries have different labelling requirements for alcoholic beverages. In 2018, in the U.K., The
Royal Society for Public Health called for a change in alcoholic drinks labelling, citing a public “awareness
vacuum” on how alcohol affects health. Their research indicated that 81% of the public wanted to see drinking
guidelines on labels, like those on food items which clearly show a low, medium or high content of fat,
saturated fat, sugars and salt. In 2022, in England, mandatory calorie labelling will be introduced in many
restaurants, cafes and takeaways. Interestingly, in New York, where calories have been listed on menus since
2008, these menus were seen to encourage some people to order dishes with more calories, not fewer,
perhaps in a bid to get value for money. A study of a large restaurant franchise highlighted a small decrease in
mean calories purchased per transaction, but this reduction diminished over one year of follow-up. So, more
generally, do you agree with this principle, and would you like to see your foods and beverages more
clearly labelled?
Tools of the trade: understanding free radicals

Free radicals are species that have one or more unpaired valence electrons and are
typically very reactive. The picture below shows all the valence electrons on the oxygen
atom in the hydroxyl radical (i.e. those electrons in the outermost shell of oxygen), which
include two lone pairs of electrons (••) – notice that there is also one unpaired electron,
making this a free radical (sometimes simply called a radical).

Free radicals are usually represented with a single dot next to the atom with the unpaired electron (like a lone
pair, but with a single electron instead of two). An example of the common representation of the hydroxyl
radical is shown below.

In the mechanisms that we studied in week one, any bonds broken occurred via heterolytic cleavage (also
known as heterolysis) and we used double-headed curly arrows. A double-headed curly arrow indicates the
movement of a pair of electrons.

Conversely, free radicals are formed by homolytic cleavage (also known as homolysis). This is the
process by which a covalent bond is broken symmetrically with one electron going to each atom.
A single-headed curly arrow is used to represent the movement of a single electron.
Homolysis generates two free radicals, as shown in the following example. The covalent O–O
bond in hydrogen peroxide undergoes homolysis to form two hydroxyl radicals. H2O2 contains a
very weak O–O bond, which can undergo homolysis at temperatures below 150 °C, or on
irradiation with UV light. The process of forming free radicals is called initiation.

Radical reactions that produce new radicals are called propagation reactions. H-O• is a highly reactive radical
and can abstract a hydrogen atom from an organic compound to form a strong O-H bond in water.

Now you try

Draw the radicals formed on homolysis of the O–O bonds in the following compounds.

(a) (H3C)3COOC(CH3)3

(b) (CH3CO2)2

(c) PhCO2OC(CH3)3
The stability of beer
Trying to maintain the flavour stability of beer begins before the bottles even leave the brewery. Due to
the presence of hop-bitter compounds, ethanol and acidic compounds (such as propanoic and ethanoic
acids), the finished beer has a high level of resistance to being spoiled by microbes; in fact, all cask
beers and some keg and bottle beers are untreated (the resistance relying on the stability of the beer).
Leaving a beer untreated can have an impact on its shelf life, but the attraction of drinking fresh beer
outweighs the shortened lifetime.

The presence of microbes can result in an off-flavour in a beer however, so mainstream commercial breweries
treat their beers in one of two different ways in order to try and extend their shelf life. This involves using very
fine filters or pasteurizing the beer by heating it to 60 °C, which eliminates any microbes before packaging in a
sterile environment.

Catty beer
After packaging, as time passes, the taste of beer diminishes and it can develop a stale quality – its bitterness
is replaced with sweet, toffee-like flavours. Eventually, as these sweet flavours continue to rise, the beer can
take on a 'catty or cat-pee like' taste, officially called ribes. When the 'catty' taste reaches its maximum, a
cardboard or 'straw-like' taste dominates, which indicates staleness. Unfortunately, little is known about how
the 'catty' taste develops in beer, but it has been linked to the presence of a smelly thiol called 3-mercapto-3-
methylbutyl formate, Me2C(SH)–CH2–CH2–O–CHO, amongst other compounds. What is known is that if there
is no oxygen in the packaged beer, then no catty taste develops.

Going stale
(E)-2-Nonenal (or trans-2-nonenal, C9H16O) is the main contributor to the stale flavour in beer. It is often cited
as the most important stale compound of lager beer, because its concentration is seen repeatedly to increase
during ageing to levels above the flavour threshold. However, other compounds with similar structures
(containing both a carbonyl (C=O) group and a carbon-carbon double bond (C=C)) will also contribute to the
cardboard or straw-like flavour.

If oxygen is present in the packaged beer it does not contribute to the production of the staling compound (E)-
2-nonenal. Instead, it is suggested that this compound is made earlier during the brewing process, perhaps
from the oxidation of fatty acids in hop, such as linoleic acid (C18H32O2). In the fermentation process, (E)-2-
nonenal may bind to amino acids, which can function like a protecting group, and stop the reduction of the C=O
bond in (E)-2-nonenal (by the yeast) to a tasteless alcohol. Then, during storage, (E)-2-nonenal can be
released, resulting in the beer gaining a stale taste. However, controversy still exists over the relative
importance of the different mechanisms in brewing practice and to what extent staling aldehydes are formed at
the various stages during beer storage.

Light-struck
You might have noticed that beer bottles are normally made of glass that is brown in colour. This is important
as it reduces the beer’s exposure to sunlight, which can result in 'light-struck' beer; when a beer becomes light-
struck, it takes on a skunky flavour. This process can only occur in beers made from hops as the mechanism
takes place on hop-derived iso-alpha acids.
The process of a beer becoming light-struck occurs through a mechanism involving free radicals –
species containing an unpaired valence electron. Energy from the sunlight is transferred to the
hop-derived iso-alpha acids, giving the compound enough energy to break the (HO)C–C(=O) bond
leading to the six-carbon side chain (in pink). This occurs through homolysis, where the bond

cleaves symmetrically.

The smaller side-chain radical can then 'kick out' carbon monoxide in a propagation reaction, commonly called a
decarbonylation reaction.

The resultant free radical, with a five-carbon chain, can then react with sulfur-containing proteins
(abbreviated below as HS-R), to make 3-methylbut-2-ene-1-thiol (or MBT), which has an odour

similar to that of skunk spray. The precise mechanism of this step is still unclear.

Using brown glass bottles helps to ensure that very little light, with the correct wavelength, can reach the hop-
compounds to produce light-struck beer. Beer is also frequently sold in cans or kegs, which eliminate the
possibility of light-struck beer altogether, as no light can penetrate the container. If clear or green glass bottles
are required for marketing reasons, then hydrogenated hop extract replaces the natural version, which
effectively 'skunk-proofs' the beer.

The use of hydrogenated hop extract results in the removal of one or more of the side chain carbon-carbon
double bonds, which can result in two different scenarios: either the light is hindered from being able to interact
with the molecule, or the breaking of the bond to the side chain can still occur, but the radical that is formed
lacks a C=C bond. The absence of a C=C bond means the radical is unstable, so only very small amounts are
formed. (A C=C bond can stabilise an unpaired electron on an adjacent carbon, by resonance, i.e. the ‘dot’ can
be spread over the C=C bond and this ‘spreading’ or delocalisation helps to stabilise the radical. For example,
the allyl radical, H2C=CH–CH2•, is more stable, and more easily formed, than the propyl radical, H3C–CH2–
CH2•.)

After the lengthy process of brewing beer, it is important to take measures to increase shelf life and the stability
of the final brew… because no one wants to drink stale or skunky beer (even if it is safe to drink)! On saying
that, if you want to try a taste test then take two beers in clear bottles. Leave one in the light for a week or two
and the other in a dark cupboard and then try them side-by-side (for more extreme results leave one in direct
sunlight).

See the downloads section below for a set of images that further illustrate the flavouring components (including
the chemical structures above).

Does beer really taste better out of a bottle?

Research, both controlled and anecdotal, suggests that most people would say that the same beer tastes
better when it’s in a bottle versus when it’s in a can. However, when you take away any knowledge of the beer
and what the original packaging format was, the preference for bottled over canned beer disappears. So, in
essence, people’s preference for bottled over canned beer is likely to be a psychological phenomenon. It appears
to be due to preconceived preferences for certain packaging types that influence consumer preference for
beer, which is a concept well known to beer marketers.

The same consumer preference for bottles over cans has been seen in other drinks like Coca-Cola – some
think the subtle variations in taste are due to polymers that line aluminium cans absorbing small amounts of
soluble flavour components from the drink. Also, a preference for glass over plastic bottles has been linked to
plastic being more permeable to carbon dioxide – so the fizz will leak out of a plastic bottle of Coke quicker
than it would from a can or glass bottle. But do you agree? Why not try this experiment at home, by taste-
testing a fizzy drink from a can versus a plastic and/or a glass bottle – is there any difference? We look
forward to hearing your results!
Tools of the trade: a further look into free radicals

The mechanism of the reaction between phenol and a H–O • radical is shown below. For the
abstraction of the hydrogen atom (in pink) by the hydroxyl radical (in green), the O–H
covalent bond in phenol must be broken and a new covalent H–O bond in water must be
formed, represented by three single-headed curly arrows.

The driving force for the reaction can be explained by bond strengths – the O–H bond in phenol is
weaker than the O–H bond in water (i.e. a weak bond is broken in a reactant to form a much
stronger bond in a product). Alternatively, you could consider the stabilities of the two radicals –
the reaction progresses to convert a reactive radical into a more stable radical. The phenoxyl
radical (PhO•) is more stable than the hydroxyl radical because the unpaired electron can be
spread around the benzene ring, through resonance. To show this, it is better to use the Kekulé
structure of the benzene ring – as shown below, notice that the unpaired electron is stabilised
by delocalisation.

Finally, radicals are destroyed when a covalent bond is formed from the reaction of two free radicals. This is
called homolytic bond formation and is known as a termination step, because it converts reactive free radicals
into non-radical products.

To show the formation of a covalent bond between the oxygen (in blue) and hydrogen (in green), a
single-headed curly arrow is drawn from each atom and they meet in the middle.

Because of their high reactivity, oxygen-centred free radicals, such as HO•, can cause damage to
cells in the human body, which can result in disease. We will learn more about this in the following
section.
In summary
Initiation reactions form radicals
Propagation reactions convert radicals into different radicals
Termination reactions convert radicals into non-radicals

Now you try

Classify the following as initiation, propagation or termination reactions.

(a) Cl• + CH4 → HCl + •CH3

(b) Cl• + •CH3 → ClCH3

(c) •CH3 + Cl2 → •Cl + ClCH3

© University of York
Antioxidants
Antioxidants are very important in maintaining the health of the human body. They play a vital role in
stopping, or reducing, the oxidation of other biomolecules.

What is oxidation?
Oxidation is the process by which a molecule is oxidised; this occurs due to the loss of electrons. Oxidation can
be helpful in certain situations: for example, oxygen-based cleaners (bleach is a solution of sodium
hypochlorite, NaOCl, dissolved in water) and hydrogen peroxide sterilisers found in many household cleaning
products use oxidation. However, it can also be incredibly destructive, like the blackening of a potato or the
browning of an apple.

An example of oxidation is the rusting of iron. When iron is in its pure, unoxidised form, it has three electrons
that can be removed easily. When iron undergoes oxidation, the three easily removable electrons are lost and
it has a 3+ charge. This oxidised Fe3+ ion is a brittle, red powder, in stark comparison to the structurally sound
unoxidized iron. It is clear why you would want to try to minimise the effects of oxidation on structures made out
of iron.

Oxidation does not only affect molecules and compounds outside the body. The presence of free radicals, within
the body, can have very destructive effects. For example, oxygen-centred radicals, such as HO •, cause very
serious damage to cells resulting in ageing and disease. This process is called oxidative damage because
reactive oxygen-containing species, such as HO•, are involved and another definition of oxidation is the
reaction of a substance with oxygen (or a reactive oxygen species).

An antioxidant: vitamin C
Antioxidants are important because they are able to react with free radicals, turning them into unreactive
species. The removal of reactive oxygen-centred radicals helps to protect our body from oxidative damage, and
nature has different ways of doing this, including using antioxidants. Vitamin C, or ascorbic acid, is a type of
antioxidant (which has the structure shown below, and the chemical formula C6H8O6).
Vitamin C is naturally found in all citrus fruits and many non-citrus fruits such as papaya, pomegranate and
raspberries. Unlike humans, many animals can make their own vitamin C; for example, goats can produce over
13 g of their own vitamin C every day. Humans need a much smaller amount of vitamin C; the daily
recommended maximum is 90 milligrams for an adult male and slightly lower for a female, but it is still an
essential vitamin in the human diet. Without consuming an adequate amount of vitamin C, humans are
susceptible to developing an illness called scurvy, which results in tiredness, fever, bleeding gums and, in
extreme cases, death.

Working in harmony
Vitamin C works in harmony with vitamin E (a group of eight compounds, including alpha-tocopherol which has
the structure shown below, and the chemical formula C29H50O2) to protect the body from oxidative damage.
Vitamin E is another antioxidant. While vitamin C is water soluble and is important for tissue growth and repair,
vitamin E is fat soluble and plays an important role in protecting cell walls from damage caused by oxygen-
centred radicals (e.g. RO•), which are formed as by-products of metabolism (the chemical processes that occur
within a living organism in order to maintain life).

The process of metabolism takes O2 and converts it into 2O2- (each O2- then accepts two H+ ions,
or protons, to form water); this means that a total of four electrons needs to be gained (two
electrons per oxygen atom). Due to a very high energy barrier, only one electron can be gained at
a time resulting in the production of various oxygen-centred radicals (e.g. RO•) as O2 is converted
into 2O2-. Here is a simplified diagram to show how vitamin C and vitamin E work together to

protect cell walls.

Vitamin E donates a hydrogen atom to a reactive oxygen-centred radical (RO•) forming an alcohol,
ROH, and a vitamin E radical (an aroxyl radical, ArO•), which is much less reactive (destructive).
The vitamin E radical can then abstract a hydrogen atom from vitamin C, forming a vitamin C
radical, which is converted back into vitamin C by an enzyme; the cycle then continues ensuring
that cells are protected from oxidative damage.

The structure of vitamin E contains a substituted phenol. A destructive oxygen-centred radical, RO •, selectively
abstracts the hydrogen atom shown in blue, in vitamin E. This is because the O–H bond in vitamin E is
relatively weak, much weaker than the RO–H bond formed in the alcohol (i.e. the reaction is driven forward by
the formation of a stronger O–H bond). Notice than when the phenolic hydrogen is abstracted the resulting
oxygen-centred radical is stabilised by resonance – the unpaired electron can move around the benzene ring (in

pink). As it is more stable than the RO• radical it is much less destructive and harmful.

Rancidity and antioxidants

Fats and oils, such as olive oil, butter and margarine, are widely used in cooking and contribute to the
flavouring of foods. However, they are susceptible to oxidation, which is responsible for the formation of off-
flavours and aromas, described as rancidity. The oxidation of such oils at room temperature is
called autoxidation, and it is influenced by factors including light and temperature (cooking oils should be
stored in a cool, dark place). Autoxidation involves radical reactions, and unsaturated fats, such as linoleic acid,
are particularly prone to autoxidation. This is because their structures include a skipped diene unit(s),
~CH=CH–CH2–CH=CH~, which is particularly susceptible to reaction with oxygen-centred radicals. An oxygen-
centred radical (e.g. RO•) abstracts a hydrogen atom on the carbon in-between the two double bonds (this is a
particularly weak C–H bond). This gives a carbon-centred radical, ~CH=CH–CH•–CH=CH~, that is stabilised
by delocalisation involving the neighbouring two C=C bonds. This radical then reacts with oxygen to give a
range of oxidised products, that are responsible for the undesirable flavours. (For details of the reaction
mechanisms see the autoxidation pdf below.)

Antioxidants are often added to foods to slow down their oxidation and extend their shelf-life (e.g. vitamin E can
be added to vegetable oils and margarine). However, fans of potato crisps (potato slices deep-fried in cooking
oil) will note there are no added antioxidants – so how do the crisps stay fresh in the packet? The answer is to
remove oxygen, and store them under nitrogen gas! However, once opened, they come into contact with
oxygen and are prone to autoxidation, hence that unpleasant rancid (stale) taste on standing.
Do packaged foods with labels promoting antioxidants boost your health?
No, not necessarily. Outside of science, it has become more of a marketing term, to attract customers, rather
than a scientific term. For example, food manufacturers may add vitamin C or E to their product, in small
amounts, to ensure shelf-life (e.g. to delay the oxidation of fats that make them go rancid), and then label it as
containing antioxidants, presumably in the hope of boosting their sales. Also, some mention antioxidant
‘power’, represented by ORAC (which stands for oxygen radical absorbance capacity). But this measures
antioxidant activity in a test tube, not in the body, and it has not been validated, so many believe it misleads
consumers. The best health advice remains to get your antioxidants from a well-balanced diet containing plenty
of varieties of fruit, vegetables, whole grains, and legumes.

Autoxidationmechanism.pdf
Antioxidants: vitamin C
When humans are suffering from vitamin C deficiency, enzymes called prolyl hydroxylase
(that catalyse important reactions in the body, called hydroxylation reactions, and aid in the production
of collagen) are oxidised and hence deactivated.

A hydroxylation reaction results in the introduction of one, or more, hydroxyl groups (HO–) into an
organic molecule. For example, the dihydroxylation of ethene is shown. Ethene is an alkene, so it
contains a carbon-carbon double bond (a so-called unsaturated organic compound) and has the
molecular formula C2H4. The hydroxylation reaction results in the breaking of the carbon-carbon
double bond and the addition of two HO– groups, one on each carbon.

In the body, enzymes catalyse this type of reaction. Oxidative stress on the enzymes leads to their deactivation
and, hence, a reduced rate of hydroxylation, resulting in disease. Taking in an adequate amount of vitamin C
reduces the oxidation of this class of enzymes and so prevents disease.

In 1970, Linus Pauling (winner of two undivided Nobel prizes, the first in 1954 for chemistry followed by the
Nobel Peace Prize in 1962) controversially claimed that by taking 1 g of vitamin C per day (more than 10 times
the daily recommended amount) a person could reduce their chances of getting a cold by up to 45%.

Over the course of several years, Pauling increased his intake to 18 g of vitamin C daily (around 300 times the
daily recommended amount), stating that, by consuming so much vitamin C, the colds he would experience
frequently throughout the year had stopped occurring. However, many double-blind studies (where neither the
doctor nor patient know if they have received a placebo or a real sample) indicated that vitamin C does not
stop a person from catching a cold.

In 1973, the Linus Pauling Institute of Science and Medicine was founded, where research into the significance
of vitamins and micronutrients in enhancing human health and preventing chronic disease, was undertaken.

In 1976, Pauling and a colleague reported that results from a study of one hundred 'terminal' cancer patients
taking 10 g of vitamin C a day showed these patients survived up to three to four times longer than those not
taking vitamin C in such high quantities. However, the results of the study were not considered to be accurate
due to the two studied groups being incomparable, with blind studies confirming that consuming vitamin C did
not increase the length of survival for 'terminal' patients. Research continues and, in 2017, mixing vitamin C
with the antibiotic deoxycycline was found to be surprisingly effective at killing off cancer stem cells.
According to the NHS website, the highest tolerable limit for vitamin C intake is 1 g a day; consuming doses
higher than this could be considered an overdose and can result in gastrointestinal irritation and diarrhoea.
Even though there is no need to consume such a vast quantity of vitamin C, there is no doubt that consuming
antioxidants (including vitamin C made in the lab), are an important part of maintaining a healthy body.

The usefulness of vitamin C continues to attract media attention, whether it be for the
treatment of the common cold or targeting cancer stem cells or reducing the time patients spend
in intensive care. Do any of you worry about not getting enough vitamins and take any
supplements?

Or E300?
A food additive that has passed safety tests to allow its legal use in the European Union (EU) is given an E
number (where ‘E’ stands for Europe) – vitamin C has the E number E300 and it falls within the ‘antioxidants’
category.

E100–E199: colours e.g. E100 is curcumin (a yellow-orange dye, from turmeric)

Role: To add colour to food, which may have lost its natural colour during cooking

E200–E299: preservatives e.g. E211 is sodium benzoate (PhCO2– Na+)

Role: To stop microbes from growing

E300–E399: antioxidants e.g. vitamin E, from natural sources, is E306

E400–E499: thickeners, stabilisers & emulsifiers e.g. E412 is guar gum, made from seeds of the guar bean
Role: For example, stopping oil and water based substances from separating, such as mayonnaise

E500–E599: pH regulators and anti-caking agents e.g. E500 encompasses sodium carbonates, including
sodium bicarbonate (NaHCO3)
Role: Controlling the acid/alkaline levels of foods and ensuring powders, such as milk powder etc, do not cake
or stick

E600–E699: flavour enhancers e.g. E621 is monosodium glutamate, used to impart an ‘umami’ taste in food
Role: Enhances flavours which were lost in cooking, or adds flavour to bland foods

E700–799: antibiotics

E900–999: glazing agents, gases and sweeteners e.g. E901 is white and yellow beeswax, and E948 is
oxygen

E1000–1599: additional additives e.g. E1510 is ethanol

So, you may be surprised that many E numbers are of natural origin. Despite the fact that every E number has
gone through a lengthy testing and analysis process, they tend to have a bad press. For example, E621
(monosodium glutamate, MSG) has been linked to ‘Chinese-restaurant syndrome’, but this has never been
scientifically confirmed. On saying this, not all E numbers are innocent, for example, the food colour sunset
yellow FCF (E110) must carry a warning on the packaging saying it ‘may have an adverse effect on activity and
attention in children’. Also, sulfur dioxide, used as a preservative in dried fruits and drinks, including wine, can
exacerbate asthma. Many argue that it’s not the Es that make them unhealthy - it’s the salt, fat and sugar
content.
E numbers have divided people’s opinions for years, but where do you stand on the
subject?

Antioxidants and apples: an experiment

0:07Skip to 0 minutes and 7 secondsAntioxidants protect cells against the damaging effects of free radicals,
which are suspected culprits in heart disease, cancer and even the ageing process. Free radicals are so
reactive because they possess one or more unpaired electrons, such as the hydroxyl radical (HO). Within the
body, the hydroxyl radical can oxidise and damage lipids, proteins and nucleic acids, including DNA. To protect
itself from the radical attack, and oxidation, our body has two basic forms of protection. One method uses
enzymes, while the other uses organic compounds, such as vitamins, including vitamins C and E. Antioxidants
are essential in the diet and are found in a range of foods and drinks, including tea.
0:50Skip to 0 minutes and 50 secondsIt has been suggested that the health benefits are due to the antioxidants
they contain, which counteract the damaging effects of oxidation caused by free radicals. This activity
compares the antioxidant activities of various multivitamins on the browning of apples. Leaving slices of apples
exposed to the air leads to the apple turning a familiar brown colour. This process, called enzymatic browning,
is due to the reaction of certain enzymes with phenolic compounds, both present within the apple, that occurs
when they are exposed to oxygen in the air. These oxidation reactions produce a number of compounds
including a brown pigment called melanin. The longer the apple is exposed to the air, the more brown pigments
are formed, and the darker the apple becomes.
1:37Skip to 1 minute and 37 secondsSo, we will use the rate of browning of apple to compare the relative
effectiveness of antioxidants - we are looking to see which antioxidants best protect the apple from browning.
First, place freshly sliced apple pieces into different glasses of water together with an antioxidant tablet ???
then monitor the rate of browning over time, by inspecting the slices. Make sure you do a control reaction, as
simply soaking an apple in water alone will temporarily reduce the level of browning by restricting the amount of
oxygen in contact with the slices. Tabulate your results, noting the relative levels of browning against time, with
the multivitamins you have chosen. Take photos and don’t forget to post your ‘radical’ results.
2:19Skip to 2 minutes and 19 secondsFinally, although enzymatic browning impairs the quality and stability of
freshly cut vegetables and fruit, it is beneficial to the colour and flavour of tea and coffee - so the appearance of
browning is not always bad news.

The following ingredients will be required:

 An apple
 A vitamin C tablet (we used 99p vitamin C tablets from a local pharmacy)
 Another type of nutrient tablet (we used an inexpensive variety of A-Z multivitamins and
minerals tablets)
 Water

The following items will be required:

 Chopping board
 Knife
 One cup for each type of tablet you are testing plus one extra for a control sample

Ensure that you read through all of the instructions before beginning the experiment to ensure that you
understand exactly what will happen at each step.
Step one
Fill each cup three quarters full with cold water. Then cut the apple into three equally sized segments and drop
one into each cup.

Step two
Carefully crush up each of the tablets that you have chosen, and place one into each glass, ensuring that you
leave one glass with just water, as a control.

Make sure to stir each glass so that the tablet powder has dissolved in the water.

Step three
Once a day, monitor the apples to see how the oxidation process is progressing. Our results, on a pink lady
apple, are shown below.

Figure 1. The three apples after twenty-four hours, from left to right: The apple in water, the apple
in water containing the vitamin C tablet, the apple in water containing the A-Z multivitamins and

minerals tablet.

Figure 2. The apples after forty-eight hours, from left to right: The apple in water, the apple in water containing
the vitamin C tablet, the apple in water containing the A-Z multivitamins and minerals tablet.
Figure 3. The apples after seventy-two hours, from left to right: The apple in water, the apple in
water containing the vitamin C tablet, the apple in water containing the A-Z multivitamins and
minerals tablet.

Figure 4. The apples after ninety-six hours, from left to right: The apple in water, the apple in water containing
the vitamin C tablet, the apple in water containing the A-Z multivitamins and minerals tablet.

Examining the results

When the apple is cut and exposed to oxygen, free radicals are released and the apple undergoes the
oxidation process and turns brown (i.e. oxygen is introduced into the injured plant tissue causing enzymes in
the apple to rapidly oxidise phenolic compounds that are naturally present in the apple tissues to form the
brown-coloured products called melanin). This is seen in the first apple, the one in water. Because the apples
are in water, the rate of oxidation is reduced slightly. This is why the control glass (the apple in plain water) is
necessary, otherwise it would not be a fair comparison.

The apple in water containing the vitamin C tablet (which contains antioxidants) has a very reduced rate of
oxidation. This is because vitamin C is an antioxidant that reacts with the very reactive oxygen-centred
radicals, turning them into less reactive species. Hence, this reduces the amount of oxidative damage to the
apple flesh. This is why the apple in the vitamin C solution does not turn brown and remains fresh looking.

The apple in water containing the A-Z multivitamins and minerals tablet goes black and looks mouldy. While
there is still some protection from oxidation due to the presence of antioxidant, the over-the-counter A-Z
multivitamins and minerals tablet contain traces of 'crude oils' and 'heavy metals', which turn the apple black.
The A-Z multivitamins and minerals tablets contain many ingredients that are used as fillers and colourants,
such as titanium dioxide, magnesium stearate and corn oil. While the vitamin C tablets also contain fillers and
sweeteners, from the results, it appears that they probably do not contain any 'heavy metal' or 'crude oil' traces.

Overall, we have determined that the vitamin C tablets are the most effective at reducing the oxidative damage
and produce the healthiest looking apple, but, we would like to see the results that you have found. Upload
pictures of your apple experiment onto our open Padlet (we have included some examples from a
previous course to help inspire you) and/or on Twitter or Instagram hashtag #FLchemistry.

Another biologically important antioxidant is vitamin E, so you could compare the effects of vitamin C and
vitamin E and see which apple browns first! Also, consider trying other commonly available vitamins and/or
different types of apples. There are around 7,000 varieties of apples, but around 20 common ones, to pick
from.

By the way, another way to stop the browning is to add lemon juice. The citric acid in the lemon
increases the acidity and when the pH drops to around 4, the enzymes involved in the browning
process stop working.
Experiment adapted from: R. Torres, The Apple Oxidation Test (USANA Mega AntiOxidant vs other brands).

In the headlines – the genetically modified Arctic Apple

One of the most famous GM foods, which hit the supermarket shelves in the US in 2017, is Arctic Apples. This
is a group of trademarked apples that don’t go brown. The apples have been engineered, using a technique
called ‘gene silencing’, to stop the production (by around 90%) of the browning enzymes (called PPO). They
decay naturally, just like any other apple, but will not turn brown from bruising, cutting or biting – not in minutes,
hours or days. Unlike the first genetically engineered crops (such as the Flavr Savr tomato) the apples are not
transgenic, that is, they do not contain a gene from another species. So, despite protests from environmental
groups, perhaps this crop may be more popular with consumers.

What do you think?

Extension activity – get your teeth into it

For those of you keen to do some further practical work - we have added an extension activity in the downloads
section below. This uses denture tablets, with a dye in them, to test for the antioxidant activity of vitamin C
tablets.

Instructions_for_Antioxidant_Testing.pdf

Measuring_the_Effects_of_Antioxidants_on_Denture_Tablets.pdf
Coffee and taste
Grown 600-1800 m above sea level, coffee is a tropical plant, which was originally indigenous of
Ethiopia. The coffee plant has now spread to India, Indonesia, Brazil, Colombia and Central America. A
coffee plant yields red fruits called 'cherries', each of which holds two coffee beans that are green in
colour. These green coffee beans are distributed to consuming countries, where beans from all over
the world are mixed together. The green beans are then roasted, resulting in the colour and flavour
found in an iconic cup of coffee.

There are two primary types of coffee beans, arabica and robusta. Arabica is considered to have a nicer flavour
and robusta is generally described as being harsh and rubbery. This is due to robusta having an increased
concentration of phenols, pyrroles and sulfur-containing compounds, such as furfurylmethyl sulfide (which is
present in robusta coffee at 2200 ppb which is much higher than the 1100 ppb present in arabica), and
furfurylmethyl disulfide (present in robusta at 650 ppb compared to the 120 ppb in arabica).

The structures of furfurylmethyl sulfide and furfurylmethyl disulfide both contain a furan ring, made up of four
carbon atoms and an oxygen atom (in pink). However, the sulfide only contains one sulfur atom, while the
disulfide contains two – hence the use of 'di' in the name disulfide.

Making a good brew

It is important to get good quality coffee if you want to end up with a fine tasting brew. Grinding has an
important role in developing the final flavour of your cup of coffee. Coffee particle sizes that are too large will
result in weak coffee due to poor extraction of the organic compounds from the coffee particles, whereas
grinding your beans too fine will lead to very fast extraction of all the flavour compounds, including the bitter
ones.

When making a cup of coffee, the water temperature plays a direct part in the overall bitterness of the final
brew; using water that is too hot (greater than 96 °C) risks burning the coffee, resulting in astringent (drying)
compounds being present in high quantities. But, using water that is lower than 94 °C results in the flavour
compounds being poorly extracted from the coffee particles. However, using much lower brewing
temperatures, combined with a very long extraction time, can lead to a pleasant cup of coffee with low bitter
compound concentrations, such as 2-furylmethanol (or furfuryl alcohol, C5H6O2), which is shown below.

When brewing coffee, the first compounds to be extracted are the acidic, fruity-flavoured compounds, such as
methyl propanoate (or propionate, C4H8O2), which has a fruity, rum-like odour. Following this, the earthy,
caramel-type compounds are extracted such as abhexone (C7H10O3), and then the bitter tasting compounds
including tannins such as chlorogenic acid (C16H18O9). Therefore, it follows that the length of the brew is
important so as to let the full flavour of the coffee develop, whilst not over-brewing and developing a brew that
is overpowered with bitter flavours.

Milk in your coffee

Studies into the role of milk in coffee have shown that it can have positive and negative effects on the health
benefits of this drink. The consumption of caffeinated drinks can result in an increased risk of osteoporosis (a
disease where the bone strength of an individual is decreased leading to the increasing likelihood of a broken
bone), however studies indicate that a daily intake of two cups of coffee, with milk, can counterbalance the
diminishing bone mineral density. On the other hand, while milk is a good source of nutrients itself, it can result
in decreased absorption of polyphenols and antioxidants in coffee.

The sweetness of milk is due to the presence of lactose (a sugar, C12H22O11). Lactose is a disaccharide, which
means that it is made up of two monosaccharides joined together (glucose, C6H12O6 and galactose, C6H12O6).
The structure of lactose can be represented in two different ways. One shows the two sugars in so-called
'chair' conformations (as the shapes resemble a garden chair), which is the lowest energy conformation (or
shape) that a sugar can have as there is the least amount of strain on the rings.

When you drink cold milk, it doesn't taste very sweet. This is because lactose is quite insoluble.
However, when you heat up milk, the solubility of lactose is increased and the lactose molecules
break down into one molecule of glucose and one molecule of galactose, in a hydrolysis reaction.
Consequently, the presence of the two monosaccharides makes the hot milk sweeter.

When milk is foamed to add to coffee (the foamed milk on a cappuccino for example), the final temperature
guideline for heating the foam is between 58 and 60 °C. This is because above this temperature the proteins in
the milk start to become denatured and you end up with a lower quality foam to add to your coffee.

Overall, it is important to get the perfect balance when making the perfect cup of coffee, though sometimes the
caffeine hit is more important than the quality of the brew.

The overall experience

Interestingly, Professor Charles Spence (Oxford University) says all the senses - taste, touch, sight, smell, and
sound - combine to contribute to our enjoyment when we eat and drink. Based on his research, coffee chain
Starbucks enlisted composers to create music, to accompany drinking coffee. His work in this area has been
recognized with awards including an Ig Nobel Prize, together with co-author Massimiliano Zampini.

https://youtu.be/t01xzhcYBqM
Another study from Professor Spence found that, more often than not, we can tell whether a liquid
is hot or cold just from hearing it being poured into a cup. So, can you tell whether water is hot
or cold solely on the basis of the sound that it makes when poured into a cup made from
glass, porcelain, paper or plastic? You may be surprised by your results (and this video will
explain why).

Tools of the trade: understanding the Maillard reaction

The Maillard reaction is a very important flavour-producing reaction, as illustrated when sugar forms
brown nutty-flavoured caramel, in a process called non-enzymatic browning. It is also an important
reaction in developing the flavour compounds in tea and coffee. Before coffee beans are roasted, they
lack the aroma of roasted coffee beans, and the Maillard reaction is very important in providing the
characteristic flavour of a brew.

Stage 1: Making a glycosylamine

The first step of the Maillard reaction is a sugar-amine condensation reaction to form an N-substituted
glycosylamine. The carbonyl group on the glucose sugar (C=O in red) reacts with the amine group (NH2 in
green) in a protein or amino acid in the food to form an imine bond (in pink) in the N-substituted glycosylamine,
together with water. Notice that the sugar (glucose) reacts when it is in an open-chain form (you may
remember that for most of the time, glucose exists in a cyclic form).

The mechanism for the first step of the Maillard reaction involves the nitrogen lone pair from the
amine group attacking the electrophilic carbon atom on the aldehyde carbonyl (C=O). Electrons
are pushed from the double bond onto the oxygen atom giving it a negative charge. A proton is
then transferred from the positively charged nitrogen to the negatively charged oxygen atom
forming a hemiaminal (in pink). The oxygen atom then donates its lone pair to a proton forming –
OH2+, which is a very good leaving group. Finally, the nitrogen can donate its lone pair to form a
new C=N bond of the imine in the N-substituted glycosylamine; in the process, it kicks out H2O as
a leaving group.
Stage 2: The Amadori rearrangement
The second step in the Maillard reaction is the Amadori rearrangement, a spontaneous reaction even at
temperatures as low as 25 °C. The Amadori rearrangement is an isomerisation reaction (the N-substituted
glycosylamine, 1,2-enaminol and the Amadori compound all have the same chemical formula, with the atoms
arranged in different ways, and so they are isomers of each other) that results in the formation of a ketosamine
called the Amadori compound – this contains both a ketose (a sugar bearing a ketone) and an amine.

The isomerisation reaction (called an imine-enamine tautomerism) that occurs between the N-
substituted glycosylamine and 1,2-enaminol is reversible. The nitrogen atom in N-substituted
glycosylamine donates a lone pair of electrons to a proton, forming a new N–H covalent bond;
following this, the two electrons in the C–H bond (in pink) are donated to form a new carbon-
carbon double bond, pushing the electrons in the C=N double bond onto the positively charged
nitrogen atom.

The second reversible isomerisation reaction occurs between the 1,2-enaminol and the Amadori compound,
and is an example of keto-enol tautomerism. In the forward reaction, two electrons from the O–H oxygen (in the
1,2-enaminol) are donated to a proton, through the C=C double bond, and this is followed by deprotonation.

Stage 3: Further reactions

The Amadori compound can then react to form several different products, including hydroxypropanone
(C3H6O2), all of which can react again to form even more products. The products that are formed depend on
whether the reaction mixture is alkaline or acidic (e.g. hydroxymethylfurfural, C6H6O3), so, this is a complicated
process.
Melanoidins are a class of brown polymers with high molecular weights, and one of the potential end products
of the Maillard reaction; melanoidins are the species that give cooked food its colour. The analysis of
melanoidins is challenging, meaning that it is hard to assign the structures to the end products of the Maillard
reaction; this difficulty stems from the vast numbers of Maillard reaction products and the difficulty in purifying
and identifying individual products.

So, there is still a lot about the Maillard reaction that remains unclear, however, its responsibility for the flavour
development of cooked or roasted foods is recognised as being very important.

As the Maillard reaction is such a complicated process, with a number of tricky reaction steps, we have
summarised the key steps in the poster below.

Alongside the Maillard reaction, the Strecker degradation (established in the 1960s) also plays an important
role in the formation of flavour compounds - take a look at the information in the downloads section below.

Does coffee cause cancer?

Perhaps you remember the ruling made by a California judge in 2018, that coffee companies must serve the drink
with a cancer warning? It arose from the presence of acrylamide (H2C=CH–C(=O)NH2) in coffee, which is
formed from a Maillard reaction involving the amino acid asparagine (HO2CCH(NH2)CH2C(=O)NH2) – evidence
has shown that acrylamide is a likely carcinogen (a cancer-causing substance).

However, each 150 mL cup contains only a trace amount of acrylamide, 0.9-2.4 micrograms – one estimate
suggests an 80 kg adult would need to consume over 208 micrograms of acrylamide a day to have an
increased risk of cancer. Interestingly, the same scientists who classified acrylamide as a carcinogen also
found no conclusive evidence that drinking coffee caused cancer (and in 2016, the World Health Organisation
took coffee off its list of possible carcinogens).

(By the way, concerns over acrylamide in foods has also made the headlines. For example, in
2017, the Food Standards Agency (FSA) warned that acrylamide is produced when starchy foods
are roasted, fried or grilled for too long at high temperatures, so browned toast and potatoes could be
a cancer risk. But, once again, because of the small quantities of acrylamide formed, it was
subsequently reported that someone would only be at risk if they consumed 320 slices of browned
toast each day!)

Now you try

(a) Draw the structure of the imine formed from the following condensation reaction.

H3CNH2 + H3CCOCH3 →

(b) Draw the structure of the enamine formed from the following condensation reaction.

(H3C)2NH + H3CCOCH3 →

(c) Draw the enol form of acetophenone, C6H5COCH3.

The_Maillard_Reaction.pdf
Strecker_Degradation_PDF_revised_.pdf
The organic flavour compounds in coffee
0:06Skip to 0 minutes and 6 secondsCoffee contains over 1000 aroma compounds, and a variety of antioxidants,
the most abundant being chlorogenic acids - a family of structurally similar esters. We will see that chlorogenic
acids are mainly responsible for coffee’s bitter taste. When coffee beans are roasted, chlorogenic acids and the
numerous other organic compounds undergo a whole host of chemical reactions. This includes Maillard
browning reactions that produce a range of compounds, which affect the taste of coffee, such as brown-
coloured and very bitter-tasting polymers called melanoidins. Interestingly, melanoidins are very poorly
characterised due to their complexity, and their chemical structures remain largely unknown. The majority of
the coffee aroma is found in the coffee oil, which makes up about 10% of the roasted beans.
0:59Skip to 0 minutes and 59 secondsThe aroma of coffee is a result of volatile (low-boiling) compounds. Non-
volatile (high-boiling) compounds in the coffee give the sour, bitter and astringent flavours. The final
temperature of the roasting process correlates directly to the colour of the beans; at higher final temperatures,
the beans have a darker colour. The roasting time can be anywhere between 90 seconds and 40 minutes. The
length of time influences the reactions that occur in the beans; roasting the beans for longer periods result in
bitter coffees, which lack a pleasing aroma, whereas a very short roasting period may not give enough time for
all the reactions happening in the roasting process to be completed, which can result in underdeveloped
coffee.
1:43Skip to 1 minute and 43 secondsThe non-volatile compounds in coffee, that gives the sourness, bitterness
and astringency, include caffeine which provides body, bitterness and strength to brewed coffee; carboxylic
acids, such as ethanoic acid and maleic acid, which provide sourness; and nicotinic acid, which has a
nutritional value (as it is a vitamin of the B complex). Alongside these, there are many other non-volatile
compounds in coffee that contribute to the taste sensation and feel of the final cup. Interestingly, caffeine is
able to bind the adenosine receptor in the body, as it has a similar chemical structure to adenosine. Adenosine
suppresses physiological activity, resulting in drowsiness as is it slows down nerve cell activity.
2:27Skip to 2 minutes and 27 secondsBy blocking the action of adenosine at its receptor, caffeine causes a
stimulant effect and helps to prevent tiredness. Volatile compounds in coffee provide the aroma. The major
mechanisms for the formation of coffee aroma include the Maillard reaction and the Strecker Degradation.
There are other contributing mechanisms and interactions, which results in a very complex system for
producing coffee aroma. Gas-chromatography-olfactometry techniques have been used to identify individual
odourants in coffee. Scientists achieved this by sniffing the eluted compounds, which allowed 18 different
aroma compounds to be identified. The mass spectrum for each compound was compared to authentic
standards, along with their odour activities and retention indices.
3:16Skip to 3 minutes and 16 secondsIn gas chromatography there is a mobile and a stationary phase, the
retention indices of compounds are the times it takes for a compound to move through the column. Different
compounds have different elution times because they will all interact differently with the mobile and stationary
phases. Identifying these odorants showed that the aroma change between roasted and brewed coffee is due
to a change in concentration of the odourants present. Using these 18 odourants, a synthetic mixture was
produced that followed the same odour profile as coffee brews. Four out of the eighteen identified aroma
compounds are shown here.
3:56Skip to 3 minutes and 56 secondsThese are sotolon, which has the smell of toast, vanillin, which smells of
vanilla, furaneol, which has a caramel smell and finally, abhexone, which smells of honey. Omission studies
have been carried out on these synthetic coffee mixtures. An omission study is when a synthetic cup of these
compounds is created but a specific compound is excluded. An expert in coffee aromas tastes the mixtures to
determine if the flavours are significantly affected by the loss of each compound. Through these emission
studies, it has been determined that the removal of certain compounds from the brew, including 2,3-butadione
and vanillin did not result in a significant loss in coffee flavour.
4:43Skip to 4 minutes and 43 secondsOverall, out of over 900 volatile components in coffee, amazingly, fewer
than 20 have actually been determined to be relevant to the aroma of the brew.

Most of us have a favourite style of coffee and every cup will taste different because of the
roast of the coffee bean, the grind, the amount of bean used, the water, the temperature and
many other factors. At home, you may have tried to reproduce the coffee goodness from
the coffee houses, but do we actually know the best method to use in order to elicit the
captivating flavours perfectly? Here are some pointers for how to make these excellent
coffees.

Cappuccino: This drink consists of equal parts of steamed milk, frothed milk and espresso. The
milk is poured on top of the coffee shot and dusted with nutmeg, cinnamon or chocolate powder.

Mochas: The easiest way to do this is to make it with quality hot chocolate, add a shot of coffee
with steamed milk poured in and top with whipped cream, then lightly dust with chocolate powder.
It looks great in a clear glass mug with a long-stemmed spoon.

Lattes: Foam and steam milk to 75 °C. The ratio is worked out as 50% coffee and 50% milk.
Slowly pour the milk down the side of the coffee cup or glass so it infuses with the coffee shot. The
main difference between a latte and a cappuccino is a latte blends the milk and coffee together,
whereas the cappuccino keeps the two apart.

Espressos: Using a finely ground good quality coffee bean and a high pressure espresso pot
gives a coffee shot with a golden-brown foam on top, to be served in an espresso cup to keep the
coffee warm as it is being drunk.

Steaming milk involves introducing hot water vapour into cold milk until it reaches the ideal
temperature for the drink. It is not as simple as it sounds as different milks (with different amounts
of proteins, carbohydrates, and fats) require different amounts of steaming time. For example, fat
molecules can stabilise the formation of foam by surrounding the air (injected during the steaming)
and entrapping it in a bubble - milks with higher fat content lead to stable foams at temperatures
below room temperature, while milks with lower fat contents (like skimmed milk) are better at
stabilising foam at higher temperatures. The temperature is key as too high a temperature can
scald the milk, which denatures the enzymes and causes curdling (as denatured milk proteins
clump together).

At the end of the day, making coffee correctly is all up to you and the way your taste buds take to
the coffee taste, or indeed, if you like coffee in the first place! A survey of our second year York
undergraduate chemists showed that water was their most popular non-alcoholic drink, followed by
tea then coffee. They preferred black tea to green or herbal teas, and almost all put milk in their
tea in preference to adding lemon juice or drinking it black. By far the most popular coffee was latte
followed by cappucino and only 5% of the students knew what bulletproof coffee was.

So, what is your favourite non-alcoholic drink, and why? Do you have a particular routine
(such as a Turkish coffee approach) for making your favourite brew and/or do you use any
unusual ingredients? For example, I wonder how many of you enjoyed an eggnog latte at
Christmas?

Rather than a handful of words, why not use over 100?

You might describe your coffee as ‘strong, black and hot’, but the Specialty Coffee Association of
America (SCAA) has taken it to a whole new level. The SCAA’s Flavour Wheel provides a full
descriptive glossary for both good coffee and bad coffee, and is standard-use for modern coffee
tasting (or coffee cupping). To describe your coffee, start at the centre and work outward. The
most general taste descriptors are near the middle, and they get more specific as the tiers work
outward.

Why not give it a go and share your findings, so we can discuss and compare your results?

Understanding decaffeination processes

Caffeine is a compound that is known worldwide due to its presence in many, widely
consumed, foods and drinks; it has many natural sources, including coffee beans, cocoa
beans and tea leaves. It has the systematic name 3,7-dihydro-1,3,7-trimethyl-1H-purine-2,6-
dione, with the chemical formula C8H10N4O2.

The effects of caffeine on the human body are well known. Caffeine in low doses is a stimulant,
allowing the drinker increased concentration and reduced tiredness. However, the consumption of
higher doses of caffeine can result in insomnia, nervousness, hostility and mood swings – among
other side effects. Commonly, the biological effects of caffeine are explained by it mimicking
adenosine (look at their similar structures). In the body, adenosine acts on a receptor that causes
a 'slowing-down' of the body, whereas when caffeine acts on the same receptor it has the opposite
effect. So, because of its similar shape, caffeine competes with adenosine and reversibly blocks

the action of adenosine on its receptor.

The negative effects of caffeine have led to an increasing demand for decaffeinated brews. In order to provide
decaffeinated coffee, various methods of removing caffeine from the beans have been developed. This
includes using organic solvents (such as dichloromethane or ethyl acetate – more than 99% of the
decaffeinated tea sold in the UK is decaffeinated using dichloromethane), water (the Swiss water method is
described in a pdf in the downloads section below) and supercritical CO2 .
Supercritical CO2
The first extraction method uses supercritical carbon dioxide (CO2). Carbon dioxide, at standard temperature
and pressure, is a gas and, when frozen, carbon dioxide forms a solid (called dry ice). It is also possible for
carbon dioxide to exist in a fluid state but, for this to occur, it has to be held at its critical pressure and critical
temperature.

The critical point is the combined critical temperature and critical pressure where, at or above which, the
molecule is no longer able to be classed as a liquid or gas, but is instead known as a supercritical fluid. Above
its critical point, there are no clear liquid or gas phases, and the carbon dioxide can take on both liquid and gas
properties. Carbon dioxide in this state is called supercritical CO2 (abbreviated sCO2) and has the ability to act
as a solvent and dissolve other materials.

Supercritical CO2 has been used in industry to extract caffeine from coffee beans since the 1970s. Caffeine
has a low solubility in supercritical carbon dioxide and, for this reason, a co-solvent (either water or ethanol) is
used which triples the caffeine solubility, hence accelerating the extraction. It is a widely used method of
decaffeination, as it has proved to be more efficient, as well as being better for the environment, than other
decaffeination methods. On top of this, it is also an easily removable, non-flammable and non-toxic solvent.
These beneficial properties have helped ensure its application in other extraction areas, such as teas and
cocoa beans.

Large-scale decaffeination takes place on green coffee beans (before roasting). The green beans are soaked
in hot water (until they have about 50% moisture content) and put into an extraction vessel, which is then
sealed. Soaking the beans in water allows the pores of the coffee beans to open as the beans swell, thus
allowing the caffeine (which converts into a mobile form) to diffuse out of the beans. At a pressure of about 300
atmospheres, the supercritical CO2 is pumped into the sealed vessel, where the CO2 solvent dissolves the
caffeine compound and draws it out of the coffee beans. Any larger odour molecules are not dissolved and
remain inside the beans. During the decaffeination process, the carbon dioxide moves through an extractor
and a scrubber, on a loop, for approximately 8-12 hours. The scrubber has a high water to CO 2 ratio and the
water carries away any dissolved caffeine before the carbon dioxide recirculates to extract more caffeine.

The CO2 and caffeine mixture is then moved into a strong tank, where the pressure on the container is
removed. As the pressure lowers, the supercritical CO2 returns to the gaseous phase – when the pressure is
lowered, it falls below the critical value. The gaseous CO2 evaporates, leaving the caffeine behind in the
container. Any remaining caffeine is removed from the gaseous CO2 by charcoal filters, allowing the clean
CO2 to be pumped back into a high-pressure vessel so it can be recycled and used again and again.
Extraction using supercritical carbon dioxide is a highly selective process. It only extracts the caffeine
compound and leaves the other flavour precursors (such as carbohydrates and peptides) in the bean.
However, there are two disadvantages to using this decaffeination method; firstly, precise processing
parameters need to be specified and secondly, the process is very capital intensive meaning that this process
is only commercially viable if the plant makes over 3000 tons of decaffeinated coffee a year.

What happens to the caffeine that is extracted?

Caffeine obtained from the decaffeination is mainly used by pharmaceutical, cola-type soft drinks and cosmetic
industries (it is a common ingredient in cellulite creams), and, more recently, in energy drinks. Although decaf
coffee amounts to around 10% of the global coffee market (in 2012), the need for caffeine by these industries
is much greater than the amount of caffeine being produced, hence caffeine is prepared in the lab (the first
reported synthesis was back in 1895). Following on from the discussion in week 1, it is interesting that ‘natural’
caffeine (mainly from coffee beans) is generally assumed to be healthier than energiser drinks (or beauty
products) containing ‘synthetic’ caffeine.

Extension activity – How much caffeine is present in a coffee bean?

If you are interested in finding out how to determine the amount of caffeine in a coffee bean, take a look at the
caffeine analysis pdf in the downloads section below. This describes a further use of titration, a common
laboratory method of quantitative chemical analysis, and it provides another opportunity to do some
calculations.

Extension activity – try this ‘stimulating’ practical

Finally, for those of you keen to do some further practical work and who have access to a laboratory (including
a fumehood and equipment such as a thermometer) - we have added an extension activity on the extraction
of caffeine from tea and/or coffee (see the downloads section below). This gives you the opportunity to
extract a reasonably pure sample of caffeine from tea leaves and/or filter coffee – as always, ensure you read
all of the instructions and risk assessment before starting (the experiment involves heating an appropriate
cooking oil, such as corn oil, to around 250 °C).

Swiss_water_method_revised_.pdf

Caffeine_extraction_revised_.pdf
Section_3.20_caffeine_analysis.pdf
Art with coffee rings
0:06Skip to 0 minutes and 6 secondsCoffee beans are the green seeds of a cherry that when roasted, changes
colour from green to brown. This change of colour is caused by the Maillard Reaction, which is commonly
called browning. The brown colour is made up largely of compounds called melanoidins, which are high
molecular weight polymers formed when sugars and amino acids combine. This is an extremely complicated
process and not much is known about the detailed structures of the melanoidins that are formed. One thing
that we do know about the melanoidins, is that these brown-coloured pigments, together with other coloured
compounds in coffee, can act as stains.
0:44Skip to 0 minutes and 44 secondsDunking paper in coffee can be used to give it an antique look, while
painting plywood with a cup of coffee can darken it. In fashion, coffee has even been used to dye fabric, such
as t-shirts and jeans for a distressed look. You may also have seen the scientific interest in coffee rings - the
phenomenon to explain the ring-shaped stain of particles leftover after coffee drops evaporate is important in,
for example, inkjet printing. Coffee rings are also important in the art world - Malaysian artist Hong Yi took the
internet by storm with a stunning painting of singer Jay Chou made simply with thousands of coffee stains,
using a coffee cup.
1:24Skip to 1 minute and 24 secondsShe had to get the coffee mixture just perfect, as the ring stains were
permanent the moment they touched the canvas and she spent around 12 hours working on this incredible
piece. Now it is your turn to utilise the staining power of coffee to positive effect! Use a coffee cup, brush and/or
anything else to paint the structure of an organic compound found in coffee. Perhaps methylpropanal, caffeine
or a more complicated structure, such as cinnamic acid. You have over 1000 compounds to pick from! It could
be an abstract design, or more detailed, showing individual atoms. You may want to experiment with different
coffees, of different strengths. Now is the time to make your mark!
2:05Skip to 2 minutes and 5 secondsDon’t forget to post a photo of your creation, including why you picked the
structure you did, and the coffee you used.

So, rather than coffee staining our teeth, or coffee table, let’s use this staining power creatively to
make a ‘molecular’ work of art.

Hints and tips for coffee art

1. Make sure that the coffee is very, very strong. We used approximately three times the amount of filter coffee
needed for a standard cup and around a third of the amount of water – it needs to be very concentrated to
colour the paper.

2. To make a background like ours (to look like parchment paper) you can use tea or coffee. If using tea, soak
the tea bag in cold water for a minute or two and then rub the soaked tea bag directly onto the paper (try not to
get it too wet), before placing the tea-soaked paper into a preheated oven at about 80 °C until it is dry. Keep an
eye on it because it will not take very long to dry. For coffee, brush a coat onto the paper and then put it into
the oven at about 80 °C until it dries. Repeat if the colour is not as dark as you'd like.

3. After the background is complete, ensure you draw dots to mark the places of atoms for your molecule. This
is an important step, because drawing the molecule freehand, without any guide marks, will most likely result in
you running out of space.

4. Using a thin paintbrush dipped in coffee, carefully trace out the shape of the molecule (once only). It is
tempting to continue to go over it multiple times quickly to try and get a more defined figure, but it is important
to allow the first coat to dry before you do the second, if you want a well-defined molecule. Putting more coats
on top of a wet coat will just result in the coffee spreading to the surrounding paper and you will get a more
abstract-looking molecule. For best results, put the paper back into the oven at about 80 °C between
coats, allowing for a quick dry.
5. In total, your molecule will probably require six to seven coats to look well-defined and, using the oven
method, you should have a perfect coffee-coloured compound in no time. If you'd like to burn the edges to
make it look more ancient, you can use a match or a lighter, holding it to the edges of the paper and blowing
out the flame once you have got the desired look. Do each little piece slowly, otherwise you risk burning too
much and losing some of your molecule.

Remember to upload your work onto our open Padlet (we have included some examples from previous years to
help inspire you) and/or on Twitter or Instagram hashtag #FLchemistry. We are eager to see what you can
create!

While you are creating your masterpiece, you might like to reflect on some interesting articles about coffee.

Coffee can be good for you

A study has found that if the coffee beans have been highly roasted then the drink is less beneficial for your
health. Concentrations of chlorogenic acid were found to be reduced on roasting and so the potential positive
effects of this polyphenol (such as improving the function of the circulatory system and reducing the risk of
developing high blood pressure) are diminished.

Also, in 2017, a large review of studies showed that moderate coffee drinking is safe, and 3-4 cups a day may
have some health benefits. This included finding a lower risk of liver disease and some cancers in coffee
drinkers, and a lower risk of dying from stroke. However, the researchers could not prove it was due to coffee,
so experts advise that you should not start drinking coffee for health reasons.

When is the best time to drink coffee?

Interestingly, the time at which we drink our coffee may alter its effect. If we consume coffee during times of peak
cortisol (a hormone that is produced in our bodies to make us feel more awake and alert)
production, research has shown that the effect of the caffeine will be greatly diminished and our bodies will build
up a greater tolerance to the effects of caffeine. For many people the optimal times for consuming caffeine
appear to fall somewhere around 9.30-11.30 am and 1.30-5.00 pm. For the coffee-drinkers among you, why
not challenge yourselves to limit your caffeine intake to the prescribed time slots over the next few
days and tell us what you loved and hated about it, and whether you noticed any difference?

Does coffee or tea stain teeth more?

Our tooth enamel is prone to staining as it contains microscopic pits and ridges that can hold particles of food
and drink. Pigments from dark-coloured drinks such as coffee and tea can become embedded in the cracks
and ridges causing yellowing staining of our teeth.

A dentist recently claimed that black tea can stain our teeth more than coffee. Many assume coffee has a greater
staining power because it is darker in colour than tea, but looks can be deceptive. The high levels of tannins (a
family of compounds containing phenolic groups) in tea, including theaflavins and thearubigins, are believed to
be responsible for the greater staining. Tannins from plants, particularly oak bark, were once used in the
process of tanning leather (they preserve the animal skin by binding to collagen proteins, making the skins
stronger, stiffer, waterproof and resistant to bacterial attack).

Comparing the staining properties of coffee versus tea makes a really interesting science project!

Watching coffee dry

You may find this (microscopic) video of the coffee ring effect interesting - watch as spherical particles get
swept to the edges.
https://youtu.be/ZaCGoSTMHyc

Finally, add a drop of whiskey

On drying, drops of whiskey have been shown to produce unique patterns that might provide a tool for
authorities investigating counterfeited products. Research showed that when American whiskey was diluted
with water down to about 20–30% alcohol content, on evaporation, the droplets began to leave web-like
patterns on the glass. The patterns arise because hydrophobic compounds within the whiskey aggregate when
the water is added – each whiskey has its own flavour profile, made up of thousands of different compounds,
including different hydrophobic compounds, so different brands gave unique motifs.

caffeine_coffee_draiwing.pdf

4e_Making_coffee_art__3rd_Draft_.pdf

Fake Science News

On a course looking at everyday chemistry, it seems very appropriate to touch on the subject of fake
science news. We have positioned this topic here as what we drink often grabs the attention of
headline makers and the internet is awash with sensationalist science news stories. One day coffee is
good for us, the next it is bad. But such headlines are based on science, and you have to know how to
read a scientific paper and take the time to do it.

Articles that make no attempt to explain methodology, make no attempt to indicate any limitations on the
conclusions, are from obscure journals, link the research to something only tangentially related, and draw
sweeping conclusions from a single study should all raise a red flag. (An academic study found fake news
articles tend to contain less complex language, an easier reading level, and longer sentences than genuine
news.) There is also the issue of confirmation bias, where information is interpreted and favoured to confirm
one’s pre-existing beliefs, leading to biased reporting. For example, should we be concerned by an
examination, by Vox in 2017, of 100 Mars-funded studies that found these studies overwhelmingly drew
positive conclusions about cocoa and chocolate?

Sharing misinformation can have serious consequences including eroding public trust in real news and the
integrity of science journalism. So, how easy is it to spot misinformation and falsehoods? Take, for example,
this slogan used in an advertising campaign:

“More than 80% of dentists recommend Colgate”

This statement is true, but it is phrased in a misleading way, and according to the Advertising Standards
Authority, must not be used again. The ad wants us to think that 80% of surveyed dentists (by the way, how
many were surveyed?) preferred Colgate above all other toothpastes. But the survey allowed dentists to
recommend more than one brand, and Colgate’s biggest competitor was named nearly as often!

Now it is your turn. Take a look at this abstract of a paper on how chocolate helps weight loss – perhaps
another reason to enjoy an extra chocolate biscuit with your cuppa! It was published in a research journal. See
if you can work out why it made the news. Ahead of this, here is some background information:

1. The p value, or probability value, is a way to determine the significance of results – by

convention, journals often say something is statistically significant if the p-value is less than
0.05, which indicates that there is less than a 5% chance that the result is due to chance.
 2. The term mg/dl stands for milligrams per decilitre, a measurement that indicates the amount
of a particular substance in a specific amount of blood.
3. If a diet is high in fats, and extremely low in carbohydrates, or carbs, the body does not have
enough glucose for energy. Instead, it starts to use fats for energy, releasing ketones (such
as propanone, or acetone) into the blood, which are eliminated in urine. As the fat stores are
broken down this can translate into weight loss. (This is the basis of the ketogenic diet,
where a low-carb, high fat diet raises blood ketone levels.)
Chocolate with High Cocoa Content as a Weight-loss Accelerator
Background: Although the focus of scientific studies on the beneficial properties of chocolate with a high cocoa
content has increased in recent years, studies determining its importance for weight regulation, in particular
within the context of a controlled dietary measure, have rarely been conducted.

Methodology: In a study consisting of several weeks, we divided men and women between the ages of 19 and
67 into three groups. One group was instructed to keep a low-carb diet and to consume an additional daily
serving of 42 grams of chocolate with 81% cocoa content (chocolate group). Another group was instructed to
follow the same low-carb diet as the chocolate group, but without the chocolate intervention (low-carb group).
In addition, a third group was asked to eat at their own discretion, with unrestricted choice of food. At the
beginning of the study, all participants received extensive medical advice and were thoroughly briefed on their
respective diet. At the beginning and the end of the study, each participant gave a blood sample. Their weight,
BMI, and waist-to-hip ratio were determined and noted. In addition to that, the researchers evaluated the
Giessen Subjective Complaints List. During the study, participants were encouraged to weigh themselves on a
daily basis, assess the quality of their sleep as well as their mental state, and to use urine test strips.

Result: Subjects of the chocolate intervention group experienced the easiest and most successful weight loss.
Even though the measurable effect of this diet occurred with a delay, the weight reduction of this group
exceeded the results of the low-carb group by 10% after only three weeks (p = 0.04). While the weight cycling
effect already occurred after a few weeks in the low-carb group, with resulting weight gain in the last fifth of the
observation period, the chocolate group experienced a steady increase in weight loss. This is confirmed by the
evaluation of the ketone reduction. Initially, ketone reduction was much lower in the chocolate group than in the
low-carb peer group, but after a few weeks, the situation changed. The low-carb group had a lower ketone
reduction than in the previous period, they reduced 145 mg/dl less ketones, whereas the chocolate group had
an average reduction of an additional 145 mg/dl. Effects were similarly favourable concerning cholesterol
levels, triglyceride levels, and LDL cholesterol levels of the chocolate group. Moreover, the subjects of the
chocolate group found a significant improvement in their well-being (physically and mentally). The controlled
improvement compared to the results of the low-carb group was highly significant (p <0.001).

Conclusion: Consumption of chocolate with a high cocoa content can significantly increase the success of
weight-loss diets. The weight-loss effect of this diet occurs with a certain delay. Long-term weight loss,
however, seems to occur more easily and more successfully by adding chocolate. The effect of the chocolate,
the so-called “weight loss turbo,” seems to go hand in hand with personal well-being, which was significantly
higher than in the control groups.
End of week summary
So, how did you find the third week? Is there anything that remains unclear?

Over the week, was there any course content that really surprised you? Perhaps the diverse range of chemistry
that takes place within a cup of tea or coffee.

What is your highlight of the week?

The importance of beverages, for example coffee, is highlighted by Starbucks’ Kelly Goodejohn’s claim that
“coffee is the second-most traded commodity after oil, and 25 million farmers around the world rely on income
generated from growing coffee”. While others may argue that the markets for several commodities including
soybeans, wheat, aluminium, copper and iron ore are larger than that of coffee, few would doubt that it is still
an enormous industry, and it even has its own World Championships!

If any of you are interested in finding out what a chemistry-related career in brewing is like, you may find this
interview with Briana Francisco useful.

Everyday Chemistry Collage – Get Involved!

Why not contribute to our course collage! Take a photo of the ingredients on the container/packet of a
product at home, which is related to beverages – perhaps your favourite fruit tea, soft drink or alcoholic
drink. We would also welcome accompaniments to your favourite brew, such as the ingredients of your
favourite biscuit or cake. Upload your photo on our open Padlet (we have included some examples from a
previous course to help inspire you) and/or use Twitter or Instagram hashtag #FLchemistry. Please include
the name of the product, the country where it was bought, and your initials. We will then collate them to
make a unique eeDc course collage with contributions from (hopefully) all of our learners! A nice end-of-
course keepsake.

Do you spot any of these additives?

Potassium metabisulfite (K2S2O5): A popular antioxidant used in the brewing industry preventing spoilage.
Potassium sorbate (C6H7KO2): A preservative that prolongs the shelf-life of foods by stopping the growth of
mould, yeast, and fungi.
Sodium benzoate (C7H5NaO2): A preservative that inhibits the growth of potentially harmful bacteria, mould,
and other microbes.
Sodium erythorbate (C6H7NaO6): An antioxidant produced from sugars derived from beets, sugar cane, and
corn. An urban myth claims that sodium erythorbate is made from ground earthworms, but there is no truth to
this.
Tartaric acid (C4H6O6): Acts as a preservative (many bacteria are unable to grow in an acidic environment) and
as a flavouring agent (due to its tartness).
Tartrazine (or yellow dye 5) (C16H9N4Na3O9S2): A bright yellow pigment that is used in many foods and drinks
as a colouring agent. Around 1 in 10,000 people suffer allergic reactions to this chemical. A notorious example
of the unfounded rumours surrounding tartrazine is that drinking ‘Mountain Dew’ affects a man’s fertility –
Mountain Dew has little or no effect on sperm count!

Finally, I thought this may interest you. Research in 2014 showed that food picked up just a few seconds after
being dropped is less likely to contain bacteria than if it is left for longer periods of time. The findings suggest
there may be some scientific basis to the ‘5 second rule’! The research team also carried out a survey that
showed 87% of people surveyed said they would eat food dropped on the floor, or already have done so.