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Espresso coffees, caffeine and chlorogenic acid intake: potential health
implications
Thomas W. M. Crozier,a Angelique Stalmach,a Michael E. J. Leanb and Alan Crozier*a
Received 29th October 2011, Accepted 13th November 2011
DOI: 10.1039/c1fo10240k
Published on 30 November 2011. Downloaded on 11/10/2017 08:17:20.

HPLC analysis of 20 commercial espresso coffees revealed 6-fold differences in caffeine levels, a 17-fold
range of caffeoylquinic acid contents, and 4-fold differences in the caffeoylquinic acid : caffeine ratio.
These variations reflect differences in batch-to-batch bean composition, possible blending of arabica
with robusta beans, as well as roasting and grinding procedures, but the predominant factor is likely to
be the amount of beans used in the coffee-making/barista processes. The most caffeine in a single
espresso was 322 mg and a further three contained >200 mg, exceeding the 200 mg day
1 upper limit
recommended during pregnancy by the UK Food Standards Agency. This snap-shot of high-street
expresso coffees suggests the published assumption that a cup of strong coffee contains 50 mg caffeine
may be misleading. Consumers at risk of toxicity, including pregnant women, children and those with
liver disease, may unknowingly ingest excessive caffeine from a single cup of espresso coffee. As many
coffee houses prepare larger volume coffees, such as Latte and Cappuccino, by dilution of a single or
double shot of expresso, further study on these products is warranted. New data are needed to provide
informative labelling, with attention to bean variety, preparation, and barista methods.

1. Introduction sleeplessness with a racing mind. For others, through tolerance

to increasing exposure, drinking ten times this amount may still
Coffee is an extremely popular beverage with more than 300 be pleasant, partly reflecting genetic variation in susceptibility.3
million cups being consumed each day in the US alone. The half-life of caffeine in adults is around 5 h, but can be up
Reflecting its popularity, in economic terms coffee is a most to 30 h, with extended retention in the body by women taking an
valuable agricultural product with exports by third world and oral contraceptive, pregnant women, the developing fetus, young
developing countries, amounting to 7.2 million metric tonnes in children, and those with liver disease. These groups are thus more
2009.1 Coffee beans are produced from the cotyledons of seeds of susceptible to the effects of caffeine toxicity.4 Current advice in
plants belonging to the genus Coffea. Commercial production the UK from the Food Standards Agency is for pregnant women
mainly exploits the seeds of Coffea arabica (so-called arabica to restrict caffeine to below 200 mg day
1, or four cups of strong
coffees) which represent 70% of the world market, while Coffea coffee each with an assumed caffeine content of 50 mg.5 For the
canephora (robusta coffees), which has a more bitter taste than general public assessing caffeine intake is difficult. Current
arabica, is used principally with instant coffees and in espresso guideline figures suggest that an 8 oz (225 mL) cup of instant
blends to promote the formation of ‘‘crema’’. coffee contains 60–85 mg of caffeine, and a 1 oz (28 mL)
The value of coffee as a human beverage was initially recog- espresso 30–50 mg.6 However, despite the increasing number of
nised from the invigorating effect of wild coffee berries on goats coffee shops on the high street and in airports, there appear to be
in Abyssinia, sometime around 850 AD.2 This action, subse- no recent publications on the caffeine contents of the various
quently attributable to its caffeine content, has led to the types of commercially prepared coffees.
extraordinary attraction of the beverage to many consumers who As well as caffeine, coffee contains substantial amounts of
exhibit increased alertness and a capacity to remain awake for a family of conjugated hydroxycinnamates collectively referred
longer periods without sleep. Caffeine can, however, have to as chlorogenic acids. The main chlorogenic acids are 5-O-
unpleasant symptoms, and, in excess, can lead to a state of caffeoylquinic acid (5-CQA) and its isomers 3-O-caffeoylquinic
excitement and anxiety. Dose-responses vary. For some people acid (3-CQA) and 4-O-caffeoylquinic acid (4-CQA) (Fig. 1) and
even a single cup may be acutely unpleasant and cause together these account for 80% of the total chlorogenic acids.7
Although the CQAs in coffee have antioxidant properties, and
a
School of Medicine, College of Medical, Veterinary and Life Sciences, in vitro are able to scavenge free radicals, which in humans have
Joseph Black Building, University of Glasgow, Glasgow, G12 8QQ, UK.
E-mail: alan.crozier@glasgow.ac.uk; Tel: +44 141 330 4613
been linked to conditions such as Parkinson’s disease and
b
University of Glasgow College of Medical, Veterinary and Life Sciences, cardiovascular disease, there is much speculation but only
Walton Building, Royal Infirmary, 84 Castle Street, Glasgow, G4 0SF, UK limited evidence of coffee consumption being linked to protective

30 | Food Funct., 2012, 3, 30–33 This journal is ª The Royal Society of Chemistry 2012
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caffeine and 17-fold less total CQA. There was also substantial
variation in the total CQA : caffeine ratio of the coffees which
ranged from 0.47 to 1.94 (Fig. 2).
It is evident from the data presented in Table 2 that the quoted
figures for caffeine content of espresso coffee in the 2008 IFIC
Review,6 which are widely cited in the popular press, do not
provide a realistic picture. The levels of caffeine per serving
varied more than 6-fold from 51 to 322 mg. At the low level,
a pregnant woman and others with a need to restrict caffeine
consumption, might safely drink 4 cups per day without signifi-
Fig. 1 Structures of caffeine and the chlorogenic acids, 5-O-caffeoyl-
cantly exceeding the recommended caffeine intake. In marked
quinic acid, 3-O-caffeoylquinic acid and 4-O-caffeoylquinic acid.
contrast, at the higher end of the scale, drinking even one cup
of espresso will be well in excess of the advised limit of 200 mg
effects on human health.8 Epidemiological evidence and some day
1.
intervention studies do, however, indicate that coffee consump- Responses to caffeine vary. Those habituated to the purine
Published on 30 November 2011. Downloaded on 11/10/2017 08:17:20.

tion may reduce the risk of type 2 diabetes8 and one recent report alkaloid suffer headaches when caffeine is withdrawn. At the
provides evidence that coffee decreases the risk of depression other extreme, doctors not uncommonly see patients with a range
among women with the effect being attributed to caffeine intake.9 of rather non-specific symptoms grouped as ‘‘caffeinism’’ which
It is against this background that HPLC analysis has revealed are resolved when caffeine is removed from the diet. These
substantial variations in the caffeine and chlorogenic acid
content of espresso coffees purchased from coffee shops in the
west end of Glasgow near the University of Glasgow.

2. Results and discussion

The caffeine and 3-, 4- and 5-CQA contents of single servings of
espresso coffee from 20 different outlets are presented in Table 1.
The cup size ranged from 23–70 mL. The amount of caffeine that
a consumer would ingest per serving ranged from 51 to 322 mg,
while the CQA content varied from 24–422 mg. The main
chlorogenic acid in all the coffees was 5-CQA with smaller
amounts of 3- and 4-CQA. Both the caffeine and total CQA
content were highest in coffee from Pattiserie Francoise and Fig. 2 Variation in the total CQA : caffeine ratio of 20 espresso coffees
lowest in Starbucks espresso which contained 6–fold less purchased from outlets in the west end of Glasgow.

Table 1 Quantities of caffeine and CQAs in servings of espresso coffee. Data expressed as mean values (n ¼ 3), standard error <7% of mean values

As a percentage of total CQA

Source Serving size (mL) Caffeine (mg/serving) Total CQA (mg/serving) 3-CQA 4-CQA 5-CQA

Pattiserie Francoise 52 322 422 23% 26% 51%

University Cafe 49 260 230 36% 23% 41%
Caf
e Cinnamon 59 242 179 23% 28% 49%
Paperino’s 50 205 207 31% 26% 43%
S’mug 32 173 294 21% 27% 52%
Costa Coffee 25 157 227 21% 27% 52%
Heart Buchanan 24 156 127 17% 30% 53%
Jellyhill 63 151 122 21% 33% 46%
Baguette Express 45 140 145 21% 28% 51%
Chapter1 26 140 215 22% 27% 51%
Peckham’s 70 140 199 13% 29% 58%
Little Italy 23 129 217 17% 27% 56%
Coffee @ 291 49 98 108 21% 29% 50%
Crepe
a Croissant 34 95 81 21% 28% 51%
Kember & Jones 43 90 175 21% 26% 53%
Beanscene 48 77 93 20% 28% 52%
Tinderbox 25 75 90 22% 27% 51%
Morton’s 35 73 56 23% 29% 49%
Antipasti 36 72 44 18% 34% 48%
Starbucks 27 51 24 21% 29% 50%
Median value 43 140 145 21 28 51
Range 23–100 51–322 24–422 13–36% 23–34% 43–58%

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Table 2 Details of coffee beans and roasting proceduresa

Coffee Origin Roasting temp. ( C) Roasting time (s) Roasting

Washed arabica Colombia — — —

Colombia 350 244 H-S
Colombia 270 595 L-L
Unwashed arabica Ethiopia — — —
Ethiopia 350 247 H-S
Ethiopia 270 612 L-L
a
H-S: high temperature, short roast; L-L: low temperature, long roast.

problems would only be suspected if caffeinated soft-drinks or results in 3-CQA and 4-CQA being destroyed more slowly than
coffee intake were high: our data show that one cup of high- 5-CQA10 while caffeine is lost through sublimation.7,11 The data
caffeine coffee could cause as much difficulty to these susceptible in Table 3, including the total CQA : caffeine ratios, indicate that
Published on 30 November 2011. Downloaded on 11/10/2017 08:17:20.

consumers as six cups of coffee to another. CQAs are lost more rapidly than caffeine, especially during L-L
This large variability in caffeine and CQA content could be roast conditions. This is in keeping with the long held use of the
due to a number of factors with the amount of coffee used to CQA : caffeine ratio as a rule of thumb index of the extent of
prepare a serving of espresso probably being substantially less for roasting.12 The CQA : caffeine ratios in Table 3 are much higher
the low-caffeine coffees than for those at the upper end of the than those obtained with the commercial espresso coffees
scale (Table 1). Other factors that could impact on the caffeine (Fig. 2), probably because the beans used to prepare the various
and CQA content, arguably to a lesser degree, are (i) batch-to- espresso coffees were roasted for longer periods of time resulting
batch differences in the arabica beans, (ii) roasting procedures, in enhanced breakdown of CQA compared to losses of caffeine.
(iii) grinding conditions and (iv) the coffee-making/barista Robusta coffee beans contain almost twice as much caffeine as
process (temperature of water/steam in the extraction vessel, its arabica13 so if any of the espressos were produced from an
duration, coffee : water/steam ratio etc.). arabica–robusta blend, as opposed to being 100% arabica, this
To investigate the possible impact of roasting techniques on would also contribute to a lower the CQA : caffeine ratio. Batch
the CQA and caffeine contents of coffee, infusions were prepared to batch variation in coffee beans is also likely to have an impact
from two batches of espresso coffees. One was a washed on this ratio.
Columbia coffee and the other unwashed beans from Ethiopia.
Unroasted coffees were included along with samples which had
been roasted (i) at a high temperature for a short time (H-S) and 3. Experimental
(ii) at a low temperature for a longer time (L-L) as outlined in
3.1. Coffees
Table 2. After grinding the beans, 100 mL of boiling water was
added to 5 g of coffee and brewed for 5 min before filtering. The Single shot espresso coffees were purchased from 20 different
CQA and caffeine contents of the infusions prepared in this outlets in the west end of Glasgow. The volume of the coffee
manner are presented in Table 3. The caffeine content of brews servings was measured after which aliquots were diluted 50-fold
from both coffees declined by 80% with both the H-S and L-L with methanol and stored at
80  C prior to analysis of caffeine
roasts. There was a bigger loss of CQAs in the infusions with and CQA levels. In addition, in order to explore one possible
11.0% and 13.3% recoveries after H-S and 8.0% and 6.8% reason for variation in the micronutritient contents of coffees, six
following L-L roasting conditions. This was associated with samples of ground espresso arabica coffee, prepared from beans
reduced CQA : caffeine ratios of the coffees. subjected to different roasting profiles, as outlined in Table 2,
During roasting, the chlorogenic acids are subjected to were supplied by Finlay Beverages (South Elmsall, London WF9
a complex series of reactions including acyl migration which 2XS). In Glasgow, 100 mL of boiling water was added to 5 g of

Table 3 Effect of washing and roasting conditions on the CQA and caffeine content of arabica coffee beans. One hundred mL of boiling water was
added to 5 g of ground beans and after 5 min samples were filtered and the caffeine and CQA content of the filtrate analysed by HPLC. Data expressed as
mean values in mg mL
1 (n ¼ 3). Standard error <7% of mean values. Figures in parentheses represent mean values for total CQAs and caffeine as
a percent of the unroasted valuea

As a percentage of total CQA

Coffee Roast 3-CQA 4-CQA 5-CQA Total CQAs Caffeine Total CQA : caffeine ratio

Washed arabica — 11% 16% 71% 11.3 (100%) 1.90 (100%) 5.9
H-S 22% 27% 51% 1.45 (13.3%) 0.39 (20.6%) 3.7
L-L 23% 27% 50% 0.92 (8.0%) 0.37 (19.6%) 2.5
Unwashed arabica — 8% 10% 81% 14.6 (100%) 1.94 (100%) 7.5
H-S 22% 25% 51% 1.59 (11.0%) 0.37 (19.2%) 4.3
L-L 24% 26% 50% 1.06 (6.8%) 0.35 (18.6%) 3.0
a
H-S - high temperature, short roast; L-L - low temperature, long roast (see Table 2).

32 | Food Funct., 2012, 3, 30–33 This journal is ª The Royal Society of Chemistry 2012
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the ground beans which was brewed for 5 min before filtering, graduate laboratory class in Plant Science taught by A. C. at the
dilution 50-fold with methanol and storage at
80  C prior to University of Glasgow, was carried out as part of T. W. M. C.’s
analysis. Hyndland Secondary School (Glasgow) Advanced Higher
Chemistry research project.
3.2. Analytical procedures
Five mL volumes of the diluted coffee infusions were analysed in 6. References
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according to procedures previously outlined.14,15 Caffeine was exporting countries. http://www.ico.org.
2 T. J. Bond, in Tea, Cocoa and Coffee: Plant Secondary Metabolites
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4 M. E. J. Lean, H. Ashihara, M. N. Clifford, and A. Crozier, in Tea,
Our data represent only a snap-shot of the caffeine contents of Cocoa and Coffee: Plant Secondary Metabolites and Health, ed. A.
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espresso coffees, but the range and scale of the results is sufficient Crozier, H. Ashihara and F. Tom^as-Barb
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to demonstrate that there is a problem, unlikely to be restricted Publishing, Oxford, 2011, pp 25–44.
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coffees, by dilution a single or double shot of expresso, further Barb
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9 M. Lucas, F. Mirzaei, A. Pan, O. I. Okereke, W. C. Willets,
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5. Acknowledgements 14 D. Del Rio, A. J. Stewart, W. Mullen, J. Burns, M. E. J. Lean,
The authors wish to thank Thomas Blackwall of Findlay F. Brighenti and A. Crozier, J. Agric. Food Chem., 2004, 52, 2807–
2815.
Beverages for kindly providing the samples of roasted coffees 15 A. Stalmach, W. Mullen, C. Nagai and A. Crozier, Brazilian Journal
listed in Table 2. This project, which has its origins in under- of Plant Physiology, 2006, 18, 253–262.

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