Republic of Iraq

Ministry of Higher Education & Scientific research

Al-Nahrain University
College of Pharmacy

Antibacterial effects of Green Coffee extract

Done by:
Zahraa Maan Abd AL Rasool
Mariam Kanaan Maher
Zainab Dheyaa Sabah
Taif Reydh Abd AL Satar
Rusul Mohammed Khazal

Graduation Research Project

Submitted To the College of Pharmacy / Al-Nahrain University in
Partial Fulfillment of The Requirement for the Bachelor’s degree in
pharmacy.

Supervised by:
Dr. Rafal Shakeeb AL Anee

2023-2024 AD 1445 AH
 ‫بسم هللا الرحمن الرحيم‬

‫ب ا ْل َعالَ ِمين)‬ ‫آخ ُر َدع َْوا ُه ْم أَ ِن ا ْل َح ْم ُد ِ ه ِ‬

‫َلِل َر ِّ ِ‬ ‫( َو ِ‬
‫صدق هللا العلي العظيم‬

‫سورة يونس‬
‫االية )‪(10‬‬
 ‫االهداء‬

‫درب‪ ،‬وال ُختم‬

‫ٌ‬ ‫الحمدهلل عند البدء وعند الختام‪ ،‬فما تناهى‬
‫جهد‪ ،‬وال تم سعي إال بفضله ‪..‬‬
‫سر البدايات وبلغنا النهايات‬
‫الحمدهلل الذي ي َ‬

‫الى نفسي الطموحة اوالً ابتدت بطموح وانتهت بنجاح‬

‫نور يُضيء عتمتي عندما تُطفئني االيام والضروف…‬ ‫الى ٍ‬

‫الى غيم ٍة تضلُّني وتسقيني دون رغب ٍة بردِّي لجميلها…‬
‫الى االيدي التي تم ُّد لي العَون عندما اتعثر ‪ ،‬وتدفعني‬
‫ل ُمقاومة كل هذه االشياء التي تستدعي السقوط ‪.‬‬
‫الى عائلتي …‬

‫الى رفاق الخطوة االولى والخطوة ما قبل االخيرة الى من‬

‫كانوا في السنوات العُجاف سحابا ً ممطرا ً‬

‫الى الذين حملوا اقدس رسالة في هذه الحياة ومهدوا لنا‬

‫طريق العلم والمعرفة‬
‫الى الدكتورة المحترمة رفل شكيب ‪..‬‬

‫‪I‬‬
 List of Contents
Subject Page
‫االهداء‬ I

List of Contents II

List of Figures III

Abstract 1

The Aim of the Study 1

Chapter One

1.1.Introduction 2

1.2.History 3

1.3.Uses 3-4

1.4.Chemistry 4

1.5. Plant active Compound & Pharmacological activities 5

1.6. Organisms Used 6-7

Chapter Two

2.1.Material and Methods 8-10

2.2. Culture Media 11

Chapter Three

3.1. Results and Discussion 12

3.2. Conclusion 12

3.3. Recommendations 12

List of Figures
II
Number Page
Figure 1.1 1

Figure 1.2 2

Figure 1.3 4

Figure 1.4 5

Figure 1.5 6

Figure 2.1 8

Figure 2.2 8

Figure 2.3 9

Figure 2.4 10

Figure 2.5 11

III
Abstract:
Green coffee is unroasted coffee beans from Coffea fruits (Coffea
arabica, Coffea canephora). It contains more chlorogenic acid than
roasted coffee.

The chlorogenic acid in green coffee is thought to have health benefits. It

might affect blood vessels so that blood pressure is reduced. It might also
affect how the body handles blood sugar and metabolism. Green coffee
also contains caffeine, but in lower amounts than regular coffee.
(2005-2024 WebMD)

The Aims of The Study:

1. To collect and to identify green coffee medicinal plant.
2.To extract green coffee plant.
3. to evaluate the antibacterial activity of pure green coffee bean extract
on E. coli and staphylococcus and candida albican

Figure (1.1)

1
 Chapter One

1.1 Introduction
A medicinal plant is any plant that includes chemicals that can be utilized
for therapeutic.
reasons or that are precursors for the manufacture of valuable
pharmaceuticals in one or more.
of its organs. This description distinguishes between medical plants who's
therapeutic.
activities and ingredients have been scientifically verified and plants
recognized as medicinal.
but have not yet been submitted to a full scientific examination. (Jun21,
2023 by Franziska Spritzler)
Green coffee beans are simply regular coffee beans that haven’t been
roasted and remain completely raw.
Their extract is popular as a dietary supplement, but green coffee can also
be purchased in whole-bean form and used to make a hot beverage, much
like roasted coffee.
Bear in mind that a mug of this light green drink will not taste like the
roasted coffee you’re used to, as it has a much milder flavor. It’s said to
taste more like herbal tea than coffee.
What’s more, its chemical profile is quite different than that of roasted
coffee, though their origins are similar.
It boasts an abundant supply of chlorogenic acids — compounds with
potent antioxidant and anti-inflammatory effects that may provide many
health benefits.
Roasted coffee products also contain small amounts of chlorogenic acid,
but most of it is lost during the roasting process.
The leaves of the coffee tree
The coffee tree is an evergreen with spear-shaped leaves, which are green
and shiny on the upper side. As with all Rubiaceous plants, the leav grow
in pairs on either side of the stem. The leaves of the Robusta trees are
much larger than those of the Arabica.(Aug14, 2017)

Figure (1.2)

2
1.2 History:
Coffee beans come from coffee bean plants which are a type of shrub or
bush where the seeds are harvested from coffee cherries and processed to
obtain green coffee beans before they are roasted into beautiful brown
beans.

A coffee bean resembles a berry bush or grapevine but they are a lot
taller.
Coffee plants have rich, dark, and shiny green leaves, but some may be
more purple or yellow.
Coffee beans come from two main types of coffee plants: Arabica, and
Robusta. There are many different types of coffee beans out there, so
don't be fooled by their simplicity.
(1995to 2024 Culinary Depot)

1.3 Uses:
1) IMPROVED DIGESTIVE HEALTH
Metabolic acceleration and improved digestive health in general have
been associated with the consumption of both caffeine and chlorogenic
acid. Both of which are more prolific in green coffee beans than in beans
that have been roasted.
2) ELEVATED ANTIOXIDANT LEVELS
The same can also be said for the antioxidant content of raw coffee beans,
which in their unprocessed form contain far higher levels of antioxidants.
This has become one of the primary points of appeal for those who now
regularly consume green coffee, due to the broad and diverse health
benefits associated with antioxidants.
3) WEIGHT LOSS AND WEIGHT CONTROL
This particular claim is open to interpretation, but there’s growing
evidence to suggest that drinking coffee (green or regular) can assist with
weight loss and weight control. Unroasted coffee beans in particular are
theorized to be an organic appetite suppressant, making it easier to make
sensible dietary decisions while benefiting from improved metabolic
health.
4) A SERIOUS KICK OF CAFFEINE
For many, drinking coffee is all about that characteristic energy-kick that
accompanies caffeine intake. In which case, green coffee beans with their
elevated caffeine content are often in a league of their own. Going OTT
with caffeine consumption is inadvisable, but there really is nothing like
the kick that comes with a quality cup of green coffee!
5) GENERAL HEALTH BENEFITS

3
Lastly, numerous studies conducted over the past few years have linked
regular coffee consumption with a long list of general health benefits.
Research suggests that by drinking 3 to 5 cups of coffee a day, you may
be statistically less likely to develop certain diseases and conditions. If
this is the case, the same most certainly applies to green coffee beans.
which unlike their roasted counterparts have all their beneficial
compounds intact.
(Apr 13, 2018)

1.4 chemistry:
Green coffee beans contain a variety of chemical compounds, including
Polysaccharides, lipids, and proteins appear to be the major constituents
of the green beans, while minor components such as:

1- Caffeine.
2- Trigonelline, (contributes to the aroma and flavor of coffee).
3- Chlorogenic acids (are the primary phenolic compounds found in
green coffee beans and are responsible for their antioxidant properties
and others. Chlorogenic acids also contribute to the flavor profile of
green coffee).

free sugars (mainly sucrose), free amino acids, minerals, and others are
also relevant. (2008-2024 ResearchGate GmbH)

The minor compunds are the particularly important and considered

important sources of coffee aroma.

The exact composition can vary depending on factors such as the coffee
plant variety, growing conditions, and processing methods.

Figure (1.3)

4
1.5 Plant active Compound & Pharmacological
activities
Among the various herbal products, green coffee bean extract has
received greater attention due to its antimicrobial effect against both
Gram‐positive and Gram‐negative bacteria.
Some components in coffee such as caffeine, volatile and nonvolatile
organic acids, phenols and aromatic compounds are reported to have
antimicrobial activity. Chlorogenic acid (CGA) and caffeic acid, which
are nonvolatile organic acids found in coffee, inhibit the growth of some
Gram‐positive microorganisms such as Staphylococcus aureus, Bacillus
cereus, Lactobacillus bulgaricus, Streptococus lactis and Streptococcus
faecalis and Gram‐negative bacteria like Escherichia coli, Salmonella
typhi and Pseudomonas auerginosa (Fardiaz S . Antimicrobial activity)

Figure (1.4)

5
1.6 Organisms used:
One Gram Negative and One Gram Positive were used and yeast:

Gram-positive bacterial strains:

• Staphylococcus aureus.

Gram-negative bacterial strains:

• Escherichia coli.
• Yeast.
• Candida.

Figure (1.5)

Staphylococcus aureus is a Gram-positive spherically shaped bacterium,

a member of the Bacillota, is a usual member of the microbiota of the
body, frequently found in the upper respiratory tract on the skin. is often
positive for catalase and nitrate reduction and is a facultative anaerobethat
can grow without the need for oxygen

E. coli (Escherichia coli) is a type of bacteria that normally lives in your

intestines. It’s also found in the gut of some animals.

6
Most types of E. coli are harmless and even help keep your digestive tract
healthy. But some strains can cause diarrhea if you eat contaminated food
or drink polluted water.

Candida parapsilosis or C. parapsilosis is a yeast that's common on the

skin and often harmless. It also lives in soil and on the skin of other
animals. A healthy immune system can prevent C. parapsilosis infection
as well as having intact skin, or skin that doesn't have open nicks,
scrapes, or cuts.

7
 Chapter Two
2.1.Material and Methods:
1. green coffe bean grinded to fine powder.

Figure (2.1)

2. in the Lab. we weight (25 g) of green coffe.

Figure (2.2)

8
3. 25 grams of powder were extracted with 70% ethanol (350 ml of
ethanol and 150 ml DW.) using soxhlet apparatus.

Figure (2.3)

9
4. The extract solution was concentrated to dryness under reduced
pressure in a rotary evaporator, then let it dry in room temperature
for one day to yield a dried extract to used for antimicrobial testing.

Figure (2.4)

10
2.2 Culture media:
1. Muller Hinton.
2. Sabroud dextrose agar.

Culture media preparation:

1. Muller Hinton agar for E. coli isolation and staphylococcus aureus
isolation and Sabroud dextrose agar for candida isolation were
prepared.
2. Each media were sterilized by autoclaving at 121 Cº for 15 min.

Antibacterial activity crude extract by agar well diffusion method:

1. All bacteria were refreshed by culturing it overnight in nutrient broth.

2. Bacterial lawn was made for all types of bacteria on agar by using
sterile swabs.

3. Wells were made on pre inoculated agar 3 extract concentration were

added into well using micropipettes 50, 100 and 250 mg/ml.

4. The plates were kept for incubation for 24 hours at 37 C° after the
incubation the zone of inhibition were observed and measured.

Figure (2.5)

11
 Chapter Three
3.1 Results and Discussion:
Concentration E. Coli Staphylococcus Candida
50 R R R
100 R R R
250 R R R

Green coffee extract was taken to study the anti-microbial effect against
Staphylococcus aureus, Escherichia coli and Candida parapsilosis the
whole plant was extracted using, ethanol. The ethanol extract of green
coffee showed no anti-microbial effect on S. aureus, E.coli and Candida
with MIC value of (50,100,250 mg/ml) so ethanol extract of green coffee
of50, 100, 250 mg/ml showed no anti-microbial activity against both Gram
positive and Gram negative bacteria tested and candida. This result may be
belong to low concentration of plant and if we have higher concentration
the result may be different.

3.2 Conclusion:
1. The current study demonstrates that the green coffee extract exerts no
anti-microbial activities.
2. Anti-microbial activities reveals that this plant has no anti-microbial
activities against Staphylococcus aureus, and has no effects on
Escherichia coli and no effect on candida.

3.3 Recommendations:
1. Further studies on extraction of plant using different types of alcohol.
2. Further studies on anti-microbial of green coffee but using higher
concentration from extract.

12