1.

INTRODUCTION

1.1.Background
Nowadays, the brewing industry has applied a whole spectrum of technical, biochemical,
microbiological and genetic innovations. Furthermore, in today’s competitive beer-market
it is very important to develop different types of beers, for example beers obtained by the
use of new ingredients or new adjuncts (Carvalho et al., 2008). There are many kinds of
commercial fruit beer in the market. Fruit beers are any beers using fruit or fruit extracts as
an adjunct in either primary or secondary fermentation, providing obvious, yet harmonious
fruit qualities. Fruits have been used as a beer adjunct or flavoring for centuries. Modern
breweries may add only flavored extracts to the finished product rather than fermenting the
fruits. It was shown that the fruit beer contained more flavors and had better constitution
proportions compared with conventional beers (Liang, 2007). Fruit juice beers mostly
popular in occident market mainly contained the type of pineapple taste, lemon taste,
strawberry taste, etc. More and more researches are made to study the technology on juice
beer. In this research, fruit juice beer was improved by using lime juice as an adjunct. Lime
is common citrus fruit in Thailand. They are usually smaller and content more vitamin C
than lemons. Lime usually used to improve flavor in food because it has a strong flavor.
Lime also has a higher acid content (Nelson, 1996).

1.2.Literature Review
1.2.1.Lime
Lime (Citrus aurantifolia (Christm.) Swingle), is a highly polyembrionic species with them
fruit surface smooth, greenish-yellow in color and thin-skinned, core solid at maturity, and
juice highly acidic. Lime is used for the extraction of juice, preparation of squash,
concentrates, beverages and by- products such as citric acid and pectin, etc (Yadav et al.,
2003). Refrigerate uncut limes can be stored in a plastic bag for up to 10 days. Lime is
common citrus fruit in Thailand. They are usually smaller and content more vitamin C than
lemons. Lime usually used to improve flavor in food because it has a strong flavor. Lime

1
also has higher acid content and higher antioxidant content (Nelson, 1996 and
Thirugnanavel, 2007).

Lime is a good source of vitamin C. Vitamin C is a water soluble vitamin essential for the
formation of collagen and the healthy functioning of tissues containing collagen. It is
required for the repair of joint tissues which are often damaged during high levels of
physical activity. Vitamin C acts as a stimulant for body defense mechanisms and protects
vitamin A, E and dietary fats from oxidation. Vitamin C also plays an important role in the
absorption of iron from plant foods. Mild deficiencies can cause fleeting joint pains, poor
tooth and bone growth, poor wound healing, and an increased susceptibility to infection.
The nutrient values of raw lime juice are shown on Table 1.

Table 1. Nutrient Values of Raw Lime Juice / 100 gram

Nutrient Units Value per 100 gram
Water gram 90,79
Energy Kcal 25
Protein gram 0,42
Total Lipid (fat) gram 0,07
Carbohydrate gram 8,42
Total Dietary Fibre gram 0,4
Total Sugar gram 1,69
Vitamin C mgram 30
Source: USDA Nutrient Database, 2007

1.2.2.Beer
Beer is generic term for low-alcohol beverages brewed from a mash of malted barley and
other cereals, flavored with hops and fermented with yeast. It is produced by employing
either a bottom-fermenting yeast, which falls to the bottom of the container when
fermentation is completed, or a top-fermenting yeast, which rises to the surface. Lager
beers are bottom-fermented and stored at a low temperature for several months; most are
light in color, with high carbonation, medium hop flavor, and alcohol content of 3-5%by
volume. Top fermented beers, include ale, stout and porter, are characterized by prominent
head of released carbon dioxide, a sharper and more strongly hopped flavor than lagers, and
an alcohol content of 4-6,5% by volume (Hardwick, 2005).

2
1.2.2.1.Raw material of beer
Traditionally the raw materials of brewing are water, malt, hops and yeast. Many brewers
use in addition adjuncts. Each ingredient can affect flavor, color, carbonation, alcohol
content, and other subtle changes in the beer.

1.2.2.1.1.Water
Beer is composed mostly of water. The quality and composition of the water from different
beer-making regions contributes greatly to the character of the finished product. The
amount and composition of dissolved salts in water govern hardness and alkalinity. These
two properties influence the efficiency with which brewing processes operate and the way
the water extracts the desirable and undesirable substances from brewing raw materials
(Hardwick, 1995).

1.2.2.1.2.Malts
Malt, which is made from germinated grain (usually barley), provides beer with a slightly
sweet character. How malt is treated-dried but not roasted, lightly roasted, heavily roasted,
and so on impacts a beer’s flavor. Malt is derived from special varieties of barley by
controlled germination of the grain in the malting process. Grain is malted by soaking it in
the water, allowing it to begin germination, and then drying the partially germinated grain
in a kiln. Germination changes the physical and chemical structure of the barley, so that the
malt is friable product which provides a source of enzymes able to break down the
polymers of malt. Different roasting times and temperatures are used to produce different
colors of malt from the same grain. Darker malts will produce darker beers. Ground malt is
extracted with warm water in brew house to give a sweet wort. Arguably, malt is the most
important brewing raw material.

1.2.2.1.3.Hops
Hops are female flower cones of the hop plant (Humulus lupulus). They are dried in an oast
house before they are used in the brewing process. Hops resins are composed of two main

3
acids: alpha and beta acids. Alpha acids are responsible for the bitter flavor in beer. Beta
acids do not isomerize during the boil of wort and give a negligible effect on beer flavor.
Instead they contribute to beer’s bitter aroma, a high beta acid hop varieties are often added
at the end of the wort boil for aroma. Hop pellets are used because they are readily stored,
less bulky to store than whole hops, deteriorate less quickly and can reduce boiling time.
The effect of hops on the finished beer varieties by type and use, though there are two main
hop types: bittering and aroma. Bittering hops have higher concentration of alpha acids and
are responsible for the large majority of the bitter flavor of a beer. Aroma hops usually have
lower concentration of alpha acids and are the primary contributors of hop aroma and (non-
bitter) flavor. Bittering hops are boiled for a longer period of time, typically 60-90 minutes,
in order to maximize the isomerization of the alpha acids. They often have interior aromatic
properties, as the aromatic compounds evaporate off during the boil.

1.2.2.1.4.Yeast
Yeast is the microorganism that is responsible for fermentation in beer. Yeast metabolites
the sugars extracted from grains, which produces alcohol and carbon dioxide, and thereby
turns wort into beer. In addition to ferment the beer, yeast influences the character and
flavor. The dominant types of yeast used to make beer are ale yeast (Saccharomyces
cerevisiae) and lager yeast (Saccharomyces uvarum); their use distinguishes ale and lager.

Saccharomyces cerevisiae is knowm as top-fermenting yeast, so called because during the

fermentation process its hydrophobic surface causes the flocks to adhere to CO 2 and rise to
the top of the fermentation vessel. It is one of the major types of yeast used in the brewing
of ale. All strains of Saccharomyces cerevisiae can grow aerobically on glucose, maltose,
and trehalose, and fail to grow on lactose and cellobiose. It was shown that galactose and
fructose were two of the best fermenting sugars. The ability of yeast to use different sugars
can differ depending on whether they are grown aerobically or anaerobically.

4
1.2.3.Process of Beer
The process of making beer is known as brewing. Brewing process includes boiling mashed
malt to produce a wort, flavoring the wort with hops, fermenting this mixture with yeast,
and drawing off the fermented wort for bottling.

1.2.3.1.Milling
Milling of any substrate must achieve two objects: partial size reduction and particle size
control. Roll mills are used for milling of malt for brewing, because this best suits one
primary objective of milling: to leave the malt husk as intact as possible. An intact husk,
including absence of a shredded husk, helps hop separation in lautering and may reduce
extraction of tannins and other undesirable components. The objective of milling is to
produce crashed malt with the ideal spectrum of particles for extract production and
recovery. Small particles more easily yield extract, but lager one allow faster wort
separation. The way malt is milled depends on the expectations of the brewer, the quality of
malt and the equipment in which it will be processed (Lewis, 2002).

1.2.3.2.Mashing
Mashing is an important step in the brewing process and influences the type and quality of
the produced beer. The aim of mashing is to produce a wort containing suitable amounts of
fermentable sugars, yeast nutrients and flavor compounds. The final wort composition
depends on the temperature-time profile adopted which changes according to the type and
quality of the finished beer. The goal when choosing a suitable temperature-time profile is
to produce wort with the desired properties. Resting the mash at a specific temperature is
important for inducing certain enzymatic changes (Lewis, 2002). Mashing involves:
dissolving substances which are directly soluble in water; enzymic hydrolysis followed by
dissolving a series of substances important for the type and character of beer and separation
of the dissolved substances.

Wort is boiled to be sterilized and prepared hops are added. The objects of wort boiling are
arrest further enzyme action, precipitate some of the proteinaceous material of the wort as a

5
hot trub, sterilize the wort and hasten certain desirable chemical changes. The boiling is
typically last 45-60 minutes. Brewers add hops at prescribed times during boiling. The
addition of the hops is important because they contribute to the bitterness of the beer.
Aroma hops added shortly before the end of boil can impart a unique late hop flavor
character. When hops are boiled in wort the single most important reaction of hop
component is the isomerization of α-acid to iso-α-acid. At the end of the boil the wort must
be clarified, cooled, aerated, pitched with yeast and then sent to fermentation cellar
(Montanari et al., 2005).

1.2.3.3.Fermentation
The cooled aerated wort made in the brew house is fermented by yeast to make green beer.
During this process, yeast reproduces. The main fermentation (primary) in which green
beer and yeast are produce is often followed by a slower process at lower temperature in the
presence of lesser amount of yeast. This is referred to as secondary fermentation. It is
traditional practice to ferment infusion-mashed wort with top yeasts. The yeast is intimately
mixed with the wort at temperature around 16° C for top yeasts. About 1-2 gram of pressed
yeast is used for every liter of wort. The yeast grows using amino acids of the wort as the
main source of carbon and energy. Fermentation is complete in 2-10 days depending on the
conditions, particularly temperature (Briggs et al., 1981).

1.2.3.4.Finishing Process
Storage or aging practice follow a somewhat prolonged primary fermentation which yields
mature beer. This is achieved by lengthening the time at primary fermentation temperature
in a stage called the “diacetyl rest”. Storage and aging of beer comprises relatively short
term cold storage of up to a week or 10 days at -2 to 4° C. The objective is to permit yeast
to scavenge the oxygen, to encourage the formation of chill haze particles and to allow time
for yeast sedimentation (Briggs et al., 1981).

Carbonation is important to contribute the mouth feel of the beer. Generally beer will gain
about 1.5 volumes of CO2 per volume of beer as a result of fermentation at atmospheric
pressure. Carbonation therefore is used to increase the nature level of carbonation to that

6
required in the market place, which is above 2.0 volumes of gas per volume beer. Most
beer is carbonated by 10% (Briggs et al., 1981).

Beer is packaged under pressure of CO 2 into a metal keg. More recently the use of sterile
filtration rather than pasteurization of beer before packaging carbonated beer into kegs have
been introduced (Lewis, 2002). For most beer, it is considered desirable that they remain
brilliantly clear for several months stored at any temperature in the range 0-25° C, they
develop stable white foam on pouring into a glass, and the flavor and gas content remain
stable (Briggs et al., 1981).

1.3.The Objectives of Research

The objectives of this research is to determine the optimum condition of lime juice beer
using ale yeast and to evaluate the important property of lime juice beer.

7
2.MATERIALS AND METHODS
The research project was done during February 2010 to April 2010 and it was done in the
Chemical Laboratory and Pilot Plan Laboratory of Faculty of Biotechnology Assumption
University Bangkok Thailand.

2.1.Materials
2.1.1.Equipments
The equipments used in this research are frying pan, filter cloth, boiling pan, plastics,
grinding machine, vacuum filter, digital scale, keg, knife, fermentation tanks, thermometer,
bottles, hand refractometer, ebulliometer, pH meter, and baker glass.

2.1.2.Materials
The materials used in this research are malts, aroma hops, bitter hops, water, sugar,
Diatomaceous Earth (DE), ice, ale yeast (Saccharomyces cerevisiae), iodine, starch and
lime

2.2.Methods
2.2.1.Lime Juice Preparation
Whole limes were washed and dried, and then sliced each one in half. After cut, limes were
squeezed and the lime juice stored in a tightly sealed container in the refrigerator.

2.2.2.Brewing Process
2.2.2.1.Malt Preparation
Commercial malts were grinded, and then mixed with water to achieve the desired solid-
liquid ratio. Every kilogram of malt needs 4L of water.

2.2.2.2.Mashing
Since there is no adjunct used in this research, the mixture will go through a single
temperature infusion mash process. The process is described in Table 2.

8
Table 2. Mashing Process
Temperature (° C) Holding Time (minutes)
50 30
50-65 30
65 40
65-75 20

After boiled, the wort were filtered with filter cloth.

2.2.2.3.Wort Boiling
0,11% of ground bitter hops were added to the wort, then boiled the wort to 90° C for 60
minutes. After that, 0.05% of ground aroma hops were added and then boiled vigorously.
The whirlpool boil was done and the wort let to be precipitated. The wort and then lacked
into a fermentation container and cooled down by using ice.

2.2.2.4.Yeast Pitching
Dry yeast of Saccharomyces cerevisiae is used in this research. About 0,05 gram per Litre
yeast starter culture s were pitched and added into wort. The container and then shaken to
entrap air.

2.2.2.5.LIime Juice Addition and Fermentation

Different ratios of lime juice to wort were tested. 0.3%, 0.6%, 1%, 2% and 3% juice
additions were tested. 2 groups were set. For group 1, lime juice was mixed with wort and
then fermented under room temperature for around 5 days, and after that transferred into
the containers and put to the cold room for about 2 weeks. For group 2, after the wort was
fermented in a room temperature for 5 days, the wort then transferred into the containers
and the lime juice was added and put to the cold room for about 2 weeks.

2.2.2.6.Fitration, Carbonation, Bottling, Sanitation

Beer was filtered by powder filtration using the DE, and then carbonated. The beer and then
filled into bottles and sealed, and then pasteurized at 65° C for 30 minutes. The beer and
then kept in the cold room (Bamforh, 2002). The beer samples will be analyzed the alcohol

9
 content, degree of acidity (pH), sugar degree, vitamin C and also sensory. The result
presented in ten codes, A (0.3% lime juice addition before primary fermentation), B (0.6%
lime juice addition before primary fermentation), C (1% lime juice addition before primary
fermentation), D (2% lime juice addition before primary fermentation), E (3% lime juice
addition before primary fermentation). F (0.3% lime juice addition after primary
fermentation), G (0.6% lime juice addition after primary fermentation), H (1% lime juice
addition after primary fermentation), I (2% lime juice addition after primary fermentation),
J (3% lime juice addition after primary fermentation). The process of beer making can be
seen in Figure 1.

Malt Grinded

Mixed with water and then mashed

10
Wort
0,11% of bitter hops
 Whirlpool boil

Cooled

Yeast pitching

Primary fermentation
+0,3% +0,6% +1% +2% +3%
lime lime lime lime lime
juice juice juice juice juice
+0,3% +0,6% +1% +2% +3%
lime lime lime lime lime
juice juice juice juice juice

Primary fermentation

Secondary fermentation

Filtered

Carbonated

Bottled

Pasteurized

Lime juice beer

= Process

=Raw Material

= Product

Figure 1. The process of beer making

2.2.3.Measurement

11
2.2.3.1.Degree of Acidity (pH) Measurement
First the sample preparation was done. For the homogenous liquid sample, the pH
measurement can be done directly without dissolving the sample. Before measuring pH of
samples using pH-meter, temperature of samples were measured. Before used, pH-meter
stabilized for ± 15-30 minutes. After that, the electrode of pH-meter washed by samples
aliquots continued with aquadest. After washed, electrode soaked in the samples and leave
for a while until stable. The results were collected (Apriyantono et al., 1989).

2.2.3.2.Alcohol Measurement
In this measurement, ebulliometer was used to measure the alcohol content. First, the
boiling chamber washed using aquadest. After that, the aquadest were put in and boiled.
The boiling point was collected after it’s stabil. After the boiling point of aquadest was
collected, the boiling chamber washed by lime juice beer aliquots for around 30 seconds.
The beer soon was put in the chamber and some antifoam were added to avoid excess foam.
After the boiling point was stable, the boiling temperature was collected and the alcohol
content was counted using a calibration standard.

2.2.3.3.Vitamin C Measurement
First, samples were taken 10 mililiters and then added with 10 ml H 2SO4 1M. After that,
samples were added with 3 mililiters of 1% starch solution as an indicator just right before
titration. Samples after that titrated with 0,01 N iodin solution (Apriyantono et al., 1989).

2.2.3.4.Test of Beer Clarity

The clarity of all lime juice beer were tested by pouring the beers in the glasses and seen
visually with a brigth background.

2.2.3.5.Sugar Degree
Sugar degree of the samples were tested using hand refractometer.

12
2.2.3.6.Sensory Evaluation
Sensory evaluation of all samples were done with some parameters. Those paramaters were
clarity, flavor, taste, general impression. The evaluation is done using quistionaire and it’s
done by 10 trained panelist. The method which used in this evaluation is hedonic rank test
(Meilgaard et al., 1999). Each panelist were asked to fill the quistionaire with the scores
that have been given in the scoresheet.

3.RESEARCH RESULT

13
The effect of lime juice addition in beer before fermentation and after fermentation on
alcohol content, degree of acidity (pH), sugar degree, vitamin C and also sensory can be
seen in this chapter.

B D F H J
A C E G I

Figure 2. Lime juice samples

Annotation:
 A (0.3% lime juic
fermentation), C (1% lime juice addition before primary fermentation), D (2% lime juice addition before
primary fermentation), E (3% lime juice addition before primary fermentation)
 F (0.3% lime juic
fermentation), H (1% lime juice addition after primary fermentation), I (2% lime juice addition after
primary fermentation), J (3% lime juice addition after primary fermentation)

Visually, lime juice beers can be seen the difference very clearly. Some lime juice beers
have darker colors. The right side lime juice beers have darker colors than the left ones.

3.1.pH

14
The effect of lime juice addition before fermentation and after fermentation on degree of
acidity (pH) can be seen in Table 3 below.

Tabel 3. pH of Lime Juice Beers

Treatment pH
A 4.22
B 4.18
C 3.86
D 3.77
E 3.68
F 4.36
G 4.25
H 4.18
I 4.00
J 3.77
Annotation:
 All values are bas
 A (0.3% lime juic
fermentation), C (1% lime juice addition before primary fermentation), D (2% lime juice addition before
primary fermentation), E (3% lime juice addition before primary fermentation)
 F (0.3% lime juic
fermentation), H (1% lime juice addition after primary fermentation), I (2% lime juice addition after
primary fermentation), J (3% lime juice addition after primary fermentation)
 Measurement usin

Shown from Table 3, the pH of the first group beers (A-E) which were mixed with lime
juice beers before primary fermentation significantly decreased while in the second group
beers (F-J) which were mixed with lime juice beers after primary fermentation the pH were
also significantly decreased. The highest pH was F (0.3% lime juice addition after primary
fermentation), while the lowest pH was E (3% lime juice addition before primary
fermentation).

3.2.Alcohol Content
The effect of lime juice addition before fermentation and after fermentation on alcohol
content can be seen in Table 4 below.

Tabel 4. Alcohol Content of Lime Juice Beers

15
 Treatment Alcohol Content
(%)
A 5.12
B 4.20
C 5.82
D 5.55
E 6.40
F 4.20
G 4.60
H 4.52
I 6.50
J 6.40
Annotation:
 All values are bas
 A (0.3% lime juic
fermentation), C (1% lime juice addition before primary fermentation), D (2% lime juice addition before
primary fermentation), E (3% lime juice addition before primary fermentation)
 F (0.3% lime juic
fermentation), H (1% lime juice addition after primary fermentation), I (2% lime juice addition after
primary fermentation), J (3% lime juice addition after primary fermentation)
 Measurement usin

Shown from Table 4, the highest alcohol content was I (2% lime juice addition after
primary fermentation, while the lowest alcohol content was B (0.6% lime juice addition
before primary fermentation) and F (0.3% lime juice addition after primary fermentation).

3.3.Sugar Degree
The effect of lime juice addition before fermentation and after fermentation on sugar degree
can be seen in Table 5 below.

Tabel 5. Sugar Degree of Lime Juice Beers

Treatment Sugar Degree
(°Brix)
A 6.20
B 6.00
C 6.40
D 6.40
E 6.00
F 6.60
G 6.20
H 7.80

16
 I 7.20
J 7.40
Annotation:
 All values are bas
 A (0.3% lime juic
fermentation), C (1% lime juice addition before primary fermentation), D (2% lime juice addition before
primary fermentation), E (3% lime juice addition before primary fermentation)
 F (0.3% lime juic
fermentation), H (1% lime juice addition after primary fermentation), I (2% lime juice addition after
primary fermentation), J (3% lime juice addition after primary fermentation)
 Measurement usin

Shown from Table 5, the highest sugar degree was H (1% lime juice addition after primary
fermentation), while the lowest alcohol content was B (0.6% lime juice addition before
primary fermentation) and F (0.3% lime juice addition after primary fermentation).

3.4.Vitamin C Content
The effect of lime juice addition before fermentation and after fermentation on sugar degree
can be seen in Table 6 below.

Tabel 6. Vitamin C Content of Lime Juice Beers

Treatment Vitamin C (mg/L)
A 1.83
B 0.23
C 0.30
D 0.30
E 0.45
F 0.17
G 0.25
H 0.33
I 0.35
J 0.35
Annotation:
 All values are bas
 A (0.3% lime juic
fermentation), C (1% lime juice addition before primary fermentation), D (2% lime juice addition before
primary fermentation), E (3% lime juice addition before primary fermentation)
 F (0.3% lime juic
fermentation), H (1% lime juice addition after primary fermentation), I (2% lime juice addition after
primary fermentation), J (3% lime juice addition after primary fermentation)
 Measurement usin

17
Shown from Table 6, the highest vitamin C content was A (0.3% lime juice addition before
primary fermentation), while the lowest vitamin C content was F (0.3% lime juice addition
after primary fermentation).

3.5.Sensory Evaluation
The effect of lime juice addition before fermentation and after fermentation on sensory
evaluation can be seen in Table 7 below.

Tabel 7. Sensory Evaluation of Lime Juice Beers

Treatment Clarity Flavor Taste General Impression
A 2.43 4.82 5.00 5.00
B 4.14 3.39 3.85 3.13
C 3.36 5.00 4.18 3.93
D 1.00 3.75 3.48 3.93
E 2.71 1.00 1.00 1.00
F 2.43 3.85 4.33 3.67
G 5.00 4.55 4.12 3.80
H 1.71 4.88 2.48 4.06
I 3.36 2.70 3.55 3.00
J 4.43 2.56 3.48 3.67
Annotation:
 All values are bas
 A (0.3% lime juic
fermentation), C (1% lime juice addition before primary fermentation), D (2% lime juice addition before
primary fermentation), E (3% lime juice addition before primary fermentation)
 F (0.3% lime juic
fermentation), H (1% lime juice addition after primary fermentation), I (2% lime juice addition after
primary fermentation), J (3% lime juice addition after primary fermentation)
 Evaluated by 10 t
 All scores have be

Shown from Table 7, the highest score for lime juice beers clarity was achieved by G (0.6%
lime juice addition after primary fermentation), while for flavor the highest score was
achieved by C (1% lime juice addition before primary fermentation). For the taste, the
highest score achieved by A (0.3% lime juice addition before primary fermentation), while
for general impression the highest score was achieved by A (0.3% lime juice addition
before primary fermentation).

18
Shown also from Table 7, the lowest score for lime juice beers clarity was achieved by D
(2% lime juice addition before primary fermentation), while for flavor, taste and general
impression the lowest score was achieved by E (3% lime juice addition before primary
fermentation).

19
4.DISCUSSION

From Figure 2, can be seen that the lime juice beers have different colors. Some of the lime
juice beers have darker colors. According to Hardwick (2005), different roasting times and
temperatures of malts are used to produce different colors of malt from the same grain.
Darker malts will produce darker beers. So, the different color of final lime juice beers can
probably be affected by the roasting times and temperature of the raw malts before used to
make the lime juice beers.

4.1.pH
From Table 3, can be seen that the results of pH of the lime juice beers are around 3-4. This
means that the pH of the lime juice beers have reached to acid condition. This acid
condition can be obtained as the result of lime juice addition. According to Yadav et al.,
(2003), lime (Citrus aurantifolia (Christm.) Swingle), has a highly acidic juice and usually
used for the by- products making such as citric acid. The addition of lime juice can
decrease the pH level into acidic one and make the final beers become acid beers. Higher
additions of lime juice (higher concentration of lime juice) cause lower pH (more acid) of
the beers in both additions before primary fermentation and after primary fermentation.
Beside the addition of lime juice, the acid pH of the beers can probably be affected by the
hops. According to Hardwick (2005), hops resins are composed by two main acids: alpha
and beta acids and the acidity of hops is a preservative. The acid composed in hops can be
the other reason of the acid pH obtained by the beers.

4.2.Alcohol Content
From Table 4, can be seen that the results of alcohol content of the lime juice beers are
around 4.20-6.5 %. In this beer making process, the yeast which was used to make the beers
was Saccharomyces cerevisiae which is ale yeast. Consequently the type of the beers is ale
beers. According to Hardwick (2005), top fermented beers, include ale, stout and porter, are
characterized by prominent head of released carbon dioxide, a sharper and more strongly
hopped flavor than lagers, and an alcohol content of 4-6,5% by volume. The addition of

20
lime juice has no effect to the alcohol content of the beers since the alcohol content of all
beers is still in the range.

4.3.Sugar Degree
From Table 5, can be seen that the results of sugar degree of the lime juice beers are around
6-7 °Brix. From Table 5, shown that after primary fermentation (F-J) the sugar degree
results are higher than before primary fermentation (A-F). This probably because the
additions of lime juice after primary fermentation cause the sugar degree of the beers
increased. Addition of lime juice before primary fermentation cause the yeast to break
down all the sugars contained in beers during fermentation, while addition after primary
fermentation, cause the yeast to break down not all the sugars contained but just sugars
contained in primary fermentation.

4.4.Vitamin C Content
From Table 6, can be seen that the results of vitamin C content of the lime juice beers
increase with the increase of lime juice concentration addition. According to Nelson (1996)
and Thirugnanavel (2007), lime is common citrus fruit which is usually smaller and content
more vitamin C than lemons. Lime is a good source of vitamin C. Therefore, higher
addition of lime juice, the vitamin C content will also higher.

4.5.Sensory Evaluation
From Table 7, can be seen the results of sensory evaluation on lime juice beers. For clarity,
the highest score has achieved by G (0.6% lime juice addition after primary fermentation)
which means that the beer has the clearest color while the lowest score has achieved by D
(2% lime juice addition before primary fermentation) which means that the beer has the
most turbid color. This clarity is probably affected by filtration process. During filtration,
some beers need to be filtrated in two or three stages because there are suspended materials
still remained and make the beers turbid. According to Hardwick (2005), the first filtration
stage removes the bulk yeast and suspended materials and the second stage produces a

21
brilliantly clear beer. The stage of filtration and the suspended materials remained in beers
cause different clarity.

For flavor, the highest score has achieved by C (1% lime juice addition before primary
fermentation) which means that the beer has the strongest flavor. This probably affected by
the addition of lime juice in the right proportion. According to Nelson (1996) and
Thirugnanavel (2007), lime usually used to improve flavor in food because it has a strong
flavor. And C probably has the most balanced flavor composition between lime juice
flavor and hops flavor. For taste, the highest sore achieved by A (0.3% lime juice addition
before primary fermentation). This probably because A has the lowest concentration of
lime juice and generating a suitable bitter sensation from hops which is liked by most of the
panelists. For the general impression, the highest score has achieved by A (0.3% lime juice
addition before primary fermentation). This result is probably affected the most by taste
where A is liked by most of the panelists. The panelists consider taste as the strongest
parameter which determines the quality of the beers which means acceptance from the
consumers in wide market. While flavor and clarity considered not the first parameter
which will be seen by consumers in wide market.

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5.CONCLUSION

 The results of pH of the lime juice beers are around 3-4 which means that lime juice
beers are acid.
 The results of alcohol content of the lime juice beers are around 4.20-6.5 % in range
which means the addition of lime juice has no effect to the alcohol content of the beers.
 The results of sugar degree of the lime juice beers are around 6-7 °Brix.
 The results of vitamin C content of the lime juice beers increase with the increase of
lime juice concentration addition.
 The highest clarity score for lime juice beers was achieved by G (0.6% lime juice
addition after primary fermentation).
 The highest flavor score was achieved by C (1% lime juice addition before primary
fermentation).
 The highest score for the taste achieved by A (0.3% lime juice addition before primary
fermentation).
 The highest general impression score was achieved by A (0.3% lime juice addition
before primary fermentation).

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6.REFERENCES

Apriyantono, A; Dedi Fardiaz; N.L. Puspitasari; Sedarnawati; Slamet Budianto. Analisis

Pangan. (1989). Institut Pertanian Bogor Press. Bogor.

Bamforh, C.W. (2002). Standard of Brewing. Brewers Publication 73, 124, 126-127.

Briggs, D.E., J.S. Hough, R. Stevens, dan T.W. Young. (1981). Malting and Brewing
Science Volume 1 Malt and Sweet Wort. Chapman & Hall. London.

Carvalho, G., Daniel S., Antonio V., Maria das Gracas, Jose T, dan Joao B. (2008). A New
Strategy Using Banana as an Ingredient In the Brewing Process.
http://repositorium.sdum.uminho.pt/bitstream/1822/9195/1/Carvalho_G%5B1%5D.pdf

Hardwick, W. A. (1995). Handbook of Brewing. Marcel Dekker, Inc.

Lewis, M. dan Tom W. Young. (1995). Brewing. Chapman & Hall. London.

Liang, Zhang and Zhao Changxin. (2007). Study of Different Compositions of Fruit Beer
and Beer. China Brewing, Serial Number 167, 37-40.

Meilgaard, M; G.V. Civille and B. T. Carr. (1999). Sensory Evaluation Techniques 3 rd

Edition. CRC Press. ASA.

Montanari, L., Simona Floridi, Ombretta Marconi, Michela Tironzelli, dan Paolo Fantozzi.
(2005). Effect of Mashing Procedures on Brewing. Eur Food Res Technol 2005.

Nelson, G. (1996). Fruits of Warm Climates.

http://www.fs.fed.us/global/iitf/pdf/shrubs/Citrus%20aurantiifolia.pdf

Thirugnanavel, A., R. Amutha, W. Babby, K. Indira, P. Mareeswari, S. Muthulaksmi, dan S

Parthiban. (2007). Studies on Regulation of Flavouring in Acid Lime (Citrus aurantifolia
swingle.). Research Journal of Agriculture and Biological Science. ISInet Publication.

USDA Nutrient Database. (2008).

http://www.nal usda.gov

Yadav, Ramesh A., A.S. Chauhan, M.N. Rekha, L.J.M. Rao, R.S. Ramteke. (2004). Flavor
Quality of Dehydrated Lime. Journal of Food Chemistry 85, 59-62.

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7.APPENDIX

Appendix 1. Process of Making Lime Juice Beer

Curing the Malt Yeast Pitching, Lime Juice Addition and

Fermentation

Milling Filtering

Mashing and Wort Boiling Bottling (Lime Juice Beer)

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