Table of Content

1. Abstract
2. Introduction

2.1 History of Rye cultivation

2.2 History of Pilsner cultivation

2.3 History of Rice cultivation

2.4 Geographical Distribution of Rye, Pilsner, and Rice

2.5 Nutritional facts of rye, pilsner and rice

3. Review of Literature
4. Materials and Methods

4.1 Procedure

5. Evaluation of brewing characteristics and their analysis

5.1 Determination of Ethyl Alcohol Content - Pycnometer Method or Hydrometer

Method

5.2 Determination of Apparent extract

5.3 Determination of Ph

5.4 Determination of Real extract

5.5 Determination of carbohydrates by Anthrone method

5.6 Estimation of protein by biuret method

5.7 Estimation of iron in beer

5.8 Estimation of Polyphenols in Beer

5.9 Estimation of Vitamin E in Beer by Gas Chromatography

6. Result and Discussion

1
 6.1 Sensory Evaluation and Comparison with commercial beer

7. Effect of storage period on shelf-life and nutritional quality of rye beer

7.1 Off flavours in beer

8. Conclusion

2
Abstract:

This research paper explains the creative development and characterization of a beer
formulation by incorporating with Rye. The Pilsner malt and rice as alternative
adjuncts to beer. The incorporation of Rye, Pilsner mat and rice into beer formulations
presents an creative approach to diversifying the beer landscape while enhancing its
nutritional value.This Abstract provides a comprehensive overview of beer,
combining historical insights, technical knowledge, culture context, sensory
exploration, and contemperory developments. Also this study aimed to investigate the
impact of incorporating rye, pilsner malt and rice on the physicochemical, sensory,
and nutritional properties of the resulting beer.

The research employed a series of controlled brewing experiments, where in varying

proportions of rye, pilsner and rice were used in conjunction with malted barley. The
brewing process involved mashing, lautering, boiling, fermentation, and conditioning,
adhering to standard practices. Physicochemical analyses, including specific gravity,
pH, color intensity, and bitterness level, were conducted to assess the impact of the
adjunct grains on the beer's chemical composition and appearance. Additionally,
sensory evaluations were performed by a trained panel to discern any perceptible
differences in aroma, flavor, mouthfeel, and overall acceptance between the
alternative grain-incorporated beers and traditional barley-based beer.

3
STUDY AND DEVELOPMENT OF BEER FROM RYE AND PILSNER MALT

2.Introduction

RYE Scientifically named as Secale cereale, one of the cereal grain related to wheat and
barley. And is also called as Winter rye chiefly used for making Bread, Whisky, Beer and for
animal fodder. It is high in carbohydrates and dietary fibre and provides small quantity of
protein, potassium, and B vitamins. Rye can meet the height of 1 to 2 metres, depending on
variety. Rye became a staple food in Europe during the Middle Ages, where is was cultivated
in Central and Eastern Europe and was used to develop the bread and alcohol. Leading
countries produces Rye are Russia, Germany, Austria, USA, Hungary. And in India Punjab,
Haryana, UP cultivate Rye crop.

Figure – 1 Rye grains

Also there are some hybrids in Rye such as Abruzzese Rye. Danko Rye. Brasetto Rye used by
distillers. Brewers, and bakers.

Pilsner malt is type of lager malt made from two-row spring barley primarily grown as
animal fodder and as a source of malt for alcoholic beverages especially beer. Pilsner malt is
always highly modified (i.e., good protein degradation) during malting, also a very light
coloured base malt, with a neutral flavor and high enzyme content. Typically has enough
amylase enzyme to convert a significant percentage of unmalted adjuncts.

Fig – 2 Pilsner malt

4
Rice (Oryza sativa) is one of the most important cereal crop in developing world belongs to t
grass plant (family poaceae) and it is the staple food of over half the world´s population. And
it is commonly considered as semi-aquatic annual grass plant and grows to about 1.2 metres
in height. The leaves are flattered, long, and are borne on hollow stems. The fibrous root
system is often board and spreading. Varieties differ greatly in the length, shape, and weight
of the panicle and the overall productivity of a given plant.

Fig – 3 Rice

Alcoholic beverages which occur throughout the world in many forms and taste results from
the action of microorganisms or enzymes on a wide range of agricultural products such as
grapes, grains, and soybeans ( Rose et al., 1989). Such biological action was associated with
biochemical changes that gave rise to significant organoleptic improvements to the final
products. These products which are more nutritious and more easily digestible are
toxicologically and microbiologically safer (Smith et al., 1996). The process of fermenting
raw materials is of wide diversity, using technology from the most primitive to the most
advanced achieving an outstanding range of sensory and textural quality in the final product.
Alcoholic beverages and potable spirit industries represent one of the most economically
stable sectors in the present day commerce. Materials for alcoholic beverages normally
comprise either sugary material (fruit juice, plant sap) or starchy materials (grains or roots)
which need to be hydrolyzed to simple sugar before fermentation. When these substrates are
incubated with suitable microorganism and allowed to ferment, the end product is a liquid
containing from a few percentages to about 16% alcohol with a slightly acidic pH.

Rye beer is one of the beer style that originated in Bavaria, in Southern Germany. Ingredients
that are used for rye beer recipe will have a combination of rye grains, water and hops. Rye

5
Beer carries the spice taste, and flavor of rye beer is with added sweetness and aroma of
honey. The Alcohol percentage of Rye Beers depends on the beer type, ale or lager, but on
average tends to fall between the 4-6% range. (Chermaine Chee et al., 2022)

Pilsner malt is used as adjunct in rye beer made from Hana, a common barley variety that
grown in Bohemia and Moravia. × It is a modified malt with excellent processing
characteristics ,containing total protein and glucan levels , also have outstanding enzymatic
strength. Also with good lautering properties with high extract. × Pilsner malt provides the
final beer with delicate maltiness ,mouthfeel as well as good foam development , also tend to
be soft by giving more colour with toasty note. (Thomas Kraus – Weyermann et al.,2003).

Addition of rice along with hull will reduce the stickiness during mashing by giving bulk
nature. Rice is the least allergenic of all grains and naturally gluten free with complex
carbohydrates composition, also contain more vitamins such as B-complex and minerals such
as , iron, zinc and etc. (Jonas Rembert et al., 1997)

2.1 History of Rye cultivation

Rye cultivation probably originated in Southwestern Asia about 6500 bce across the Balkan
peninsula and over Europe ana also originates from the area around black sea and is primarily
grown in an area known as Rye Belt. It has been harvested for more than 6000 years, but it
was around year 500A.D. that cultivation of rye became important in Central Europe. In the
12th and 13th centuries rye had became the major grain and placed as major cereal until the
Second World War in Germany. It is primarily grown in an area denominated as the Rye Belt.
This comprises countries in North and East Europe, i.e. Germany, Poland, The Nordic
Countries, The Baltic Countries, Belarus, Ukraine, Austria, Czech Republic and Slovakia.
Some rye is also produced in Canada and the USA. In the countries of the Rye Belt the
grains are primarily used for bread flour, it gives a good taste to the bread and is considered a
healthy grain.

Within the Rye Belt, rye has different applications. The most important are: Bread, animal
feed and bio energy. In the Nordic and the Baltic countries, rye is traditionally used for
bread. In Denmark rye production for animal feed has increased in the last years as a result

6
of a policy of more self-sustained feed production. An increased production of rye for both
biogas and animal feed is foreseen.
2.2 History of Pilsner cultivation ( 2-row barley)

Most countries outside of North America exclusively use two-row barley for malt (six-row, if
produced, is used for feed), but six-row varieties have played an important role in US
brewing. Barley was first introduced by European colonists in the early 1600s who quickly
found that six-row types were better suited to the new world climate compared to the
two-row types. With the pass of time barley production has been pushed west cut to
displacement by disease pressures and increased production of other major crops such as corn
and soybean. Relatively higher yields than two-row and high demand from the big brewers
kept six-row as the dominant barely type as late as the ‘90s. Today that has changed and
according to surveys conducted by the American Malting Barley Association (AMBA) about
85% of reported acreage was planted with two-row in 2018.

Breeding efforts primarily focus on improvement of two-row varieties such as pilsner and
etc., and in fact AMBA, which publishes guidelines for breeding based on end usage, no
longer provides recommendations for six-row. This is matched by AMBAs 2019 list of
recommended varieties which includes more than 25 two-row types but only 7 six-row
options.

2.3 History of Rice cultivation

Many cultures have a evidence of early rice cultivation, including China, India, and Southeast
Asia. However the earliest archeological evidence comes from the central and eastern China
and dates to 7000-5000 BCE. Almost 90 percent of the world’s rice is grown in Asia,
principally in China, India, Indonesia, and Bangladesh, with smaller amounts grown in Japan,
Pakistan, and various Southeast Asian nations. Rice is also cultivated in parts of Europe, in
North and South America, and in Australia.

Rice is fundamentally important to various cultures. It is often directly associated with

prosperity and much folklore and legend surrounds the grain. In many cultures and societies,
rice is integrated directly into religious belief. In Japan rice enjoys the patronage of its own
god, Inari, and in Indonesia its own goddess, the Dewie Srie.
Rice is also linked to fertility and for this reason the custom of throwing rice at newly
wedded couples exists. In India, rice is always the first food offered by a new bride to her

7
husband, to ensure fertility in the marriage, and children are given rice as their first solid
food.
How easy to see that from its early beginnings to the present day, rice continues to play an
integral role in sustaining both the world's appetites and cultural traditions.

2.4 Geographical Distribution of Rye, Pilsner, and Rice

In India Rye grain is grown in a local villages around Kullu, and locals mostly call it “Neela
Jau” and “Russi Gehu”, owing its color and prevalent geographical production respectively.
Also some have been growing it in upper Himalayan region near Mukteshwar.

According to FAOSTAT in 2022 around 15.32 million tonnes of rye grains was harvested
globally, out of which 81% was grown in Europe and 13% in Asia. In the world it was
cultivated in area of 16.3m.ha with a productivity of 40.m.t.

Top 10 Rye producers

Countries Metric tonnes

Germany 3,237,600
Poland 2,415,640
Russia 1,428,421
Denmark 883,510
Belarus 755,547
China 511,363
Ukraine 334,680
Canada 333,400
Turkey 310,000
United States 269,810
table - 1

Pilsner malt( 2-row) is type of barley and grown in around 70million hectares of land across
the world includes 6-row barley. Europe is the most leading continent growing Barley
followed by Asia. Other barley producing countries are Russian federation, China, Canada,
USA, Spain, France, Australia, UK and India. In India, Uttar Pradesh, Rajasthan, Madhya
Pradesh, Haryana, Punjab and Himachal Pradesh are the major producers of barley crop. In
India.

8
 ● Rajasthan (53% of total Barley)
● Utter Pradesh (26% of total Barley)
● Madya Pradesh (11% of total Barley)

According to the US Department of Agriculture’s World Agriculture Supply and Demand

Estimates (WASDE) November 2022 report, the global production of rice is projected to
reach 503.69 million tonnes in 2022-23. Of that total, USDA expects 52.83 million tonnes
will be exported to world markets.

China is the largest producer and importer of rice to meet its projected 2022-23 domestic
consumption of 155 million tonnes. Exports are seen at 2.2 million tonnes. India, the world
second populous country with 1.39 billion people, will harvest 124 million tonnes of rice in
2022-23. India is expected to consume 109 million tonnes while exporting a world leading
19.5 million tonnes.

2.5 Nutritional facts

Rye

Nutrients Amount (100g)

Carbohydrates 26.84g
Protein 10.98g
Fats 1.3g
Calcium 24mg
Iron 2.93mg
Sodium 6mg
Potassium 510mg
Fibre 13.9g
table - 2

Pilsner (2-row barley)

Nutrients Amount (100g)

Carbohydrates 25.8g
Protein 11g
Fats 2,2g

9
 Potassium 452mg
Calcium 67mg
Fibre 17g
Sodium 12mg
table - 3

Rice

Nutrients Amount (100g)

Carbohydrates 27.9g
Protein 2.64g
Fats 1.07g
Potassium 35mg
Calcium 10mg
Fibre 0.4g
Sodium 368mg
table - 4

Benefits of Rye ,Pilsner, and Rice

Rye

Rye has been shown to offer a range of health benefits, particularly for people with conditions
such as PCOS, diabetes, and thyroid problems, and those looking to lose weight. For PCOS
(Polycystic Ovary Syndrome), rye is a low-glycemic food, meaning that it has a low impact
on blood sugar levels. This makes it a good choice for women with PCOS, who are often at
higher risk for insulin resistance and other blood sugar-related issues.

For weight loss, rye can be a helpful addition to a weight loss diet because of its fibre and
protein content. The fibre in rye helps to fill you up and regulate digestion, while the protein
helps to support muscle growth and repair.
Vitamins & Minerals in Rye
Rye is a good source of several vitamins and minerals that are important for overall health
and wellness. Some of the key vitamins and minerals in rye include:

10
 ● Thiamine (Vitamin B1): This B vitamin is important for energy production
and brain function.
● Niacin (Vitamin B3): This B vitamin supports healthy skin, nerves, and
digestion.
● Magnesium: This mineral is important for heart health, bone strength, and
muscle function.
● Phosphorus: This mineral is important for strong bones and teeth, and helps
the body store and use energy.
● Manganese: This mineral is important for healthy bone development, wound
healing, and antioxidant defense.
Pilsner Malt

Reduce Hunger; Barley may reduce hunger and promote feelings of fullness both of which
may lead to weight loss over time. Barley lessens hunger largely through its high fiber
content. A soluble fiber known as beta-glucan is particularly helpful. That’s because soluble
fibers, such as beta-glucan, tend to form a gel-like substance in gut, which slows the digestion
and absorption of nutrients. In turn, this curbs appetite and promotes fullness.

Reduce Cholesterol; The beta-glucans found in barley have been shown to reduce “bad”
LDL cholesterol by binding to bile acids.

Reduce Heart Disease; Whole grains are consistently linked to better heart health.
Therefore, it should come as no surprise that regularly adding barley to diet may lower risk of
heart disease.

Prevent colon cancer; A diet rich in whole grains is generally linked to a lower likelihood of
many chronic diseases, including certain cancers especially those of the colon. Its insoluble
fiber specifically helps reduce the time food takes to clear your gut, which appears
particularly protective against colon cancers. Additionally, soluble fiber may bind to harmful
carcinogens in your gut, removing them from body.

Rice

11
Gluten free; Rice is a hypoallergenic food and also a blessing for people who are allergic to
gluten or have celiac disease. A gluten-free grain, rice can also be easily turned into flour,
noodles, and bread for people who opt for gluten-free options.

Aids Heart health; When you opt for wholegrain rice, you help your heart health out. Even
the American Heart Association agrees that whole grains improve blood cholesterol levels
and further reduce the risk of heart disease and strokes.

Objectives of study

● Formulation of beer production from Rye, Pilsner, and Rice

● Evaluation of brewing characteristics and their analysis

● Effect of storage period on shelf-life and nutritional quality of rye beer

3. Review of Literature

Segobia Munoz et al., (2022) evaluate the addition of malted and unmalted rye to brew a
Belgian Pale Ale style craft beer. This research is to innovate and explore the attributes
provided by this cereal, in addition to evaluating the phytochemical (color, pH, alcohol
content and turbidity), microbiological (molds and yeasts) and sensory (flavour, odour, color
and texture parameters. An experimental design with a bifactorial A*B arrangement was
applied with 3 replicates per treatment and 1 control using the Dunnett test. Regarding the
sensory test, the Kruskal Wallis test of contrasts was applied using SPSS Statistics software.
Thus is was found that the addition of addition of malted and unmalted significantly
influenced the characteristics evaluated in the craft beer, giving rise to a profile typical of
Belgian Pale Ale, with the distinctive touch provided by the rye.

Most outstanding treatments in terms of flavour and texture were T5 (80% base malt +
malted rye); in terms of odour, T1 (90% base malt + malted rye) stood out, and T4 (85% base
malt + unmalted rye) stood out in terms of color.

12
The result of this study show which quality parameter of rye malt can be optimised by
varying germination conditions. The most limiting factor for the use of malt for brewing
purposes was very high wort viscosities caused by high amount of gluten. However, the use
of malt as raw material for functional food appears to be more promising. Health promoting
properties of the malts, especially of nutrients and their breakdown products can be utilised.
Choosing favourable germination. (Hubner et at., 2020)

Elisabeth Steiner et al., (2019) shows the comparison of beer quality attributes between
beers brewed with 100% barley malt and 100% barley raw material, and it was shown that by
brewing beer with 100% barley and an appropriate addition of exogenous Ondea Pro
enzymes , it was possible to efficiently brew beer of a satisfactory quality. The production of
beers brewed with 100% barley resulted in good process efficiency and to be a final product
whose sensory quality was described as light, with little body and mouthfeel, very good foam
stability and similar organoleptic qualities compared to the conventional malt beer. In spite of
the sensory evaluation differences could still be seen in protein content and composition.

In this study, the aptness of Korean wheat varieties, Anzunbaengi and Keumkang, for wheat
beer was investigated by analysing the characteristics of malt and beer making. The Korean
wheat malt samples (AM and KM) made of the Anzunbaengi and Keumkang were compared
with two other wheat malt samples (UM and GM) from the USA and Germany, which were
used as control samples. The results of this study showed that most malt quality parameters
satisfied the recommendations of EBC for beer brewing; these were generally similar, except
for those factors related to protein. The Korean malt samples were higher in SN, TN, and
FAN than the control malt samples. KM had a lower Kolbach index than the other malt
samples. However, any compounds responsible for the discrimination of the four beer
samples were not related to the sensory attributes perceived by the consumers (n = 102). The
consumers preferred AB to KB in terms of appearance and flavour. They reported that AB
was lower in turbidity and higher in alcohol aroma, sweetness, and fruity flavour than KB. In
terms of consumer acceptability, the overall acceptance was not significantly different among
the four wheat beer varieties, but each consumer group by AHC clustering had a different
trend to prefer beer products. This report is the first study to perform a comparative
characterisation of Korean wheat malt with commercial ones for beer brewing. These might
be useful to small breweries attempting to use Korean wheat malt and/or develop new

13
distinctive local products. However, considering that this study was done by laboratory scale,
variations in brewing conditions should be considered before applying these results directly
to industries. Therefore, further research is required to apply these results at the industrial
scale. (YS Byeon et al., 2021)

In this study a series of proteomic procedures has been applied to analyze beer and wort
proteomes including two-dimensional gel electrophoresis combined with mass spectrometry
(MS), pre-fractionation followed by gel electrophoresis and MS, and gel-free based shotgun
liquid chromatography. These approaches have detected a number of protein species in beer
and wort, from barley and yeast, and occasionally from rice and maize. Of these beer
proteins, barley dimeric alpha-amylase inhibitor-1, barley trypsin inhibitor precursor and
yeast thioredoxin have been identified as factors responsible for beer quality traits of foam
and haze. In addition, a number of hordein derived polypeptides involved to celiac disease
have also been identified. Among these proteins and peptides, the modifications such as
glycation and partial digestion in several proteins such as lipid transfer protein (LTP) and
protein Z have been characterized. These findings may provide novel tools to improve quality
control during beer processing and they may enhance selection in malting barley breeding.
(Takashi et al., 2020)

In this study the malting industry in Ethiopia is often challenged by the availability of barley
variety that meets the quality and quantity requirements set by the industry. Recently, the
demand for malting barley has increased significantly in Ethiopia because of the increase in
the number of beer industries in the country, and more attention is now being given to
meeting the demand for the barley crops. Selection of improved malt barley variety in
different location is important to boost to the grain yield and to fill the demand of malt barley
for the users. Based on the results, three malting barley varieties, EH1847, IBON174/03, and
Bahati, are found to be relatively high -yielder and best quality malt barley varieties.
Moreover these varieties fulfilled quality parameter requirements set by the National
Standard Athority for malting barley. Therefore, these varieties are recommended for the
production for their high yield, kernel size and protein content. (Essien et al., 2019)

In this study all proportions of malt/ adjunct ratio produced beer of desirable physico
chemical parameters however 60:40 malt adjunct ratio produced beer with low pH which
could have deleterious effect on the beer and the yeast cells used in fermentation. The
emergence of deformed yeast after fermentation showed that the 60:40 formulations did not

14
contain enough nutrients for yeast nutrition and metabolism. Since the aim of every brewing
outfit is to be able to compete favorably in the highly competitive Nigerian beer market as
well as to produce beer of optimum quality, it is advisable to stick to beer produced with malt
/adjunct proportion of up to 70:30 malt /adjunct proportion whose yeast behaviour can be
scientifically predicted to avoid deleterious side effects like fusel oil production, acid, sulfide
and other volatile compounds which could affect the quality of the beer. Also good beer yeast
is expected to be used up to 8 – 10 generations. The emergence of deformed yeast in the
60:40 malt / adjunct ratio in the first generation showed that the yeast used might not be able
to withstand the stress of long usage. However the use of external enzymes like amylase,
protease and filtrase could assist in bringing out optimum activity when using sorghum
adjunct in a higher proportion as in 60:40 malt adjunct proportion. The use of higher
percentage of adjunct should therefore be explored putting in consideration the use of
external enzymes which although would give desired end product but will be more finance
intensive. (Daniel et al., 2019)

In brewing the basic principles remain the same over the centuries. This includes malting,
mashing, boiling and fermentation. During malting, the grains are allowed to steep in water
for 48-72 h at 10-15°C. The grains are allowed to germinate. However, as soon as the
enzymes are produced and before the young seedling can make an appreciable in road into
the nutrient reserve of the grain, development of the seedling is halted by drying at a
temperature which will not completely inactivate the enzymes. These enzymes are reactivated
during mashing to hydrolyse starch and protein and release nutrients for the nourishment of
the yeast during fermentation. (Okafor et al., 1987)

After malting the grains are milled and mixed with water. The mash after several processes of
brewing is filtered and boiled before fermentation. Wort is usually inoculated with the
fermenting organism, yeast. The sugar in the wort is fermented by yeast to alcohol. For this
purpose yeast specie such as Saccharomyces carlsbergensis are used. Selected strains are
systematically isolated and grown. (Pollocket et al., 1981)

In this study, hop variety and yeast strain have been chosen to evaluate their influences on the
volatile compound profile in beer. Volatile compounds were analysed by
Gas-Chromatography. Among the 39 volatile compounds detected, 9 of them were influenced
only by hop variety, 2 by yeast only and the 28 remaining by both hop and yeast together. As
expected, terpenes concentration in beer is directly linked to the choice of hop variety used,

15
but it appears the yeast strains can also influence that parameter. Surprisingly, in this study
hops variety influenced the final concentration of esters.

Among all the volatile compounds produced by the yeast, it has been shown in this study that
the
Base Ingredients Control RBS-1 RBS-2 RBS-3
Sample
Barley malt- 80% _ _ _

Rye malt _ 40% 50% 60%

Pilsner Malt _ 35% 30% 25%

Rice 20% 25% 20% 15%

compounds like ether esters, acetate esters and medium chain fatty acids are dependent on the
yeast strain but also on the hop variety used. This interaction between the hop and the yeast
strain could lead to better control the production of yeast derived compounds in the beer.
(Damien steyer et al., 2017)

Formulation of beer production from Rye, Pilsner and Rice

Composition of Rye beer

table - 5

Other ingredients; Water, Hops, Yeast (saf-ale S – 33), Silica powder, zinc sulphate.

16
Among all the volatile compounds produced by yeast, it has been shown in this study that
compounds like ethyl esters, acetate esters and medium chain fatty acids are dependent on the
yeast strain but also on the hop variety used. This interaction between hop variety and yeast
strain could lead to better control the production.

4. METERIALS AND METHODS

Raw materials

Raw materials used for brewing were collected from the Sattvic Foods. Rye (Secale cereale)
was the chief raw material used. Others include pilsner malt and rice used as adjunct, brewing
water, yeast, hop, and zinc sulphate to provide enough energy to yeast.

Sample collection

Pilsner malt with moisture content of 5.01% was used as adjunct in brewing. These malt was
collected form the Brewmart India. And rice was collected from the local field from the
farmers, also used as the second adjunct.

Brewing process

Initially milling machine was used to reduce the particle sizes of the grain and also to expose
the starch in the grain in readiness for enzymatic action during brewing. In the development
of rye beer, it was planned for 2 litres of each sample.

4.1 Procedure

Malting;

Rye grains were undergoes malting process includes soaking, germination and kilning. Rye
grains were soaked in water for 24 hours for germination (activation of enzymes). After
germination rye grains were allowed for sun drying to reduce the moisture content.

17
 Fig - 4

Milling;

Rye malt, pilsner malt, and rice was milled in the milling machine to reduce the grain size
that makes easy to dissolve the grains during mashing.

Mashing:

Milled grains of rye, pilsner and rice (complex carbohydrates) were boiled to convert the
complex carbohydrates into simple sugars.

Masing temperature & holding profile.

Phase Start temp (ºC) End temp (ºC) Minute

Rest – 1 52 51 10
Rest – 2 71 72 25
Rest – 3 81 83 10
table - 6

Fig - 5

Wort separation
When conversion is complete, the mash was consist of a sugar solution called wort and was
confirmed by the iodine test. Separation of wort (sugar solution) has been done through the

18
muslin cloth. The objectives of effective wort separation are the removal of unwanted
material while at the same time extracting all the available wort.

Boiling

After the wort has been filtered, it is then subjected to boiling at elevated temperatures to
bring significant changes in its characteristics properties.

Hops addition; During the boiling of wort, addition of hops were taken which are responsible
for flavour and also it has plant antiseptic properties. Hops contain bitter resins has α-acids
will modify into isomerised α-acids during boiling process.

Fig - 6

Wort chilling and Aeration

In this process the wort of 100 ºC converts into 11.2ºC. Because yeast cannot survive under
the higher temperature.

Yeast propagation

A genetically modified strain saf-ale yeast was used as the fermenting organism. 5gms of
yeast was taken into the round bottom flask along with 50ml of wort and allowed for 24
hours. The inoculated wort was transferred into the 200ml of wort and again allowed for 24
hours in cold temperature. And later it was mixed with 2liters of wort for fermentation.

Yeast propagation Flow chart

19
 5ml yeast + 50ml wort

Store at 26ºc for 24hours

Transfer into 500ml

Store at 24ºc for 24hours

Transfer into 2lires

Fermentation

Fermentation was started as soon as the wort transferred to fermenting vessel. Fermenting
vessel was placed in refrigerating incubator. Ater 7hours Co2 was released, confirmed that
fermentation was started, and was allowed for 15 days.

The over all reaction for fermentation is;

C6H1206 2 (C2 H5 OH) + 2 (CO2) + Energy

(Ethanol)

Beer Maturation

Fermentation occurs in the wort is of two types, one is primary fermentation (aerobic
fermentation) and another one is secondary fermentation (anaerobic fermentation). Beer
maturation was done through the air lock system. Temperature was observed daily through
the digital thermometer. This method was allowed for 13 – 15 days for proper fermentation.

20
 Fig - 7

Parameters monitored during fermentation temperature and Ph.

Days of fermentation Temperature (º C)

RBS – 1 RBS- 2 RBS – 3

Day – 1 11.1 10.5 10.8

Day – 3 10.5 9.4 9.2
Day – 5 9.2 8.3 8.5

Day – 7 8.6 7.2 7.5

Day – 9 8 6.4 6.8
Day – 11 7.4 5.9 5.9
Day – 13 6.1 5.2 5.3

table - 7

Filtration:

Beer was filtered through a whatman filter paper.

Packaging: Packaging was done through manual crimper , which is particularly used for beer
in laboratory.

21
 Fig – 8

5. Evaluation of brewing characteristics and their analysis:

Analysis methods:

S.no Parameter Method

1. Ethyl Alcohol Pycnometer or Hydrometer method
2. Apparent extract By SOP of Apparent extract
3. PH PH meter
4. Carbohydrates Anthrone method
5. Protein Biuret Metod
6. Iron Atomic absorption spectrometry
7. Real Extract By SOP of Real extract
8. Polyphenols By SOP of polyphenol estimation
9. Vitamin E Gas chromatography
table - 8

5.1 Determination of Ethyl Alcohol Content - Pycnometer Method or

Hydrometer Method:
The determination of ethyl alcohol content in alcoholic beverages can be carried out using
either the Pycnometer Method or the Hydrometer Method after distillation. The principle
involves measuring the specific gravity of the distilled alcoholic beverage. This method is
universally applicable to all types of alcoholic beverages.

The required apparatus includes a distillation unit comprising a 500 mL capacity flask
connected to a water-cooled condenser, along with a pycnometer of 50 mL capacity or a
short-range hydrometer (0.96 – 1.00), a thermometer (0-100 °C), and a 200 mL capacity
volumetric flask.

The procedure begins by transferring precisely 200 mL of the alcoholic drink into the
distillation flask containing about 25 mL of distilled water and some pumice stones. The
contents are then distilled for approximately 35 minutes, and the distillate is collected in a

22
200 mL volumetric flask. Once the distillate reaches room temperature (20 °C), it is made up
to volume with distilled water and thoroughly mixed. The specific gravity of the distillate is
then determined using the pycnometer or hydrometer.

For the Pycnometer Method, a clean and dry pycnometer is weighed empty along with the
stopper at 20 °C. It is then filled with the liquor sample distillate to the brim, and the stopper
is gently inserted. After wiping away any spilled liquid, the pycnometer is weighed again at
20 °C. Following this, the liquor sample distillate is removed, the pycnometer is washed with
distilled water, and then filled with distilled water to determine its weight at 20 °C.

Calculation:

Specific gravity = 𝑊1−𝑊2

𝑊2−𝑊
W: Weight of Empty Pycnometer

W1: Weight of Empty Pycnometer with liquor

W2 : Weight of Empty pycnometer

% Alcohol (v/v) = 8610.6 – (16584× SG) + (7973.3 × SG 2)

5.2 Dertermination of Apparent extract:

The apparent extract in beer, often referred to as the original or final gravity of fermented
beer plays a significant role in determining the taste and flavor of the brew. This parameter
represents the concentration of dissolved solids in the liquid, primarily comprising sugars and
other non-volatile compounds. These substances contribute to various aspects of the beer,
including its flavor profile, body, and overall composition. The measurement of apparent
extract provides valuable insights into the brewing process and helps brewers maintain
consistency and quality in their products.

Calculation:

AE = (ABV x 2.0665 + RE)

x 100 (ABV x
1.0665 + 100)
Where ABV is alcohol by volume, RE is real extract.

23
5.3 Determination of Ph:
Determining the pH of beer is crucial for quality control, as it directly impacts flavor,
stability, and overall consumer satisfaction. Employing a pH meter for precise measurement
is a widely adopted and efficient approach.

The procedure entails gathering a sample of room temperature beer, calibrating the pH meter
using standard buffer solutions, and submerging the electrode until a stable reading is
obtained. Brewers document the pH value and assess it against predetermined acceptable
ranges for the beer.

5.4 Determination of Real extract:

The real extract in beer denotes the genuine concentration of dissolved solids, primarily
sugars and other non-volatile components, found in the finished brewed product. It offers a
more precise depiction of the beer's actual composition, considering both the initial sugars
from the ingredients and those sugars converted into alcohol during fermentation. This
parameter provides valuable insight into the complete makeup of the beer, accounting for all
components contributing to its flavor, body, and overall character.

Calculation:

RE = (0.198 x OG) + (0.8052 x AE)

Where, OG original gravity of wort

AE is the apparent extract of final fermented beer

5.5 Determination of carbohydrates by Anthrone method:

Carbohydrates undergo initial hydrolysis into simple sugars with the use of dilute
hydrochloric acid. In a heated acidic environment, glucose undergoes dehydration, leading to
the formation of hydroxymethyl furfural. When combined with anthrone, this compound
produces a green-colored product with an absorption peak at 630 nm.

Procedure:

For sample preparation, begin by taking 100 mL of the alcoholic beverage and completely
remove the alcohol through distillation on a water bath at 80 °C. Note down the weight (A

24
mg) of the residue obtained. Weigh 100 mg of the residue into a boiling tube. Proceed to
hydrolyze it by placing it in a boiling water bath for three hours with 5 mL of 2.5 N
hydrochloric acid and then cool it to room temperature. Neutralize the solution with solid
sodium carbonate until the effervescence ceases. Make up the volume to 100 mL and
centrifuge, followed by collecting the supernatant and taking 0.5 and 1 mL for analysis as the
test sample.

Moving on to the method of analysis, prepare standards by taking 0, 0.2, 0.4, 0.6, 0.8, and 1
mL of the working standard glucose solution, with '0' serving as a blank. Make up the volume
to 1 mL in all tubes, including the sample tubes, by adding distilled water. Then add 4 mL of
anthrone reagent to each tube. Heat the tubes for eight minutes in a boiling water bath,
followed by rapid cooling to room temperature and making up to 5 mL with distilled water.
Read the green to dark green color at 630 nm. Draw a standard graph by plotting the
concentration of the standard on the X-axis versus absorbance on the Y-axis. From the graph,
calculate the amount of carbohydrate present in the sample tube.

5.6 Estimation of protein by biuret method:

The biuret method relies on the reaction between peptide bonds in proteins and copper ions.
When exposed to the biuret reagent, containing copper ions, peptide bonds form a complex
that causes a color change from blue to violet. The intensity of this color change is directly
proportional to the protein concentration in the sample, facilitating its estimation.

For this method, the required reagents include the biuret reagent, prepared by dissolving
copper sulfate, sodium potassium tartrate, and potassium iodide in sodium hydroxide
solution, and a standard protein solution, typically bovine serum albumin dissolved in
distilled water to create a concentration of 5mg/ml.

To determine protein concentration, a calibration curve is constructed using standard

solutions of known protein concentrations. The sample is mixed with the biuret reagent, and
after allowing for color development, the absorbance of the sample and a blank is measured
at 540 nm. By plotting absorbance values against protein concentrations, a calibration curve
is generated. Extrapolating the sample's absorbance value from this curve allows for accurate
determination of the protein concentration.

5.7 Estimation of iron in beer:

25
The Atomic Absorption Spectrophotometric (AAS) method is employed to determine the iron
content in alcoholic beverages. The beer samples are aspirated into an AAS flame, and the
absorbance of iron is measured and compared with standard readings.

In terms of apparatus, acetylene and nitrogen of ultra-pure grade, along with distilled water
and Iron Standard Reference Material (SRM) traceable to NIST, are required. To prepare the
reagents, suitable aliquots from Copper or Iron SRM are taken to create 0.25, 0.50, and 1.00
µg/mL Cu/Fe solutions, which are then adjusted to a known volume with 1N nitric acid.

Procedure:

The procedure begins with following the manufacturer's instructions for optimal gas flow,
wavelength settings, and beam alignment. For clear samples, the liquor sample filtered
through a 0.45 µm filter can be directly injected into the AAS for copper quantity
determination. However, in cases of samples with high residues, direct injection of the
filtered sample may cause clogging of the AAS burner head, necessitating wet ashing.

The process involves aspirating the blank into the AAS flame to set the instrument for zero
absorbance, followed by aspirating the Cu/Fe standard solutions sequentially for absorbance
data acquisition. Finally, the liquor sample or nitric acid-digested wine sample solution is
aspirated into the AAS flame to record the absorbance, and the displayed concentration of
Cu/Fe in µg is noted down. The concentration in the test sample is then calculated,
considering any dilutions made.

Iron content in wine (in µg/mL or mg/L) = Reading in µg displayed × Dilution

Volume of sample
5.8 Estimation of Polyphenols in Beer:

Polyphenols are crucial compounds that significantly contribute to the flavor, color, and
antioxidant properties of beer. To estimate polyphenols in beer, a standardized method is
followed:

Sample Preparation:

Begin by selecting representative beer samples for analysis. If necessary, filter the beer
sample to remove any particulate matter that might interfere with the analysis. Ensure that the
sample is brought to room temperature before proceeding further.

26
Preparation of Reagents:

Prepare a standard solution of a known polyphenol compound, such as catechin or gallic acid,
at a concentration within the expected range of polyphenol content in the beer samples.
Additionally, prepare a blank solution using the same solvent used for sample dilution.

Calibration Curve:

Prepare a series of standard solutions by diluting the stock standard solution with an
appropriate solvent to obtain concentrations covering the expected range of polyphenol
content. Measure the absorbance of each standard solution at the characteristic wavelength
for polyphenols, typically in the UV-visible range. Plot a calibration curve with absorbance
on the y-axis and concentration on the x-axis.

Analysis Procedure:

Dilute the beer sample appropriately to ensure that the polyphenol concentration falls within
the linear range of the calibration curve. Measure the absorbance of the diluted beer sample at
the same wavelength used for the standards. Repeat the measurement for multiple dilutions if
necessary to ensure accuracy.

Calculation of Polyphenol Content:

Using the calibration curve, interpolate the concentration of polyphenols in the beer sample
based on its absorbance value. Express the polyphenol content in the beer sample as
milligrams per liter (mg/L) or another suitable unit.

Following this standardized procedure ensures accurate estimation of polyphenols in beer,

providing valuable information for quality control and product development in the brewing
industry.

5.9 Estimation of Vitamin E in Beer by Gas Chromatography:

Sample Preparation:

Begin by selecting representative beer samples for analysis. Extract the vitamin E from the
beer sample using a suitable solvent, such as hexane or ethanol. Filter the extract to remove
any particulate matter that may interfere with the chromatographic analysis. Concentrate the
filtered extract to reduce the solvent volume, if necessary.

Standard Preparation:

27
Prepare a series of standard solutions of known concentrations of vitamin E in the chosen
solvent. These standards will serve as references for quantification during GC analysis.

Gas Chromatography Analysis:

Inject the prepared beer sample extract and standard solutions into the gas chromatograph.
Separate the components of the sample extract and standards based on their respective
retention times and volatility. Detect the vitamin E peak(s) using a suitable detector, such as a
flame ionization detector (FID) or a mass spectrometer (MS). Record the chromatogram and
measure the peak area(s) corresponding to vitamin E in both the sample and standard
solutions.

Quantification:

Calculate the concentration of vitamin E in the beer sample by comparing the peak area(s) of
the sample extract with those of the standard solutions. Use an appropriate calibration curve
or external standard method to determine the concentration of vitamin E in the beer sample.
Express the results in terms of the amount of vitamin E per unit volume or mass of beer (e.g.,
micrograms per milliliter or milligrams per liter).

28
 6. RESULT AND DISCUSSION

Beer's chemical composition is a complex amalgamation of various constituents, each

playing a crucial role in shaping its flavor, aroma, appearance, and overall quality. Water acts
as the primary ingredient, influencing mineral content and pH levels. Malted barley provides
fermentable sugars, leading to alcohol and carbon dioxide production during fermentation.
Hops contribute bitterness, aroma, and microbial stability. Yeast, crucial for fermentation,
converts sugars into alcohol while imparting diverse flavors and aromas. Additionally, trace
elements, proteins, lipids, and other compounds influence mouthfeel and stability.
Understanding this composition is vital for brewers to craft diverse beer styles and
consistently deliver enjoyable experiences.

29
Comparing the chemical composition of prepared beer with traditional barley-based beer is
essential. However, the chemical composition of beer available in the market is not typically
disclosed. Hence, we obtained the chemical composition of a control beer sample from the
laboratory of Crown Breweries India Pvt Ltd. Prior to analyzing the chemical composition, it
is crucial to decarbonate the beer samples to eliminate any potential inaccuracies. This is
necessary because CO2 may interfere with other beer components, leading to misleading
results. To achieve this, the beer samples underwent treatment in an ultrasonic bath operating
at 80kHz for 30 minutes. This effectively removed CO2, as indicated by the disappearance of
foam. This preparatory step ensured accurate testing of the beer's composition, devoid of
CO2 interference.

Alcohol content:

Alcohol is the primary byproduct produced during fermentation, facilitated by yeast

metabolism. Yeast converts sugars into ethanol and carbon dioxide. The percentage of
ethanol is indirectly proportional to the percentage of dissolved solids in beer, resulting in a
gradual decrease in specific gravity during fermentation. The RBS1 beer having 5.12%
ethanol, RBS 2 beer having 4.88% and the RBS 3 beer having 4.41% of ethanol whereas the
controlled beer having 6.22% of ethanol, which is higher than the beer is made from pilsner
malt and rye. This is because the unavailability of fermentable sugars in pilsner malt and rye
beer. The availability of fermentable sugars is indirectly propotional to the amount of pilsner
malt and rye.

Apparent extract:

The apparent extract of the control beer stands at 1.5 which is in ideal range for this
parameter in beer production. However, the prepared beer samples shows a deviation in each
type, firstly the RBS 1 beer is having 2.1, RBS 2 beer is having 2.4 and RBS 3 having 2.5o
plato. This higher than desired apparent extract is likely due to the presence of
non-fermentable sugars originating from rye and pilsner malt incorporated into the brewing
process. These grains contribute additional sugars that resist fermentation, ultimately
affecting the final apparent extract of the beer. In this case the apparent extract is directly
proportional to the amount of rye and pilsner taken.

Ph:

30
 Days of fermentation PH
Controlled beer RBS – 1 RBS- 2 RBS – 3

Day – 1 5.30 5.41 5.39 5.36

D ay – 3 5.11 5.25 5.21 5.23
Day – 5 4.98 5.04 5.00 5.05
Day – 7 4.62 4.74 4.79 4.72
Day – 9 4.13 4.30 4.28 4.31
Day – 11 3.97 4.08 4.11 4.03
Day – 13 3. 92 3.97 3.95 3.98

The significance of pH in beer preparation plays a critical role in shaping the flavor, aroma,
and overall quality of the final product. This slightly acidic pH level not only contributes to
the beer's refreshing tartness but also acts as a crucial factor in inhibiting the growth of
harmful microorganisms, thereby ensuring the stability and safety of the beer.

The above table clearly demonstrate the Ph value during the time of fermentation from day 1
to day 13. At the end of fermentation, the controlled beer having 3.92, whereas the RBS 1,
RBS 2 and RBS 3 having Ph values of 3.97, 3.95 & 3.98. The Ph values which are falling
under optimal range.

Real extract:

The real extract, encompassing total soluble proteins, fats, and non-fermentable sugars, is a
crucial measure for assessing the beer's body, residual sweetness, and flavor potential. In the
crafted rye and pilsner malt beer, the real extract is determined different values, representing
the overall dissolved solids present in the beer before fermentation.

Analysis of the real extract in the control beer yields a concentration of 3.93, falling within
the recommended ideal range for this parameter in beer production. However, examination of
the prepared beer sample samples RBS 1, RBS 2 & RBS 3 reveals a slightly elevated level of
4.05, 4.11, 4.13 surpassing the ideal range. This difference may be attributed to the
ingredients used in the brewing process. The interaction between insoluble proteins and

31
polyphenols from rye and pilsner malt could have contributed to the higher real extract value,
potentially influencing the beer's overall quality and characteristics.

Carbohydrates:

The crafted beer sample having approximately 30.1 grams of carbohydrates in controlled
beer, a notable increase in carbohydrates found in RBS 1, RBS 2 and RBS 3 to the typical
carbohydrate content range from 32.22, 32.67, 32.39 grams. This rise in carbohydrate content
is primarily attributed to the presence of complex starch molecules that remained unutilized
by yeast cells during fermentation. Additionally, the inclusion of rye and pilsner malt, known
for its high carbohydrate content, further contributes to the overall increase in carbohydrates
observed in the beer.

Protein:

Beer typically contains a relatively low amount of protein compared to other beverages and
food products. The protein content in beer primarily arises from the grains used in the
brewing process, such as rye, wheat, pilsner malt. While these grains contain proteins that
contribute to the beer's overall composition, most of these proteins are removed or
precipitated during brewing. As a result, the final beer typically contains only trace amounts
of protein, usually less than 0.62 grams per 100 milliliters. The control beer exhibits a protein
content of 0.39 grams.

In contrast, the prepared beer samples shows a slightly higher protein content, amounting
from 1.2, 1.31, 1.40 gm in RBS 1, RBS 2, RBS 3. This increase can be attributed to the
presence of rye, which are rich in proteins. However, the elevated protein content may impact
the quality of the final beer, potentially causing haziness. Hence, careful consideration of
protein levels is necessary to maintain the desired clarity and quality of the beer.

Iron:

Beer typically contains very low levels of iron, primarily because the ingredients used in
brewing, such as malted barley, hops, yeast, and water, do not contribute significant amounts
of iron. Additionally, the brewing process itself does not introduce substantial iron into the
final product. While the iron content of beer can be influenced by the iron levels in the water
used for brewing and the equipment employed, these levels generally remain low.
Consequently, beer is not considered a significant source of dietary iron.

32
In the prepared beer samples RBS 1, RBS 2, RBS 3 & controlled sample exhibits 1.24, 1.16,
1.20 and 0.63 (ppm) of iron was detected, this increase in iron content can be attributed to the
raw materials used in the beer preparation, which are rich in iron. As a result, the beer sample
exhibits a higher iron content compared to traditional barley beer.

Polyphenols:

The controlled beer demonstrates a polyphenol range of 25 mg per 100 ml, indicating a
moderate level of these compounds. In contrast, the beer crafted from a blend of rye and
Pilsner malt exhibits a notably higher range of polyphenols, measuring at 37mg, 35mg and
40mg per 100 ml in RBS 1, RBS 2 and RBS 3 samples. This elevated polyphenol content
suggests a richer concentration of these compounds, which contribute to the beer's flavor,
aroma, and antioxidant properties.

Vitamin E:

In comparing the vitamin E content between the controlled beer and the beer crafted from rye
and Pilsner malt, notable differences emerge. The controlled beer exhibits a relatively low
level of vitamin E, measuring at 0.14 milligrams per 100 milliliters. In contrast, the beer
made from rye and Pilsner malt showcases a significantly higher concentration of vitamin E,
registering at 3.93mg, 3.82mg and 4.01mg per 100 milliliters in RBS 1, RBS 2 and RBS 3
samples. Such variations in vitamin E content highlight the potential for diversifying the
nutritional profile of beer through thoughtful ingredient choices and brewing techniques.

6.1 Sensory Evaluation and Comparison with commercial beer

The sensory analysis of our beer, crafted with a blend of Rye . Pilsner malt, and rice, reveals a
dense of flavors. Compared to traditional barley-based beers. The rye brings a light spice note
and clove flavour while the rice contributes a delicate smoothness. Pilsner malt add a layer of
complexity with their smooth, seamlessly integrated into the overall taste. In contrast to
traditional barley-based beers, this beer provides a low caloroie option that doesn't
compromise on flavor. For this sensory evaluation study, we have chosen 10 panel members
who are trained in beer flavor profile. The Ram kumar, Shivaraj, Akhilesh kumar, Rajeev,
Venkat, Arun kumar, shiva kumar, Somaiah, Keshav, Vijay kumar. The sensory panelists
voted the rye beer because of its unique flavour profile, whereas the few panelists chosen
conventional beer for its mouthfeel and plain texture. Out of 10 panelists 8 panelists liked
this unique beer.

33
Beer sensory evaluation relies on a specialized sensory analysis table, guiding tasters through
a systematic assessment of appearance, aroma, flavor, mouthfeel, and overall impression.
This structured approach ensures a thorough examination of the beer's characteristics,
facilitating consistent and precise analysis.

The score reveals how the prepared beer tastes. Usually, the score would be given from 1 to
10. Highest number suggests high acceptability and low number denotes the low
acceptability.

34
 SENSORY ANALYSIS SCORE CARD

Name of the sample:

Name of the panel member:

Gender:

Age:

Sensory Control RB 1 RB 2 RB 3
attributes sample

Appearance

Colour

Texture

Flavour

Taste

Other
attributes

Faculty:

Hedonic scale reading:

9. Like extremely, 8. Like very much, 7. Like moderately, 6. Like slightly, 5. Neither like or
dislike, 4. Dislike slightly, 3. Dislike moderately, 2. Dislike very much, 1. Dislike
extremely.

Any other comments:

35
 Sensory Appearance Colour Flavour Taste Overall
Attributes Acceptance

Control 9 9 9 9 9
sample

RB 1 7 8 6 7 7

RB 2 8 8 7 8 7.5

RB 3 8 9 8 9 8.5

7. Effect of storage period on shelf-life and nutritional quality of rye beer

In a recent shelf life study comparing prepared rye beer to barley-based beer, it was
discovered that the prepared beer have a shorter shelf life. The flavor profile of the rye beer
showed changes in terms of odour and taste comparison to its barley based beer. Despite
these variations and finding the both sampled beers doesn’t showed any noticeable colour
variation and haziness. This suggests that the prepared rye beer may have a reduced shelf
life and altered flavor, their visual characteristics remain consistent with the traditional
barley-based beer.

There are several reasons behind the shorter shelf life of prepared beer when compared to
market beer. The presence of high amonut of dissolved oxygen and some other micro
organisam may lead to oxidation of beer results in off flavoured beer.

The another reason is that the packaging condition also greatly effect the shelf life of beer.
The compressed Co2 in beer will help in prevention of oxidation reaction as well as
protecting the beer structure and flavour. Whereas during packing of prepared beer doesn’t
contain enough Co2 for limiting oxidation reaction.

7.1 Off flavours in beer:

36
The data given by the trained panalists shows that the present beer sample contains some off
flavour caharcteristics. These flavours may effect the flavour profile of beer and some off
flavours give unpleasant flavour to beer. Lets have a look on each off

flavour found in our beer sample.

Diacetyl:

Out of 10 panalists 8 panalists found buttery flavour in prepared beer sample. This is
because the presence of diacetyl which is generated during the fermentation process
aids by the yeast mechanism. Where as only 3 panalists observed light diacetyl flavour
in controlled beer. The amount of dicetyl present in the beer is usually determined by
the Gas chromatography technique. The presence of diacetyl leads to the lower shelf
life of beer.

Acetaldehyde:

This flavour represents the incomplete fermentation or oxidation of beer. Approximately 10

panalists found that the prepared beer is slightly oxidised due to the inappropriate measures
taken in some procedure. This is also leads to instability of beer. Usually, the acetaldehyde
flavour is identified by the wood smell or wet cardboard smell expels after immediate
opening of the bottle.

Other:

Few of the panalists found phenolic and sulphatic flavours in the beer. This is may be due to
the wort boiling temperature and ph. However, most of the panalists doesn’t find these off
flavours in the beer.

Carbohydrates: There is no change in CHOs and remains same 32.22, 32.67, 32.39 grams in
RB1 RB2 RB3 respectively.

Proteins: Also no change in proteins that is 1.2, 1.31, 1.40 gm in RBS 1, RBS 2, RBS 3
respectively.

37
8. CONCLUSION

38
In conclusion, the development of beer with the infusion of rye, pilsner malt and rice
has ushered in a transformative era in the brewing industry. These alternative grains
have not only expanded the palate of flavors available to beer enthusiasts but also
addressed crucial issues of dietary inclusivity and sustainability.

Through their distinct flavor profiles, rye, pilsner malt and rice have pushed the
boundaries of what beer can be. From rye mild spice to pilsner smoothness and rice's
mild undertones, these grains have added a wealth of diversity to the beer landscape,
captivating taste buds and challenging conventions. Equally important is the role these
grains have played in enhancing dietary inclusivity. By offering low calaories with
high vitamins ( E) and polyphenols options. they have made beer more accessible to
individuals with sensitivities, ensuring that no one is left out from enjoying this
beloved beverage. Moreover, the sustainable and drought-resistant qualities of these
grains contribute to the brewing industry's commitment to environmental
responsibility, addressing issues like water scarcity and climate change. Looking
ahead, the trajectory of beer development with these alternative grains appears
promising. It symbolizes an industry that continually adapts to changing consumer
preferences, embraces innovation, and strives for inclusivity and sustainability. As
consumers seek new taste experiences and more responsible choices, the infusion of
rye, pilsner malt and rice into beer production represents not just a trend but a
meaningful and enduring shift in the brewing landscape.

References:

Willaert, R. (2007). The beer brewing process: Wort production and beer. Handbook of
food products manufacturing, 2, 443.

Hardwick, W. A. (1994). An overview of beer making. Handbook of brewing. USA:

Marcel Dekker, 87e96.

39
Stewart, G. G. (2006). Beer stability. In Handbook of brewing (pp. 730-743). CRC
Press.

Wannenmacher, J., Gastl, M., & Becker, T. (2018). Phenolic substances in beer:
Structural diversity, reactive potential and relevance for brewing process and beer
quality. Comprehensive Reviews in Food Science and Food Safety, 17(4), 953-988.

Willaert, R. (2012). Biochemistry of beer fermentation. Food biochemistry and food

processing, 627-653.

Grassi, S., Amigo, J. M., Lyndgaard, C. B., Foschino, R., & Casiraghi, E. (2014). Beer
fermentation: Monitoring of process parameters by FT-NIR and multivariate data
analysis. Food chemistry, 155, 279-286.

Parker, D. K. (2012). Beer: Production, sensory characteristics and sensory analysis. In

Alcoholic beverages (pp. 133-158). Woodhead Publishing.

Langstaff, S. A., Guinard, J. X., & Lewis, M. J. (1991). Sensory evaluation of the
mouthfeel of beer. Journal of the American Society of Brewing Chemists, 49(2), 54-59.

Elgaard, L., Jensen, S., Mielby, L. A., & Byrne, D. V. (2019). Performance of beer
sensory panels: A comparison of experience level, product knowledge, and
responsiveness to feedback calibration. Journal of sensory studies, 34(6), e12540.

Colgrave, M. L., Goswami, H., Howitt, C. A., & Tanner, G. J. (2012). What is in a
beer? Proteomic characterization and relative quantification of hordein (gluten) in beer.
Journal of proteome research, 11(1), 386-396.

Lodolo, E. J., Kock, J. L., Axcell, B. C., & Brooks, M. (2008). The yeast
Saccharomyces cerevisiae–the main character in beer brewing. FEMS yeast research,
8(7), 1018-1036.

Gibson, B. R., Lawrence, S. J., Leclaire, J. P., Powell, C. D., & Smart, K. A. (2007).
Yeast responses to stresses associated with industrial brewery handling. FEMS
microbiology reviews, 31(5), 535-569.

40
Verbelen, P. J., Dekoninck, T. M. L., Saerens, S. M. G., Van Mulders, S. E., Thevelein,
J. M., & Delvaux, F. R. (2009). Impact of pitching rate on yeast fermentation
performance and beer flavour. Applied microbiology and biotechnology, 82, 155-167.

McMurrough, I., Madigan, D., Kelly, R. J., & Smyth, M. R. (1996). The role of
flavanoid polyphenols in beer stability. Journal of the American Society of Brewing
Chemists, 54(3), 141-148.

de Lima, A. C., Aceña, L., Mestres, M., & Boqué, R. (2022). An Overview of the
application of multivariate analysis to the evaluation of beer sensory quality and
shelf-life stability. Foods, 11(14), 2037.

Palmer, J. J. (2001). How to Brew: Ingredients, Methods, Recipes and Equipment for
Brewing Beer at Home. Defenestrative Publishing, Company.

Berner, T. S., & Arneborg, N. (2012). The role of lager beer yeast in oxidative stability
of model beer. Letters in applied microbiology, 54(3), 225-232.

Gibson, B., Dahabieh, M., Krogerus, K., Jouhten, P., Magalhães, F., Pereira, R., ... &
Vidgren, V. (2020). Adaptive laboratory evolution of ale and lager yeasts for improved
brewing efficiency and beer quality. Annual review of food science and technology, 11,
23-44.

41