You are on page 1of 33







MSC (IBT)4th sem

My project dealing with ALCOHOL PRODUCTION. I owe a heart-felt gratefulness
to some of the people for being present all the time whenever any help was required in
completion of the report.
At the outset, I would like to thank Mr. K.P.SINGH (Director Production-O) & Mr. S.
Pande (AGM - HR) for giving me the approval to this project in the organization. I wish
to acknowledge the entire HR Team at Radico Khaitan Ltd. Special appreciation
extended to Mr. Ashish Kumar Singh (Executive - HR) who helped me to shape the
direction of my research work. This project would not have been possible without his
A heartfelt thanks to the respondents surveyed whose ideas, critical insights and
suggestions have been invaluable in the preparation of this report.


Radico Khaitan is one of India's oldest and largest liquor manufacturers. Formerly known
as Rampur Distillery, which was established in 1943. It was only in 1999, that Radico
decided to launch and market its own brands, thereby embarking on a period of
phenomenal growth. To further boost its production capacity of bottled and branded
products, the company has tied up with bottling units in various parts of the country.
Radico Khaitan Ltd today has three millionaire brands in its portfolio. Radico's flagship
brand, 8 PM Whisky, launched in 1999, was a runaway success. In the first year alone, it
sold one million cases - a record for any Indian or foreign brand operating in India. This
also made it the first brand in the liquor industry to make it to the Limca Book of
Drinks International, the acclaimed international liquor magazine has rated 8 PM whisky
as the fastest growing whisky in the world in the regional category (2004-05). The other
millionaire brands are: Contessa Rum has won the prestigious Monde Selection award for
its overall quality for the past three executive years.
It has a large market share in the defense market... Old Admiral Brandy has also been
rated by Drinks International as the fastest growing Brandy in the world in the regional
category (2004-05 and 2005-06) also it has won the Monde Selection award for its
overall quality in 2004-05.
Today, Radico Khaitan has brands that straddle almost every market segment - whisky,
rum, brandy, vodka & gin - and price category. Our fine blends, consistent quality,
distinctive packaging and superior value have resonated with customers.

Rampur Distillery is one of the largest distilleries in India and a leading manufacturer of
Extra Neutral Alcohol (used in manufacturing Indian Made foreign Liquor) it also
manufacturers Rectified Spirit (used in manufacturing of lower segments Country
Liquor) and manufacturing of Anhydrous Alcohol or Ethanol or Gasohol (used in Petrol
Mixing) and the recent addition of grain distillery . Today with a production capacity of
60 million liters p. a and with the recent addition of the grain distillery which has taken
the capacity up to 90 million lit p.a. it is one of the largest distilleries in the country The
Unit has a series of firsts to its credit:
It is the first Indian distillery to obtain ISO 9001:2000 certifications. It has achieved
capacity utilization of over 100% in the alcohol plant. It is the first environment-friendly
distillery in the country.

Molasses Distillery 60 million liters per annum.

Grain Distillery 30 million liters per annum.
Malt Distillery 720 thousand liters per annum.


The effluent Treatment Facility in Rampur Distillery is unique in nature when compared
among and in the Industry. The Distillery complies with Zero Discharge concept set up
by CPCB. The treatment has varied by products, which not only improves operational
stability of the plants but also adds on to company's profitability. Primary Treatment of
the Effluent yields Bio Gas, which is used as fuel in Cogen Boiler to generate steam and
then Power through a backpressure Turbine. The backpressure steam is used again in the
Distillation Plant to produce Extra Neutral Alcohol and Rectified Spirit.


Meeting out 100% Pollution Control norms, the Treated Effluent is not discharged
outside and in turn is mixed and cured with organic mass like Press Mud of Sugar Mills
and suitable organic manures to manufacture Bio Manure or Bio Compost, a bio fertilizer
used successfully in growing the crop of sugar canes etc.

The cogeneration plant of Rampur Distillery consist of 26 MT capacity India's first stand
alone Bio Gas fired steam boiler and 2 MW Turbine Generator in tandem to make Radico
Khaitan self reliant on its requirement for power for its normal operation.


The base for Indian whisky is ENA. The other ingredients used are matured Indian malt
sprit, Scotch whisky concentrate, added flavors, and caramel & dm water. All these
ingredients are not added at a time but different stages.
1. Special appointment whisky
2. 8pm Royale whisky
3. Whitefield whisky
4. 8pm whisky
5. Radico gold supreme whisky
6. Radico gold whisky
7. Contessa deluxe whisky
8. Rampur no.1 whisky

9. Masters Stroke Deluxe whisky

10. Royal Cambridge Grain Whisky
Brandy is a Dutch word means (Brunt Wine). In India we make brandy by blending with
ENA, matured grape spirit, added flavoring.
1. Contessa Brandy
2. Old Admiral Brandy
3. White Field Brandy

8pm Excellency Brandy

Rum is distilled liquor made from sugarcane products, cane juice spirit along with base
ENA and flavoring.
1. Contessa rum
2. Largest Rum
3. 8pm Bermuda
4. Black Cat Rum
5. Rampur No.1
6. Big Hit Rum
7. Contessa White Rum


Gin is alcoholic liquor obtained by distilling grain mash or redistilling sprit with
botanicals such as juniper berries, angelica, lemon and orange peels, cassia bark,
coriander and cardamom.
1. Contessa Gene
2. Big Hit Gene
3. Magic Moment Gene

For the production of alcohol, materials containing sustaining amount of glucose Content
can be used as a raw material. But for commercial production, common raw materials
used are as follows:
1. Molasses
2. Cane Juice
3. Sweet Potato
4. Sugar Beet
5. Grain (Barley, Rice)


1 Molasses as a raw material

The basic raw material for manufacturing alcohol is from Saccharine waste such as
Molasses, Sulphite waste Liquor Hydro wood waste, Mohowa, Bagasses etc. Some other
starchy materials like potato; Maize Barley etc. are used in the production of alcohol
which has already been discussed earlier.
Molasses is the by product of sugar Factories. It is the mother liquor which is left after
removal of sugar crystals and is used as a raw material for the manufacture of alcohol.
It is a dark brown colored viscous substance. The composition of molasses depends upon
a number of factors such as quality and variety of cane, soil and climate conditions and
the mode of processing cane juice. The average chemical composition of molasses is
given as below. However it depends mainly on the factors such as quality and variety of
cane, agricultural practices and the process adopted for manufacturing of cane sugar in
sugar factories:(Table 3.11 showing compositions of molasses)

Total reducing sugar after inversion (%Wt.)

Moisture (%)
Total Nitrogen (%Wt.)
Calcium as CaO (%Wt)
Phosphorus as P505(%Wt)
Magnesium MgO (%Wt)
Potassium K20 (%Wt)
Zinc, Zn (Mg/100gm)
Total ash (%Wt)
Biotin mg.100gm
Thiamine mg/g/100gm
Pyridoxine mg/10/gm
Riboflavin mg./100/gm
Nicotinamide mg.100mg/
Folic Acid mg/100gm

Black Strap Cane Molasses


2 Characteristics of molasses
Molasses is a dark reddish colored jelly like viscous material. The pH of molasses
varies from 6.8 to 8.5. The high osmotic pressure of molasses protects it from
microbial spoilage, and it can be easily transported by large tankers etc. Molasses can
be pumped easily. For production of ethanol in non sugar producing areas, and
particularly in areas with suitable waterways for barge shipment, molasses may be the

best suitable raw material for fermentation.

Classification of Molasses: Molasses are classified into following three grades:
a) First grade Molasses: It contains more than 50%TRS.
b) Second grade Molasses: It contains 40%to 50% TRS.
c) Third grade Molasses: It contains less than 40% TRS.

3 Consumption of Molasses
In Rampur Distillery the consumption of molasses is about 5,500 to 6,000 quintals/day
when all the three plants viz. Two Copper RS. Plants and one RS. Steel Plant runs.
Laboratory is the controlling section of a Distillery. It is the most important section of
every chemical industry where samples of raw material and final products are analyzed to
maintain the quality and recovery of the product. This is the section where records are
also maintained to know the working efficiency of the process.

Table for Fermentation conditions

Temperature during fermentation
Settling gravity of yeast vessels
Settling pH of yeast vessels
Settling gravity of prefermenters
Final gravity after complete fermentation
Duration of fermentation
% Alcohol after completion

28-34 0C
4.5 4.9
24 hours

4 Scale up Process for increasing the biomass of yeast.

In laboratory pure culture is raised from single yeast cell. Pour plate method is used for
propagation of yeast (pure Culture). This method is based on propagation of yeast (pure
culture. In this method six test tubes plugged with cotton containing 9 ml. of sterilized
distilled water is taken. Then 1 ml. of yeast culture is added in the first test tube. It is
mixed well by rolling the test tube between the hands. 1 Now 1 ml. of this is transferred
in the 2nd test tube for more dilution. Again 1ml of is transferred in the third test tube for
more dilution. The process is repeated till the test tube (i.e. test tube No. 6) has sufficient

Take a cleaned and sterilized flasks of 250-ml. plugged with cotton and transfer the
weighed quantity of the following in it.


2% 10gm

Yeast extract

0.5% 3gm


0.5% 3gm

Agar Agar

3% 15gm



The mixture is kept into an autoclave and sterilized for about 15-20 minutes at a pressure
of 15 PSI. After cooling it to 50C it is transferred to petridishes. A solid media is formed
in the petridishes when the temperature of media falls below 40C.
To this solid media in the petridish the dilution sample of the 6th test tube is poured under
aseptic condition. In the course of 48 hrs very tiny white or yellowish colonies of yeasts
are found. Now these colonies are successfully transferred in fresh media of glucose for
propagation in the test tube. There are 32 buds in one chain. these are used for making
streaking. The slant is kept for 12 hours in incubator. After 12 hrs it is transferred to
conical flask for 12 hours liter agar media (Sp. gr. 1.025). Similarly transfers are made to
5 liter and then 20 liter flask containing gur of sp. gravity 1.025.
Once the yeast cell gets fully propagated in the 20 liter container it is then poured into
yeast vessel for further propagation at Fermentation House.

The good cells are those which are dark at the center and bright at
circumference. The good cells are thick and have strong outer cell wall.


It the cells are dark at the circumference and bright at the centre then the
cells are supposed to be sick.


If the cells are totally dark then those cells will be dead.

Molasses used in process is daily analyzed for Brix and T.R.S. Spent wash and spent less
are analyzed to ensure that it has no alcohol in it. Fermented wash back is analyzed to
know the alcohol % contained in it for easier plant operation.

5 Fermentation House Process

The Fermentation House has:

(i) Yeast Vessels

(ii) Pre Fermenters (or Bubs) and

(iii) Fermenters

5.1 Part - 1 Yeast Vessels

Batch fermentation is done in Rampur distillery and fermentation house working may be
elaborated as under in four parts:Preparation of wort as per requirement by means of diluter in which coils are arranged in
such a way that proper mixing of molasses and water takes place. Specific gravity
checked by means of sampler, which is fitted with online system.
They have two numbers diluters, whcih could be operated at the same time. This diluted
molasses is named as worth. Wort are collected in wort tank and diverted to
prefermenters as per requirement.

5.2 Part 2 Yeast vessels:

Yeast vessels are used to propagate maximum yeast cells. The yeast vessel has aeration
steaming and cooling arrangements within it. We have four yeast vessels. The capacity of
each yeast vessels is follows:*


Vessel No. 1

5001 liters


Vessel No. 2

25001 liters


Vessel No. 3

5001 liters


Vessel No. 4

25001 liters

Yeast of yeast vessel No. 1 is out to yeast vessel No. 2 for further propagation of yeast
cells. Similarly yeast of yeast vessel No. 3 is out to yeast vessel No. 4 in rotation. Urea
and sulphuric Acid is used as nutrient and to maintain pH of the medium. The pH is kept
between 4.5 - 4.8

5.3 Part-3 Prefermenters (Bubs)

Here we are not concerned with the alcohol production, we are concerned mainly with
the growth of yeast culture and adaptation of cells in a large cylindrical metallic vessels
having capacity 86230 BL. Thats why low dilution is maintained to avoid alcohol
formation for 7 to 8 hours in anaerobic conditions. The specific gravity of the medium is

1.050 and optimum temperature for the growth of S.cerevisiae is 300C but somewhat
higher temperatures (35 - 380C) are tolerated. Yeast growth is inhibited by solution of
high osmotic pressure and high concentration of ethanol. Molasses contain variable
amounts of nitrate which can be reduced to nitrate by bacterial action during the
production of yeast. A considerable loss in yield has been reported for concentrations of
0.004 to 0.001% nitrite.
Many products have been reported to be activators or inhibitors of yeast growth. Flour
milling waste, sludge from aerobic digesters etc. are considered as activators while SO 2
inhibits yeast growth but concentrations up to 800 ppm molasses can be well tolerated.
S.cerevisiae adapts well to the presence of even higher concentrations of SO2 as is known
from the use of this species in the wine industry where fermentation is often carried out in
the presence of 80 to 100 ppm of SO2.
In Bub maximum propagation of yeast on large scale takes place. The prefer enters have
also the accretion and streaming system We have also the aeration steaming system We
have five pre-fermenters whose capacities are so follows:*

Bub No. 1

20868.4 Liters

Bub No. 2

20959.2 Liters

Bub No. 3

21143.0 Liters

Bub No. 4

43600.0 Liters

5.4 Part 4 Fermenters

Fermentation applies to both the aerobic and anaerobic metabolic activities of microorganisms in which specific chemical changes are brought about in an organic substrate
due to the enzymes secreted by those microbes.
Fermentation is the core part of distillery which is carried out in large cylindrical vessel
generally made of stainless steel having thickness 5-6 mm and capacity (46300 BL) is

called as fermenter. The media which has been prepared is introduced in the prefermenter
and after 7 8 hours it is transferred into fermenter where fermentation is carried out.
Here batch fermentation is carried out without establishing pure culture conditions and
without maintenance of complete sterility of equipment. However, this pre supposes a
rapid start of the yeast fermentation. This fermentation inhibits the growth of other
microbes by depleting the available nutrients, by lowering the pH and most importantly
by the formation of ethanol. To reduce the pH H2SO4 is usually added.
The formation of ethyl with the evolution of Co2 takes place in Fermenters. The
fermenters are provided with cooling coils so as to maintain the temperature during the
process of fermentation. The fermenters have cone shaped bottom so that yeast sludge
may settle in the bottom after the completion of fermentation.
There are eleven fermenters in Rampur distillery with the following capacity:Fermenter

No. 1

251697 Liters


No. 2

251575 Liters


No. 3

252252 Liters


No. 4

299103 Liters


No. 5

300211 Liters


No. 6

299103 Liters


No. 7

299103 Liters


No. 8

299103 Liters


No. 9

303072 Liters


No. 10

301400 Liters


No. 11

303072 Liters

Except this part fermentation house also uses 2 settler tanks and one wash charger.
5. Settler tank no. 1------ 91925liters
6. Settler tank no. 2------91925liters
7. Wash charger 3 -------99030liters
Most of the fermented wash is transferred to distillation house, but the bottom part is not
transferred to distillation house because sludge is present there and it might create
problem in distillation plant while remaining sludge is sent to methane recovery plant to
obtain suspended organic matter.

5.5 Fermentation Problems

Fermentation may sometimes get slow and the resultant yield may be disrupted. There are
many reasons associated with the facts. During fermentation may be attacked by certain
microbes and harmful microorganisms which are acid forming bacteria and yeasts. The
molasses used sometimes may not supply the required pH and concentration difference
required for the survival of yeasts cells.

Presence of high amount of raff nose sugar

and caramel substance in raw material. . Over heating molasses causes presence of 5
hydroxyl methyl furfural and subsequent decomposition of the sugar.

5.6 Controlling infections of yeast

Some times the yeast gets infected due to which less alcohol is formed in the fermented
wash. This infection can be checked successfully by using antibiotics such as Pencillin or
Benzillin Penciling from yeast Vessel stage to fermentation stage. The doses suggested
are as under:

For yeast Vessel (1000 Liter)

: 100000 Units

For yeast Vessel (400 Liter)

: 2500000 Units

For yeast Vessel (40000 Liter)

: 2500000 Units

However the dosages mentioned above can be determined on trial basis by each distillery
because the condition of each distillery and contamination varies.
The main aim of such processes and control is to obtain quality alcohol in the form of
ENA (extra neutral alcohol). It requires proper fermentation and proper processing to
generate spirit of good quality.

6 Distillation for recovery of alcohol

6.1 General Principle
Distillation is a process by which different components of a substance are separated by
virtue of their difference in boiling point.

Their are three Distillation Assemblies in Rampur distillery to give the production of
130000 litters Rectified spirit/day

55000 Liters/Day

(Made of Copper)



(Made of)



(Made of Steel)

6.1.1 Details of the Distillation plant

Various parts in the distillation plants are as follows:1. Analyzer Column


Degasifying Column

3. Aldehyde Column


Rectifier Column

5. Exhaust Column


Beer Heater

7. Condensers



9. Heat Exchangers



11. Fusel Oil Decanter

1. Analyzer Column
The Analyzer column analyses the more volatile vapor (Alcohol) from the fermented
wash leaving behind less volatiles liquid at a certain temperature.
The analyzer column has 18 plates. The plates in the column are tunnel cap type. Pressure
of ascending vapors within the column prevents the liquid from falling through the
perforation of the plates. There are down pipes (or down comers) in alternate positions on
the plates. The pipes are located in the centre and around the periphery on alternate plates
due to which there is a good distribution of the wash. The upper opening of the down
pipe enters into a cup of the next plate. This cup serves as a liquid seal and prevents
vapors from entering the down pipes.
The fermented wash is feeded to the top of the column while steam is applied in the
bottom of the column through a sparger. The temperature in the bottom of the Analyzer
column in about 105C. Since alcohol has a higher vapour pressure and evaporated from

each plate more easily than wash, so the alcohol concentration of the rising vapours
increases from plate to plate. The concentration of liquid on each plate remains
unchanged because it receives a continuous supply of fresh feed of the next above plate.
The concentration of alcohol at the top of the Analyzer column is about 40%.
2. Degasifying or Water Purifying Column
There is no separate degasifying column. The analyzer column has been extended upward
giving three segments. The main function of Degasifying column is to carry impure and
uncondensed gas (More volatile) to Head column, for improving the quality of spirit.
The Alcohol vapours are drawn from the top of the Analyzer to the Ratification column.

3. Head Column or Aldehyde Column

The vapours coming from the top of the degasifying column are introduced in the middle
of the Head column. Steam is applied to the bottom of the column. From the top of the
Head column the alcohol vapors rises and gets condensed in the condenser the
condensate is obtained as Head spirit which is impure. Thus Acetaldehyde Sulphur
Dioxide and Carbon dioxide gas get removed through the vent. The aldehyde column has
Bubble cap type of plates.

4. Rectifier Column
The Rectifier rectifies the different product of the vapour (mixture) at their boiling points.
The vapors separating from a particular layer (i.e. plate) in the rectifier are well saturated
with the liquid on the plate at that temperature. Thus there is a difference of temperature
varying from plate to plate.
The plates in Rectifier column ranges from 44 to 48. The plates are bubble cap type. The
spacing between the two plates is 7.5" to 10". The spacing in the rectifier plates is less
than that Analyser column plates.
The vapours from the top of the analyzer are introduced at the bottom of the Rectifier

through from Breaker. The form Breaker or catcher does not allow the adhering wash to
alcohol vapours to pass through it to the Rectifier bottom. It only allows the alcohol
vapours to pass through it. The concentration of alcohol at the bottom of Rectifier is 40%
V/V at a temperature of 94 to 90C. While at top it is 94% V/V at a temperature of
In rectifier high boiling fraction remains at the lower plate while tower boiling fraction
gradually ascends plate to plate. Thus high boiling fraction gradually ascends plate to
plate. Thus high boiling fraction which is known as "Fusel Oil" is separated from the
bottom of the Column at a temperature of 90 to 90C. Fusel oil is a very important and
costly material. It is very pungent bad smelling substance. Fusel oil separation helps to
improve the quality of spirit.
The vapours from the top of the Rectifier are quite rich in alcohol and get condensed in
condensers. The condensate of all the condensers (viz. Beer heater, Primary, secondary
and Vent Condenser) is know as "Reflux'. It is taken back to the top of the Rectifier to
increase the strength of alcohol and also to maintain the temperature (i.e. 78C.) at the
top. The final product (i.e. Rectified spirit) is drawn at a rate of 1/4th of the total reflux
from the 3rd or 4th plate.

5. Exhaust Column
It is just like a small Rectifier column with lesser diameter and vapourisation area.
Actually the liquid of rectifier contain some alcohol in it. So a steam connection is given
from the bottom of the Rectifier to the top of the exhaust. The temperature of exhaust is
maintained at 104 to 150C. So as to evaporate the alcohol vapours to the Rectifier.
Spent less from the bottom of the exhaust is drained to spent less tank from where it is
pumped to the analyser top of the column, to recover the last drop of alcohol (if there is

6. Beer Heater
Beer heater is hollow metallic cylindrical vessels having metal tubes indisde it. Wash is
taken inside the tubes while vapours are passed in the shell (i.e. out side the tubes).
Partial condensation of following where coolant used in wash. Thus the use of beer

Heater economizes the steam by heating the wash. It raises the temperature of wash from
32-34C to 62-65C.

7. Condensers
The function of condensers is same as beer Heater but instead of wash water is passed
through the tubes. The vapours are passed in the shell of condenser. The condenser is
primary, secondary andvent. First the alcohol vapours gets condensed in primary
condenser. The remaining vapours are then passed to secondary are passed to the vent
condenser having vent pipe in the bottle provided to take out the product. From this pipe
the incondensable gases gets escaped to the atmospheres.

8. Coolers
The condensate from the beer heater and condensers is taken back to the rectifier to plate
as reflux, from the top of the rectifier (i.e. from 3rd and 4th plate) the final product (i.e.
Rectified sprit) taken out. Since the product obtained is quite hot, so it is brought to room
temperature by the use of coolers. The coolers are also a kind of condensers having tubes
in it in which water flows.

9. Heat Exchangers
In this factory there are eight (9) numbers of plate type heat Exchangers, which are made
up of stainless steel.
Heat Exchangers play an important role in the economy of steam. The wash preheated in
the Beer Heater at a temperature of about 60C-65C is passed in the other side spent
wash (105C) is introduced. Heat transfer taken pace and the wash gets heated to 88C90C. This heated wash in the feeded at the top of the Analyser column.

10. Syphon

The spent wash, which leaves the bottom of, the analyser column passes through syphon.
The dimension of syphon determines the maximum pressure in the column. syphon is
used to keep a very steady flow of wash are steam conomy.

11. Fusel Oil Decanter

The fusel oil tends to collect at the plates having temperature of 89C-92C The fused Oil
is tapped from the 3rd, 4th, 6th and 7th plate of the rectifier column. The fusel Oil tapped
is cooled in the Decanter in which the connection of water is also provided this fusel Oil
separates as a separate layer from the water in the Decanter. Finally it is decanted to the
Storage tanks.

7 Recovery of alcohol in the form of (ENA)

There were three ENA plant in Rampur Distillery having following capacity:*



25,000 Ltr./day




10,000 Ltr./day




50,000 Ltr./day

Thus the total ENA production hen all the three plant run is 85,000 liters/day
Extra Neutral Alcohol (ENA) is the pure spirit, which is made by double or triple
distillation. This type of spirit is used for potable purpose. ENA is free from all the
impurities like Aldehyde and other substances creating bad taste and smell in it.

1. Purifying Column
It is a long column with number of bubble cap type plate. The number of plates varies
from 32-46 depending upon the capacity of the plant. The diluted RS is fed to the 15th
plate and steam is charged from the bottom keeping the temperature of the top plate to
81-82C. The vapours from the top of the purifying column are condensed in the Main
and Vent condensers. The reflux thus obtained is again introduced to the top plate. The
draw is taken from the plate just below the top plate. This spirit obtained is called/dead
spirit and is impure. The liquid from the bottom of this column is feeded to he 17th-20th
plates of another column known as Rectification column.

2. Rectifying Column
It is also a long column having a number of bubble cap type plate. The number of plates
varies from 60 to 65. The liquid from the bottom of purifiers is fed to the Rectifying
column and steam is applied from the bottom. The temperature of the bottom is kept at
105C-106C. While that of 23rd plate is 82C-83C. The temperature at the top of the
column 78C from where the draw of ENA is obtained. The fusel Oil tapped from 19th to
28th plate.

8 Lab Analysis
Laboratory is the controlling section of the distillery .It is the most important section of
the chemical industry where samples of the raw materials are analyzed.
In Rampur distillery only Grade 1 and grade 2 molasses are used for processing ,Grade
3 molasses are rejected .Molasses are analyzed by checking total reducing sugar (TRS)
,Brix , and Sludge content.
Standard test procedure for determination of total reducing sugar (TRS) and Brix in

1. Calculation for Brix


Stirred well the molasses sample with glass rod.

8. Weighed 100 gm molasses from the sample.

9. Dissolved it in 1000 ml water and measured Brix by Brix Hydrometer.
10. Noted the reading of the hydrometer and also noted the temperature of the
11. Read Brix from table, add correction factor corresponding to the Brix result.
Total Brix in molasses = (Hydrometer reading + Correction factor) x 10
(8.50+0.06) x 10 = 85.6%

2. Calculation for TRS

12. Now transferred the above solution into 1000ml volumetric flask and the
solution is maintained at 1000 ml.
13. Sucked 10 ml of above solution into a 100ml flask.

14. Added 20 ml water and 5 ml HCl.

15. Kept at water bath for inversion at 67C for 5- 7 minutes.
16. Cooled it and then added 2-3 drops of phenolphthalein indicator and neutralized
the solution with 6N NaOH till red colour appears.
17. Allowed to cool and made up the final volume 100 ml.
18. Took 5ml of each Fehling solution A and B in 250 ml flask.
19. Added 25 ml of water and put it on a hot plate for boiling.
20. Run in from the burette the whole prepared solution required to effect reduction
of all the copper so that if possible not more than 1 ml is required to complete
21. Gently boiled the content of the flask for 2 min. At the end of boiling added 1
ml of methylene blue indicator solution without interrupting boiling.
22. Completed titration in one min and take end point at the disappearance of blue
colour and appearance of brick red colour.
23. Noted the burette reading as titre value (TV).
Calculation =

Fehling Factor x Dilution x 100

Titer volume

=43.52 gm/l


3. Alcohol percentage in the fermented wash.

Sample from processed fermented wash of 48 hrs is taken which shows gravity almost
near to 1. 250 ml of sample is collected for processing.
1. Sample is added in a round flask with 250 ml water.
2. Heated the above solution on a heater and collected 180 ml of condensate in a
measuring cylinder.
3. After 180 ml of solution has been collected added 70 ml water to make it 250 ml
4. Immersed hydrometer.
5. Reading of hydrometer reflects the alcohol percentage in the fermented wash.

4. Procedure for checking sludge %

24. Stirred well the molasses sample with glass rod.
25. Weighed 125 gm .molasses from the sample.

26. Dissolved it in water and made up the volume up to 250 ml.

27. Boiled the solution up to 70C then allow cooling.
28. After 1hr. checked the sludge which was settled down at the bottom.
Sludge% = sludge volume100
Wt. of molasses
=34 x 100 =27.2%


The moisture content inside the grain should be 11 12 % as taken for every 10 gm of
sample otherwise it will create a problem in slurry preparation and proper water addition
that is to flour for making slurry. Dust % should be between 3 4 % for every 100 gm of
sample otherwise it will allow choking of the instruments and equipments. Moisture % in
flour should be not more than 10 12 % otherwise it will create problem in slurry
formation. Starch presence should be above 50% otherwise that sample is neglected. 8
9 % of alcohol in the fermented wash shows that the whole fermentation has been a

healthy one. The spent grain wash should exhibit almost negligible amount of alcohol
content around 0.02 % otherwise if alcohol content is more it shows that the process has
not been meant properly. The whole set up of the process has been designed in such a
way to give 80 KLPD of alcohol per day in the form Extra neutral alcohol after
distillation is over.

Residual Starch percent is analyzed by taking a certain amount of sample and iodine is
added to if the colour changes to blue it shows that some starch is still present and that
material is again recycled. The specific gravity of the fermented wash should be below
1.000 which shows that fermentation has occurred successfully. The total fermentation
should result in 8 9 % of alcohol, if the analysis shows less percentage then it means
that the process is lacking proper control. Saccharifaction time is noted which is 5 10
minutes. Colour change is observed and time is noted. Grist analysis helps in
determentation of the various materials generated after grinding in the form of middle,
coarse and fine. If middle and fine are present in appropriate percentage then it shows
that wort preparation will be easy. The whole process has been designed and adjusted in
such a way to yield almost 2.5 KLPD of alcohol in the form of RS (Rectified Spirit). The
spirit thus obtained is sent for maturation in barrel made of sap and oak wood.

Table 4.3 Lab analysis results for molasses


Alcohol Percentage
Sludge %

Sample Amount Taken

100 gm molasses
100 gm molasses
450- 500 ml
125 gm of molasses

Range and limit

85 % and above
40 %and above
25 40 % sludge

85.6 %
43.52 %
27.2 %

Determination of the total solid content inside the molasses is brought about by brix
calculation which indicates that more sugar is present inside the molasses. Brix of 85 and
above suggests the molasses sample is a good one and molasses of low Brix 80 and
below are rejected and not taken for processing. Similarly TRS of 40% and above are
valid to be processed as it shows the reducing sugar present that will be reduced to

fermentable sugar. A healthy fermentation should result in 8 9 % of alcohol in the

fermentation tank of every 450ml sample taken. Similarly sludge or impurities should not
be more than 40%, as such the sample is rejected if sludge percent is more. After
distillation is over the obtained alcohol in the form of RS (rectified spirit) is sent for
recovery of alcohol in the form of ENA (extra neutral alcohol ). The whole process yields
almost 85 KLPD of alcohol per day.


Fermentation processes utilize microbiological processes in producing chemical
compounds but have an industry similar to all natural product processing. Those
processes yielding simple structural chemicals, e.g. Ethanol, Butanol and Acetone are
gradually being replaced by synthetic processes with chief and abundant raw materials.
However, fermentation processes are still useful for production of complex structure,
such as Citric Acid, Lactic Acid derived from low cost carbohydrate sources.

The Starchy materials are usually grains like wheat bajra and sorghum which are not fit
for human consumption are utilized since it saves a lot of economic burden from the
company. These are primarily grain materials which are either infected or have got low
quality or have been defected due to certain diseases. Even though they still contain
starch which can be converted to fermentable sugar during fermentation. But in the case
of Barley malt these materials are carefully analyzed before milling to give a decent
output as such analysis is done to check the moisture percentage and dust percentage in
the materials.
In conversion of the grain materials requires a series of processes which are; charging
milling, liquefaction, saccharifaication, fermentation and distillation for recovery of
alcohol. In milling process the grain is charged in huge filling sections known as silo.
After charging the grain is sent for further processes known as liquefaction .In this
process the hot steam treatment is given which helps in breaking up of the starch
molecules bond. The starch molecule contains a complex structure of amylose and
amylopectin which are linked through 1-4 linkage and 1-6 linkage. Hence these are
very complex structures which are very difficult to be broken. The slurry of the material
is prepared by mashing at different levels and as such the process is completely exposed
to varying temperature difference following which the material completely turns to easy
fermentable sugar. The yeast employed for the purpose is Saccharomyces cerevisiae
which is known as top fermenting yeast as it reaches to the top column of the fermenter.
The complete fermentation lasts for about 40 -42 hours and the specific gravity comes
around 1.010 to 1.001. The complete process yields 80 KLPD of alcohol. The obtained
result after fermentation is almost 8-9 % in the fermented wash .This wash is then sent
for distillation in the distillation column to obtain almost 95 % of ethyl alcohol known
ENA (extra neutral alcohol).The obtained alcohol is then sent to the blending unit of the
distillery to get the required products. Grain generated products are usually more
expensive owing to the fact that the whole process requires much more quality and
generated spirit is of very high quality.
In malt spirit plant the process in some manner is similar to grain but in malt spirit plant
grains are allowed to sprout and then their moisture is removed to obtain moisture free
grains .This process is known as malting in which soaking of the seed allows embryo
activation and as such during mashing their is no need to add enzyme alpha amylase.

After malting the material is sent to the milling sections to obtain raw material in the
form of middle, coarse and fine. Only amyloglucosidase is added to obtain mashed wort.
This wort is collected by treatment of grains by varying levels of hot water treatment
which helps in breaking of the starch bonds. This process is known as cooking. The
collected wort is sent for fermentation which gets over in 40 - 42 hours and results in 8- 9
% of alcohol in the fermented wash. This wash is then sent to the distillation column for
obtaining malt-generated spirit.
The Molasses of 86-90C Brix with an average PH of 5.5 is received from different sugar
factories. This Molasses is first received in pucca pit and from it the molasses is pumped
to the storage M.S. steel Tank.
Molasses is allowed to settle in the storage steel tank for 15 days and from here it is
pumped to the Fermentation House. In the Fermentation House the Molasses is diluted
with water to a required gravity thought a semi-automatic diluter. In the yeast vessels and
bubs specific gravity of 1.040 is maintained and the fermenters are set up at about 1.100
The yeast from the laboratory is taken in a 4 gallons capacity yeast vessel to
Fermentation House. It is first developed in sterilized molasses solution of wort of 1.040
degree sp. gravity for 12 hours.
Before adding the yeast the molasses is sterilized in the yeast vessels, by steam coils
inside the vessel. Air is supplied throughout the period for propagation of yeast From the
yeast vessel the yeast it developed in Bub having wort of sp. gravity 1.040 Aeration is
done in the Bubs.
Sulphuric acid (commercial goods) Ammonium sculpture and urea are added in the yeast
vessel and Bubs.
After 12 hours yeast culture is transferred from the bubs to the fermenters. High gravity
fermentation is being followed. The set up gravity is about 1.100. The duration of
fementation in the Fermentors is about 20 to 24 hours.
After the Feermentation, the wash is pumped to the Distillation House with the help of
wash pumps. First the wash is received in an overhead wash charger. Then the wash is
passed in Beer Heater. The wash gets heated by the vapours coming from the rectifier top.
The temperature of wash becomes 62-64C. This heated wash is now passed through the
plate type heat exchangers due to which it's temperature rises to 88-90C and this heated
wash is now fed to the top of the analyzer column.
Steam is applied to the bottom of the column. The rising vapours are then feed to the
bottom of Rectification column.
From the top of the Analyser column alcohol vapours are fed to the Aldehyde or Head

column from where Head spirit (impure) is obtained after its condensation in the
In the Rectification column further rectification takes place and the rich alcohol vapours
are sent to the preheater or Beer Heater from the top of the Rectifier. The vapours get
condensed in it. The remaining vapours are passed to Main secondary and vent
condensers. The condensate of all the condensers (i.e. reflux) including Beer Heater is
again sent to the top of the Rectifier to increase the strength of alcohol and also to
maintain temperature at top plate. The draw of RS is then taken from the second last plate
(i.e. from the plate cost below the reflux plate) at a temperature of 78C. The RS is then
brought to the room temperature by passing it in the cooler.
Thus RS spirit, Heat spirit and Vent spirit are obtained as our desired product. In addition
to this fusel Oil from 2nd to 9th plate at a temperature or 90-95C is also obtained in the
fusel oil Decanter.
The liquid left in the bottom of the Analyser column, which is without any alcohol is
drain away as spent wash (.e. effluent) similarly the liquid left at the bottom or rectifier
column is taken in an exhaust column so as to recover the last drop of alcohol present in
the liquid. Finally from the exhaust column the liquid is drained as spent lesse. Spent
wash is sent to the effluent treatment plant to produce biogas by methane recovery plant
and also to produce biocompost by biocomposting.

To conclude we can say that alcohol in the coming era will be a good source in the form
of biofuel when petroleum and other fuels become a problem for mankind .Also in the
coming time the myths regarding consumption of alcohol will be shattered as now the
researches are proving that alcohol helps in curing heart diseases and other diseases
related to heart.
It is apparent from the foregoing discussion that the selection of feedstocks for ethanol
production will vary from region to region, and even from farm to farm. The results of
development work now being carried out will influence choices but, most significantly,
the additional choices open to farmers resulting from the opportunity to produce
feedstocks for ethanol production from a large variety of crops will alter the patterns of
farming. It is not possible to predict what new patterns will evolve. However, it is clear
that there will be benefits from the creation of choices in the form of new markets for









In the near future, ethanol is likely to be produced primarily from grain. However, the
development of processes for the effective use of other crops should yield results in the
near term which could bring about a rapid increase in the use of nongrain feedstocks.
The above study done was to obtain the idea and principles behind the fermentation
process that yields alcohol. This project allowed me to venture and get a knowledge of
the various methods of preparation of alcohol production through starchy and saccharine
materials while it also allowed me to get an industrial exposure in an alcohol producing
industry which helped me to grasp knowledge regarding biotechnological aspects and the
techniques, methods that are adopted for production of alcohol on large scale.


Amore, T., Panchal, CJ., Stewart GG.(1986). An intercellular accumulation of

ethanol in Saacharomyces cerevisiae, fuel biotechnology,9, 324 345.
Arnold, L., Demain and Davies, J.E. (1999). Manual of Industrial Microbiology
and Biotechnology. ASM Press Washington, D.C. 435 436.
Ansany, V., Dequin, S., Camarassa., Schaeffer, C., Grivet, V., Blondin, JP.,
Salmon, B., JM and Barre, P. (1995). Ability of yeast strains to perform both

alcoholic and malolactic fermentation in winemaking. Industrial Microbiology 4: 455

Beretta, A., Micheli, E., Tagliabue, L. and Tronconi (1998). Development for a
process for higher alcohol production via synthesis gas. American Chemical Society
37: 3896.
Buchner, E., (1907). Nobel Lectures.Chemistry.103 120, Elsevier
Cabras, P., Giovanni, A., Maric, G.F. and Pusino (2003). Interaction between
Fenhexamid and Yeasts during the Alcoholic Fermentation of Saccharomyces
Cerevisiae. Agri Food Chem 51: 5612 5615.
Carlsen, H.H., Degan, H., Lolyd D (1991). Physiology Of Saccharomyces
Cerevisiae. Physiology Of Yeast 7: 376 389.
Chamy, R., Nunez, M.,J., Lema JM.(1989). Energetic of Saccharomyces
cerevisiae , Bioenergetics 3 : 170 187.
Chandrakant, P. and Bisaria, V.S. (2000). Metabolic engineering of Saccharomyces
cerevisiae for ethanolic Fermentation. Metabolic Engineering. 27
Ciesarova, Z., Smogrovicova, D. and Domney Z. (1997). Ethanol is formed by
bakers yeast. Baking Technology 34.
Davidson and Dennis, M. (1989). Cardiovascular Effects of Alcohol. Western
journal of Medicine 4: 430.
Dequin, S. and Barre, P. (2004). Microbiological and Technological Fermentation.
Mixed lactic Acid Fermentation by Saccharomyces cerevisiae.
Dietler, M. (2007) Alcohol Anthropology Archeological Perspectives. Annual Review
of Anthrolpology 35: 229 249.
Figueroa, L.I., Cabada, M.A. and Broock, M.R. (2002). Pilot Plant and Process
and Industrial Microbiology. Agri Food Chemicals 51: 5612 5615.
Guide to Small Scale Alcohol Production(1988). The United States Department of
energy, Washington, D.C. 20: 545.
Heagler and Fontenot (1986). Fuel Alcohol Benefits to Agriculture. Lousiana Farm

Reporter. 13: 148

Iconomopoulou, I.,Kanellaki, K., Psarianos and Koutinas, A.A. (2000).
Delignified Cellulosic Material Supports as Freeze Dried Product in Alcoholic
Fermentation. Food Biotechnology Group, Department of Chemistry , University of
Patros. Agri Food Chemicals 51: 5012 5015.
I.S.I. Prescribed Lab Manual.,(1998). Rampur Distillery For Molasses.
Jain, J.L. (2003). Furdamentals of Biochemistry S. Chand Company Limited : 367385 :
Madan, R.,D., Bisht,B.,S.,(2001) . ISC Chemistry Book I S.Chand Company, : 602632 .
Manchester, K.L. (1999). New Beer in Old Bottle. The growth of biochemical
knowledge,Trends in Biochemical Sciences 24:125
Marin, M.R. (1996). Alcoholic Fermentation Modeling and Current State and
Perspectives. Area and Technology, University Publication , Spain 42: 452 455.
Munden, C. P. (2006). Joseph S. Fruton Fermentation Vital Or Chemical Process.
Boston Brill 15: 141.
Murata, Y and Ado,Y. (1987). Technical rep. Japan Yeast Association 57: 1 5.
Palnitkar, S.,S., Lachke, A., H.(1988). Ethanol Production by yeast from agricultural
residue, .2: 14 23.
Raymond, P. (1926). Alcohol and Longevity. NY: Knopf.
Praj Industries lab manual., (2001). Operational Manual for Starch Based industries. Praj
Rimm., Eric B., Williams, Paige., Fosher K., (1999). Moderate Alcohol intake and lower
risk of coronary heart disease. British Medical Journal 319: 1523 1528.
Satroutdinov, A. D., Kuriyama, H., Kobayashi, H. (1994). The immediate effects of
externally added alcohols on Co2 production. Journal of fermentation 2: 12 16.

Singh, B.D., (2005). Biotechnology . Kalyani Publishers, , 422-552

Singh, K.K. and Norton R.S.(1991) . Glucose consumption during fermentation, food

biotechnology.3, 27 46.
Soares, E. V.and Mota, M. (1996). Signal transduction pathways leading to trancriptional
activation of the glycolysis in Saccharomyces cerevisiae. Kinetics of fermentation 14: 109
Small Scale Fuel Alcohol Production(1988). The United States, Department of

Agriculture, Mother Earth Alcohol Washington, D.C. 20 250.

Tankana, T., and Taniguchi, M.(1990). The effect of gamma irradiation on the

fermentive capacity of strains of yeast cells. Introduction to mutation, 4, 324 356.

Verbakel, J.M. and Verrips, C.T (1993). Glucose Repression in Saccharomyces

Cerevisiae. Journal Of Applied Microbiology 24: 9 17.

Verudyn, C., Postma, E., Scheffers, W.A and Dijken, V.JP. (1992). Analysis of

Saccharomyces Cerevisiae under defined physiological conditions. Applications of

Microbiology 11: 56 54.
Waites, J. Michael., Morgan, N.L., Rockey, J.S. and Higton Gary. (2001)

Introduction to Industrial Microbiology . Blackwell Sciences London 86 93.


1 Fehling A

34.64 gm Cu So4 .5H2O

Dissolve in 1000 distilled water make up.

2 Fehling B

Sodium Potassium Tartarate 173 gm.

And sodium hydroxidein 50 gm

173 gm + 50 gm dissolve in DM water make

up to 1000 ml.

3 Methylene Blue

9,3,7-Bis(dimethyl amino) phenolthaiazin-5-ium

chloride. Dissolve the above in 3.5 gm /lt of
Distilled water. C16H18 N3SCl.3H2O

4 Phenolphthalein

C20H14 O4
Add Phenol on phthalic anhydride

5 Na OH (N/10)

Add 40 gm sodium hydroxide crystal to 1000 ml

distilled water. Make up and dissolve in 1000ml

6 Na OH (6N)

Add 120 gm of sodium hydroxide to 1000ml

distilled water. Make up and dissolve in 1000ml.

7 Culturing Media

Take the appropriate quantatity of the following

in a clean and dried flask of 250 ml.
Glucose 10 gm, Yeast Extract 3 gm, Peptone
3gm, Agar Agar 15 gm. Dissolve and sterlize it
autoclave for 20 min at 15 PSI at 120C