AN INDUSTRIAL TRAINING REPORT SUBMITTED TO

KADI SARVA VISHWAVIDYALAYA, GANDHINAGAR

FOR THE DEGRE OF

M.SC IN MICROBIOLOGY
BY

KAMALABEN NATHABHAI CHAUDHARY

22MSCMB22011

CARRIED OUT AT: ELITE PHARMA PVT. LTD

SITUATED AT: VATVA, AHMEDABAD, GUJARAT

1
2
3
 DECLARATION

hereby declare that I have undertaken industrial training at "Elite Pharma Pvt. Ltd."
during a period from "8th weeks" in partial fulfillment of requirements for the award
of degree of M.Sc. (Masters of Science) a Shri Maneklal M Patel Institute Of
Sciences & Research. which is being presented in the training report submitted to
Department Of Microbiology at Shri Maneklal M Patel Institute Of Sciences &
research. Is an authentic record of training work and it has not formed the besis
for the award Of any Deegree/ Fellowship or other similar tital to any candidate
of any university.

Place: Phase 3, Vatva G.I.D.C, Ahmedabad-382445, Gujarat

Date: 29/02/2024

CHAUDHARY KAMALABEN N"

Roll No:22MSCMB22011
Shri Maneklal M Patel institute of science & Research
Department Of Microbiology
Gandhinagar, Gujarat

4
 ACKNOWLEDGMENT

It is a matter of pleasure and happiness to make and submit this industrial

training report during Course of completion of the industrial work. Many of the
person have offered their valuable time And support.

I would like to express my sincere gratitude to Jatin P. Patel for their inspirimg
guidance & Invaluable suggestions that have been the driving force in the success
of my report. I obtained All the information needed and even the extra knowledge
regarding subject. I can never Forget everyone for their positive nature and
helping hands.

I am also grateful to P.G. Incharge_for providing great support at all level of work
During curriculum.

would like to thank all teaching & non-teaching staff of chemistry department for
providing Med environment. Me co-operative.

Chaudhary Kamalaben N.
M.Sc Microbiology Shri Manek M patel Institute Of
Sciences & Research,
Gandhinagar, Gujarat

5
 Index

Contents. Page no.

1. Introduction of company 10
2. Introduction of work 14
3. Lab equipment 18
4.Materials & Methods 32
5. Result 42
6. Conclusion 45
7. Bibliography 47

6
 Abbreviation

1.QC Quality control

2.QA. Quality Assurance

3.API Active Pharmaceutical Ingredient

4.SQC. Statistical Quality Control

5.CP Control plan

6.CQI. Continuous Quality Improvement

7.APQP. Advanced Product Quality planning

8.AQL Acceptable Quality Limit

7
 List of figures

Contents. Page no.

1. Microbiology Lab 13
2. Microscope 18
3. Analytical balance 20
4. Deep freezer 21
5. Vortex 22
6. Incubator 24
7. Autoclave 25
8. Heating Plate 26
9. Colony Counter 27
10. Ph meter 28
11. Spectrophotometer 29
12. Hot air Oven 31
13. Gel clot method 41

8
 List of tables

Contents. Page no.

1.Composition of Cultural Media 33

2. Composition Simple Media 35
3. Pepton water 35
4. Growth Promotion test 37
5. Membrane filtration method 38
6. Validation 39
7. Gel clot method 40
8. Control standard endotoxin 41

9
INTRODUCTION OF COMPANY

10
Elite Pharma Pvt. Ltd. a WHO-GMP certified company Elite Pharma Pvt. Ltd is a
well-reputed Pharma Manufacturing House always on a move for rapid progress
& fast growth. We have our own Manufacturing Plant is located at Vatva,
Ahmedabad in the State of Gujarat, which is the home of major Pharma units.
The Company is promoted by a group of entrepreneurs with considerable
experience in purchasing, manufacturing and marketing of formulations, viz.
General Category Tablets, Capsules and Oral Liquids as well as Beta Lactam
Category Tablets, Capsule & Dry Powder. The main promoter of the company.
All departments are being computerized gradually. Already the operation of Stores
Department is controlled using Computerized Pharma Software, which ensures
flawless and meticulous Transactions The Excise Section is also automated with
Custom-made Excise Software. We are committed to excel in our mission of
producing quality and we strongly believe that our success rest in continued
patronage and support of the medical fraternity, highly motivated and dedicated
work force and teamwork.

Address:

Location-: 1210, Phase 3, G.I.D.C., VATVA

AHMEDABAD-382.445. GUJRAT.INDIA
PHONE: ++91-79-25831668
Email: eppl@live.in

11
 Pharmaceutical Microbiology:

Microbiology is the biology branch involved with the study of microscopic organisms
e.g. bacteria, fungi, algae, viruses

Pharmaceutical Microbiology is an applied sub-branch of microbiology concerned with

the study of microorganisms associated with the manufacture of pharmaceuticals

Pharmaceutical Microbiology Scope:

Study of microorganisms associated with the industry

control and monitor microorganism through the process to

ensure the safe manufacturing of pharmaceutical products Microbiological Analysis

Tests Environmental monitoring

Clean areas:
Washing
Media preparation
Sterilization
Testing areas
MLT area

Live cultural areas:

Growth promotion
Strain Maintains
Microbial Identification
Decontamination area

12
Figure 1 microbiology lab

13
INTRODUCTION OF WORK

14
microbiological testing plays an important role in the production of pharmaceutical
drug substances and drug products Pharmaceutical microbiological testing is
essential for patient safety as the patients consuming the medicines might already
be in a compromised position and easily susceptible to infections pharmaceutical
microbiology testing ensures that the raw materials used in drugs match the
standard quality requirements before they are processed in the production
environment. Our microbiologists also validate the methods used for testing
finished products as well as monitor the quality of air and water from the
microbiological perspective.

Microbial Limit Tests:

Microbial contamination testing is performed for non-sterile products in

which harmonised pharmacopoeia or client-supplied methods are used that
determine the bioburden within the sample.

Total Bacterial Counts:

Total bacterial count indicates the number of microorganisms present in a

sample. The number of microorganisms should not be greater than the specified
guide values that are expressed in CFU (colony-forming units) per gram
or milliliter.

Total Fungal Counts:

It indicates fungal count present in a sample. Monitoring this is important to

know about the fungal contamination in pharmaceuticals.

15
 Sterility Testing:

Sterility testing is an essential microbiology testing requirement that

ensures sterile pharmaceuticals, medical equipment and substances
are safe for use.

We offer two sterility testing methods:

• Direct Inoculation
• Membrane Filtration

Bacterial Endotoxin Testing by LAL Test:

Limulus Amoebocyte Lysate (LAL) test is performed to check and quantify

bacterial endotoxins that are extracted from the products.

Microbiological Water Testing:

Microbiological testing of water is done during different phases of water system

validation.

Area Monitoring:

We use plate exposures and air sampling methods for environmental

monitoring.

16
 Procedure for the entry in gowning room:
1.All company employees, visitors, and auditors shall access the intended area
through the respective change.

2. Open the door and enter into the change room.

3. Remove the street footwear and keep it in the designated cupboard/designated

place under lock and key.

4. Keep the entire personal ornaments e.g. a key ring, bindi, etc. in the designated
place under lock and key.

5.Collect the shoes or shoes cover from the designated place.

6.Collect the fresh gown from the cleaned cabinet.

Procedure for the exit in gowning room:

1.All personnel moving out from the respective department shall enter into the
change room.

2.Open the door and enter into the change room.

3 Remove the gowning and safety shoes or shoes cover and keep in the
designated place.

4. During the break, the used gown shall be kept in the cupboard/designate
place and can be used for the rest of the day or after the break.

5.Used gown shall be dropped into the used gown cabinet after the end
of shift and housekeeping personnel shall collect them in a plastic bag and
send for washing with the label to cleaned.

6. Ensure all personnel gowning shall be kept in the designated place.

17
 Microbiology lab equipment:

1.Microscope:

The microscope is a device that magnifies. objects (or) organisms that are to
small to see with the naked eyes.

Part:

Eye lens,
Objective lens,
Condenser,
Beam of light,
Specimen stage,
Aperture diaphragm

Figure 2 Microscope
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 Principle:

•Light is produced from either an internal external Light source and passes
through the iris diagram.

•The light then passes through the condenser which focuses the light
onto the specimen.

•The objective lens magnifies the image of the specimen before the light
travels through the barrel of the microscope

Uses:

•In biological field, microscopes are used to study bacteria, cells and many more.

•This device helps biologists in their study of living organisms and their cell
structures.

•used for visual observation of morphology, motility, staining and fluorescent

reactions of bacteria.

19
 2. ANALYTICAL BALANCE:

• An analytical balance is a type of balance that is commonly used for the

measurement of mass in the sub milligram range.

• It uses the force necessary to counteract the mass rather than measuring the
mass itself.

• An electromagnet is used to create a force required to achieve a balance with

the mass of the substance, and the resulting force Is displayed.

Figure 3 analytical balance

20
 Principle:

• Modern electronic laboratory balances work on the principle of magnetic

force restoration.

• In this system, the force exerted by the object being weighed is lifted by
an electromagnet. A detector measures the current required to oppose the
downward motion of the weight in the magnetic field.
Uses:

• Analytical balances are used in laboratories for weighing test materials and
sampling amounts, formulation, density determination, purity analysis quality
control testing, and material and conformance testing.

3.DEEP FREEZER:

•A freezer is used to preserve foods between 25 & 10degree fahrenheit for

use usually within a few weeks (or) months at most.

Figure 4 deep freezer

21
Principle:

• Deep freezers are based on the principle that under extremely low temperatures
there is minimum microbial growth which allows for the protection and preservation
of different substances.

•Based on this principle, we can even preserve cultures over a long period of
time without any change in the concentration of the microorganisms

Uses:

•A deep freeze can be used for the preservation of different things used in the
laboratories for a very long period of time.

•Deep freezers are used in laboratories to store and preserve medical equipment,
food items, blood samples, medicines, And injection etc. for a more extended
period of time.

4. VORTEX MIXTURE/VORTEXER:

Figure 5 vortex

22
 •A vortex mixture is one of the basic technologies used for the mixing of
samples in glass tubes or flasks in laboratories.

Principle:

• It is based on the simple principle of causing reactions and homogenization by

agitating the mixture.

• Motorized draft shafts present on the mixer oscillates and transfers the movement
to the sample tubes causing the sample fluids to undergo turbulent flow.

Uses:

• Vortex mixer is mostly used for the mixing of various sample fluids in the sample
tubes and also allows for the homogenization of cells and cell organelles.

5.INCUBATOR:

• An incubator is a device that is used in the laboratories for the growth and
maintenance of microorganisms and cultures.

• Incubator provides an optimal temperature, pressure, moisture among other

things required For the growth of microganisms

23
 Figure 6 incubator

Principle:

• The incubator is based on the principle of maintaining a proper atmosphere for

the growth of microorganisms.

• Incubators have a heating system that allows for the temperature within the
incubator to be adjusted according to the type of organism cultivated inside.

• Similarly, they are provided with adjustments for maintaining the concentration
of CO2 to balance the pH and humidity required for the growth of the organisms.

• Variation of the incubator like shaking incubator is also available which allows
for the continuous movement of the culture required for cell aeration and solubility
studies.

24
Uses:
• Incubators have a wide range of applications including cell culture
pharmaceutical studies, hematological studies, and biochemical studies

• Incubators can also be used in the steam cell research area

6. AUTOCLAVE:

•An autoclave is a pressurized chamber used for the process of sterilization and
disinfection by combining three factors: time, pressure and steam

Figure 7 autoclave

25
Principle:

• Autoclaves use steam as their sterilization agent. The basic principle of an

autoclave is that all the items within the autoclave come in direct contact with
the steam for a particular period irrespective of the nature of the material- whether
it is liquid, plastic ware, or glassware.

• The amount of time and the temperature depends on the type of material being
sterilized and the increase in temperature of the cycle allows for shorter periods

Uses:

• Autoclaves are mostly used for the sterilization of medical or laboratory equipment
with the capacity of sterilizing a large number of materials at once.

• They are commonly used for the preparation of culture media during laboratory
applications

7.HEATING PLATE:
• A hot plate is a stand-alone appliance used in microbiology laboratories as a
tabletop heating system.

Figure 8 heating plate

26
 Principle:

• Unlike the traditional ways of producing heat through the fire, a hot plate
produces heat by the flow of electricity.

• On a hot plate, electricity runs through the coils which have a high level of
electrical resistance. The resistance in the coils converts the electrical energy
into heat energy which causes the coils to release heat.
Uses:

• In a laboratory, hot plates are used to heat glassware and their components.

• They are used over water baths as in water baths might be hazardous in case
of any spills or overheat.

8. COLONY COUNTER:

• A colony counter is used to estimate the density of a liquid culture by counting the
number of CFU (colony forming units) on an agar or culture plates.

Figure 9 colony counter

27
 Principle:

• This instrument can accommodate different sizes of plates which are scanned
on top with UV, white light and/or fluorescent illuminatio.

• One can accomplish the counting either manually with the touch pressure or with
a digital counter.

Uses:

• A colony counter is primarily used for counting the number of colonies present
on a culture plate to estimate the concentration of microorganisms in liquid culture.

9.PH METER:

• pH meter is a device used in laboratories that measure the H-ion concentration in water-
based solutions to determine the acidity / alkalinity of the solution.

• A pH meter is often termed as "potentiometric pH meter" as it measures the difference

in electric potential between the reference and a pH electrode.

Figure 10 ph meter

28
Principle:

• In a potentiometric pH meter, single or multiple glass electrodes, connected to a

bulb selective to hydrogen ions, are attached to a metal rod.

• When the bulb with the electrodes is dipped into a solution, hydrogen ions in the
solution exchange with positive charges on the electrode generating an electrochemical
potential which is displayed in terms of ph units on Display.

Uses:

• A pH meter is primarily used to measure the acidity of pharmaceutical chemicals,

cultures, soil, and water treatment plant.

• It can be used to measure the acidity level in wine and cheese during
their production.

10.SPECTROPHOTOMETER:

• The spectrophotometer is an optical instrument for measuring the intensity of light in

relation to the wavelength.

• Based on the amount of light absorbed by a colored solution, a quantitative analysis

of the solution canbe done.

Figure 11 spectrophotometer
29
Principle:

• Spectrophotometry is based on the Beer-Lambert Law, which states the

absorbance of light by a solution (of a particular wavelength) is directly proportional
to the concentration of the substance.

• Different wavelengths of lights are passed through a solution as different substances

have better absorbance at different wavelengths Based on the absorbance of a
particular wavelength, the quantitative analysis of a solution can be done.

Uses:

• In a microbiology laboratory, a spectrophotometer is applied for the measurement

of substance concentration of protein nucleic acids, bacterial growth, and enzymatic
reactions.

11.HOT AIR OVEN:

• A hot air oven is an electrical device that is used for sterilization of medical
equipment or samples using dry heat.

• Hot air oven is a type of dry heat sterilization which is performed on dry materials
and on substances that do not melt or catch fire under high temperature.

• inside the oven is distributed throughout the oven with a fan. This prevents the
rising of hot air towards the top while keeping the cold air at the bottom This allows
for the adequate heating Of materials inside the oven.

30
 Figure 12 hot air oven
Principle:

• Static air hot air oven: In this type of oven, the heat is produced by coils present
at the bottom of the oven with no fan. The hot air rises and doesn't allow the
effective sterilization of the materials.

Uses:

• Hot air oven can be used to sterilize materials like glassware metal equipment,
powders, etc

• It allows for the destruction of microorganisms as well as bacterial spores

31
MATERIALS AND METHODS

32
Media:

• A growth medium or culture medium is a solid, liquid or semi-solid designed to

support the growth of microorganisms or cells.

• It is either an organic or a synthetic substance that provides both the biophysical

and the biochemical factors necessary for the growth of bacteria.

WHY DO WE REQUIRE A MEDIA?

• Microorganisms need nutrients, a source of energy and certain environmental

conditions in order to grow and reproduce. In the environment, microbes have
adapted to the habitats most suitable for their needs, whereas in laboratory,
these requirements must be met by a culture medium.

NEED FOR CULTURE MEDIA:

• is usually essential to obtain a culture by growing the organism in an artificial

medium.

• If more than one species or type of organism are present each requires to be
carefully separated or isolated in pure culture.

Composition of culture media:

Water
Energy source
Carbon source
Nitrogen source
Mineral salts
Special growth factors

33
Types of culture media:

1.Classification based on physical state:

a) solid medium
b) semi solid medium
c) liquid medium

2.Classification based on the ingredients:

a) simple medium
b) complex medium
c) synthetic or defined medium
d) Special media

Solid medium:
agar is the most commonly used solidifying agent

What is agar?

Golden-yellow granular powder.

Prepared from seaweeds.
Not affected by the growth of the bacteria.
Melts at 98°C & sets at 42°C.

Semi solid media:

Such media are soft and are useful in demonstrating bacterial motility and separating
motile from non- motile strains.

34
 Simple Media:

Simple or basal media are culture media which contain the minimum adequate
nutrition for non fastidious organisms.

Example- Nutrient broth/agar

Peptone water

Composition:

Beef extract. 3.0 g.

Peptone. 5.0 g
Agar. 20.0 g
Dist. Water - 1000ml
PH. 7.0 - 7.2

When 2-3% agar is added, then we have it as nutrient agar. For semisolid media -
agar concentration is 0.2-0.4.

PEPTONE WATER:

TYPE: Basic liquid media

APPEARANCE: clear, colorless, watery, usually in test tube

Composition:

SODIUM CHLORIDE

NaCI-5g WATER-1 litter

35
Solid media:

contains 2% agar,
Colony morphology,
pigmentation,
hemolysis can be appreciated.

Eg: Nutrient agar, Blood agar

Sterility Test:

•The sterility of a product is defined by the absence of viable and actively

microorganisms when tested in specified culture media.

• The test is applied to substance, preparations or articles which, according

to the Pharmacopoeia, are required to be sterile.

• Test is performed on the end -product and is one of the quality control tests
specified for release of a batch of sterile product.

Media types:

Fluid Thioglycollate medium (FTM)

Soybean Casein Digest Medium (SCD or TSB)

36
 1.Prior to test, make sure that:

Media is sterile

Media supports growth of microorganisms

2 components in Media validation:

1.Media sterility Test

2.Growth Promotion Test

1.Media sterility:

Negative Control - may be used to identify a "false positive" test result

Incubate for 14 days prior to use, may be conducted concurrently with test

30-35°C for Fluid Thioglycollate medium (FTM).

20-25°C for Soybean Casein Digest Medium (SCD/TSB

2.Growth Promotion Test:

• To test the ability of media to support the growth of micro- organisms.

• The media should be inoculated with <100 cfu of challenge organisms.

• The challenge inoculum should be verified by concurrent viable plate counts.

• Growth promotion challenge organisms should show clearly visible growth

in the test media within 3 days for bacteria and 5 days for fungi.

37
Methods are defined in Pharmacopoeia:

1.Membrane Filtration Method (open or a closed system)

2.Direct Inoculation Method

Membrane Filtration Method: (Open Funnel Method)

Sample been filtered and rinsed
|
Membrane filter is cut into half

|
Membrane into medium Incubate

Membrane Filtration Method: (Closed System Method)

Sample been filtered and rinsed

|
Adding medium (FTM/SCD) into apparatus
|
Incubate

Incubation Method:

Period: At least 14 days incubation

Temperature: 30-35°C for FTM 20-25°C for SCD/TSB

Incubation and Examination:

1.All test & sterility control containers - incubated for at least 14 days (unless
microbial contamination detected earlier.

2. Examine for evidence growth.

38
 3.Preparation not readily seen (turbid/cloudy due to its nature) - after 14 days
of incubation transfer a suitable portion (2-5% of contents) to fresh, same
medium incubate for 7 days.

Validation (bacteriostasis & fungistasis) Test:

1 The test should be validated by inoculation with <100 cfu of challenge organism
strains to the media/product container at the beginning of the test
incubation period.

2. The challenge inoculum should be verified by concurrent viable plate counts.

3. Validation done should mimic the test proper in every detail.

4. Perform a growth promotion test as a positive control. Incubate all the containers
containing medium for not more than 5 days.

Bacteria endotoxin test:

To detect and quantify the endotoxins from gram-negative bacterial origin

using amoebocyte lysate from horseshoe crab(limulus polyphemus and
tachypleus tridentatus, tachypleus gigas, carcinoscropius rotundicauda.

Sterilization does not remove any endotoxins as they are heat stable

Methods:
1.Gel clot:

a) Gel clot (Limit test)

b)Gel clot (Semi-quantitative test)

39
2.Photometric:

a) Chromogenic (Kinetic)

b) Turbidimetric (Kinetic)

c) Chromogenic (End-point)

d) Turbidimetric (End-Point)

Gel clot method:

1.Take 5 ml glass tubes, cover them with aluminum foil, and depyrogenate the
tubes (depyrogentaion: 250°C for half hour or 200 C for 3 hours in the oven).

2.Take 0.1 ml lysate in each glass tube.

3.Add 0.1 ml (100 µl) sample in each of the tube.

4. Prepare the relevant positive and negative controls.

Control:

1.Positive control = Lysate + CSE.

2. Negative control = Lysate + LRW.

3. Positive product control = Lysate + CSE + Product.

4. Test Sample = Lysate + sample (Three replicates).

40
Preparation of CSE (Control Standard Endotoxin):

CSE is provided with the kit, it has different endotoxin units/ml.

Reconstitute the CSE vial as per manufacturers recommendations and dilute

up to the required level.

Prepare the sample by the following formula

Endotoxin limit X Product concentration

MVD ===_____________________________________
Lysate sensitivity

Figure 13 gel clot method

41
Result

42
 After the incubation and during the incubation period

growth is not observed

Passes the test of sterility (preparation is sterile)

If growth is not observed
(passes the test)

If they are not readily distinguishable from those growing in containers reserved
in the first test

Preparation fails the test

If growth is not observed

Preparation passes the test

If growth is observed

Containers are reserved & re-test is performed as in the original test

If growth is observed

microorganisms are isolated & identified

If they are readily distinguishable from those growing in containers reserved

in the first tes

Second re-test is performed using twice the no. of samples

If growth is observed

Preparation fails the test 43

 After incubation, gently pick each of the vial and invert it upside down and
observe the gel formation.

1.Positive control = Gel formation

2.Negative control = No gel formation

3.Positive product control = Gel formation

4.Sample if there is gel formation = endotoxins above the permissible limit

5.Sample = no gel formation = endotoxins are in the permissible limit

44
CONCLUSION

45
In the end I am glad to tell you that training in "ELITE PHARMACEUTICALS"
Ahmedabad very good and fabulous experience. During the training I actually
learned about the company and above its working the theoretical knowledge is
worth for getting a degree, and it is accessible in the book. During My training
period, I had seen the various instruments and apparatus in the industry. The
highly sophisticated instruments that work precisely must be operated with
intense care for optimum use. We could acquire a lot of information regarding
the latest instruments and their working procedures. Industry staff is very good
and supportive Similarly from practical point of view a pharmaceutical company
is very difficult. During the training session I tried to my level best to gain
practical knowledge.

46
BIBLIOGRAPHY

47
https://elitepharma.com/welcome/

https://www.elite-pharma.com/

https://elite-pharmaskills.com/most-frequently-asked-
questions-in-pharma-inteviews/

https://pubmed.ncbi.nlm.nih.gov/15191214/

https://www.elite-pharma.com/profile.html

https://vibrantdirectory.com/company-profile/?
ELITE-PHARMA-PVT.-LTD.&firm=NjcwMA==

https://elite-pharmaskills.com/

https://www.elitepharma.net/

https://in.investing.com/equities/elite-pharma-inc-
company-profile

https://www.medindia.net/directories/pharma/elite-pharma
-pvtltd-ahmedabad-gujarat-32641-1.htm
https://www.tofler.in/elite-pharma-private-limited/company/
U24231GJ1987PTC010062

https://www.zaubacorp.com/company/ELITE-PHARMA-
PRIVATE-LIMITED/U24231GJ1987PTC010062

https://www.drugtodayonline.com/companies/company_
info/elite-pharma-pvt-ltd

https://company.pharmahopers.com/elite-pharma-pvt-ltd
49