INTRODUCTION

Quality is important in every product or services but it is vital in medicine as it

involves life. The pharmaceutical industries, as the vital segment of health care
system, conduct research and manufacture and markets pharmaceuticals and
biological products and medical devices used for the acute /chronic treatment and
diagnosis of disease.
Recent advances in drug discovery primarily in the field of Biotechnology and in
the required control over manufacturing processes, are presenting the new
challenges to the control of quality and to the system that operate internally in the
industry and by external regulations established by the federal food and drug
administration (FDA). Quality Assurance (QA) and Quality Control (QC) develop
and follow standard internal operating procedures directed toward assuring the
quality safety, purity and effectiveness of the drug supply.
The FDA has issued a primary regulation to industry titled current Good
Manufacturing Practice for finished pharmaceuticals (commonly referred to as
CGMPs or GMPs). Numerous guidelines have been issued relative to specific
dosage form and operation, which give increased guidance and direction to the
industry for them to plan for the remain in compliance.
These guidelines also serves as the basis for compliance investigation conducted
by FDA and are used in their inspections of facilities and operation.
CONTROL OF QUALITY VARIATIONS
Good raw material specification must be written in precise technology, must be
complies , must provide specific details of test methods , type of instrument and
manner of sampling and must be properly identified . the testing is done according
to the pharmacopeia monograph either IP/USP/BP.
Any raw material not meeting specifications must be isolated from the acceptable
material, snickered as rejection and returned to the supplier or disposed off
promptly.
Any raw materials may be classified into two groups :-
1. Active Material
2. Excipients
OTHER ACTIVE MATERIALS
There is such a wide variance in the nature of the ingredients used in
manufacturing, it is impossible to summaries briefly the testing of those raw
materials . One of the most important disicions to be made in raw material control
is the degree of purity to be maintained for each material.
Raw material specification normally include solubility, identification, melting
range, loss on drying , residue on ignition, special metal testing, specific impurities
that are pertinent to the method of synthesis of each individual raw material and
assay. The methods of assay are usually chemical in nature.
ACTIVE OR INERT MATERIAL
Inactive or inert material usually makes up the major portion of the final dosage
form. Therefore , their physical characteristics such as colour, odour and foreign
matter are as their chemical purity. Among other important specifications of
inactive or inert materials are particle size, heavy metal content, arsenic, selenium,
water content, microbial limit, forign metter, residue onignition and pH.
If a flavored dosage form is desired, flavors or volatile oils may be used. Flavour
and volatile oils are usually tested for refractive index, specific gravity and alcohol
content if any.
IN PROCESS CONTROL
It is important function of the in process in quality assurance program to ensure
that finished dosage have uniform purity and quality within a batch and between
batches. This is accomplished by identifying critical steps in the manufacturing
process and controlling them within limits.
A variable group of tests that are widely used for in- process controls measures
characteristics including physical appearance, colors, odour, thickness, diameter,
hardness, weight variations, disintegration time, volume check, viscosity and pH.
If derivation from the specified limits occurs the necessary corrective action taken
and a resamble is taken and tested to determine whether the quality attribute of the
product is now within the limits.
FINISHED PRODUCT CONTROL
Final testing of finished product is made in the quality control lab. These tests are
designed to determine compliance with predetermined standards prior to the
release of product for packaging and subsequent distribution is a critical factor for
quality assurance . This testing along with in process testing assure that each unit
contain the amounts of drug claimed on the label that all of the drug drug in each is
available for complete absorption that the drug is stable in the formulation in its
specific final container closure system for its expected shelf life and that dosage
units themselves contain no toxic forign substances.
ROLE AND OBJEECTIVE OF Q.C
As the name indicates this department deals with analysis of (chemical &
microbiological) of raw material and finished product.
In pharmaceutical industries Q.C department deals with analysis and formulation
of tablets and syrups etc. by checking the formulation we can find out the finished
product is of same kind the industry declare.
Formulation recheck in the industry provides the valuable data to the industry for
its production unit.
Objective
1. To screen out the Raw quality of material and finished product.
2. To screen out the Quality of purified water, which is the basic need of every
industry.
REVIEW OF LITERATURE
The technique of recombinant DNA (rDNA) was invented by Cohen and boyar in
1973, based on the Watsen, Crick and Franklin hypothisis concerning the double
helix model of DNA. As pinpointed by Bud (1998), no connection at all was
established by the early 1980s between traditional biotechnological techniques and
genetic opportunities, whereas at the end of the 1980s, at least in the USA,
incumbents from the Pharmaceutical and agro sciences industries were including
biotechnological resources based on an extensive use of rDNA techniques. The
spectacular degree of influence that biotechnology has on the pharmaceutical
industry is with no doubt a structural change that must be more precisely depicted,
especially with regards to the implications for knowledge generations
accumulation, at the least, molecular biologists have penetrated an industrial
knowledge regime that was mainly.
Dominated by traditional synthetic and/or organic chemistry and that
process have resulted on a significant shock to the knowledge regime of the
pharmaceutical industry. The pervasive entry of biotechnology into the
pharmaceuticals industry can be depicted by stressing a few stylized facts that
reveals themselves essential to the characterization of this evolution.








IMPULSE FROM THE US FINANCIAL THE MARKETS
The transition from an old to a new biotechnology industry is largely due to the
support financial investors that speculated on the success of a few academics
startups at the end of the 1970s. The transition occurred because of a simplistic
change in the business vision of biotechnology. Some extremely positive
expectations were based on an analogy with infectious diseases, whereas
treatments for bacterial infections resulted from progress in antibiotics, it was also
to be expected that genetic diseases should be cured by progress in genetic
methods. Even if such an expectations was not sheared by the scientific
community, it appears that some large pharmaceuticals corporation ( Such as
syntax corporation at that time), as well as some financial resources for analyst,
launched a new biotechnology industry by providing a blast of financial resources
for human insulin via r-DNA technology by Genentech in June 1979, marketing
the starting point of a new knowledge regime for biotechnology as well as a
redefinition of the division of labour between biotechnology and pharmaceuticals
applications. As a consequences, the new biotechnology is very interesting
example of the structuring of an industry where biotechnologys transition from
being the subject of anxious comment to wonder-boy status can be pinned down
exactly()Hud 1998;14). From this date, the role of venture capital firms has been
crucial in the development pf the so called dedicated biotechnology firms (DBFs)
and thereby, in their increasing presence in the pharmaceutical industry. Around 20
DBFs went public between 1980 and 1986, following the Genentech IPO in 1980,
and raised a cumulate gross amount of approximately US$580 million (Robbins-
Roth 2000:21).
That first generation of biotech firms actually launched the industry,
just as they had changed the dominant business vision (even if this was largely
speculative at that time). The working of financial markets contributed quite
importantly to the boom of the DBFs were founded by academic researches and a
high percentage of these firms still have academic scientists among their founders.
But the role of financial capital has gradually shifted away from funding scientific
ventures, which existed only if the form of ideas and plans, toward the guidance of
young. Already established firm towards their stock market listing (Ostro and
Esposito 1999). In that respect, the working of financial markets has a very
powerful role in the reduction in the number of small firms that characterizes the
current biotechnology industry.
MYOPIC INNOVATIVE BEHAVIORS FROM LARG
PHARMACEUTICALS CORPORATION
Achillaclelis and Antonakis (2001) provided a very interesting historical
innovation in the pharmaceutical industry. As pharmaceutical firms developed
successive generations of drugs, a few large multinational corporations were
induced to focus on product innovation and consequently, developed strategies
based on a high level of R&D expenditures, vertical concentrations and horizontal
diversifications. As pointed out by Bud (1993), all the major technological
breakthroughs of the 20
th
century have been incorporated in in-house capabilities
(from fermentation to organic chemistry and biological engineering) and this
contribute to the ongoing concentration effects experienced by large
pharmaceuticals corporations. These large firms were perusuing increasing in
market share, an objective that is still prevalent, as witnessed by their recent
mergers. Consequently, merger and acquisitions have been an essential means of
obtaining innovation and assuming control of any major technical changes
occurring in the dynamics of a particular industry. This also applies to the first
stage in the development of biotechnology (rDNA techniques). This stage has
mainly been thought of as reflecting a set of new techniques that could be easily
integrated into pharmaceutical corporations by mean of acquisitions, even if these
pharmaceutical firm had no significant expertise in molecular biology since they
were issued from chemistry and biology traditions.
In the current phase of the evolution of the pharmaceutical
industry, R&D is still a crucial determinant of firms competitiveness (McKelvey &
Orsenigo 2001). However, large pharmaceutical corporation are now contracting a
growing part of their R&D activities to DBFs. It would be difficult for large
pharmaceutical industries to survive without benefiting from the innovative
capabilities of DBFs. Through alliances with DBfs , several large pharmaceutical
corporations succeeded in internalizing the new biotechnology during the
1980s(Grabowski & Vernon 1994), although successful

Results are not yet significantly available and those corporations still have
considerable difficulties in updating their knowledge bases. Thus, one can
reasonably wonder to what extent the second stage development of the
biotechnology industry will be more difficult for large pharmaceutical firm as they
are becoming increasingly dependents on DBFs. The former have no real
absorptive capacities to fully benefit from a strategy of merging with and acquiring
DBFs (Gal ambos and Sturchio 1998).In any case, what is impressive is the
significant increase in cooperative agreement that occurred in the 1990s between
DBFs large pharmaceutical corporations.











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MATERIAL AND METHODS










EQUIPMENT/ INSTRUMENTS USED:-
IN CHEMICAL LAB. OF OC DEPARTMENT
Friability apparatus
UV Spectrophotometer
Dissolution apparatus
Karl fischer titrator
Disintigration apparatus
HPLC
GLC
Hot Plate
Sonicator
IN MICROBIOLOGY LAB
Laminar Air Flow
Autoclave
Air sampler
Filter assembly
Weighing balance
Refrigerator
Hot plate
Incubator
pH meter
Conductivity meter
Colony counter
IN MICROBIOLOGY LAB.
1). LAMINAR AIR FLOW

In fluid dynamics, laminar flow (or streamline flow) occurs when a fluid flows in
parallel layers, with no disruption between the layers. At low velocities, the fluid
tends to flow without lateral mixing, and adjacent layers slide past one another like
playing cards. There are no cross-currents perpendicular to the direction of flow,
nor eddies or swirls of fluids. In laminar flow, the motion of the particles of the
fluid is very orderly with all particles moving in straight lines parallel to the pipe
walls. Laminar flow is a flow regime characterized by high momentum diffusion
and low momentum convection. When a fluid is flowing through a closed channel
such as a pipe or between two flat plates, either of two types of flow may occur
depending on the velocity of the fluid: laminar flow or turbulent flow. Laminar
flow tends to occur at lower velocities, below a threshold at which it becomes
turbulent. Turbulent flow is a less orderly flow regime that is characterised by
eddies or small packets of fluid particles which result in lateral mixing. In non-
scientific terms, laminar flow is smooth while turbulent flow is rough.

Laminar flow barriers:-

Laminar airflow is used to separate volumes of air, or prevent airborne
contaminants from entering an area. Laminar flow hoods are used to exclude
contaminants from sensitive processes in science, electronics and medicine. Air
curtains are frequently used in commercial settings to keep heated or refrigerated
air from passing through doorways. A laminar flow reactor (LFR) is a reactor that
uses laminar flow to study chemical reactions and process mechanisms.

2). AUTOCLAVE

Industrial autoclaves are pressure vessels used to process parts and materials which
require exposure to elevated pressure and temperature. The manufacture of high
performance components from advanced composites often requires autoclave
processing.

Principle of operation

An autoclave applies both heat and pressure to the workload placed inside of it.
Typically, there are two classes of autoclave. Those pressurized with steam process
workloads which can withstand exposure to water, while circulating heated gas
provides greater flexibility and control of the heating atmosphere. Processing by
autoclave is far more costly than oven heating and is therefore generally used only
when isostatic pressure must be applied to a workload of comparatively complex
shape. For smaller flat parts, heated presses offer much shorter cycle times. In
other applications, the pressure is not required by the process but is integral with
the use of steam, since steam temperature is directly related to steam pressure.
Rubber vulcanizing exemplifies this category of autoclaving. For exceptional
requirements, such as the curing of ablative composite rocket engine nozzles and
missile nosecones, a hydroclave can be used, but this entails extremely high
equipment costs and elevated risks in operation. The hydroclave is pressurized
with water; the pressure keeps the water in liquid phase despite the high
temperature. The key component of the industrial autoclave is the fast-opening
door; this is also the critical component in cost of autoclave construction. On one
hand, the operator must be able to open and close the door quickly and easily; on
the other, the door must satisfy stringent safety requirements. Such is the quality of
autoclave door design that the US experiences as few as an estimated five or six
autoclave failures annually. Autoclave design is driven by various safety standards,
foremost among which is the ASME Pressure Vessel Code. While most nations use
the ASME code, some have developed their own. The CE standard in Europe
applies to vessels as well as to electrical controls, and China requires that pressure
vessels comply with their domestic code. All codes specify conservative
requirements intended to maximize safety. Local governments may also impose
licensing requirements related to autoclave operation.

Design and construction

Pressure vessel
Pressure vessel design involves Barlow's formula, used to calculate the required
wall thickness. However, the design of a complex pressure containment system
involves much more than the application of this formula. For almost all pressure
vessels, the ASME code stipulates the requirements for design and testing. Prior to
delivery, the pressure vessel is hydrostatically tested at 130% of its rated pressure
under the supervision of an ASME code inspector. It is filled with water, and a
small pump raises the pressure to the necessary test value, at which it is held for a
specified time (30 minutes according to the ASME code). The inspector checks for
leaks as well as evidence of flaws or inadequacies in the welding. The design of
small autoclaves need not take into consideration the possibility of drawing a
vacuum inside the pressure vessel, but this assumption must not be made in larger
ones. Steam autoclaves, for example, can be exposed to an internal vacuum if the
steam fully condenses while the vessel remains sealed. Although external
pressure cannot exceed one atmosphere, that can suffice to collapse the vessel in
some cases. Thus, stiffening may be required.
In unusual situations, the autoclave itself might have to be
square or rectangular instead of round, or it might be vertical instead of horizontal.
If the autoclave is unusually large, it may have to be set into an excavation in the
floor if there is to be floor level loading, as is generally the case.

3). AIR SAMPLER


The instrument selected for monitoring is the Lumsden-Lynch vertical elutriator. It
should operate at a flow rate of 7.4 + or - 0.2 liters/minute. The samplers should be
cleaned prior to sampling. The pumps should be monitored during sampling.





4). COLONY COUNTER

In microbiology, colony-forming unit (CFU) is a rough estimate of the number of
viable bacteria or fungal cells in a sample. Viable is defined as the ability to
multiply via binary fission under the controlled conditions. In contrast in a
microscopic evaluation, all cells, dead and living are counted. The visual
appearance of a colony in a cell culture requires significant growth - when
counting colonies it is uncertain if the colony arose from one cell or 1,000 cells.
Therefore results are reported as CFU/mL (colony-forming units per milliliter) for
liquids, and CFU/g (colony-forming units per gram) for solids to reflect this
uncertainty (rather than cells/mL or cells/g).The purpose of plate counting is to
estimate the number of cells present based on their ability to give rise to colonies
under specific conditions of nutrient medium, temperature and time. Theoretically,
one viable cell can give rise to a colony through replication. However, solitary
cells are the exception in nature, and most likely the progenitor of the colony was a
mass of cells deposited together. In addition, many bacteria grow in chains (e.g.
Streptococcus) or clumps (e.g. Staphylococcus). Estimation of microbial numbers
by CFU will, in most cases, undercount the number of living cells present in a
sample for these reasons. The plate count is linear for E. coli over the range of 30 -
300 CFU on a standard sized petri dish.[1] Therefore, to ensure that a sample will
yield CFU in this range requires dilution of the sample and plating of several
dilutions.
Typically ten-fold dilutions are used, and the dilution series is plated in
replicates of 2 or 3 over the chosen range of dilutions. The CFU/plate is read from
a plate in the linear range, and then the CFU/g (or CFU/mL) of the original is
deduced mathematically, factoring in the amount plated and its dilution factor. An
advantage to this method is that different microbial species may give rise to
colonies that are clearly different from each other, both microscopically and
macroscopically. The colony morphology can be of great use in the identification
of the microorganism present. A prior understanding of the microscopic anatomy
of the organism can give a better understanding of how the observed CFU/mL
relates to the number of viable cells per milliliter. Alternatively it is possible to
decrease the average number of cells per CFU in some cases by vortexing the
sample before conducting the dilution. However many microorganisms are delicate
and would suffer a decrease in the proportion of cells that are viable when placed
in a vortex.

5). INCUBATOR

In biology, an incubator is a device used to grow and maintain microbiological
cultures or cell cultures. The incubator maintains optimal temperature, humidity
and other conditions such as the carbon dioxide (CO2) and oxygen content of the
atmosphere inside. Incubators are essential for a lot of experimental work in cell
biology, microbiology and molecular biology and are used to culture both bacterial
as well as eukaryotic cells. Incubators are also used in the poultry industry to act as
a substitute for hens. This often results in higher hatch rates due to the ability to
control both temperature and humidity. Various brands of incubators are
commercially available to breeders. The simplest incubators are insulated boxes
with an adjustable heater, typically going up to 60 to 65 C (140 to 150 F), though
some can go slightly higher (generally to no more than 100 C). The most
commonly used temperature both for bacteria such as the frequently used E. coli as
well as for mammalian cells is approximately 37 C, as these organisms grow well
under such conditions. For other organisms used in biological experiments, such as
the budding yeast Saccharomyces cerevisiae, a growth temperature of 30 C is
optimal. More elaborate incubators can also include the ability to lower the
temperature (via refrigeration), or the ability to control humidity or CO2 levels.
This is important in the cultivation of mammalian cells, where the relative
humidity is typically >80% to prevent evaporation and a slightly acidic pH is
achieved by maintaining a CO2 level of 5%. Louis Pasteur used the small opening
underneath his staircase as an incubator.




























IN CHEMICAL TESTING LAB.

1). FRIABILITY TEST


A friable substance is any substance that can be reduced to fibers or finer particles
by the action of a small pressure or friction, such as inadvertently brushing up
against the substance. The term could also apply to ska any material that exhibits
these properties, such as:
Ionically bound substances that are less than 1 kg/L in density
Clumps of dried clay
Chalk
Stone
Tablets
Friability testing is a laboratory technique used by the pharmaceutical industry to
test the likelihood of a tablet breaking into smaller pieces during transit. It involves
repeatedly dropping a sample of tablets over a fixed time, using a rotating wheel
with a baffle, and afterwards checking whether any tablets are broken, and what
percentage of the initial mass of the tablets has been chipped off. A typical
specification will allow a certain percentage to be lost by chipping, and will not
allow any broken tablets.

2). DISINTEGRATION TEST


This method shall be used to measure the disintegration time of uncoated, plain
coated and enteric coated tablets, intended to be swallowed whole, as described in
Section C.01.015 of the Food and Drug Regulations.

1. A device for raising and lowering the basket-rack assembly, vertically along its
axis, in the immersion fluid at a constant frequency between 28 and 32 cycles per
minute through a distance of not less that 5.0 cm and not more than 6.0 cm.
2. A basket-rack assembly (see Figures I and II) containing six open-ended glass
tubes, each 7.75 0.25 cm long and having an inside diameter of not less than 21.0
mm and not more that 22.5 mm and a wall approximately 2 mm thick. The tubes
are held in a vertical position by two superimposed plastic plates each about 9 cm
in diameter and 6 mm in thickness, with six holes, each about 24 in diameter,
equidistant from the centre of the plate and equally spaced from one another. These
holes are bored completely through the lower plate, but only through the lower 4
mm of the upper plate. The upper 2 mm of the plate contains six 21 mm diameter
holes concentric with the 24 mm holes in the bottom portion of the plate, thereby
forming a lip to retain the glass tubes. Alternatively the holes may be bored
through the upper plate which is then covered with a stainless steel disk perforated
by six holes about 22 mm in diameter which fit over the tubes. Attached to the
underside of the lower plate in woven stainless steel wire cloth made from wire
0.635 mm in diameter having mesh apertures of about 2.0 mm and which retains
the tubes. The plates are held rigidly in position, 77.5 mm apart, by means of 3
metal bolts, about 6.0 mm in diameter, passing through the upper and lower plates.
A metal rod, about 8 cm in length and 7 mm in diameter, is also fixed to the centre
of the upper plate and provided with a suitable means to suspend the assembly
from the raising or lowering device.1
3. Six slatted and perforated cylindrical disks 9.5 0.15 mm thick and 20.7 0.15
mm in 9/25/2014 Official Method: Determination of the Disintegration Time of
Tablets diameter. Each disk is made of a suitable transparent plastic material
having a specific gravity of between 1.18 and 1.20. Five 2 mm holes extend
between the ends of the disk, one of the holes being through the cylinder axis and
the others parallel with it, equally spaced on a 6 mm radium around it. Equally
spaced on the sides of the disk are four V-shaped notches. The dimensions of each
notch are such that the openings on the bottom of the disk are 1.6 mm square and
those on the top are 9.5 mm wide and 2.55 mm deep at the centre (and about 1.3
mm deep at the ends). All surfaces of the disk are smooth.
4. Six plungers each consisting of two plastic disks and a 3.2 mm diameter
stainless steel rod approximately 9 cm in length. The lower disk is cylindrical and
smooth, 7.5 0.15 mm thick and 20.7 0.15 mm in diameter. Six holes, 4.0 0.1
mm in diameter, are bored through the disk symmetrically distributed in a circle
around the axis of the disk. One end of the stainless steel rod is permanently
embedded in the centre and flush with the lower edge of the disk. The upper disk is
smooth and approximately 7.5 mm thick. The lower half has a diameter of 20.7
0.15 mm and the upper half has a diameter of approximately 24 mm. This provides
for an intent to enable the seating of the plunger in the glass tube. Twelve holes 2.4
0.1 mm in diameter, are bored through the disk symmetrically in two circles
around the axis. A 3.2 mm hole is bored through the axis of the disk, through
which the stainless steel rod is inserted such that when the apparatus is assembled
the lower edge of the bottom disk is 2.8 0.1 cm from the bottom of the glass tube.
5. A cylindrical glass jar having an outside diameter of about 15 cm and a height
of 20-21 cm.
6. A water bath or other suitable means of maintaining the text fluid in the jar at
37 2oC.

3). DISSOLUTION TEST

Tablets or capsules taken orally remain one of the most effective means of
treatment available. The effectiveness of such dosage forms relies on the drug
dissolving in the fluids of the gastrointestinal tract prior to absorption into the
systemic circulation. The rate of dissolution of the tablet or capsule is therefore
crucial. One of the problems facing the pharmaceutical industry is to optimize the
amount of drug available to the body, i.e. its bioavailability. Inadequacies in
bioavailability can mean that the treatment is ineffective and at worst potentially
dangerous (toxic overdose). Drug release in the human body can be measured in-
vivo by measuring the plasma or urine concentrations in the subject concerned.
However, there are certain obvious impracticalities involved in employing such
techniques on a routine basis. These difficulties have led to the introduction of
official in-vitro tests which are now rigorously and comprehensively defined in the
respective Pharmacopoeia. Tablet Dissolution is a standardized method for
measuring the rate of drug release from a dosage form. The principle function the
dissolution test may be summarized as follows:
Optimization of therapeutic effectiveness during product development and
stability assessment.
Routine assessment of production quality to ensure uniformity between
production lots.
Assessment of bioequivalence, that is to say, production of the same
biological availability from discrete batches of products one or different
manufacturers.
Prediction of in-vivo availability, i.e. bioavailability (where applicable).

Although initially developed for oral dosage forms, the role of the dissolution test
has now been extended to drug release studies on various other forms such as
topical and transdermal systems and suppositories.

4). KARL FISCHER TITRATOR

Karl Fischer titration is a classic titration method in analytical chemistry that uses
coulometric or volumetric titration to determine trace amounts of water in a
sample. It was invented in 1935 by the German chemist Karl Fischer. The
popularity of the Karl Fischer titration is due in large part to several practical
advantages that it holds over other methods of moisture determination, including:
High accuracy and precision - typically within 1% of available water, e.g.
3.00% appears as 2.97 - 3.03%.
Selectivity for water
Small sample quantities required
Easy sample preparation
Short analysis duration
Nearly unlimited measuring range (1ppm to 100%)
Suitability for analyzing:
Solids
Liquids
Gases
Independence of presence of other volatiles
Suitability for automation
Linearity - single-point calibration, no calibration curves necessary In contrast, loss
on drying will detect the loss of any volatile substance. The major disadvantage is
that the water has to be accessible and easily brought into methanol solution. Many
common substances, especially foods such as chocolate, release water slowly and
with difficulty, and require additional efforts to reliably bring the total water
content into contact with the Karl Fischer reagents.






WATER FOR PHARMACEUTICAL USE
Water is one of the most widely and abundantly used in pharmaceutical industry. It
is required for purposes ranging from manufacturing processes to the preparation
of the final dosage forms. The quality of water therefore assumes considerable
importance.
The pharmacopeias has monograph for the following type of water.
RAW WATER
Water is one of the main substance for any pharmaceutical product ,it may provide
by municipal supply or by any local agency. The water as in its natural form is
termed as the raw water. Raw water used in the manufacturing of pharmaceutical
products. This water is pretreated for its transfer in purified water by multi-grade
filtration or quartz filtration, softening of water, chlorination and many other
dosing by chemical .
PURIFIED WATER
Method for preparation of purified water
The microbiology of water is of great importance in pharmaceutical industry due to
it multiple uses as a constituents of many products as well as for various washing
and cooling purpose. Two main aspects are involved in the quality of raw water
and any processing it receives and and the distribution system. Both should be
taken into consideration when reviewing the hazardous to the finished product and
any critical point.
Microorganisms indigenous to fresh water include pseudomonas spp.,
flavobacterium spp., E. coli., staphylococcus, salmonella, such bacteria are
harmful and often have a relatively low optimum growth temperature.


1).Chlorine treatment :-
Chlorine in the form of hypochlorite act as disinfectant and 5-12% solution of
sodium hypochlorite act as a house hold disinfectant. The reaction of chloride is as
given below:-
Cl
2
+ H
2
O HCL + HCLO (HYPOCHLOROUS ACID)
The hypochlorous acid is further decompose
HCLO HCL + [O] (Nascent oxygen)
The oxygen released in this reaction is strong oxidizing agent through its action on
cellular constituents microorganisms are destroyed. The killing of microorganisms
by chlorine and its compounds is also due in parts to direct combination of
chlorine with proteins of the cell membrane and enzymes. The chlorine dosing
done in the dilutions, after disinfection of raw water, water comes into the raw
water storage tank.
MULTIGRADE SAND FILTER
The water comes into the MGF from the RWS tank and comes into the columns.
There are four partitions occur in the column in which gravel of different size
occur in the partitions of TDS & TSS (Total dissolved solids & Total suspended
solids) are removed from the water when it passes through the column. 95% TDS
& TSS are removed at this point.
ACTIVATED CARBON FILTER
In this column adsorption is generally used to remove chlorine.
ADSORPTION
Adsorption is generally used to remove chlorine and organic impurities. It is
accomplished typically with granular activated carbon. Efficiency of the removal
depends on the activated carbon, and the operating conditions. In general, organic
adsorption efficiency is inversely proportional to solubility and may be inadequate
for the removal of low molecular weight polar compounds.
SOFTENER
Softener produce softened water because it removes the total hardness of the water.
This usually prepared by either a base or by addition of sodium
hexametaphosphate.
REVERSE OSMOSIS SYSTEM
In reverse osmosis water is forced by an osmotic pressure through a semipermiable
membrane which acts as a molecular filter. The diffusion of this soluble desolved
in the water is impended and those with molecular weight excess of 250 do not
diffuse at all. The process is the reverse of natural process of osmosis. Thus
removes microorganisms and their pyrogens. Post RO contamination occurs if the
storage vessel or distribution system is not kept free from microorganisms. In the
reverse osmosis there are two modules, three membranes occur at each module.
The membrane size is .001m. the water is inlet in RO through CF (cartridge filter)
which has pore size 5m in RO membrane 75% water hardness and TDS (Total
Dissolved Solids) & minerals are removed from the water. In the RO membrane
inlet pressure is 10 kg /cm
2
. First of all water comes in to the RO-I membrane and
circulation purified water goes to the RO storage tank and the rejected came into
the RO-II membrane and here also purification process occurs and 25% water goes
to the drain and 75% water in RO storage tank. CIP (Claning In Place) tank is used
for washing a membrane by NaOH, Citric acid and HCl at a time interval, now
water is collected in RO tank. RO has pH 6 - 7.5 and conductivity 25 - 30sim/cm.
DM WATER (Dematerialized Water Or,
Deionized water )
Deionized water is prepared by passing main water through anion and cation
exchangers resins beds to remove the ions . Thus any bacteria present in main
water will also be present deionized water and beds which are not regenerated
frequently with strong acid or alkali are often problem has prompted the
development of resins able to resist microbial contamination. One such resins a
large pore , strong base quaternary ammonium ions exchange resins which permits
microorganisms to enter the cavity and then electrostatically binds them to the
cavity surface is currently being marketed. Deionized water is used in
pharmaceuticals formulation, for washing containers and plant. Mixed bed is used
to prepare DM water and DM water has a pH 5 7 and conductivity less then
1.2sim/cm. The bed in DM plant which holds cations and anions. Anions are
lighter in weight and is placed all upper side, while cation are placed on lower side.
HCl and NaOH are used to charge the DM plant. HCl 33% and NaOH 40%
solutions are required and resin is also used. Resins binds with cations and gives
the active site to reaction and removes the minerals from water.
ULTRA FILTRATION PLANT
Now the water comes into ultra filtration plant where all the impurities are
seperates and microorganisms are also separated. UF plants have three modules
and the size of modules is .0001m which is able to separate microbes and other
TDS, TSS and other minerals. In this system water comes in to the membrane and
the membrane size is so minute and then all the impurities are entrapped in the
membrane which is washed by the hot water(60
0
C) by the help of CIP
tank(Cleaning In Place Tank) at the time interval . In the UF plant on 60% water
comes into further use , it is called permit flow, and remains 40% water goes to the
wastage. Another filter also found in the system, its name is conical filter, pore size
is 0.001m and it is helpful in removing impurities. The water rotates into the
pipelines and when not in further use then comes into main loop and this loop is
called Return Loop. Return loop is helpful in avoiding the germination of microbes
that cannot germinate in the flowing water. Now this water is called Purified water
and it is stored in the PWS(Purified Water Storage Tank) which have pH 5-7 and
conductivity less then 1. It is dematerialized water and have TOC limit 500ppb.
PURIFIED WATER SYSTEM
UV LIGHT TREATMENT:-
UV light at a wavelength of 254nm is a useful for the disinfection of water of good
optically clarity. Such treatment has an advantage over chemical disinfection, as
there is no order or flavor problem and, unlike membrane filters, is not subject to
microbial colonization. The sanitary fittings downstream of the unit will
recontaminate the water. One of the most useful techniques for checking the
microbial quality of water is by membrane filtration, since this permits the
concentration of a small number of organisms from a large volume of water. When
chlorinated water supplies are tested it is necessary to add an inactivating agent
such as sodium thiosulphate. Although an incubation temperature of 37
o
C may be
necessary to recover some pathogens or fecal contaminants from water, may
indigenous species fail to grow at this temperature and it is usual to incubate at 20
26
o
C for there detection.
TABLE:- WATER SYSTEM IN A VIEW
(OPERATING SYSTEM)
SAND FILTER
Operating flow rate 7.5m
3
/hr
Operating pressure 3.5kg/cm
2
Suspended solids <5ppm

CARBON FILTER
Operating flow rate 7.5m
3
/hr
Operating pressure 3.5kg/cm
3
SOFTNER
Operating flow rate 7.5m
3
/hr
Operating pressure 3.5kg/cm
2
Treated water hardness <5ppm
REVERSE OSMOSIS
Permeate flow rate 5.2m
3
/hr
Reject folw rate 1.8m
3
/hr
Recycle flow rate 4m
3/
hr
Permeate conductivity 2sim/cm
System Pressure 5kg/cm
2

MIX BED DM
PLANT
Operating flow rate 5m
3
/hr
Operating Procedure 3.5kg/cm
2

Treated water conductivity <sim/cm
ULTRAFILTRATION
Permeate flow rate 4.5m
3
/hr
Reject flow rate 0.5m
3
/hr
Inlet pressure 0.8kg/cm
2
Permeate pressure 0.5kg/cm
2

Rejected pressure 0.2kg/cm
2

MICROBIOLOGICAL TESTING FOR RAW
MATERIAL & CAPSULES
TESTS FOR E.COLI
E.coli belongs to the enterobacteriaceae. E.coli is the best studied organism and
the experimental organism of choice for many perposes. It is an inhibitant of the
colon of humans and other warm blooded animals and is quite uesfull in the
analysis of the water for fecal contamination. E.coli is gram negative rods(coco -
bacilli) and motile or non motile with having peritrichous flagella , aerobic or
facultative anaerobic, non endosporeforming capable of fermenting lactose with
the production of acid and gas in 24 hrs of incubation at 34
o
C. The biochemical
test for E.coli is IMVIC test which is differentiation between the
enterobacteraceae.
PRIMARY TEST
Prepare Mac Conkey Agar (MCA) and distributed 100ml in test tube and
sterilized it .
Transfer 1ml of the pre-enriched medium into the sterile MCA and
incubated at 32 2
o
C for 48 hrs.
Streaked the growth over MCA, incubated it for 24 hrs at 32 2
o
C.
E.coli ON MAC CONKEY AGAR
Gram stain Charecterstics colonial morphology
Gram negative Brick red may have surrounding zone of
ppt bile



CONFORMATIVE TEST
IMVIC TEST
a). INDOLE TEST

Method
Add a loop full of saline suspension in to 1% peptone water and incubatednat 30
35
o
C for 24 hrs. After 24 hrs of incubation added few drops of xylene, shaked
thoroughly and added 1.0ml Kovacs reagent.
b). METHYL RED TEST
Method
Add a loop full of saline suspension into tubes containing sterile glucose phosphate
broth and incubate at 30 35
o
C for 24 hrs. After incubation of 24 hrs, added 1.0ml
of barrits

reagent and observe change in colour.
c). VOGUES PROSKOUR TEST
Method
Add a loop full of saline suspension in to tubes containing sterile glucose broth and
incubated at 30 35
o
C for 24 hrs. After incubation, add 1.0ml of vagues
proskours reagent and observe change in the colour.
CITRATE UTILIZATION TEST
Methods
Add a loop full of saline suspension into tubes containing sterile kissers broth and
incubated at 30 35
o
C for 24hrs
Response of E.coli for IMVIC test :-


Indole test +ve
Methyl red test +ve
Vogues proskours -ve
Citrate utilization test -ve

TEST FOR SALMONELLA
A). Primary test
Prepare tetrathionate brilliant broth and distribute in a test tube.
Transfer 1 ml preriched sample medium to 10 ml of tetrationate brilliant
broth and incubate at 30 35
o
C for 48hrs.
If growth observed in the inoculated medium, streaked out growth on BGA
and incubate for 24 hrs at 36 2
o
C.
Morphological charecterstics of Salmonella
species on selective agar media
Selective media Charecterstic colonal
morphology
Brilliant Green Agar Small, Transparent, pink to white
opaque.
Bismuth Sulphate Agar Red without black centers.


Secondary or confirmative test
Prepared triple sugar iron Agar and distributed it in the 10 ml test tube
having a cap opening and sterilized it.
Stabs were prepared.
Subcultured colony showing specific characteristics on triple Sugar Iron
Agar slants surface and deep inoculated (stab) with the same inoculating
loop and incubated at 35 2
o
C for 24 hrs.
The formulation of acid and gas in the stab and the absence of acidity
from the surface growth in triple Sugar Iron Agar indicated the presence
of salmonellae. If acid but no gas produced in the stab culture, the
identification of the organisms is done by agglutation test.
Test for Psudomonas aeruginosa
Subculture the pre-enriched medium to cetrimide agar base incubated at
30 35
o
C for 24 48hrs. Then pigment and oxidase tests were done.
Pigment test
Prepare Psudomonas agar medium for detection of fluorescein and
pycocyanin.
Streaked a colony on the agar plate.
Incubated the petri dishes in an inverted position at 35 37
0
C for 48 72
hrs.
Examine the colony.
Oxidase

Test
Prepared 1% solution of (N,N,N,N- Tetra methyl-4-phenylene diamene
dichloride)
Took 2 drops of solution on the filter paper and smear with colony.
Test for Stephylococcus aureus
Subcultured the sample on the Baired Parker Agar, Vogel Johnson Agar and
Manitol salt Agar media. Incubated at 30 35
o
C for 18 24 hrs.
Coagulase test
Transferred the colony from the agar media to a tube containing 0.5 ml of the
rabbit plasma with or without additives. Incubated in water bath at 37
o
C and
examined the test tube after 3 hrs and subsequently at suitable intervals up to 24
hrs.
METHODS FOR ESTIMATION OF Vit- B12
CYNOCOBALAMINE
PROCEDURE:-
a). Inoculums prepration:- 6 hrs culture of E.coli mutant was dissolved
in sterile saline solution(10% transmittance).
b). Basal medium used :- dehydreate Vitamin B12 assay medium.
c). Buffer solution:- Pottasium Phosphate buffer (pH 3.0)( Adjust the pH
with orthophosphoric acid).
d). Standard stock solution:- 50mg of Vitamin B12 reference standerd
is dissolved in 100ml distilled water.
e). Standerd prepration:- diluted the stock solution sufficiently with
buffer to give final concentration of
O.025 mcg/ml
0.035 mcg/ml
0.050 mcg/ml
0.075mcg/ml
0.100mcg/ml
TEST PREPARATION
Any one concentration in relation with one of the standard concentration in buffer
solution considering the average added in formulation.

METHOD
1. Sterilized the basal medium as per instruction given on the label of the
container.
2. Inoculated the medium with the inoculums at a temp. of 40 50
o
C and
immediately poured 25 ml each in to the petridishes with the help of
sterilize measuring cylinder.
3. Made 4 cavities with the help of borer equidistant to each other.
4. The volume of solutions added to each cavity was uniform.
5. Arranged the solution of standard

preparation and assay preparation to be
examined on each dish so that they alternate around the dish and so that
highest concentration of standard test preparation one not adjacent.
6. Solution is prepared in randomized block design.
7. Kept the plates for about 45 minutes or 1 hr room temperature.
8. Incubated the plates for about 18 24hrs at 37
o
C.
9. Measured the diameter of incubation

zones and calculated the % potency
with the following formula.

%Potancy = antilog(2.0+ a log I)
Where,
a = Positive or negative value
I = ratio of dilution
T
H
= Higher concentration of sample
T
L
= Lower concentration of sample
S
s
= Higher concentration of standerd

A= (T
H
+T
L
)-(S
H
+S
L
)
(T
H
-T
L
)+(S
H
-S
L
)

CHEMICAL ANALYSIS
PURIFFIED WATER(IP\EP)
1). DISCRIPTION :- A clear, colorless, odorless and tasteless liquid.
METHOD:- Checked the color, clarity, odour of the sample by comparing in a
Nesselrs cylinder against freshly distilled water.
2). OX DISABLES SUBSTANCE:-The solution remains faintly pink.
METHOD:- Transferred 100ml of sample to a dry 250ml conical flask
previously rimed with sulfuric acid and 0.10ml of 0.02ml potassium per magnate.
And boiled for 5 min . The test passes, if the solution remains faintly pink.
3). ACIDITY AND ALKALINITY
The sample is neutral to solution of methyl red or bromothymole blue indicator
METHOD
To 10ml freshly boiled and cooled in a borosilicate glass flask, add 0.05ml of
methyl red solution. The resulting solution of not red to another 10ml sample, add
0.1 ml of bromothymole blue solution. The resulting solution is not blue.
4). CALCIUM & MAGNESIUM
Took 100ml of solution in a conical flask. Add 2ml of ammonia buffer solution
(pH 10.0) add 50mg of mordent black mix and 0.5ml of 0.01 M Sodium edentate.
A pure blue color is produced.
5).CHLORIDE
Transferred 10ml of sample to a test tube. Add 1 ml of 2 M nitric acid and 0.2ml of
0.1 ml silver nitrate.
Test

passes if the appearance of the solution does not change for at least 15 min.
6). SULPHATE
Transferred 10ml of sample to a test tube . Add 0.1ml of 2M HCl and 0.1 ml of
barium chloride solution.
The test passes if the appearance of solution does not change for at least 60 min.
ENVIRONMENT MONITRING
1) PROCEDURE
Settle plate
a). Prepared Soyabean Casein Digest Agar (SCDA) medium and sterilized it in an
autoclave at 121
o
C temperature for 25min.
b). Sterilized the glassware used in the test at 121
o
C for 25 hrs.
c). Transferred the medium and the glassware through the pass box in the LAF
room.
d). Retained the test article within the dynamic pass box for at least 15 min.
e). Placed the article on the laminar air flow work station and prepare the sattle
plates of soyabean casein digest agar.
f). Marked the plates as per sampling plan and location as defined in annexure.
g). Carried the petri dishes in the container to manufacturing area in the
manufacturing area expose the plates on the floor as per sampling plan keeping the
cover on the periphery of the bottom.
h). leave the plates for at least 30 min. Now close the petri dishes with the cover
and placed the petri dishes in the container and carried to the microbiological lab
for incubation.
i). Incubated the plates at 30 35
o
C for 48 hrs and 20- 25
o
C fo next 72 hrs.
j). After incubation take out the plates and count the colonies forming units with
the help of colony counter.
k). Record the results on plate count record of manufacturing area.
SWAB SAMPLING
a). Prepared the settle plates of soyabean casein digest agar medium and
preincubate for 24 hrs before stab testing.
b). After preincubation check the plates for any contamination and discard the
contaminated plates found and use remaining plates for stab testing.
c). Prepared 0.9% w/v normal saline solution and stab. Plug the flask with cotton
and sterilize it by autoclaving . Use sterilized cotton swabs for test.
d). For swab testing take sterile 0.9% w/v normal saline solution in a sterile test
tube and put it in a closed container along with sterile cotton swabs and pre
incubated other plates and go to the manufacturing area from where swab samples
to be collected.
e). Dipped the swab in sterile 0.9% w/v normal saline solution and squeeze the
swab against the wall of the test tube to remove excess normal saline from it.
f). Now wipe the moistened swab on 25cm
2
surface of wall and floor unidirectional
(horizontally and vertically).
g). Without touching the swab head, streak it on the plateof soyabean casein digest
agar(SCDA).
h). Each medium plate should be detailed with sampling location and sampling
date.
i). Incubated the plates of SCDA at 30 35
o
C for 48 hrs for bacterial count and 20
25
o
C for yeast and mould count.
j). After incubation, observe the plates and count the colonies and record the
results.


ANTIBIOTIC ASSAY
Sample Name:- NEOMYCINE SULPHATE(I.P)
1). TEST ORGANISM
Freshly prepared microbiological culture Bassilus pumilus.
2). Standerd prepration
Accurately weighed 100mg of sample 100ml of volumetric flask. Add 50ml of
buffer solution and mixed well to dissolve completely. Make the volume upto
100ml with buffer solution. Diluted 1ml of solution to 100ml and 25ml and made
the volume with th buffer solution to get low(10 mcg/ml) & high (40 mcg/ml) of
standerd dilutions respectively.
3). Sample prepration
Accurately weighed 100mg of sample in 100ml of volumateric flask. Added 50 ml
of buffer solution and mixed well to dissolved completely. Made the volume upto
100ml with buffer solution. Diluted 1 ml of solution to 100ml and 25 ml and made
the volume with the buffer solution to get low(10 mcg /ml) & high (40 mcg/ml) of
sample dilution respectively.
4). Prepration of buffer solution:-
Dissolved 16.73 gm of di potassium hydrogen phosphate (KH
2
PO
4
) and 0.523
gm of potassium dihydrogen phosphate (KH
2
PO4) in sufficient purified water add
dissolved completely. Made the final volume to 1000ml with distilled water.
Adjusted the pH to 8 0.1 with 8 gm phosphoric acid or 10M KOH.
5). Procedure
1). Took freshly prepared Bacillus Pumilus (ATCC 14884) microbial culture slant,
aseptically transfer 10ml normal saline (0.9%) mix well to obtain suspension, then
add 1ml culture suspension in 100 ml freshly prepared media and mix well.
2). Poured 25ml of sterile antibiotic assay medium in four sterile petri plates and
made well with the help of sterile borer (5 7 mm).
3). Load 100 l of test and standard concentration on each of four plates,
alternating high and low concentration of standard and test dilutions. Kept the plate
at room temperature for 15 min and incubate at 30 to 35
o
C for 24 hrs.

CALCULATION
Calculated the % potency with the following formulae:-
Where,
A = Positive or negative value
I = Ratio of dilution
T
H
= Higher concentration of sample
T
L
= Lower concentration of sample
S
H
= Higher concentration of standard
S
L
= Lower concentration of standard
A = (T
H
+ T
L
) - (S
H
+S
L
)
(T
H
-T
L
) + (S
H
-S
L
)
% potency = antilog 2 + a log I




TOTAL AEROBIC COUNT
Sample Ferikind syrup (Batch no. FRK 09003)
a). Total bacterial count
1) Total 1ml sample and dilute in 9ml of Normal saline (NS) to get a
dilution (1:10).
2) Poured 1ml of sample in sterile duplicate petri plates of Soyabean
casein digest agar (SCDA) medium.
3) Incubate the plates at 30 35
o
C for 48 72 hrs in an inverted
position.
Count the number of colonies after incubation.
b). Total fungal count(yeast and mold)
1) Took 1ml sample and dilute in 9ml NS to get a dilution(1:10).
2) Poured 1ml sample in sterile duplicate petri plates of (SCA) medium.
3) Incubated the plates at 20 25
o
C for 48 72 hrs in an inverted position.
4) Counted the number of colonies after incubation.