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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 caresystem, 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) developand 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 :-


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1. Active Material

2. Excipients

OTHER ACTIVE MATERIALSThere 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 andassay. 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.


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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 takenand 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 unitcontain 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 everyindustry.


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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 agrosciences 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.


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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 simplisticchange 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 resourcesfor 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 ahigh 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


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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 20th

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 largepharmaceutical 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


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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 realabsorptive 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.

s


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MATERIAL AND METHODS


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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

IncubatorpH meter

Conductivity meter

Colony counter


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IN MICROBIOLOGY LAB.

1). LAMINAR AIR FLOW

In fluid dynamics, laminar flow (or streamline flow) occurs when a fluid flows inparallel layers, with no disruption between the layers. At low velocities, the fluidtends 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 thefluid is very orderly with all particles moving in straight lines parallel to the pipewalls. Laminar flow is a flow regime characterized by high momentum diffusionand 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 occurdepending on the velocity of the fluid: laminar flow or turbulent flow. Laminarflow tends to occur at lower velocities, below a threshold at which it becomesturbulent. Turbulent flow is a less orderly flow regime that is characterised byeddies or small packets of fluid particles which result in lateral mixing. In non-scientific terms, laminar flow issmooth while turbulent flow is rough.


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Laminar flow barriers:-

Laminar airflow is used to separate volumes of air, or prevent airbornecontaminants from entering an area. Laminar flow hoods are used to exclude

contaminants from sensitive processes in science, electronics and medicine. Aircurtains are frequently used in commercial settings to keep heated or refrigeratedair from passing through doorways. A laminar flow reactor (LFR) is a reactor thatuses laminar flow to study chemical reactions and process mechanisms.

2). AUTOCLAVE


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Industrial autoclaves are pressure vessels used to process parts and materials whichrequire exposure to elevated pressure and temperature. The manufacture of high

performance components from advanced composites often requires autoclaveprocessing.

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 processworkloads which can withstand exposure to water, while circulating heated gas

provides greater flexibility and control of the heating atmosphere. Processing byautoclave is far more costly than oven heating and is therefore generally used onlywhen isostatic pressure must be applied to a workload of comparatively complex

shape. For smaller flat parts, heated presses offer much shorter cycle times. Inother applications, the pressure is not required by the process but is integral withthe use of steam, since steam temperature is directly related to steam pressure.Rubber vulcanizing exemplifies this category of autoclaving. For exceptionalrequirements, such as the curing of ablative composite rocket engine nozzles andmissile nosecones, a hydroclave can be used, but this entails extremely highequipment costs and elevated risks in operation. The hydroclave is pressurizedwith water; the pressure keeps the water in liquid phase despite the hightemperature. The key component of the industrial autoclave is the fast-openingdoor; 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; onthe other, the door must satisfy stringent safety requirements. Such is the quality ofautoclave door design that the US experiences as few as an estimated five or sixautoclave failures annually. Autoclave design is driven by various safety standards,foremost among which is the ASME Pressure Vessel Code. While most nations usethe ASME code, some have developed their own. The CE standard in Europeapplies to vessels as well as to electrical controls, and China requires that pressurevessels comply with their domestic code. All codes specify conservativerequirements intended to maximize safety. Local governments may also impose

licensing requirements related to autoclave operation.

Design and construction

Pressure vessel


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Pressure vessel design involves Barlow's formula, used to calculate the requiredwall thickness. However, the design of a complex pressure containment systeminvolves much more than the application of this formula. For almost all pressurevessels, the ASME code stipulates the requirements for design and testing. Prior todelivery, the pressure vessel is hydrostatically tested at 130% of its rated pressureunder the supervision of an ASME code inspector. It is filled with water, and asmall pump raises the pressure to the necessary test value, at which it is held for aspecified time (30 minutes according to the ASME code). The inspector checks forleaks as well as evidence of flaws or inadequacies in the welding. The design ofsmall autoclaves need not take into consideration the possibility of drawing avacuum inside the pressure vessel, but this assumption must not be made in largerones. Steam autoclaves, for example, can be exposed to an internal vacuum if thesteam 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 thefloor if there is to be floor level loading, as is generally the case.

3). AIR SAMPLER


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


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4). COLONY COUNTER

In microbiology, colony-forming unit (CFU) is a rough estimate of the number ofviable 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 amicroscopic evaluation, all cells, dead and living are counted. The visualappearance of a colony in a cell culture requires significant growth - whencounting 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) forliquids, and CFU/g (colony-forming units per gram) for solids to reflect thisuncertainty (rather than cells/mL or cells/g).The purpose of plate counting is toestimate the number of cells present based on their ability to give rise to coloniesunder 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 amass 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 asample for these reasons. The plate count is linear forE. coli over the range of 30 -300 CFU on a standard sized petri dish.[1] Therefore, to ensure that a sample will


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yield CFU in this range requires dilution of the sample and plating of severaldilutions.

Typically ten-fold dilutions are used, and the dilution series is plated inreplicates of 2 or 3 over the chosen range of dilutions. The CFU/plate is read froma plate in the linear range, and then the CFU/g (or CFU/mL) of the original isdeduced mathematically, factoring in the amount plated and its dilution factor. Anadvantage to this method is that different microbial species may give rise tocolonies that are clearly different from each other, both microscopically andmacroscopically. The colony morphology can be of great use in the identificationof the microorganism present. A prior understanding of the microscopic anatomyof the organism can give a better understanding of how the observed CFU/mLrelates to the number of viable cells per milliliter. Alternatively it is possible todecrease the average number of cells per CFU in some cases by vortexing thesample before conducting the dilution. However many microorganisms are delicate

and would suffer a decrease in the proportion of cells that are viable when placedin a vortex.

5). INCUBATOR

In biology, an incubator is a device used to grow and maintain microbiologicalcultures or cell cultures. The incubator maintains optimal temperature, humidityand other conditions such as the carbon dioxide (CO2) and oxygen content of theatmosphere inside. Incubators are essential for a lot of experimental work in cell

biology, microbiology and molecular biology and are used to culture both bacterialas well as eukaryotic cells. Incubators are also used in the poultry industry to act asa substitute for hens. This often results in higher hatch rates due to the ability tocontrol both temperature and humidity. Various brands of incubators arecommercially available to breeders. The simplest incubators are insulated boxeswith an adjustable heater, typically going up to 60 to 65 C (140 to 150 F), thoughsome can go slightly higher (generally to no more than 100 C). The mostcommonly 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 wellunder such conditions. For other organisms used in biological experiments, such asthe budding yeast Saccharomyces cerevisiae, a growth temperature of 30 C isoptimal. More elaborate incubators can also include the ability to lower thetemperature (via refrigeration), or the ability to control humidity or CO2 levels.This is important in the cultivation of mammalian cells, where the relativehumidity is typically >80% to prevent evaporation and a slightly acidic pH is


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achieved by maintaining a CO2 level of 5%. Louis Pasteur used the small openingunderneath his staircase as an incubator.

IN CHEMICAL TESTING LAB.

1). FRIABILITY TEST


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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 upagainst the substance. The term could also apply to ska any material that exhibitsthese 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 totest the likelihood of a tablet breaking into smaller pieces during transit. It involvesrepeatedly dropping a sample of tablets over a fixed time, using a rotating wheelwith a baffle, and afterwards checking whether any tablets are broken, and what


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forming a lip to retain the glass tubes. Alternatively the holes may be boredthrough 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 theunderside of the lower plate in woven stainless steel wire cloth made from wire0.635 mm in diameter having mesh apertures of about 2.0 mm and which retainsthe tubes. The plates are held rigidly in position, 77.5 mm apart, by means of 3metal 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 centreof the upper plate and provided with a suitable means to suspend the assemblyfrom the raising or lowering device.13. Six slatted and perforated cylindrical disks 9.5 0.15 mm thick and 20.7 0.15mm in 9/25/2014 Official Method: Determination of the Disintegration Time ofTablets diameter. Each disk is made of a suitable transparent plastic materialhaving 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 andthe others parallel with it, equally spaced on a 6 mm radium around it. Equallyspaced on the sides of the disk are four V-shaped notches. The dimensions of eachnotch are such that the openings on the bottom of the disk are 1.6 mm square andthose on the top are 9.5 mm wide and 2.55 mm deep at the centre (and about 1.3mm 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 diameterstainless steel rod approximately 9 cm in length. The lower disk is cylindrical andsmooth, 7.5 0.15 mm thick and 20.7 0.15 mm in diameter. Six holes, 4.0 0.1mm in diameter, are bored through the disk symmetrically distributed in a circlearound the axis of the disk. One end of the stainless steel rod is permanentlyembedded in the centre and flush with the lower edge of the disk. The upper disk issmooth 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 providesfor 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 circlesaround the axis. A 3.2 mm hole is bored through the axis of the disk, throughwhich the stainless steel rod is inserted such that when the apparatus is assembledthe 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 heightof 20-21 cm.6. A water bath or other suitable means of maintaining the text fluid in the jar at37 2oC.


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3). DISSOLUTION TEST

Tablets or capsules taken orally remain one of the most effective means oftreatment available. The effectiveness of such dosage forms relies on the drugdissolving in the fluids of the gastrointestinal tract prior to absorption into thesystemic circulation. The rate of dissolution of the tablet or capsule is thereforecrucial. One of the problems facing the pharmaceutical industry is to optimize theamount of drug available to the body, i.e. its bioavailability. Inadequacies in

bioavailability can mean that the treatment is ineffective and at worst potentiallydangerous (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 suchtechniques on a routine basis. These difficulties have led to the introduction ofofficial in-vitro tests which are now rigorously and comprehensively defined in the


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respective Pharmacopoeia. Tablet Dissolution is a standardized method formeasuring the rate of drug release from a dosage form. The principle function thedissolution 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 differentmanufacturers.

Prediction of in-vivo availability, i.e. bioavailability (where applicable).

Although initially developed for oral dosage forms, the role of the dissolution testhas 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 usescoulometric or volumetric titration to determine trace amounts of water in asample. 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 practicaladvantages that it holds over other methods of moisture determination, including:


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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, losson drying will detect the loss of any volatile substance. The major disadvantage isthat the water has to be accessible and easily brought into methanol solution. Manycommon substances, especially foods such as chocolate, release water slowly andwith difficulty, and require additional efforts to reliably bring the total watercontent 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


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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:-


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Cl 2 + H2O 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 FILTERThe 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


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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 SYSTEMIn 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 /cm2 . 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 regeneratedfrequently 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


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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 otherTDS, 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(600C) 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


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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 37oC may be

necessary to recover some pathogens or fecal contaminants from water, mayindigenous species fail to grow at this temperature and it is usual to incubate at 20

26oC for there detection.

TABLE:- WATER SYSTEM IN A VIEW

(OPERATING SYSTEM)

SAND FILTEROperating flow rate 7.5m3/hr

Operating pressure 3.5kg/cm2

Suspended solids


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TESTS FORE.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 theanalysis 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 oC. 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 2oC for 48 hrs.

Streaked the growth over MCA, incubated it for 24 hrs at 32 2oC.

E.coli ON MAC CONKEY AGAR

Gram stain Charecterstics colonial morphology

Gram negative Brick red may have surrounding zone ofppt bile

CONFORMATIVE TEST

IMVIC TEST

a). INDOLE TEST


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Method

Add a loop full of saline suspension in to 1% peptone water and incubatednat 30

35oC for 24 hrs. After 24 hrs of incubation added few drops of xylene, shakedthoroughly 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 3035oC 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 35oC 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 3035oC for 24hrs

Response ofE.colifor IMVIC test :-

Indole test +ve

Methyl red test +ve

Voguesproskours -ve


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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 3035oC for 48hrs.

If growth observed in the inoculated medium, streaked out growth on BGA

and incubate for 24 hrs at 36 2oC.

Morphological charecterstics of Salmonella

species on selective agar media

Selective media Charecterstic colonalmorphology

Brilliant Green Agar Small, Transparent, pink to whiteopaque.

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 IronAgar slants surface and deep inoculated (stab) with the same inoculating

loop and incubated at 35 2oC 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


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of salmonellae. If acid but no gas produced in the stab culture, the

identification of the organisms is done by agglutation test.

Test forPsudomonas aeruginosa

Subculture the pre-enriched medium to cetrimide agar base incubated at

3035oC for 2448hrs. 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 370C for 4872hrs.

Examine the colony.

OxidaseTest

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 3035oC for 1824 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 oC and

examined the test tube after 3 hrs and subsequently at suitable intervals up to 24

hrs.


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METHODS FOR ESTIMATION OF Vit- B12

CYNOCOBALAMINE

PROCEDURE:-

a). Inoculums prepration:-6 hrs culture of E.coli mutant was dissolvedin 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 standerdis dissolved in 100ml distilled water.

e). Standerd prepration:- diluted the stock solution sufficiently withbuffer 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


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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 50oC 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 standardpreparation 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 1824hrs at 37o

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

TH= Higher concentration of sample

TL= Lower concentration of sample

Ss= Higher concentration of standerd

A= (TH+TL)-(SH+SL)(TH-TL)+(SH-SL)

CHEMICAL ANALYSIS


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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 aNesselrs cylinder against freshly distilled water.

2). OX DISABLES SUBSTANCE:-The solution remains faintly pink.

METHOD:- Transferred 100ml of sample to a dry 250ml conical flaskpreviously 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.

Testpasses if the appearance of the solution does not change for at least 15 min.

6). SULPHATE


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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 121oC temperature for 25min.

b). Sterilized the glassware used in the test at 121oC 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 3035oC for 48 hrs and 20- 25oC 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.


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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 25cm2surface 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 35oC for 48 hrs for bacterial count and 20

25oC for yeast and mould count.

j). After incubation, observe the plates and count the colonies and record the

results.


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ANTIBIOTIC ASSAY

Sample Name:- NEOMYCINE SULPHATE(I.P)

1). TEST ORGANISM

Freshly prepared microbiological culture Bassi lus pumi lus.

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) ofstanderd 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 (KH2PO4) and 0.523

gm of potassium dihydrogen phosphate (KH2PO4) 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). Procedure1). 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.


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2). Poured 25ml of sterile antibiotic assay medium in four sterile petri plates and

made well with the help of sterile borer (57 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 plateat room temperature for 15 min and incubate at 30 to 35oC for 24 hrs.

CALCULATION

Calculated the % potency with the following formulae:-

Where,

A = Positive or negative value

I = Ratio of dilution

TH= Higher concentration of sample

TL= Lower concentration of sample

SH= Higher concentration of standard

SL= Lower concentration of standard

A = (TH + TL) - (SH+SL)

(TH-TL) + (SH-SL)

% potency = antilog 2 + a log I


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TOTAL AEROBIC COUNT

SampleFerikind 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 35oC 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 2025oC for 4872 hrs in an inverted position.

4) Counted the number of colonies after incubation.

tirupati medicare

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