Parenteral products preparation and their characterization.

1. Presented by
Mr. Sree Prakash Pandey
M.Pharm (Pharmaceutics)
Central University of South Bihar, Gaya
PARENTERAL
PRODUCTS

2. CONTENTS
Introduction-Definition
Advantages & Disadvantage
Preformulation factors & Essential requirements for Parenteral
Classification of Parenteral preparation
Sterile Powder & Lyophilized products
Physiological consideration
Formulations consideration
Production Procedure or Manufacturing process
Filling & Sealing of Ampoules, Vials & Infusion fluids
Quality Control Test of Parenteral
References
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3. Introduction
DEFINITION:
 Parenteral preparations are pyrogen-free preparations intended to be
administered other than oral routes.
 The term parenteral is derived from two Greek words,
Para outside ; enteron intestine (i.e. beside the intestine).
 An injection is a preparation in the from of sterile drug, solution,
suspension or emulsion intended for parenteral administration either
directly or after constituting or diluting prior to administration.
 Route of administration for parenteral such as – i.v. i.m. s.c. commonly
used.
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4. Advantages
 Quick onset of action.
 Useful for unconscious or vomiting patients.
 Duration of action can be prolonged by modifying formulation.
 Suitable for nutritive like glucose and electrolyte.
 Suitable for the drugs which are inactivated in GIT or HCL (GI fluid)
Disadvantage
Once injected can not controlled (retreat).
Injection may cause pain at the site of injection.
Only trained person is required.
Difficult to save patient if overdose.
Sensitivity or allergic reaction at the site of injection.
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5. Essential Requirements For Parenteral Dosage Form
1. Container: Containers used for packing parental are classified as –
 Well -closed containers: A well- closed
container protects the contents from loss
during transportation, handling, storage or sale.
 Single dose containers: These container are
used to supply only one of medicament and hold
generally Parenteral products. E.g. Ampoules & Vials
 Multidose containers: Hold a quantity of the
material that will be used as two or more doses.
E.g. multiple dose vial or the plastic tablet bottles.
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6. 2. TYPES OF GLASS USED IN PARENTERALS
Glass type General description Test method Uses
TYPE 1 Highly-resistant
borosilicate glass
Powdered
Glass test
For buffered and
unbuffered aqueous
solution , powders
TYPE 2 Treated (sulphur dioxide
fumes) soda lime glass
Water attack
test
For buffered aqueous
solution with ph
below 7 and for dry
powders
TYPE 3 Soda lime glass Powdered
Glass test
For dry powders and
oleaginous solutions,
not for aqueous
preparations
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7. 3. Closures
• Closures for parenteral preparation containers should be equipped with a firm
seal to prevent entry of microorganisms and other contaminants.
• They should not be made of components that react with the contents , nor
should they allow foreign substances to diffuse into the preparation.
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8. 4. Aseptic area:
• The parenteral preparation are filtered , filled into final container & sealed
should be in aseptic area.
• Ceiling, wall & floor of that area should be sealed & painted.
• The High Efficiency Particulate Air filters (HEPA) is used for air .
• The air in the aseptic area should be free from fibers, dust &
microorganism.
Classification of Clean Room
8
Grade Maximum permitted no of particles/m3
At Rest At Operation
0.5µm 5µm 0.5µm 5µm
A 3,520 20 3,520 20
B 3,520 29 352,000 2,900
C 352,000 2,900 3,520,000 29,000
D 3,520,000 29,000 Not defined Not defined

9. Vehicles & Additives
9
1. Vehicles
A) Aqueous vehicle - water is used as vehicle for majority of injections because water
is tolerated well by the body and is safest to administer .The aqueous vehicle used
are ;-
a. Water for injections.
b. Water for injection free from CO2 (carbon dioxide)
c. Water for injection free from dissolved air, water for injection is sterile water,
which is free from volatile, non- volatile impurities and from pyrogens.
B. Non -aqueous vehicles -The commonly used non-aqueous vehicles are oils and
alcohols. Fixed oil, such as arachis oil, cottonseed oil, almond oil and sesame oil
are used as vehicle.
The oily vehicles are generally used when a depot effect of drug is required, or the
medicaments are insoluble or slightly soluble in water or the drug is soluble in oil
example dimercaprol injection by using arachis oil as vehicle.
Ethyl alcohol is used in the preparation of hydrocortisone injection hydrocortisone is
insoluble in water, hence the solution is made in 50% alcohol. Alcohol causes pain and
tissue damage at the site of injection. Therefore, it is not used commonly.
Propylene glycol is used as a vehicle in the preparation of digoxin injection .it is
relatively non-toxic, but it causes pain on S/C or I/M injection.

10. 2. Additives
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a) Solubilizing agents
b) Stabilizers
c) Buffering agents
d) Antibacterial agents
e) Chelating agents
f) Suspending, emulsifying and wetting agents
g) Tonicity factors

11. Importance of Isotonicity
11  Isotonicity is important for parenteral preparation because if the solution is
isotonic with blood, the possibility of product penetrating the RBC and
causing hemolysis is reduced.
 A solution having the equal
osmotic pressure as a
specific biological fluid.
 For hypertonic solution
crenation, and
 For hypotonic solution
hemolysis will occur.
References: Difference between Isotonic, Hypertonic,Hpotonic solutions?
PHARMACEUTICAL MICROBIOLOGY; Small World but Big Opportunities.
 An isotonic solution is one that exhibits the equivalent/same effective
osmotic pressure as blood serum.

12. 12
References: Difference between Isotonic, Hypertonic,Hpotonic solutions?
PHARMACEUTICAL MICROBIOLOGY; Small World but Big Opportunities.

13. Classifications of Parenteral preparation
Based on volume:
 Small Volume Parenterals (SVP) :
According to U.S.P “an injection that is packaged
in containers labeled as containing 100 ml or less”.
 Large Volume Parenterals (LVP) :
They are given in multi-dose and volume 100ml
or more than .
Generally, these provides electrolytes nutrition to the body.
E.g. dextrose(5-50%), body fluids.
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14. COMPARISON
Parameters SVP LVP
Volume ≤ 100 ml 101- 1000ml
Route i.m, i.v, s.c. i.v.
Dosage unit Single / multiple Multiple
Preservative Used Not used
Buffer Used Not used
Isotonicity Not essential Must
Uses As therapeutic agent,
diagnostic agent
As Nutrition,
Detoxification, Aid
during surgery.
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15. Based on States of products:
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Injection: Injections contain sterile solutions and are prepared by dissolving the
active ingredient and other substances in Water for Injection or other suitable
non-aqueous base or a mixture of both. The solution to be injected may show
sediments which can be dispersed easily by shaking the container. The
suspension should remain stable in order to deliver a homogenous dose
whenever withdrawal is made from the container.
Infusions: These parenteral preparations are composed of sterile aqueous
solution with water as its continuous phase. The preparations are free from
bacterial endotoxins or pyrogens and are made isotonic with blood. They do
not contain any antimicrobial preservatives.
Powder for Injection: These are sterile solid compounds that are distributed in
their final volume when the vial or container is shaken to form a clear particle-
free solution.

16. 16
Concentrated Solutions for Injections: The concentrated solutions are diluted
with water for injection before they are administered through injection or
through intravenous infusion.
Implants: These solid sterile preparations are implanted in the tissue in order to
release the active ingredient for long periods. They are stored in sterile
containers individually.
Injectable Emulsion: These are liquid preparations in which the drug
substances are dissolved or dispersed in a suitable emulsion medium. These
products provide essential fatty acid and vitamins.
Oily Injection: These are used to prepare parenteral controlled release dosage
forms.

17. Sterile Powder & Lyophilized products
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Formulation of sterile powders:- Due to instability in water, many drugs
are formulated as drug powders to be reconstituted prior to
administration. eg. Penicillins, barbiturates, benzocain. Sterile water for
injection is supplied with dry powders to make “solutions / or
suspensions for injections”.
The obtained solution / suspension will meet with all the requirements of
solution /suspension for parenteral. IV or IM route can give
reconstituted solutions, however suspension is forbidden for IV
administration. Sterile powers are prepared by following methods.
1. Sterile recrystallization
2. Lyophilization
3. Spray drying

18. Sterile Re-crystallization: The drug is dissolved in a solvent and the
obtained solution is sterilized through 0.22 μm membrane filter.
A sterile anti-solvent is then added to crystalize the drug particles,
which is filtered and dried aseptically.
Advantages: This method is Flexible and economic.
Disadvantage: This method represents variations from batch to batch
and contamination.
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19. 2. Lyophilization: In this method, a solid substance is separated from
solution by freezing the solvent and evaporating the ice under vacuum.
The obtained drug solution is sterile filtered into sterile trays, which are
aseptically loaded into a freeze dryer.
The solution is then frozen at -50°C and then dried by vacuum to
separate the drug powder.
Advantage: This method involves removal of water at low temperatures.
Disadvantage:
i) In this method, the biological molecules are damaged by the stress
associated with freezing, and drying.
ii) ii) This method is expensive and time consuming
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20. 3. Spray drying: In this method, the solution of the drug is sprayed into a dry
chamber where it comes in contact with a hot steam of a sterile gas 80-100 °C
temperature.
Advantage:
This method is Simple, Economical, scalable and faster.
This method involves Coating of particles during drying prolonged release.
Disadvantage:
In this method, the high processing temperatures and high shear forces can
easily damage drugs.
In this method, higher levels of drugs are lost in comparison to freeze-drying.
This method has a limited solvent choice for a given drug.
In this method, product cannot be prepared directly in vials or plates.
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21. PHYSIOLOGICAL CONSIDERATION
1. pH: The main step in selection of pH in a formulation. For the maximum
Physiological acceptability, the target pH is approx pH 7.4.
2. Buffer: Important for parenteral dosage form because due to alteration of
pH in formulation. E.g, Potassium phosphate, tartaric acid, dibasic
sodium benzoate.
3. Tonicity: All parenteral product should be isotonic ,due to risk of
hemolysis. Tonicity adjustment agent such as gelatin, manitol, sorbitol,
NaCl etc.
4. Stabilizer: To prevent the degradation of the drug substance i.e. glycine ,
sodium saccharine, etc.
5. Antimicrobial: Against microbes and prevent i.e. Benzalkonium
chloride, Benzethonium chloride, Propyl paraben, etc.
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22. FORMULATIONS CONSIDERATION
Parenteral preparation contains – solution, suspension, emulsion, powder of
injection, gels injection.
Required for parenteral preparation :
1. Active drugs
2. Solvent :
i. Water : WFI, SWFI, Bacteriostatic WFI.
ii. Aqueous vehicle : i.e, Ethyl alcohol, PEG
iii. Non-aqueous vehicles: i.e. Fixed oils (corn oil, peanut oil)
3. Surfactants: Surface active agents enhance drug solubility to the required
concentration to achieve solution clarity. e.g. Tweens, Poloxamers,
Sorbiton monooleate.
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23. 4. Polymers: Hydrophilic polymers to enhance drug solubility by complexation
(polyvinyl alcohol) , PEG, Pectin, Gelatin, CMC.
5. Preservatives: As a bacteriostatic to inhibit any microbes accidentally
introduced while doses are being withdrawn. e.g. benzalkonium chloride,
benzyl alcohol, methyl & propyl paraben, phenolic compound.
6. Buffer: Parenteral formulation should not vary significantly from physiological
pH (about 7.4) e.g. acetates citrates phosphates.
7. Tonicity adjusters: An isotonic solution is one that exhibits the same effective
osmotic pressure as blood serum. Eg, Sodium Chloride(0.9%).
8. Antioxidant: Ascorbic acid, sulfur dioxide, metabisulfite and sulfite are the most
common antioxidant used in aqueous parenteral.
9. Chelating agents: Disodium EDTA and metalic salts.
10. Inert gases : Inerts gases enhancing the product intregity of Oxygen sensitivity
medicaments is by displacing the air solution with Nitrogen or Argon.
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24. Production Procedure/Process/Step
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1) Cleaning and washing of containers and closures
2) Collection of materials
3) Preparation of solutions
4) Filling and sealing
5) Sterilization
6) Evaluation of parenterals
7) Packaging and labeling

25. MANUFACTURING PROCESS
Following steps are involved in the processing of parenteral preparation:
1. Cleaning of containers, closures & equipments :Thoroughly cleaned with
detergents with tap water → distilled water →finally rinsed with water for
injection.
• Rubber closures are washed with o.5% sodium pyrophosphate in water.
2. Collection of materials: All raw material of preparation should be collect from
warehouse after accurate weighed.
water for injection should be pyrogens free.
3. Preparation of parenteral products: The parenteral preparation must be
prepared in aseptic conditions.
• The ingredients are accurately weighed separately and dissolved in vehicle as per
method of preparation to be followed.
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26. 4. Filtration : The parenteral preparation must be filtered by bacteria proof filter
such as filter candle, membrane filter .
5. Filling the preparation in final container : The filling operation is carried out
under strict aseptic precautions .
6. Sealing the container : Sealing should be done immediate after filling in aseptic
environment.
7. Sterilization: For thermostable substances the parenteral products are sterilized
by autoclaving method at different temperature & pressure.
• Heat sensitive or moisture sensitive material are sterilized by exposure to
ethylene oxide or propylene oxide gas .
8. Evaluation of the parenteral preparation: Sterility testing, Clarity test, Leakage
test, Pyrogen testing and Assay.
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27. Process Flow Chart
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Ingredients
vehicles additives
Compounding Filteration
Washing
Packing
material
STERILIZATIO
N
Packing &
Labeling
Finished
making
product
Storage
Equipment
container s
closures Sterilization
WFI
Containers
Equipments
Finished
Product
Aseptic
filling &
sealing

28. Filling & Sealing of Ampoules, Vials & Infusion fluids
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 Ampoules are thin-walled glass containers, which after filling, are sealed by
either tip sealing or pull sealing.
 There are wide variety of ampoule types from 0.5 to 50ml volume.
 Ampoules may be closed by melting a portion of the glass of neck to either
form tip-seals or pull seals.
 Tip-seals are made by melting sufficient glass at the tip of the ampoule neck
to form a bead of glass and close the opening. This is performed in a high
temperature gas oxygen flame.
 Pull-seals are made by heating the neck of a rotating ampoule below the tip,
then pulling the tip away to form a small, twisted capillary just prior to being
melted closed. Pull sealing process is slower one, but the sealing done by
this is more secure than that of tip sealing.
Ampoules

29. Storage :
Store in a sterile, airtight , tamper- proof container.
Labeling:
The term labeling designates all labels and other written, printed or
graphic matter on in any package or wrapper in which it is enclosed.
Materials used in labeling
The following types of labels are, such as –
 paper
 foil
 Fabric
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30. Quality Control Test of Parenteral
1. Sterility testing
2. Clarity test (Particulate matter monitoring)
3. Leakage test (Faulty seal packaging)
4. Pyrogen testing
i. Rabbit test
ii. LAL test
5. Assay
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31. Sterility Testing
Definition:
 Sterility Testing: It is a procedure carried out to detect and conform absence
of any viable from of microbes in or on pharmacopoeial preparation or
product .
 Principle: If the microorganism are present in the product can be indicated
by a turbidity in the clear medium .
Objective of sterility testing :
• For validation of sterilization process.
• to check presence of microorganisms in preparation which are sterile .
• To prevent issue of contaminated product in market .
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32. Steps involved in sterility testing :
1. Sampling
2. Selection of the quantity of the product to be used
3. Method of sterility testing
i. Membrane filtration method
ii. Direct inoculation method
METHOD OF STERILITY TESTING
A. Membrane filtration method : Membrane filtration appropriate for
(advantage)
• Filterable aqueous preparation
• Alcoholic preparations
• Oily preparations
 All steps of this procedure are performed aseptically in a Laminar flow.
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33. B. Direct inoculation method :
 Direct inoculation of the culture medium suitable quantity of the preparation to be
examined is transferred directly into the appropriate culture medium & incubate
for not less than 14 days .
Clarity test:
 Performed to ensure that the parenteral product are free from foreign particles .
• Method :
1. Visual Method
2. Coulter Counter method
3. Filtration method
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Particle Size (µm)
equal to or large than
Max. no. of particles
per ml
10 50
25 5
50 nil

34. Leakage testing :
The sealed ampoules are subjected to small cracks which occur due to rapid
temperature change or due to mechanical shocks.
filled & sealed ampoule
Dipped in 1% Methylene blue solution under negative pressure in vacuum
chamber
Vacuum released colored solution enter into the ampoule
Defective sealing
 Note : vials & bottles are not suitable for this test because the sealing
materials used is not rigid.
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35. Pyrogen testing
 Pyrogen = “Pyro” (Greek = Fire)+ “gen”(Greek = beginning)
 Fever producing metabolic by- product of microbial growth and death.
 Bacterial pyrogen are called “Endotoxins” . Gram negative bacteria produce
more potent endotoxins than gram+bacteria and fungi.
 Endotoxins are heat stable lipopolysaccharides (LPS) present in bacterial
cell walls.
 Stable at least 1750 C; steam sterilization ineffective
 Source: Water (main), raw materials, equipment, process environment,
people and if using gram negative bacteria.
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36. Biological properties of endotoxin :
 Pyrogen elevated the circulatory levels of inflammatory cytokines which may be
followed by fever, blood coagulation , hypotension
 Low doses of pyrogen: asymptomatic inflammation reaction
 Moderate doses: Fever $ changes in plasma composition
 High doses : cardiovascular dysfunction, Vasodilation, vasoconstriction
• Elimination of pyrogens:
 Dry heat sterilization : for glass wares, metal equipments, powder ,waxes, oils,
heat stable drugs.
 650C temp- 1 min
 350C temp- 30 min
 180 C temp -240 min
 Ultra filtration
 Distillation
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37. Rabbit Pyrogen Test:
 Rabbit are used to perform this test because their body temp. increase when
pyrogen are introduced into their bodies by parenteral route
 3 healthy adult rabbits of either sex, each weighing 1.5kg are selected
 having a temp higher than 37.8 ̊C
 Showing temp variation >0.2 ̊C between two successive reading in the
determination of initial temp
 Same test is performed within 7 days of actual test
 Animal showing temp increase over 0.6 ̊C should be removed from pyrogen
testing
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38. Method:
 Dissolve the subs being examined in, or dilute it with a pyrogen free saline
solution
 warm the liquid being examined to approx. 38.5 ̊C temp before injection
 The volume of injection is 0.5 ml & 10ml/ kg of body weight
 Withhold water during test
 Clinical thermometer is inserted into the rectum of rabbit to record body temp
 2 normal reading of rectal temp are should be taken prior to the test injection
at an interval of half an hr& its mean is calculated- initial temp
 The solution under test is injected through an ear vein
 Record the temp of each rabbit in an interval of 30 min for 3 hrs
 The difference between initial temp & maximum temp is recorded – taken as
response
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39. NOTE: If above test not passes perform the test again
If above test not passes, the sample is said to be pyrogenic or go through
the source of contamination of pyrogen.
No . Of
Rabbits
Individual
temp. Rise C
Temp. Rise
in group
(0C)
Test
3 rabbits o.6 1.4 passes
If above not
passes 3+5=8
rabbits
o.6 3.7 passes
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40. Bacterial endotoxin (LAL) test:
 To detect or quantify endotoxins of gram – ve bacterial origin.
 Reagent: amoebocyte lysate enzyme from horseshoe crab (Limulus
polyphemus or Tachypleus tridentatus).
• The name of the test is also Limulus amebocyte lysate (LAL)
Mechanism of LAL Test :
 The test is based on the gelling properties of enzyme extracted from the
horseshoe crab of Limulus polyphemus.
 Enzyme when come in contact with bacterial endotoxin.
Gelling
Degree of Gelling related to amount of endotoxin present.
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41. Test:
1. Equal volume of LAL reagent and test solution (usually 0.1ml of each )
are mixed in a test –tube
2. Incubation at 37 ̊C, 1 hrs depyrogenated
3. Remove the tube –invert at (180) observe the result
4. Pass or otherwise fail test
Three different techniques:
1. The gel-clot technique- gel formation
2. The turbidimetric technique- the development of Turbidity after cleavage
of an endogenous substrate
3. The chromogenic technique- the development of color after cleavage of a
synthetic peptide- chromogen complex
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42. Advantages of LAL test
 Fast-60 min vs. 180min
 Greater Sensitivity, Less variability
 Less expensive
 Particularly useful for:
• Blood products
• Cytotoxic agents
• Water for injection
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43. REFERENCES
 Lachman Leon; Liberman Herbert A.; Kanig Joseph L.; “The Theory and Practice
of Industrial Pharmacy”; Third Edition; Indian Edition; Varghese Publishing
House; Dadar Bombay 400114, 1987; Page No. 639.
 Fox Shelley Chambers; “Pharmaceutics”; Remington Education; Published by
Pharmaceutical Press;1 Lambeth High Street, London SE1 7JN, UK; Royal
Pharmaceutical Society of Great Britain, 2014; Page No. 189-216.
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