PARENTERAL PRODUCTS

Parenteral products;
 Definition of parenteral products,
 Types of parenteral products,
 Preformulation and formulation of parenterals,
 Release of medicament from parenterals,
 Sterilization and validation,
 Containers and closures,
 Intravenous admixtures,
 Total parenteral nutrition,
 Pyrogens, definition, sources, nature and properties of pyrogens,
removal of pyrogens from water and pharmaceutical formulations,
 Preparation of water for injection.

INTRODUCTION
Parenteral (Gk, para enteron, beside the intestine) dosage forms differ from all other drug dosage
forms, because they are injected directly into body tissue through the primary protective systems
of the human body, the skin, and mucous membranes.
They are sterile solutions or suspensions of drugs in aqueous or oily vehicles meant for
introduction into the body by means of an injection under or through one or more layers of the
skin or mucus membranes.
They must be exceptionally pure and free from physical, chemical, and biological contaminants.
These requirements place a heavy responsibility on the pharmaceutical industry to practice
current good manufacturing practices (cGMPs) in the manufacture of parenteral dosage forms
and on pharmacists and other health care professionals to practice good aseptic practices (GAPs)
in dispensing parenteral dosage forms for administration to patients.
Parenterally-administered drugs are relatively unstable and generally highly potent drugs that
require strict control of administration to the patient.

Characteristics of parenteral dosage forms

• Parenteral products are unique from any other type of pharmaceutical dosage form
for the following reasons:
• All products must be sterile.
• All products must be free from pyrogenic (endotoxin) contamination.
• Injectable solutions must be free from visible particulate matter. This includes reconstituted
sterile powders.
• Products should be isotonic, although strictness of isotonicity depends on the route of
administration.

• All products must be stable, not only chemically and physically like all other dosage forms, but
also ‘stable’ microbiologically (i.e., sterility, freedom from pyrogenic and visible particulate
contamination must be maintained throughout the shelf life of the product)
• Products must be compatible, if applicable, with IV diluents, delivery systems, and other drug
products co-administered.

Parenteral dosage forms are preferred when there is:

i. Low oral bioavailability and/or high variability in oral drug absorption.
ii. Instability of the drug in the GI tract. For example, most protein drugs are highly
unstable.
iii. Rapid onset of drug action is desired.
iv. Ability to immediately stop drug administration is important. For example, most
emergency room medications and anesthetics.
v. ·High degree of flexibility in dosage adjustment with or without real-time patient
physiological response is needed. For example, emergency medications such as
analgesics, anticancer drugs, and fertility medications.

Advantages
 Quick onset of action
 Maximum bioavailability
 Suitable for the drugs which are not administered by oral route
 Useful for unconscious/vomiting/ Non-cooperative patients.
 Duration of action can be prolonged by modifying formulation.
 Suitable for nutritive like glucose & electrolyte.
 Suitable for the drugs which are inactivated in GIT or HCl (GI fluid)
Disadvantages of parenterals
 Only trained person is required
 If given by wrong route, difficult to control adverse effect
 Difficult to save patient if overdose
 Sensitivity or allergic reaction at the site of injection
 Requires strict control of sterility & non pyrogenicity than other formulation.
 Once injected cannot be controlled (retreat)
 Injections may cause pain at the site of administration

Parenteral routes of administration

Most injections are designed for administration
 into a vein (intravenous, IV),
 into a muscle (intramuscular, IM),
 into the skin (intradermal, ID),
 or under the skin (subcutaneous, SC).
Nevertheless, drugs may be administered into almost any organs or area in the body, including
the joints (intraarticular), joint fluid area (intra synovial), spinal column (intra spinal), spinal
fluid (intrathecal), arteries (intra arterial), and in the heart (intra cardiac).
Factors affecting selection of route
Selection of a parenteral route of administration for a new therapeutic moi0000ety depends on
several considerations, such as
·1. Desired rate of onset of action: IV route provides the most rapid onset of action, whereas the
SC, IM, and IP routes have slower rate of drug absorption into the systemic circulation. SC route
is often preferred for SR dosage forms when slow drug absorption over a prolonged period is
desired.
2. Location of drug action: Intraarterial injections are preferred for localized drug action in an
organ, whereas IP route is preferred if drug action is desired in the lymphatic system.
3. Tissue irritability: Injection of an irritant drug is likely to be more painful by the IM than the
SC route due to higher blood flow and sensory innervations in the muscles.
4. Injection volume: The volume of drug injected is lower for SC than for IM or IV routes. In
certain cases, formulation of low volume injections is not feasible, especially for protein drugs
with high doses.

Types of parenteral dosage forms

A. Small-volume parenterals versus large-volume parenterals
Injectable parenteral drug products are available as single or multiuse containers in different
container–closure systems and volumes. Small-volume parenterals (SVPs) are available in
volumes of less than 1 ml, and up to 50 ml. Large-volume parenterals (LVPs) are usually
packaged in volumes up to 1000 mL.
Small volume parenteral (SVP): volume < 100 mL

➢ Large-volume parenteral (LVP): volume ≥ 100 mL

SVPs include both unit-dose and single-dose and multidose containers. Unit dose containers
are usually hermetically sealed ampoules that are intended to be discarded after a single
injection. Multidose containers, on the other hand, are usually rubber-stoppered and sealed glass
vials that are intended for multiple injections. The drug for each injection is withdrawn by
inserting the needle through the rubber stopper, which self-seals after the needle is withdrawn.
SVPs for IV injection may not be isotonic because the large volume of blood rapidly dilutes
them. However, hypertonic solutions tend to be tissue irritants. The pH of SVPs can also vary
from the physiological pH because the blood buffering system rapidly readjusts the pH after a
small volume injection. SVPs for single-dose administration may be free of antimicrobial
preservatives, but multidose vials usually have the preservatives to ensure sterility over multiple
uses over a certain period of time.
They are packaged in single dose glass or plastic containers. Example:
i. Dextrose injection – it contains 2.5%; 20 or 50%; dextrose. They are used as a fluid
and nutrients replenishers.
ii. Dextrose and NaCI injection – it contains dextrose from 2.5 – 25% and 0.11 – 0.9%
NaCI. It is used as a nutrient and electrolyte replenishers.
 B. Injections versus infusions
Injection and infusion are the predominant methods of parenteral administration. Injection via
different routes of administration usually utilizes a SVP. An infusion involves the IV
administration of a LVP over a prolonged period of time. Infusions are commonly used for fluid
replacement, administration of drugs with a short plasma half-life, and/or dilution of a drug
immediately before administration.

C. Types of formulations
Parenteral products can be formulated as solutions, suspensions, emulsions, or lyophilized
products (solid) for reconstitution immediately before use.
1. Solutions
Most injectable products are solutions. Although usually aqueous, they may also contain
cosolvent(s), such as glycols (e.g., polyethylene glycol [PEG] or propylene glycol), alcohols
(e.g., ethanol), or other nonaqueous solvents (e.g., glycerin).
These solutions are usually filtered through a 0.22 μm membrane to achieve sterility. Solutions
that do not contain any antimicrobial agents should be terminally sterilized.
Autoclaving is the preferred method for terminal sterilization whenever drug solutions can
withstand heat. An antimicrobial agent is often added to SVPs that cannot be terminally
sterilized.
2. Suspensions
Parenteral suspensions should be easily resuspended and passed through an 18 to 21-guage
needle throughout their shelf lives. To achieve these properties, it is necessary to select and
carefully maintain particle size distribution, zeta potential, rheological properties, and
wettability.
Injectable suspensions often consist of the active ingredient suspended in an aqueous vehicle
containing an antimicrobial preservative, a surfactant, a suspending agent, a buffer, and/or a salt.
Due to the inherent long-term physical instability of suspensions, parenteral suspension dosage
forms are formulated as dry powders for reconstitution immediately before administration. The
sterile dry powder could be produced by freeze-drying, sterile crystallization, or by spray-drying.
Parenteral suspensions are prepared by mixing dry powders in sterile vehicles immediately
before administration. Examples of parenteral suspensions include penicillin G procaine
injectable suspension USP and testosterone injectable suspension USP.
Lyophilization or freeze-drying is used to prepare powder cakes for reconstitution immediately
before administration. It has inherent advantages over other methods of preparation of dry
powders, such as
·           Water is removed at low temperatures, avoiding damage to heat-sensitive drugs.
·           Freeze-dried product usually has high-specific surface area, facilitating rapid
reconstitution.
·           Freeze-dried dosage form allows drugs to be filled into vials as a solution, which can
then be freeze dried into the final, marketed dosage form. Thus, it does not require powder
filling, which is technologically more challenging than filling solutions.
Despite the advantages of freeze-drying, cautions must be taken for lyophilizing proteins,
liposomal systems, and vaccines, because they tend to get damaged by freezing, freeze-drying, or
both. These damages can often be minimized by using protective agents, such as polyols,
polysaccharides, disaccharides and monosaccharide.
The essence of the freeze-drying process depends on maintaining a critical balance between
the conversion of ice into water vapor by sublimation under vacuum and the removal of
that vapor from the frozen mass. To maintain sublimation, heat energy is applied to the
product to compensate for sublimation cooling.
There are three stages in the process: freezing, sublimation drying, and desorption drying.
The freezing phase is the most critical in the entire freeze-drying process. Annealing is a
processing step in lyophilization in which samples are kept at a determined subfreezing
temperature above the Tg', during a period of time . This process influences the size
distribution of ice crystals, leading to their growth.

3. Emulsions
Because emulsions can cause pyrogenic reactions and hemolysis, and require autoclave
sterilization in addition to their inherent physical instability, their use as IV dosage forms has
been limited. Total parenteral nutrition is often administered as an IV emulsion to enable
coadministration of both water-soluble and water-insoluble nutrients. IV fat emulsion usually
contains 10% oil. Fat emulsions yield triglycerides that provide essential fatty acids and calories
during total parenteral nutrition of patients who are unable to absorb nutrients through the GI
tract. IV lipid emulsions are usually administered in combination with dextrose and amino acids
in the aqueous phase.

PREFORMULATION
The aim of preformulation study is to develop effective, safe and stable formulation.
For that we need to assess physical, chemical and biological properties of the medicament.
 As it forms basis for the selection of suitable vehicle (aqueous or non-aqueous), excipients
(antioxidants, antimicrobials, preservatives, buffers, solubilizing agents and tonicity adjusters)
and containers and closures.
The important properties with reference to preformulation of parenterals includes solubility, pKa,
pH, solid state characteristics, chemical modification of drug and polymorphism etc.
a) Solubility /pKa/pH
Solubility (when a drug substance is dissolved in a unit volume of liquid (solvent) to form a
saturated solution under specific condition of temperature and pressure) is one of the important
parameter in preformulation of parenterals.
Most of the parenterals dosage form exist in solution form so it should maintain its properties,
consistency and stability in liquid form.
Solubility is a function of hygroscopicity, chemical nature and pKa of the salt. In general
term we state that if the pKa is at least two units lower than the pH of the medium, complete
dissolution can be achieved; the opposite holds true for basic compounds.
Other considerations like dilution prior to administration, and the rate of administration
(dilution factor) should also be simulated using in vitro techniques. Thus, by determining and
manipulating the solubility, pH and pKa one can develop a parenteral dosage form with its
stability and safety.
When a drug substance is dissolved in a liquid it gets dissociated in ionized and unionized form.
The unionized substances are lipid soluble thus dissolve in lipid material of the membrane and
transported by passive diffusion and their absorption is rapid. Whereas, the ionized substances
are a lipid insoluble therefore their permeation is slow and absorption is poor. The percentage of
ionization can be calculated as
For Acidic compounds: = PH= pKa+ log ionized drug/un-ionized drug.
For Basic compounds: PH = pKa + log un-ionized drug/ionized drug.
For acidic drugs pKa ranges from 3‐7.5. for basic drugs pKa ranges from7‐11. Beside it pH
plays a vital role in stability of solution and suspension type of parenteral products. pH value for
parenteral products:
Ideal pH 7 pH of blood
Above pH 9 Tissue necrosis
Below pH 3 Extreme pain at site of injection
The methods used to increase solubility are change in pH, cosolvency, dielectric constant,
solubilization by surfactant, complexation, hydrotropy, and chemical modification of drug, etc.
Care must be taken when using co solvents although they increases the solubility but as they are
conjugate acid base systems there may be a shift of pKa of the buffer or salt results and it may
affect the integrity of the formulation like there may be precipitation or degradation of salt .
 b) Solid state characteristics
Any active drug substance exists in two types of physical forms crystalline form or amorphous
form. Crystalline form is when it exists in crystal form and amorphous form is when it exists in
powder form. Both the physical form has its own significance with respect to absorption and
bioavailability
A crystalline solid is that in which the constituent particles are orderly arranged in a three-
dimensional pattern called the crystal lattice with uniform intermolecular forces, and the particles
intersect at angles characteristic of the crystal.
Amorphous is the shapeless, disordered, and irregular arrangement of the constituent particles of
a solid. Their inter-molecular forces are not the same, nor are the distances between the particles.
When cleaved, amorphous solids yield fragments or curved surfaces because of irregular
geometric shapes

Crystalline characteristics are analyzed by microscopy and X-ray powder diffractometry

(XRPD). Crystal habit or the external shape (cubic, tetragonal, hexagonal etc.) of a crystal may
be determined by microscopy.
So the combination of crystalline and amorphous form of a same drug may provide a dosage
form with combined benefits of rapid onset and prolonged duration of action (e,g. lente insulin, a
physical mixture of 70% crystalline ultralente and 30% amorphous semilente insulin).
Moreover, the crystalline form of the drug can withstand the high thermal stress (dry heat for
several hours) as compared to amorphous form (e.g. Pencillin G).
c) Chemical Modification of the Drug.
Chemical modification like preparation of an ester, salt, or any other form of the parent drug
sometimes increases stability, alter drug solubility, enhance depot action, avoid formulation
difficulties and possibly, to decrease pain on injection.
The preparation of salts of organic compound is one of the most important tools available to the
formulator. Compounds for both IV and IM solutions may require high solubility in order for the
drug to be incorporated into acceptable volumes for bolus administration.
Sodium and potassium salts of weak acids and hydrochloride and sulfate salts of weak bases are
widely used in parenterals requiring highly soluble compounds, based on their overall safety and
history of clinical acceptance. If the solubility of a drug is to be reduced to enhance stability or to
prepare a suspension, the formulator may prepare water insoluble salts. Ex: Procaine Penicillin
G: decrease solubility (7mg/ml) of which, when compared with the very soluble penicillin G
potassium, is utilized to prepare stable parenteral suspensions.
d) Polymorphism.
Polymorphism occurs when a chemical compound crystallizes with different internal structures.
ICH Q6A defines polymorphism as “some new drug substances exist in different crystalline
forms which differ in their physical properties.
Several crystal forms of a given chemical exhibits different physical properties. The conversion
of one polymorph to another may cause a significant change in the physical properties of the
drug and it has important formulation, biopharmaceutical, and chemical process implications.
The aspect of polymorphism that is of particular concern to the parenteral formulator is the
physical stability and bioavailability of the product.
For example, cortisone acetate exists in five polymorphic forms but first polymorphic form is the
most stable one. Indomethacin exist on three polymorphic form but two polymorphic form I and
II have higher therapeutic efficacy.
Substances that exists different polymorphs must be evaluated for stability in liquid form as well
as its thermal stability, as parenteral dosage form must be sterile.

 Apart from this we need to analyze the following factors before formulation of a parenteral
dosage form.
 Melting point
 Thermal profile
 Particle size and shape
 Hygroscopicity potential
 Ionization constant
 Light stability
 Optical activity
 pH solubility profile
 pH stability profile
 Polymorphism potential

Formulation of parenteral products

Accuracy, cleanliness and overall quality of the product should be strictly ensured.
Only a minimum number of absolutely necessary additives in the smallest possible quantities
should be added
Additives used in the formulation of parenteral products:
Vehicles
A suitable vehicle is chosen for dissolving or suspending the medicament. The most suitable
vehicle is water because aqueous preparations are well tolerated by the body and are the safest
and easiest to administer.
The water should be physically and chemically pure and pyrogen free. When water free from
dissolved gases is required, it should be freshly boiled, cooled and stored in a well-closed
container to avoid absorption of O2 and CO2
Oily vehicles are used when water is contraindicated, like;
i. To increase the stability of the preparation
ii. To prolong the duration of action of drug
The commonly used are fixed oils of vegetable origin like; cotton seed oils, sesame oil, peanut
oil, olive oil.
These oils should be free from rancid odour and taste.
NOTE: Mineral oils are never used, because they cannot be absorbed from the tissue after
injection.
Other additives
1. Solubilizing agents: -The solubility of insoluble or poorly soluble drugs in water can be
increased by:
- Co-solvents
- Complex formation
- By adding surfactants like tweens and polysorbates which act by Micellar
Solubilization
2. Stabilizers – Oxidation and hydrolysis take place more rapidly in drugs when they are in
solution.
To prevent oxidation, a suitable antioxidant is added, or the product is sealed in presence of
nitrogen or carbon dioxide to replace Oxygen in the product and minimize oxidation.
Hydrolysis is prevented by using a non-aqueous vehicle or by adjusting the PH of the
preparation.
3. Buffers: if the degradation of the drug is due to a change in PH, buffer systems are added
to maintain the pH at the desired level. Acetates, citrates, phosphates are the principle
buffer system commonly employed.
 4. Antibacterial agents: - Bacteriostatic or fungistatic agents must be present in multidose
containers. They prevent the multiplication of organisms that may be accidentally
introduced when withdrawing a dose from a multidose container.
They include:
Benzalkonium chloride 0.01%
Phenol or cresol 0.5%
Chlorocresol 0.2%
Phenyl mercuric nitrate 0.002%
Chlorbutanol 0.5%
The agent should be compatible with all other components of the solution and should not be
removed from the solution by rubber closures of the package.
Bacteriostatic agents should not be used in single dose containers because the contents remain
sterile until the solution is opened.
5. Isotonicity adjusters: - Parenteral preparation must be isotonic with blood serum and
other fluids to reduce the irritation and pain of injection in areas with nerve endings.
Isotonicity is achieved by adding sodium chloride in suitable quantities.
6. Wetting, suspending and emulsifying agents
A wetting agent is used to reduce the interfacial energy between the solid particle and the liquid
to prevent the formation of lumps.
They also act as antifoaming agents to subside the foam produced during shaking of the
preparation.
The wetting agents commonly used are Tweens 80 and sorbitan trioleate.
7. Suspending agents used:
- Methyl cellulose
- Carboxymethylcellulose
- Acacia
- Gelatin
8. Emulsifying agents are used in the preparations that are in form of emulsions Lecithin is
commonly used. Gelatin is added to aqueous vehicles to prolong the effect of the drug.
General procedure for preparation of injection
All parenteral products must be free from foreign particles and organisms.
Care must be taken regarding cleanliness and sterilization of the area, Atmosphere,
Persons involved and the materials used in the preparation of injections.
The area and the atmosphere of the room must be free from dust, fibers and microorganisms.
This is achieved with a laminar airflow or with disinfectants.
- All equipment must be thoroughly cleaned and sterilized.
- The workers should be highly trained and skilled
- They should wear sterilized special clothing including hoods and gloves.
Parenteral products should be prepared from substances of high purity that have been accurately
weighed and dissolved in oxygen free distilled water or any other suitable solvent.
The solution formed is passed through different grades of filters to remove foreign particles. The
filters are made from sintered glass, asbestos or porcelain.
Bacteria-proof filters are used to remove bacteria from solutions. The solution is packaged in
suitable containers like ampoules, vials or bottles that have been thoroughly cleaned, dried and
sterilized.
-On small scale, filling is carried out with the help of a hypodermic syringe attached with a long
needle or burette
-The sealing of the ampoule is done by fusion of glass in hot flames of a blast burner or a blow
torch specially designed for this purpose
Currently filling and sealing is done on very sophisticated automatic machines.

Sterilization and validation

Whenever possible products intended to be sterile should be terminally sterilized by heat in their
final container. Where it is not possible to carry out terminal sterilization by heating due to the
instability of a formulation or incompatibility of a pack type (necessary to the administration of
the
product, e.g. plastic eye-dropper bottles), a decision should be taken to use an alternative method
of terminal sterilization following filtration and/or aseptic processing.
Sterilization can be achieved by the use of moist or dry heat, by irradiation with ionizing
radiation (noting that ultraviolet irradiation is not normally an acceptable method of
sterilization), by ethylene oxide (or other suitable gaseous sterilizing agents), or by filtration
with subsequent aseptic filling of sterile final containers.
The microbial contamination of starting materials should be minimal and their bioburden should
be monitored before sterilization.
For dry heat sterilization, hot air ovens are used
For moist heat sterilization, autoclaves are used.
Oily and non-aqueous preparations are sterilized by dry heat at a temperature of 1600 C for 2hrs
or 1700 C for 1 hr.
Thermostable aqueous solutions are sterilized by steam under pressure in an autoclave at a
temperature of 1210c for 20 min.
Aqueous solutions of thermolabile drugs cannot be sterilized by autoclaving. They are passed
through bacteria – proof filters to remove microbes
The sterilized containers are allowed to cool and are inspected for clarity. Containers that pass
the clarity test are properly labeled and packaged into final containers.
NB: Terminal Sterilization: Final sterilization of the drug product using steam heat and/or dry
heat or radiation sterilization.

CONTAINERS AND CLOSURES

Injectable formulations are packaged into containers made of glass or plastic. Container systems
include ampoules, vials, syringes, cartridges, bottles, and bags.
Ampoules are all glass, whereas bags are all plastic. The other containers can be composed of
glass or plastic and must include rubber materials, such as rubber stoppers for vials and bottles
and rubber plungers and rubber seals for syringes and cartridges.
• Irrigation solutions are packaged in glass bottles with aluminum screw caps.
• Further, the integrity of the container/closure system depends on several characteristics,
including container opening finish, closure modulus, durometer and compression set, and
aluminum seal application force.
Small volume parenterals (SVPs)
• Ampoules
• Glass vials sealed with rubber stoppers
• Plastic ampoules (blow-fill-seal)
• Pre-filled syringes – reducing the degree of manipulation required
– facilitating administration in an emergency situation
• Needle-free injection
Large volume parenterals (LVPs)
• Glass bottles sealed with rubber stoppers
• Plastic bags

Any container for parenteral product should maintain the integrity of the product as a sterile,
pyrogen-free, high purity preparation till it is used. It should also be attractive, allow the
withdrawal of the contents and be strong enough to withstand processing and shipping; and
finally it should not interact with the product.
Glass seems to be the material of choice for containers for parenteral products. Glass containers
may either be sealed or closed with rubber stoppers. Containers of Type-I glass are best for
aqueous preparations.
Siliconization i.e. the application of a thin film of silicone to coat the inside surface of the vials
and ampoules, has been employed to prevent interaction of the product with the glass surface.
The process also minimizes adsorption of active ingredients from homogeneous solutions,
prevents adsorption of solids from suspensions and prevents aggregation at the glass surface in
colloidal preparations.
Plastics used in the packaging of parenteral products are based on polyethylene or
polypropylene. Plastic containers are much less used as compared to glass but the former are
becoming increasingly popular for intravenous fluids.
Only polypropylene containers can withstand sterilization by autoclaving. Many plastics
contain additives like plasticizers, antioxidants, antistatic agents and lubricants. These additives
may leach out from the plastic into the product. Most plastics selectively permit passage of
chemical molecules and are permeable to gases. Plastics are extensively used for containers of
administration sets particularly disposable type.
As compared to glass, plastics are light weight, less fragile and easy to handle but most of them
are not as clear as glass.
Rubber is the material of choice for closures for multi-dose vials, intravenous fluids bottles,
plugs for disposable syringes and bulbs for ophthalmic pipettes. Rubber closures permit the
introduction of a needle from a hypodermic syringe into a multi-dose vial and provide for
resealing of the vial after the needle is withdrawn.
Rubber closure is held in place by an aluminium band. Such closures are composed of several
ingredients, basic structural unit being a linear unsaturated hydrocarbon, isoprene. All or part of
the natural polymer is sometimes replaced by a variety of synthetic rubber polymers. In addition,
a vulcanizing agent usually sulphur; an accelerator e.g., 2-mercaptobenzothiazole; an activator,
usually zinc oxide; fillers such as carbon black or limestone; antioxidants and lubricants etc.,
may also be present in rubber closures. These substances may be leached into the product or may
cause chemical interaction.
Lacquer or plastic coating applied to the surface of the rubber closures in contact with the
product may partially reduce leaching and also permeation. Another most commonly
encountered problem with rubber closures is that of coring i.e., the generation of rubber particles
cut from the closures when needles are inserted; the particles are known as cores. Selection of
the proper gauge needle and its proper use may minimize the problem of coring.

INTRAVENOUS ADMIXTURE
Parenteral Admixture means a sterile preparation that is the combination of one or more
sterile products with an appropriate admixture vehicle prepared aseptically.
Aseptic technique requires sterile equipment and a sterile environment, but it also includes
manipulations that ensure sterility. Areas known as critical sites should never be touched in order
to prevent contamination. These include tips of syringes, hubs of needles, ports of bags, tops of
vials, and ends of filters or dispensing pins.
Each item used during aseptic manipulation, including syringes, needles, and medication vials,
are sprayed with 70% isopropyl alcohol (IPA) and wiped down at the edge of the LAFW.
The occurrence of admixture incompatibilities can be prevented by adhering to proper
medication administration techniques such as flushing the line using compatible fluid, through
multi-lumen catheter, through multiple IV access, using in- line infusion filters, spacing of
medication, or color coding system.
Labelling of Admixtures
A. Date of compounding;
B. Beyond-use date( expiry date);
C. Storage requirements if other than room temperature;
d. Infusion or administration rate;
E. Administration times, administration frequency, or both; and
F. Other accessory cautionary information which in the professional judgment of the pharmacist
is necessary or desirable for proper use by and safety of the patient.

TOTAL PARENTERAL NUTRITION (TPN)

Also known as parenteral nutrition (PN) is a form of nutritional support given completely via the
bloodstream, intravenously with an IV pump. TPN administers proteins, carbohydrates, fats,
vitamins, and minerals. It aims to prevent and restore nutritional deficits, allowing bowel rest
while supplying adequate caloric intake and essential nutrients, and removing antigenic mucosal
stimuli.
TPN is made up of two components: amino acid/dextrose solution and a lipid emulsion
solution. It is ordered by a physician, in consultation with a dietitian, depending on the patient’s
metabolic needs, clinical history, and blood work.
The amino acid/dextrose solution is usually in a large volume bag (1,000 to 2,000 ml), and can
be standard or custom-made. It is often yellow in colour due to the multivitamins it contains. The
ingredients listed on the bag must be confirmed by the health care provider hanging the IV bag. 
The solution may also include medication, such as insulin and heparin. The amino acid/dextrose
solution is reviewed and adjusted each day based on the patient’s blood work. Lipid emulsions
are prepared in 100 to 250 ml bags or glass bottles and contain the essential fatty acids that are
milky in appearance.
At times, the lipid emulsion may be added to the amino acid/dextrose solution. It is then called 3
in 1 or total nutrition admixture . TPN is prepared by a pharmacy, where the calories are
calculated using a formula, and is usually mixed for a 24-hour continuous infusion to prevent
vascular trauma and metabolic instability . TPN orders should be reviewed each day, so that
changes in electrolytes or the acid-base balance can be addressed appropriately without wasting
costly TPN solutions.
To prevent severe electrolyte and other metabolic abnormalities, the infusion rate of TPN is
increased gradually, starting at a rate of no more than 50% of the energy requirements.

Prepared solutions
Prepared solutions generally consist of water and electrolytes; glucose, amino acids, and lipids;
essential vitamins, minerals and trace elements are added or given separately.
Previously lipid emulsions were given separately but it is becoming more common for a "three-
in-one" solution of glucose, proteins, and lipids to be administered.
Added components
Individual nutrient components may be added to more precisely adjust the body contents of it.
That individual nutrient may, if possible, be infused individually, or it may be injected into a bag
of nutrient solution or intravenous fluids (volume expander solution) that is given to the patient.
Administration of individual components may be more hazardous than administration of pre-
mixed solutions such as those used in total parenteral nutrition, because the latter are generally
already balanced in regard to e.g. osmolarity and ability to infuse peripherally.
Incorrect IV administration of concentrated potassium can be lethal, but this is not a danger if the
potassium is mixed in TPN solution and diluted.
Vitamins may be added to a bulk premixed nutrient immediately before administration, since the
additional vitamins can promote spoilage of stored product. Vitamins can be added in two doses,
one fat-soluble, the other water-soluble. There are also single-dose preparations with both fat-
and water-soluble vitamins such as Cernevit.
Minerals and trace elements for parenteral nutrition are available in prepared mixtures, such
as Addaven.
These additional components in parenteral nutritions, however were subject to stability checks,
since they greatly affect the stability of lipid emulsions that serve as the base for these
formulations. Studies have shown differences in physical and chemical stabilities of these total
parenteral nutrition solutions which greatly influences pharmaceutical manufacturing of these
admixtures.
Emulsifier
Only a limited number of emulsifiers are commonly regarded as safe to use for parenteral
administration, of which the most important is lecithin. Lecithin can be biodegraded and
metabolized, since it is an integral part of biological membranes, making it virtually non-toxic.
Other emulsifiers can only be excreted via the kidneys creating a toxic load. The emulsifier of
choice for most fat emulsions used for parenteral nutrition is a highly purified egg lecithin, due
to its low toxicity and complete integration with cell membranes.
Use of egg-derived emulsifiers is not recommended for people with an egg allergy due to the risk
of reaction. In situations where there is no suitable emulsifying agent for a person at risk of
developing essential fatty acid deficiency, cooking oils may be spread upon large portions of
available skin for supplementation by transdermal absorption.
Another type of fat emulsion Omegaven is being used experimentally within the US primarily in
the pediatric population. It is made of fish oil instead of the soybean oil based formulas more
widely in use. Research has shown use of Omegaven may reverse and prevent liver disease and
cholestasis

PYROGEN
A Pyrogen is a substance i.e. products of the growth of micro organisms or may be parts of dead
cells
Or metabolic products which cause febrile reactions like fever, chills, back pain etc
The Pyrogen test is designed to limit the risk of febrile reaction following parentral
administration of drugs. It includes both In vitro and In vivo tests.
There are two types of natural pyrogens: (1) endogenous pyrogens that is the host's pyrogen
cytokines and (2) exogenous pyrogens that are microbial substance (e.g. lipopolysaccharides
in the cell wall of certain bacteria. Exogenous pyrogens (e.g. bacteria, viruses, toxins) initiate
fever, usually within 2 h of exposure, by interacting with macrophages or monocytes, leading to
cytokine induction. Other mechanisms to initiate fever include: Some endotoxins, produced by
bacteria, act directly on the hypothalamus to alter the set point.
Endogenous pyrogens are fever-inducing agents that arise in the body of a host when exogenous
pyrogens come in contact with certain host cell molecules such as monocytes or macrophages.
Typical examples of endogenous pyrogens include interleukins, tumor necrosis factor (TNF) and
platelet activating factor. Cytokines and prostaglandins are typical examples of endogenous
pyrogens generated by the host body.

What are some sources of pyrogens?

There can be several sources of pyrogens in parenteral and medical device products. Usual
sources are: the water used as the solvent or in the processing; packaging components; the
chemicals, raw materials or equipment used in the preparation of the product.
Depyrogenation of fluids (including organic solvents holding the lipids) is possible by
ultrafiltration through filters with cut-offs of 10 kDa.
Therefore, the manufacturer should check the quality of the raw materials, and design the
liposome formulation process in such a way that the generation of pyrogens by micro-organism
growth or contact with contaminated equipment during the production process is avoided.
Pyrogens can be destroyed by heating at high temperatures. A typical procedure for
depyrogenation of glassware and equipment is maintaining a dry-heat temperature of 250°C for
45 minutes. Exposure of 650°C for 1 minute or 180°C for 4 hours, likewise, will destroy
pyrogens.
Anion-exchange resins and positively charged membrane filters remove pyrogens from water.
Although reverse osmosis (RO) membranes will eliminate them, the most reliable method for
their elimination from water is distillation. Other in-process methods for their destruction or
elimination include selective extraction procedures and careful heating with dilute alkali, dilute
acid, or mild oxidizing agents.
Although ultrafiltration now makes pyrogen separation on a molecular-weight basis possible and
the process of tangential flow is making large-scale processing more practical, use of this
technology is limited, except in biotechnological processing.

TEST FOR PYROGENS:

1. In Vitro Test / LAL Test
2. In Vivo Test / Rabbit Test.
LAL Test: Limulus Amoebocyte Lysate Test. (In Vitro Test)
In Vitro assay used to detect the presence and concentration of bacterial endotoxins in drugs and
biological products. Limulus Amoebocyte Lysate (LAL) is an aqueous extract of blood cells
(amoebocytes) from the horseshoe crab, Limulus polyphemus.
LAL reacts with bacterial endotoxin or lipopolysaccharide (LPS), which is a membrane
component of Gram negative bacteria and forms gel which is then used for the detection and
quantification of bacterial endotoxins.
Sample + LAL (mL) -------------------------------------Gel formation (+)
Incubation at 37 oc for 1 hr

Limitations of LAL Test:

1. Disturbed by endotoxin binding components like lipids, blood components etc.
2. Difficult to correlate with rabbit test.
3. False positive for cellulose and many herbal preparations.

Rabbit Test: SHAM TEST (In Vivo)

Principle: The test involves the measurement of rise in body temperature of rabbits following IV
injection of sterile solution of substance being examined.
Animals used: Select same variety of healthy mature rabbits weighing less than 1.5Kg and should
maintain balanced diet. They should not show any loss of body weight during the preceding
week of test.
Preliminary test:
Select fresh animals / Animals not been used during 2 previous weeks.
Conditioned them for 1 to 3 days.
With hold the food from animal before 2hours of starting the test and access to water may be
allowed. Record the temperature of animals using thermometer.
After 90 min, give IV injection 10mL/Kg (Pyrogen free saline solution). Record the temperature
of animals after IV injection at an interval of 30min and continued for 3 hrs after injection.
Animals show temperature variance of 0.6°C should not be used for main test.
Note: It is carried out in room without disturbances and temperature variance must be ± 3°C.
Main test:
Preparation of Sample: Dissolve test substances in Pyrogen free saline water and warm the liquid
to 38°C before giving injection.
3 groups of selected rabbits (preliminary test passed rabbits)
With held food for 2 hrs before experiment and during the experiment
Record initial temperatures (mean of two measurements at an interval of 30 minutes)
Rabbits showing a temperature variance ≥0.2 °C between two successive readings should not be
used. Use only those rabbits that do not deviate a temperature variance 1°C between two
successive readings. Inject sample into the marginal vein of the ear of 3 rabbits. Not less than
0.5mL/ Kg and not greater than 10mL/Kg body wt.
Record the temperature of animals during a period of 3 hours at intervals of 30 minutes.
Not more than 3 rabbits shows individual raise in temperature of 0.6 °C

PREPARATION OF WATER FOR INJECTION

1. Dechlorination
This refers to the removal of chlorine from water. There are several ways of dechlorination. This
include injection of a reducing agent like sodium metabisulfite and exposure to a high dosage of
UV rays and Filtration through activated carbon media.
Carbon dechlorinates by chemically reacting with the free chlorine in water to form hydrochloric
acid and carbon dioxide or carbon monoxide..
High doses of UV light rays are widely use vgt6d in water purification systems for disinfection.

2. Ion removal
There are basically three types of ion reduction processes; Membrane processes, ion exchange
processes and distillation processes. Membranes are used in water purification system to remove
ions, remove particulate, organic compounds and living organisms. Membranes are different
from one another in terms of pore size, molecular weight and even ion rejection.

Ion removal membranes include reverse osmosis membranes and nanofiltration membranes. The
ion exchange systems provide additional ion reduction process, making the water much lower in
conductivity than required and it also provides a back up for membrane process. Distillation can
also be used to remove ion , however it is very expensive.
3. Bacterial control
Includes both prcedures and equipment. Equipment utilized are ultraviolet (UV) lights, ozone
generation systems for production of ozone, heating systems for thermal treatment, and chemical
injection and recirculation systems.
Bacterial control is usually applied during processing, storage and even distribution.. UV light is
an excellent non- chemical method of disinfecting Water for Injection.
Thermal sanitization involves the use of heat to kill the bacteria. Ozone can also be used to
oxidize bacteria. Chemicals can also be used to kill bacteria.
4. Removal of specific impurities
e.g. iron, managanese, hydrogen sulfide , hardness ions, particulate matter, high conductivity.
Filtration can be used to remove any heavy loads. Cartridge filters are also used to remove
essentially any sized particles.
The last stage is storage . Care and hygiene must be maintained during storage of WFI .

Control Tests of Parenteral products

1) Sterility  tests:- Sterility means complete absence of all viable Micro-organism. It is an
absolute term. The methods which are used to perform  sterility  tests  are
a)  Direct  transfer  method.
B)  membrane  filtration method.
A)  Direct Transfer method:– it is an traditional sterility test method which involves a direct
inoculation of required volume of a sample in two tests tube containing a culture medium that is
FTM, SCDM. This method is simple in theory but difficult in  practice  when  the  demand  for 
repetition  in  opening  container,  sampling Transferring, and mixing increases causes potential
error in operator technique.
B)    Membrane Filtration method: This method basically involves filtration of Sample
through membrane filters of porosity 0.22 micron and Diameter 47mm. The filtration is assisted
under Vacuum, after filtration completion the membrane is cut into 2 halves and one halve is
placed in two test tubes containing FTM, SCDM medium.
*Interpretation: – If no visible evidence of microbial growth in culture medium in test tube then
it is interpreted that the sample representing lot is without intrinsic contamination
2) Pyrogen Test
3)     Leaker Test: – The leaker test is intended to detect incompletely sealed ampoules, so that
they may be discarded. Tip sealed ampoules are more prone to leak than pull sealed. In addition
to that crack my present around seal or at the base of ampoule as a result of improper handling
leakers are usually detected by producing negative pressure within the incompletely sealed
ampoule usually into a vacuum chamber while those ampoules are submerged into a colored dye
solution of 0.5 to 1% methylene blue.
Vials and bottles are not subjected to such leaker test because rubber closure is not rigid however
bottles are often sealed while vacuum is pulled so that bottle remains evacuated during its shelf
life.
The presence of vacuum is detected by striking at the base of bottle sharply with the heel of hand
to produce typical water hammer sound.
Another test is to apply a spark tester probe outside to the bottle moving form liquid layer into
air space a blue spark discharge occur is air space is evacuated.
4) Particulate  matter  testing:–   Particulate  matter  is  primary  concern  in  the parenteral
products given by I.V. Route, all parenteral products should be free from insoluble particle.
Further U.S.P. states that GMP Requires that all containers be visually inspected and that with
visible particle be discarded.
The visual inspection is done by holding the ampoule by its neck against highly illuminated
screens. White screens for the detection of black particle and black screens for the detection of
white particles to detect heavy particles it may be necessary to invert container but care must be
exercised to avoid air bubble.
The instrumental methods are based on principles of light scattering, light absorption, electrical
resistance as in coulter counter. A method which utilizes a video image projection could detect a
moving particle without destruction of product unit.