plete drug availability occurs immediately; no
absorption is necessary. For all other routes, at
least a blood vessel wall, and usually one or more
tissue cell walls, must be permeated before the
drug can enter the circulation. Most often, this
occurs by passive diffusion and is most favorable
when the drug has both lipophilic and hydro-
philic properties, with the former being predom-
inant. With nonvascular injections, absorption
is also affected by such factors as the size and
number of blood vessels supplying the tissue,
the movement (exercise) of the tissue following
injection, the physical and chemical properties
of the drug, and such characteristics of the dos-
age form as whether it is a solution, suspension,
or emulsion, the nature of the vehicle, and its
pH. Once in the circulating blood, the physio-
logic effect of a therapeutic agent is affected by
the extent to which it distributes throughout the
body, by the degree of binding to plasma pro-
teins, and by its rate of elimination by hepatic
metabolism and/or renal excretion.
Intravenous and intraspinal preparations
rely are giver” xfer Omer Y jan a
“solutians The danger eof fine cay
Jaries, particularly in the brainy precludes the
use: ‘of forms other than solutions for intravenous
administration, although emulsions have been
given in which the particle size of the dispersed
phase is carefully controlled. The sensitivity of
nerve tissues generally precludes the use of any-
thing but the purest of solutions for int al
lication Preparations given intramuscularly,
istered as solutions, Suspensions, or emulsions.
Everr solid pellets may be implanted subcutane-
ously or intramuscularly. The vehicles can
range from Water for Injection, to glycols, to
fixed oils. Although care must be exercised to
avoid undue tissue irritation, mild local irritation
is permissible at these injection sites.
‘The nature of a preparation can influence sig-
nificantly the rapidity of onset of a therapeutic
effect from a drug, the duration of the effect, and
the form of the absorption pattern achieved.
Therefore, the development of the formulation
for a parenteral product must be integrated care-
fully with its intended administration in a pa-
i
‘The chemical and physical properties of a
drug must be determined, its interaction with
any desired excipients must be studied, and the
effect of each step of the process on its stability
must be studied and understood.
Solvent systems suitable for sterile products
are limited to those that produce little or no tis-
‘sue irritation; water is the most common. All
components must be of exceptional quality. Pu-
640+ The Theory and Practice of Industrial Pharmacy
ification beyond that obtainable in many com-
mercial products or by normal production proce-
ures is often necessary. Not only may chemical
or physical contaminants cause irritation to body
tissues, but extremely small quantities may
cause degradation of the product as a resuit of
chemical changes, particularly during the heat-
ing period when’ thermal sterilization is em-
ployed. For example, minute traces of copper
greatly accelerate the rate of oxidation of ascor-
bie acid in solution. These traces of copper may
come from the water vehicle, the chemical com-
ponents, or even the container. Rigid specifica
tions must therefore be developed for all ingredi-
ents,
Vehicles
By far the most frequently employed vehicle
for sterile products is water, since it is the vehi-
cle for all natural body fluids. The superior qual-
ity required for such use is described in the
monograph on Water for Injection in the USP.
Requirements may be even more stringent for
some products, however.
One of the most inclusive tests for the quality
of water is the total solids content, a gravimetric
evaluation of the dissociated and undissociated
organic and inorganic substances present in the
water. However, a less time-consuming test, the
electolye measurement of conductivity” ofthe
water, is the one most quently u: Instant
Seon measurements can be obtained by fm.
mersing electrodes in the water and measuring
the specific conductance, a measurement that
depends on the ionic content of the water.
e
conductance may be expressed by the meter)
Seal as Conductivity in mlcrombos, resistance
ii megohms, or ionic content as parts per mil-
kar (ppm) of Sodium chloride "The vaidiy of
This measurement as an indication of the purity
of the water is inferential in that methods of pro-
ducing high-purity water, such as_distillation
and revei , can be expected to remove
*e iundissociat subsancs along with those that
aie dissoclated Substances such
gens, foncrerte id be. bresent in the
senc® of ions and not be disclosed by the test.
"Pherefore:-Tor contaminants other than ions,
additional tests should be performed.
Additional tests for quality of Water for Injec-
“Barnstead Co,, Div. of Sybron Corp., Boston, MA 02132;
Beckman Instruments, Inc., Cedar Grove, NJ 07003;
Corning Glass Works, Coming, NY 14830; Foxboro Ana-
lytical, Div. Foxboro Co., “Burlington, MA 01803;
‘Vaponies, Inc., Plymouth, MA 02360.
a@@
ation with permitted limits are described in the
USP monographs. When comparing the total”
solids permitted for Water for Injection with that
for Sterile Water for Injection, one will note that,
considerably higher values ‘are permitted for
Sterile Water for Injection. This is necessary
because thé latter product has been sterilized,
usually by a thermal method, in a container that
hhas. dissolved to some extent in the water.
Therefore, the solids content will be greater than
for the nonsterilized product. On the other hand,
the 10 ppm total solids officially permitted for
‘Water for Injection may be much too high when
used as the vehicle for many products. In prac-
tice, Water for Injection normally should not
have a conductivity of more than 1 micromho a)
megohm, approximately 0.1 ppm NaCl).
Pyrogens. Water used in parenteral and irri-
gating solutions should be free of pyrogens. To
achieve this, proper controls must be main-
tained in the preparation and storage of the
water.
Pyrogens are products of metabolism of micro-
organisms. Most bacteria and many molds and
viruses have been reported as producing pyro-
gens. The gramnegative bacteria produce the
“most potent_pyrogenic >es_as endotox-
ns, Chemically, ee are Jipid substances
‘Associated with a carrier molecuile, which is usu-
ally a polysaccharide but may be a peptide
‘About I hour after injection into man, pyrogens
produce a marked rise_in_body temperature,
chills, body aches, citaneous vasoconstriction,
anda risen arterial blood pressure. Antipyretics
eliminate the fever, but not the other systemic
effects of pyrogens.
The fever response to pyrogens in rabbits is
the basis for the official pyrogen test, which is
described later in this chapter. For further infor-
mation, the reader is referred to the extensive
reviews on the nature and significance of pyro-
gens that have appeared in the literature.°?
Source and Elimination of Pyrogen Con-
tamination. Pyrogens may enter a product by
any means that may introduce microorganisms
or the products of their growth. The most likely
water, contaminated solutes, and
containers, Water is free from pyrogens if it has,
been distilled so that the condensed molecules
have gone through the vapor state protected
from inadvertent contamination, and if the dis-
tillate has been collected and stored in a sterile
condition. To be pyrogen-free, solutes must be
prepared from vehicles free from pyrogens, and
‘must be stored in a manner designed to prevent,
subsequent contamination. Opened containers
of solutes, capable of supporting the growth of
microorganisms, invite such contamination.
1°
Containers may be rendered free from pyrogens
by adequate cleaning and heating, usually at
210°C for 3 to 4 hours. Studies also have shown
that heating at 650°C for 60 sec destroys pyro-
gens; however, autoclaving temperatures do not
destroy pyrogens during a normal cycle.
Pyrogens sometimes can be removed from s0-
lutions by adsorption on the surface of select}
adsorbants,“but the often concurrent phenome-
non of the adsorption of solute ions or molecules
may prevent the use of such a method. Selective
solvent extraction methods are useful in the pro-
duction of antibiotics where heavy pyrogen con-
tamination results from the fermentation proc-
ess, New developments in ultrafiltration show
promise of moving this process from limited re-
search applications in molecular separations to
practical production processes, which may in-
clude pyrogen separation and elimination.®° For
most pharmaceutical preparations, however, it
is better to prevent pyrogenic contamination
than to attempt to remove pyrogens, a task that
is difficult to accomplish without adversely af-
fecting the product.
The product development department there-
fore must develop purity requirements for Water
for Injection which are sufficiently stringent for
its use as a vehicle in the product most sensitive
to contaminants. Tests other than those for sol-
ids and pyrogenic content might be required,
.g., qualitative and quantitative tests for the
h_as copper and iron,
Nonaqueous Solvents. In the formulation
of sterile pharmaceutical products, it is some-
‘times necessary to eliminate water entirely or in
part from the vehicle, primarily because of solu-
bility factors or hydrolytic reactions. A nonaque-
‘ous solvent must be selected with great care for
it must not be irritating, toxic, or sensitizing,
and it must not exert an adverse effect on the
edients of the formulation. The screening of
such a solvent must therefore include an evalua-
“on of ts physical properties, such as density,
Viscosity, miscibility and polarity, as well as its
stability, solvent ty, and toxicity."
~ Solvenits that are miscible with water, and thal
are usually used in combination with water as
the vehicle, include dioxolanes, dimethyl-;
acetamide, N-(B-hydroxyethyl)-lactamide, butyl-
ene glycol, polyethylene glycol 400 and 600,
propylene glycol, glycerin, and ethyl alcohol,
‘Water-immiscible solvents include fixed oils;
ethyl oleate, isopropyl myristate, and benzyl
benzoate. The most frequently used nonaqueous.
solvents aie pohethylene gyeo e,
c je5e solvents have been re-
Slewed elsewhere,!!"™ and the reader is referred
to this review for further details.
sreniLe propucts * 641Solutes *
The physical and chemical purity of solutes
used for sterile preparations must also be excep-
tional, Obviously, contaminants entering a prod-
uct with a solute have the same effect as if they
entered via the vehicle. Eyen traces of
contaminants may be 10 products,
necessitating purification of the solute. For a
few substances (for example, ascorbic acid and
calcium gluconate), special parenteral grades
are commercially available
In addition, solutes should be free from micro-
bial and pyrogenic contamination. This entails
not only proper quality of the chemical as pro-
cured but storage conditions designed to prevent
contamination, particularly after a container has
been opened. Preferably, production lots should
be designed to use the entire contents of pack-
ages of chemicals whenever possible.
‘Added Substances. Substances added to a
product to enhance its stability are essential for
almost every product.'? Such substances in-
clude solubilizers, antjoxidants, chelating
agents, buffers, tonicity contributors, antiBacte-
nial agents, antifungal agents, hydrolysis inhibi-
tors, antifoaming agents, and numerous other
substances for specialized purposes. At the same
time, these agents must be prevented from'ad-
versely affecting the product. In general, added
substances must be nontoxic in the quantity
administered to the patient. They should not in-
terfere with the therapeutic efficacy nor with the
assay of the active therapeutic compound. They
must also be present and active when needed
Taste 22-1. Excipients Used for Commercial Parenteral Products
Excipients
‘Antimicrobial Preservatives
Benzyl alcohol :
Benzethonium chloride
Butylparaben
Chlorobutanol
Metacresol
Methylparaben
Myristylgamma picolinium chloride
Phenol
Phenylmercuric nitrate
Propylparaben
‘Thimerosal
Solubilizers, Wetting Agents, or Emulsifiers
Dimethylacetamide
Dicetyl sodium sulfosuccinate
Egg yolk phospholipid
Ethyl alcohol
Ethyl lactate
Glycerin
642 The Theory and Practice of Industrial Pharmacy
throughout the useful Jife of the product. There-
fore, these agents must be selected with great
care, and they must be evaluated as to their ef-
fect upon the entire formulation. An extensive
review of excipients used in parenteral products
and the means for adjusting pH of these prod-
ucts has recently been published and should be
referred to for more detailed itiformation.'*
Table 22-1 provides a list, adapted from that re-
view, of excipients commonly used in commer-
cial parenteral products.
Antibacterial Agents. Antibacterial agents
snr bacteriostatic concentration must be included
in the formulation jucts packaged in mul-
tiple dose vials, and are often included in formu-
lations to be. stenlized By marginal processes. or
made by aseptic manipulation. The require-
tients of activity, stability, and effectiveness of
antibacterial agents in parenterals have been
ise?
reviewed in published papers.
Antioxidaats, Antoxidén’s, inchuded in
imany formulations to protect’ a therapeutic
agent susceptible to oxidation, particularly
under the accelerated conditions of thermal ster-
ilization, may function in at least two ways., i¢.,
(1) by being preferentially oxidized (seducing
its) and thereby gradually used up, or (2) by
locking an oxidative chain reaction in which
they are not uswally consumed. In addition, cer-
tain compounds have been found to act as syner-
ists, increasing ‘the effectiveness of antioxi-
Sams, paticulaiy those. blocking. oxidative
reactions. A fourth group of compounds are use-
ful in this connection in that they complex with
catalysts that otherwise would accelerate the
Concentration Range (%%)
a 05-100
0.01
ois
025-05
0.1-0.25
0.01-0.18
017.
0.065-05
0.001
0.005-0.035
0.001-0.02
0.01
ois
12°
061-49.0
ol
146-250Excipients Concentration Range (%)
Solubilizers, Wetting Agents, or Emulsifiers—continued
Lecithin 05-23
PEG-40 castor oll T0-11.5
Polyethylene glycol 300 0.01-50.0
Polysorbate 20 0.01
Polvsorbate 40 0.05
Polysorbate 80 0.04-4.0
Povidone 02-10
Propylene glycol 02-500
Sodium desoxycholate 021
Sorbitan monopalmitate 0.05
Theophylline 50
Buffers
Acetic acid
Adipic acid
Benzoic acid and sodium benzoate
Citric acid
Lactic acid
Maleic acid
Potassium phosphate
Sodium phosphate mon basic
Sodium phosphate dibasic
Sodium acetate
Sodium bicarbonate
Sodium carbonate
Sodium citrate
Sodium tartrate
Tartaric actd
Bulking Substances or Tonicity Modifiers
Glycerin 16-2.25
Lactose 0.14-5.0
Mannitol 04-25
Dextrose 375-50
Sodium chloride varies
Sodium sulfate ret
Sorbitol 20
Suspending Agents
Galatin 20
‘Methyleellulose 0.03~1.05
Pectin 02
Polyethylene glycol 4000 27-30
Sodium carboxymethylcellulose 0.05-0.75
Sorbitol solution 50.0
Chelating Agents
4 Edetate disodium 0.00368-0.05
Edetate calesum disodium 0.04
Edetate tetrasodium 0.01
Local Anesthetics
Procaine HCI 10
Benzyl alcohol 5
Stabilizers
Creatinine
Glycine
Niacinamide
Sodium acetslteyptophanate
Sodium caprylate
Sodium saccharin
‘Adapted from Wang, YJ., and Kowal, RR J, Parent. Drug Assoc, 34452, 1980
STERILE PRODUCTS + 643oxidative reaction. Because of the differences in
action, combinations of these agents are some-
times used. In Table 22-2, the more commonly
employed antioxidants are listed according to
the above four groupings. The reader is referred.
to the literature for more details concerning an:
tioxidants and their activities.’°-®
Tt should also be mentioned that for those
products in which oxygen enters into a degrada-
tive reaction, an antioxidant effect can be
achieved by displacing oxygen (air) from contact
with the product. Usually, this is accomplished
by saturating the liquid with either nitrogen or
carbon dioxide and sealing the final container
after displacing the air above the product with
the gas.
Higuchi and Schroeter have warned of the
reactivity of bisulfites with drug molecules,”!
and Halaby and Mattocks have waned of the
potential toxicity of sodium bisulfite absorbed
from peritoneal dialysis solutions.”
Buffers. Buffers are added to maintain a re-
quired pH for many products; a change in pH
may cause significant alterations in the rate of
degradative reactions. Changes in pH may occur
during storage as a result of the dissolving of
lass constituents in the product, release of con-
stituents from rubber closures or plastic compo-
“nents in contact with the product, dissolving of
ase5-and vapors from ace in the con-
tainer and diffusion through the mubber or plas-
Taste 22-2. Antioxidants Used in Sterile Prod-
ucts
Usual
Concentration
Compound (%)
Antioxidants (reductng agents)
Ascorbie acid 0.02-0.1
Sodium bisulfite 0.1-0.15
Sodium metabisulfite 0.1-0.15
Sodium formaldehyde sulfoxylate 0.1-0.15
Thiourea 0.005
Antioxidants (blocking agents)
Ascorbic acid esters 0.01-0.015
Butyl hydroxytoluene (BHT) 0.005-0.02
‘Tocopherols 0.05-0.075
Symergises
Ascorbic acid 0.01-0.05
Citric acid 0,005~0.01
Citraconic acid 0.03-0.45
Phosphoric acid 0.005-0.01
Tartaric acid 0.01-0.02
Chelating agents
Ethylenediaminetetraacetic acid salts 0.01-0.075
644+ The Theory and Practice of Industrial Pharmacy
component, or reactions within the product.
iffers must havé the capacity to maintain the
pH of the product against these influences, but
not enough to prevent the body fluids from over-
whelming the buffer following administration.
In most cases, the biologic effectiveness of the
drug is maximum at or near the biologic fluid pH
rather than at the stabilizing pH of ‘he injected
product
Acetates, citrates, and phosphates are the
principal buffer systems used, but buffer sys-
tems making use of other ingredients in the for-
mulation are often used to reduce the total num-
ber of ingredients in the product. Buffer systems
must be selected with consideration of their ef-
fective range, concehitration, and chemical ef-
fect_oni the total product. These factors have
been reviewed by Windheuser.”*
Tonicity Contributors. Compounds con-
tributing to the isotonicity of a product reduce
the pain of injection in areas with nerve end-
ingS-Buffers may serve as tonicity contributors
as well as stabilizers for the pH. Other added
substances also contribute to the colligative
properties of the preparation. Whenever possible
such dual activity is desirable.
Although the freezing point depression of the
solution is most frequently used to determine
whether a solution is isotonic, isotonicity actu-
ally depends on the eability ofa living sem-
fpemeible membrane that separates the solu
fon from a biologic cell system. Most frequently,
for~sterile pharmaceutical preparations, the
membrane concemed is the one enclosing the
red blood cells. Therefore, a preparation cannot
be considered to be isotonic until it has been
tested in a biologic system. A hemolytic method,
4usingsred blood cells, has been described.*5
Tsotonicity values for various drugs have been
recorded. Testing by such a method be-
Comes even more important when all or part of
the water is replaced with another solvent, since
dissociation is different when water is displaced
by another solvent. a
A wt
Containers yd
ch
Containers are in intiniate contact with the
product. No container presently available is
totally nonreactive, particularly with aqueous
solutions. Both the chemical and physical char-
acteristics affect the stability of the product, but
the physical characteristics are given primary
consideration in the selection of @ protective
container.
Glass containers traditionally have been used
for sterile products, many of which are closed
with rubber stoppers. Interest in plastic con-