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@@ ation 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 * 641Solutes * 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 146-250Excipients 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 + 643oxidative 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-