Parenterals

Formulation development

 Parenteral products are products that are administered to the body by injection Because this route of administration bypasses the normal body defense mechanisms, it is essential that these products are prepared with a high degree of care and skills than utilized in preparing conventional oral or topical products. The finished product must be sterile, nonpyrogenic and free from extraneous insoluble materials. These products must satisfy a number of requirements for parenteral products.

Introduction and importance of the study

Sterile formulations must meet a number of special criteria such as

Sterility Particulate material Pyrogen free Stability pH Osmotic pressure

During the formulation of parenteral products the following factors are critical
The vehicle in which the drug is dissolved or dispersed Volume (dose) of the injection Adjustment of isotonicity Adjustment of pH Stabilisers Preservatives Adjustment of specific gravity (for spinal anaesthesia) Concentration units

 Limits to the level of pyrogens present and of particulate matter The injection route dictates the volume of formulation. Hence the solubility of the drug in the selected vehicle is critical in the formulation

Difference between parenterals and other products

Vehicle
The preferred vehicle is water as it is well tolerated by the body, easy to administer and a large solvent capacity Water for injection must be sterile and free from pyrogens Cellulose, glass, rubber cores, cloth or cotton fibres may constitute the contaminants list. Suitable filtration media for removal of particulate material are sintered glass filters or membrane filters with a pore size of 0.45-1.2 microns

Ph and Buffers
As parenteral products are administered directly to tissues and systemic circulation, formulations prepared should not vary significantly from physiological pH, which is about 7.4. The acceptable pH range is 3-10.5 for i.v preparations and 4-9 for other routes. Buffers are included in injections to maintain the pH of the packaged product. The buffers used in the injection must allow the body fluids to change the product pH after injection. Acetate, citrate and phosphate buffers are

Osmotic pressure
The osmotic pressure of the blood is approx. 300 milli Osmoles/L and ideally any sterile solution would be formulated to have the same osmolarity For eg: 0.9% w/v NaCl i.v solution has an osmolarity of 308 milli Osmoles/L . 5% w/v Dextrose i.v solution has an osmolarity of 280 milli Osmoles/L . NaCl, Mannitol or glucose can be

Antimicrobial agents
Aqueous preparations which are prepared using aseptic preparations and which cannot be terminally sterilized may contain a suitable antimicrobial preservative in an appropriate concentration. Antimicrobial agents are added to multiple dose vials to inhibit the growth of microbial organisms which may occur accidentally and contaminate the product during use.

Antimicrobial preservatives
S.NO ANTIMICROBIAL AGENT

1 2 3 4 5 6 7

Benzalkonium chloride Benzyl alcohol Chlorobutol Phenol Chlorocresol Phenyl mercuric salts Methyl hydroxy benzoate

CONCENTRATION (%w/v)

0.01 1- 2

0.25 0.5 0.5 0.1 0.3 0.002 0.1- 0.2

Atioxidants
Aqueous solutions are more susceptible to oxidation Bisulphites and metabisulphites are commonly used antioxidants in aqueous injections. Injection formulations may in addition also contain chelating agents, such as EDTA or citric acid, to remove trace elements, which catalyse oxidative degradation.

Sterilization
Sterility : Absence of life or absolute freedom from biological contamination. Sterilization : inactivation or elimination of all viable organism and their spores. Disinfectant : substance used on non- living objects to render them non- infectious; kills vegetative bacteria, fungi, virus but not spores. Eg: Formaldehyde

Advantages and disadvantages

Moist Heat
Spore are killed by moist heat. Culture media & other liquids required to retain their content of water. Not applicable to waterproof materials such as oils and greases or dry materials.

Pasteurization
Moist heat at temperature below 1000 C. Heat labile fluids may be disinfected not sterilized by heating at 560 C for 30 min. Sufficient to kill mesophilic bacteria but not spores. For serum, or other body fluids containing proteins, temp to rise above 59o C. UHT; 140o C less than 1 sec. Cold Pasteurization. High pressure pasteurization.

Washer Disinfectors
Washing machines using hot water, steam and detergents, may be used. Washing action at 710C for 3 min or 800 C for 1 minute sufficient to kill vegetative organism. Recommended for instruments contaminated with HBV and HIV. Accordingly it is set at 93o C for 10 min.

Boiling at 100o C
Heating in boiling water at 100o C for 5 minutes sufficient to kill all vegetative bacteria, HBV and some bacterial spore. Used only in case of emergencies to sterilize medical and surgical equipment. Heat labile articles and hollow or porous items where water will not penetrate lumen cannot be disinfected this way.

Steaming at 100o C
To prevent glass from cracking which happens when it is heated directly. Also in case of heat labile culture media. Pure steam in equilibrium with boiling water at normal atmospheric pressure Exposure to this temperature for 5 minutes will kill microorganism.

Free Steaming
Koch and Arnold steamers are used. Useful for selective heat labile media like DCA, XLD, TCBS and Slenite F broth. These media do not support the growth of heat resistant bacteria. Tyndallization- 20 min for 3 succesive days.

Steam sterilization
Irreversibly coagulates and denatures microbial enzymes and proteins. Important parameters for effectiveness 1. Exposure time 2.Temperature of process 3.Level of moisture. 4.Pressure

1) Advantages Non-toxic Cycle easy to control and monitor Inexpensive Rapidly microbiocidal Least affected by organic/inorganic soils Rapid cycle time Penetrates medical packing, device lumens

2) Disadvantages Deleterious for heat labile instruments Potential for burns

Autoclaves
Invented by Charles Chamberland in 1879. Precursor was the Steam digester invented by Denis Papin in 1679. At correct temp lethal to all bacteria, viruses, fungi & protozoa.

 Example of usage of autoclaves are: Hospitals & OPD- Porous load autoclaves. Mortuary- Bench top autoclaves. Microbiology Lab: Media preparators or fluid cycle steam sterilizers. Pharmaceutical- Fluid Cycle Sterilizer
:

Types of autoclaves
Simple laboratory autoclave Transportable bench top autoclaves Large simple autoclave Downward displacement autoclave Multipurpose laboratory autoclave Pre vacuum High security autoclave. Porous load sterilizer. Low temperature steam. Steam flash pressure pulsing steam sterilization autoclaves

 Sterilization Hold Time Heat Penetration Time Condensation of steam: 3 effects 1.Wets microorganisms. 2.Liberates latent heat. 3.Significant contraction of steam.

 Time and temperature with pressure required for sterilization by steam under pressure in autoclave. 121-124o c at 1.1 bar for 15 min. 134-138o c at 2.2 bar for 3 min. l1 bar=1 atm pressure=14.7 pounds per square inch 1.

 Steam which is present must be 1. Saturated: 2.Dry. 3.Pure. lSteam supply: 1. Superheated steam ( Dry gas) 2.Wet steam l

 Removal of air: 1. Simple transportable (pressure-cooker) autoclaves. 2. Downward displacement autoclaves. 3. Porous load autoclaves. Items to be put in autoclave 1. Unwrapped non porous items: 2. Porous loads 3. Discard loads 4. Fluids in sealed containers. 5. Nutrient media.

Simple laboratory autoclave

1. Considered unsatisfactory. 2.Not monitor temperature of load and therefore cannot prevent spoiling nutritive value. 3.No safety interlock.

Transportable bench top autoclaves.

Sophisticated version of air displacement autoclave. Possess automatic cycle control with indicator for cycle failure and thermal safety. Disadvantages of this are: 1. Not fitted with vacuum assisted air removal. 2. No assisted drying. 3. Cannot be used for porous loads, packaged items or discard loads. 4. Cant handle liquid loads

Large simple autoclave

Larger version of simple pressure cooker. No means of assisted air removal. Size too large to permit removal of air. Unsuitable for wrapped articles and make safe loads.

Downward displacement laboratory autoclaves

Provision for removal of air from chamber{ balanced pressure steam trap} Other devices assist the drying of the load. No vacuum assistance for air removal. Air removal was found to be inadequate. Only value is sterilization of unwrapped nonporous metallic items. Cooling may take many hours to be below 80o C

Multipurpose laboratory autoclaves

Different types of load require different cycles required for these different loads. Special feature Presence of efficient means of assisted air removal and drying, assisted cooling and a temperature sensitive probe reading directly from the load.

Multipurpose laboratory autoclaves ( Contd)

Aqueous media
1. Accelerated cooling required to avoid damage to nutrient properties of culture medium. 2. Cooling to below 80o C before autoclave is opened minimize risk of breakage and explosions. 3. Volume of DIN bottle which is used should be not over 80%. 4. Duration of heating up period should be controlled automatically with a thermocouple placed in one of the largest bottle or simulators, which reproduces thermal characteristic of bottle.

Multipurpose laboratory autoclaves ( Contd)

121o c for 15 min. Cooling of load should be assisted so that media are cooled to under 80o c in 30 min. Methods of cooling. Cooling time duration set by a thermocouple placed in simulator Air blasting. Glasswares are satisfactorily processed as part of a fluid cycle.

1.High security autoclave 2. 3.Porous load sterilizer 4. 5.Low temperature steam

Gas sterilization
For delicate heat labile equipments. Monitoring their efficacy by biological test.

1. Low temperature steam formaldehyde

Steam at sub atmospheric pressure (temp below 100o C) kills the spores of thermophilic bacteria very slowly. Sporicidal at high concentration in presence of moisture. Synergism between formaldehyde and steam. Difficulty in combining the two.

2. Ethylene oxide sterilizer

Useful also for small proportion of medical and surgical devices which cannot withstand autoclaving. Kills micro organisms by altering their DNA by alkylation only materials with documented ethylene oxide penetration and dissipation properties should be used as wrappers.

Low temperature sterilization by ozone

The 125 l ozone sterilizer uses medical grade oxygen water and electricity to generate ozone within the sterilizer to provide efficient sterilants without producing toxic chemicals or using high temperatue. End of cycle O2 and H2O are formed.

Liquid Sterilization
To sterilize immersible devices like endoscopes etc with 35% liquid Peracetic acid. Done using STERIS system 1 Acid is diluted with sterile filtered water. Commercially available spores can be used for monitoring sterilization. Disadvantage is high cost.

Sterilizing Filter
Aqueous liquids sterilized by forced passage through filter of porosity small enough to retain any microorganism. 1 ) Membrane filters : Manufactured from variety of polymeric material such as cellulose diacetate, polycarbonate and polyester, as disc.

Membrane Filter (Contd)

Membrane made in 2 ways 1. Capillary pore membranes: For viruses 2. Labyrinthine pore membranes: Bacteria & yeast Exact procedure for use varies with form in which filter is supplied. Filters may be supplied with plastic holders pre sterilized for single use, or mounted in re-usable holders and fitted to filtration vessels.

Sterilizing Filter
2)Syringe filters: Membrane 13-25mm diameter. Fitted in syringe like holders of stainless steel or polycarbonate. Used for sterilization of small volumes of fluid. 3)Vacuum and in- line filters: Membranes of 25-45 mm diameter are used either with in line filter holders of Teflon or stainless steel and aluminum. Used for sterilization of large volumes of air and liquid.

4) Pressure filtration: Large membranes, 100-293 mm in diameter, housed in pressure filter holders. Production of pure water.

5) Air Filters: Large volume of air rapidly freed from infection by passage through HEPA (High efficiency particle arrester)

Air Filters (Contd)

Principal use is to render safe the air withdrawn from an exhaust ventilated safety cabinets used for work with pathogens. To decontaminate air input into laminar flow cabinet. Fitted with disposable pre filter which reduces load collected by main filter.

STERILIZATION BY RADIATION
IONIZING radiation - radiation from radioactive elements ,usually Co60.,

E.g.. Sterilization of Disposable Syringes. Bacillus pumilis used for testing.

ULTRAVIOLET RAYS: Mercury vapor lamps emitting radiation in the range of 250-

References
Mackie & McCartney Practical Medical Microbiology 14 th edition Murray, Scot et al Text book of medical microbiology Bailey & Scot, Diagnostic Medical Microbiology Twelfth Edition Ananthnarayan and Paniker, Textbook of microbiology, 8 th edition.