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STERILIZATION

Tarun K. Mandal, Ph.D.

Tarun K. Mandal, Ph.D.

STERILIZATION

STERILITY: Absence of life or absolute freedom from biological contamination. STERILIZATION: Inactivation or elimination of all viable organism and their spores.

Tarun K. Mandal, Ph.D.

STERILIZATION

DISINFECTANT: Substance used on non-living objects to render them non-infectious; kills vegetative bacteria, fungi, viruses but Not Spores. e.g. Formaldehyde

Tarun K. Mandal, Ph.D.

STERILIZATION

BACTERICIDE (GERMICIDE): Substance that kills vegetative bacteria and some spores BACTERIOSTAT: Substance which stops growth and multiplication of bacteria but does not necessarily kill them. Growth usually resumes when bacteriostat is removed.

Tarun K. Mandal, Ph.D.

STERILIZATION

ANTISEPTIC: Substance used to prevent multiplication of microorganism when applied to living systems. An antiseptic is bacteriostatic in action but not necessarily bacteriocidal.

Tarun K. Mandal, Ph.D.

STERILIZATION

VEGETATIVE CELL: Bacterial cell capable of multiplication (as oppose to spore form which cannot multiply). Less resistant than the spore form. SPORE: Body which some species of bacteria form within their cells which is considerably more resistant than the vegetative cell.

Tarun K. Mandal, Ph.D.

STERILIZATION

Methods: 1. Steam Sterilization 2. Dry heat sterilization 3. Filtration 4. Gas sterilization 5. Irradiation NOTE: End products must pass sterility tests.

Tarun K. Mandal, Ph.D.

STERILIZATION

DRY HEAT STERILIZATION: Equipment: Oven Method: Dry heat sterilization is carried out at 160 deg C. to 170 deg C. for 2 to 4 hrs. Application: Glassware Fixed oils Thermostable powders
Tarun K. Mandal, Ph.D.

STERILIZATION

Advantages & Disadvantages: Sterilization by means of heat requires higher temperatures and longer exposures than sterilization by steam. Heat transfer is slow, small volumes of oil and thin layers of powder should be used.

Tarun K. Mandal, Ph.D.

STERILIZATION

STEAM STERILIZATION: Equipment: Autoclave Method: In the presence of moisture, microorganisms are destroyed at a lower temperature than in dry heat. This is the method of choice when product can withstand such treatment.
Tarun K. Mandal, Ph.D.

STERILIZATION

10 lb Pressure (115.5 deg. C)...30 minutes 15 lb Pressure (121.5 deg. C)...20 minutes 20 lb Pressure (126.5 deg. C)... 15 minutes

Tarun K. Mandal, Ph.D.

STERILIZATION

Application: 1. Solutions sealed in containers ampuls, vials 2. Bulk Solutions 3. Glassware 4. Surgical Dressing 5. Instruments

Tarun K. Mandal, Ph.D.

STERILIZATION

Advantages: Rapid, Inexpensive, Effective, Large volumes Disadvantages: 1. Cannot use for oily preparation (oil base ointment) 2. Cannot use for moisture sensitive preparations

Tarun K. Mandal, Ph.D.

STERILIZATION

FILTRATION: Physical removal of microorganisms by adsorption on the filter medium. Used for heat sensitive materials.

Tarun K. Mandal, Ph.D.

STERILIZATION

BACTERIAL FILTRATION: Equipment: 1. Porcelain filters 2. Siliceous earth filter 3. Sintered glass filters 4. Asbestos filters 5. Membrane filters
Tarun K. Mandal, Ph.D.

STERILIZATION

Method: Direct filtration 1. Positive pressure 2. Negative pressure Application: Thermolabile solutions of low viscosity. Advantages & Disadvantages: 1. Depend on filter media 2. Thermolabile solutions can be sterilized.

Tarun K. Mandal, Ph.D.

STERILIZATION

GASEOUS STERILIZATION Equipment: Special oven, for admission of gas and humidity & hermetic Method: Humidity of less than 20% RH Ethylene Oxide

Tarun K. Mandal, Ph.D.

STERILIZATION

Ethylene Oxide-Carbon dioxide Pressure 30 psi Temperature 20-55 deg. C Application: Thermolabile powder plastic\polymers ophthalmic prep. subcutaneous, vaginal inserts, plastic syringes, tubing sets

Tarun K. Mandal, Ph.D.

STERILIZATION

Advantages & Disadvantages: 1. Explosive hazard 2. Toxic 3. Not appropriate for solutions

Tarun K. Mandal, Ph.D.

STERILIZATION

RADIATION STERILIZATION Equipment: Ultraviolet Lamp Ionization (Beta Rays, Gamma Rays, X-Rays) Application: Thermolabile Drugs (Powdered)

Tarun K. Mandal, Ph.D.

STERILIZATION

Disadvantages: 1. Highly specialized equipment required 2. Effect of irradiation on products and their containers.

Tarun K. Mandal, Ph.D.

STERILIZATION

STERILITY TESTS (A) Microorganisms: USPXXll recommends the use of biological indicators. 1. For liquid preparations-add directly to the preparations. 2. For solid preparations or equipments- add the culture to strips of filter paper.
Tarun K. Mandal, Ph.D.

STERILIZATION

Different organisms for different methods of sterilization. The organisms that are resistant to a particular sterilization method should be chosen as the marker organism

Tarun K. Mandal, Ph.D.

STERILIZATION

Sterilization Method
Steam sterilization Dry-heat sterilization Ethylene oxide sterilization Ionizing radiation sterilization

Marker organisms
Bacillus stearothermophyilus Bacillus subtilis Bacillus subtilis
Bacillus pumilus

Tarun K. Mandal, Ph.D.

STERILIZATION

(B) Pyrogen and Pyrogen Testing Pyrogens are fever producing organic substances arising from microbial contamination. The causative material is thought to be a Lipopolysaccharide from the outer cell wall of the bacteria. This is Thermostable

Tarun K. Mandal, Ph.D.

STERILIZATION

TESTS: 1. RABBIT TESTS a) Render the syringes, needles and glassware free from Pyrogens by heating at 250 deg. C for not less than 30 minutes. b) Warm the product to be tested to 37 deg. 2 deg. C. c) Take three healthy rabbits
Tarun K. Mandal, Ph.D.

STERILIZATION

d) Inject into an ear vein of each of three rabbits 10 ml of the product per kg body weight. e) Record the temperature at 1,2,and 3 Hrs.

Tarun K. Mandal, Ph.D.

STERILIZATION

CASE I
Results: (i) No rabbit shows an individual rise in temperature at 0.6 deg. C or more above its respective control temp. (ii) Sum of the three individual maximum temp. rises does not exceed 1.4 deg. C. Conclusion: The material meets the USP requirements for the absence of Pyrogen.
Tarun K. Mandal, Ph.D.

STERILIZATION

CASE II Results: (i) If any rabbits show a temp. rise of 0.6 deg.C or more or (ii) If sum of the temp. rises exceeds 1.4 deg. C Conclusion: Repeat the tests using five other rabbits.

Tarun K. Mandal, Ph.D.

STERILIZATION

Results:
(i) If not more than three of the eight rabbits show individual rises in temp. of 0.6 deg. C or more (ii) If the sum of the eight temp. rises does not exceed 3.7 deg.C Conclusion: The material meets the USP requirements for the absence of Pyrogens.

Tarun K. Mandal, Ph.D.

STERILIZATION

2) LAL TESTS: Limulus Amebocyte Lysate (LAL) Tests Extract from the blood cells of the Horse Shoe Crab (Limulus Polyphemus) contains an enzyme and protein that coagulates in the presence of low levels of Lipopolysaccharides.

Tarun K. Mandal, Ph.D.

PARENTERAL DRUG DELIVERY


Tarun K. Mandal, Ph.D.

Tarun K. Mandal, Ph.D.

PARENTERALS

PARENTERALS Injections: These are sterile, Pyrogen free preparations intended to be administered parenterally (outside alimentary tract). Parental Routes Of Administration

Tarun K. Mandal, Ph.D.

PARENTERALS

Most Common: 1. Subcutaneous (SC;SQ;Sub Q) 2. Intramuscular (IM) 3. Intravenous (IV) Others: 4. Intracisternal 5. Intradermal (ID) 6. Intraspinal 7. Intraarterial (IA)
Tarun K. Mandal, Ph.D.

PARENTERALS

PARENTERAL ROUTE IS USED FOR: 1) Rapid action 2) Oral route can not be used 3) Not effective except as injection

Tarun K. Mandal, Ph.D.

PARENTERALS

Official Types of Injections: 1. Solutions of Medicinal Example: Codeine Phosphate Injection Insulin Injection 2. Dry solids or liquid concentrate does not contain diluents etc. Example: Sterile Ampicillin Sodium
Tarun K. Mandal, Ph.D.

PARENTERALS

3. If diluents present, referred to as.....for injection Example: Methicillin Sodium for injection 4. Suspensions "Sterile....Suspension" Example: Sterile Dexamethasone Acetate Suspension

Tarun K. Mandal, Ph.D.

PARENTERALS

5. Dry solids, which upon the addition of suitable vehicles yield preparations containing in all respects to the requirements for sterile suspensions. Title: Sterile....for Suspension Example: Sterile Ampicillin for Suspension

Tarun K. Mandal, Ph.D.

PARENTERALS

The form into which a given drug is prepared for parenteral use by the manufacturer depends on the nature of the drug. 1. physicochemical characteristics 2. therapeutic consideration

Tarun K. Mandal, Ph.D.

PARENTERALS

Onset of Action\Duration 1. Chemical form of the drug 2. Physical state of the injection (a) Solution (b) Suspension 3. Vehicle used

Tarun K. Mandal, Ph.D.

PARENTERALS

Most rapid onset of action: Drugs that are very soluble in body fluids. Drugs in aqueous solutions > Drugs in oleaginous solution. Drugs in aqueous suspension > Drugs in oleaginous suspension. "Repository" or "Depot" Type injections - Long acting
Tarun K. Mandal, Ph.D.

PARENTERALS

Requirements: Solvents or vehicles used must meet special purity and other standards. Restrictions on buffers, stabilizers, antimicrobial preservative. Do not use coloring agents. Sterile and Pyrogen - Free.
Tarun K. Mandal, Ph.D.

PARENTERALS

Must meet compendial standards for particular matter. Must be prepared under aseptic conditions. Specific and high quality packaging.

Tarun K. Mandal, Ph.D.

PARENTERALS

Vehicles: Aqueous: Sterile water for injection. Nonaqueous: Fixed oils Glycerin PEG Alcohol
Tarun K. Mandal, Ph.D.

PARENTERALS

Restrictions on Fixed Oils: Remain clear when cooled to 10 deg. C. Not contain Paraffin or Mineral oil. Must meet the requirement of iodine number and Saponification number.

Tarun K. Mandal, Ph.D.

PARENTERALS

Iodine Number (Value):


It represents the number of g of iodine absorbed, under the prescribed conditions, by 100g of the substance.

Saponification Value (Number):


It represents the number of mg of Potassium Hydroxide required to neutralize the free acids and saponify the esters contained in 1.0g of the substance.

Tarun K. Mandal, Ph.D.

PARENTERALS

Must specify the oil used e.g. corn oil, cottonseed oil, peanut oil, sesame oil. Must be free from rancidity.

Tarun K. Mandal, Ph.D.

PARENTERALS

Solvents used must be: Non-irritating Non-toxic Non-sensitizing No pharmacological activity of its own Not affect activity of medicinal

Tarun K. Mandal, Ph.D.

PARENTERALS

Added Substances -preservatives -buffers -antioxidants -solubilizers -thickeners -materials to adjust tonicity
Tarun K. Mandal, Ph.D.

PARENTERALS

Do Not Use Color Preservatives: Multidose containers must have preservatives unless prohibited by monograph.

Tarun K. Mandal, Ph.D.

PARENTERALS

ASEPTIC TECHNIQUE: An aseptic technique is one which is designed to prevent contamination of materials, instruments, utensils, containers, during handling.

Tarun K. Mandal, Ph.D.

PARENTERALS

Sources of Contamination -The Air -The Breath -The Skin -The Hair -Clothing -Working surfaces
Tarun K. Mandal, Ph.D.

PARENTERALS

Methods of minimization of contamination: apply common sense Airborne contamination--use laminar airflow Horizontal Vertical

Tarun K. Mandal, Ph.D.

PARENTERALS

HEPA filter (High efficiency particulate air filter) Contamination from the breath--use masks Contamination from the skin: Nails should be scrubbed Hands and forearms should be washed thoroughly with detergent solutions
Tarun K. Mandal, Ph.D.

PARENTERALS

Hair and Clothing: Always wear sterile gown over normal clothing Long hair should be tied back Wear a cotton cap Working surfaces: Clean the working surface with a bactericidal solution or ethyl alcohol
Tarun K. Mandal, Ph.D.

PARENTERALS

PACKAGING:
1) Single dose: Hermetic container holding a quantity of sterile drug intended for parenteral administration as a single dose. Example: ampuls sealed by fusion 2) Multiple dose: Hermetic container permits withdrawal of successive portions of the contents without changing the strength, quality, or purity of the remaining portion.
Tarun K. Mandal, Ph.D.

PARENTERALS

LABELING: Name of product % of drug or amount of drug in specified volume of amount of drug and volume of liquid to be added Manufacturer/Distributor Lot number Name and quantity of all added substances
Tarun K. Mandal, Ph.D.

PARENTERALS

Expiration date Veterinary product should be so labeled Must check each individual monogram for: Type of container Type of glass Package size Special storage instructions

Tarun K. Mandal, Ph.D.

PARENTERALS

AVAILABLE INJECTION: Small volume parenterals (svp) solutions Table 8-3 suspensions Ansel Pg 280 Various insulin preparations Table 8-5 Ansel Pg 282
Tarun K. Mandal, Ph.D.

PARENTERALS

ONSET OF ACTION: Regular insulins > zinc suspension, prompt > isophane suspension (NPH) > zinc suspension *(lente) > protamine zinc suspension > zinc suspension, extended

Tarun K. Mandal, Ph.D.

PARENTERALS

DURATION OF ACTION: Regular insulin < zinc suspension, prompt < isophane suspension (NPH) < zinc suspension (lente) < protamine zinc suspension < zinc suspension, extended

Tarun K. Mandal, Ph.D.

PARENTERALS

Amorphous form- semilente insulin Larger crystalline form- ultralente insulin Lente insulin or insulin zinc suspensionphysical mixture of 70% crystalline form & 30% amorphous form Insulin injection--intravenously Suspension--subcutaneously

Tarun K. Mandal, Ph.D.

PARENTERALS

INSULIN INJECTION (REGULAR) sterile aqueous solution of insulin Prepared from beef or pork or both or through biosynthetic means The source must be stated on the labeling.

Tarun K. Mandal, Ph.D.

PARENTERALS

INSULIN ZINC SUSPENSION: Insulin is modified by the addition of zinc chloride mixture of crystalline and amorphous insulin (7.3) EXTENDED INSULIN ZINC SUSPENSION: Sterile suspension of zinc insulin crystals in an aqueous medium
Tarun K. Mandal, Ph.D.

PARENTERALS

PROMPT INSULIN ZINC SUSPENSION: Sterile suspension of insulin PROTAMINE ZINC INSULIN SUSPENSION: Sterile suspension of insulin INSULIN INFUSION PUMPS: Allow patients to achieve and maintain blood glucose at near normal levels on a constant basis.
Tarun K. Mandal, Ph.D.

PARENTERALS

Protamine is prepared from the sperm or the mature testes of fish belonging to the genus oncorhynchus and others.

Tarun K. Mandal, Ph.D.

PARENTERALS

LARGE VOLUME PARENTHERALS (LVP'S): Generally administered by intravenous infusion to replenish body fluids, electrolytes, or to provide nutrition--100ml-1L These solutions should not contain: *Bacteriostatic agents *Other pharmaceutical additives
Tarun K. Mandal, Ph.D.

PARENTERALS

IRRIGATION AND DIALYSIS SOLUTIONS: Irrigation solutions: Intended to bathe or wash wounds, surgical incisions, or body tissues. Example: Sodium chloride irrigation, USP 0.9% NaCl Use: Employed topically to wash wounds, rectally as an enema

Tarun K. Mandal, Ph.D.

PARENTERALS

DIALYSIS SOLUTION: Dialysis is a process whereby substances may be separated from one another in solution by taking advantage of their differing diffusibility through membranes. Example: Peritoneal dialysis solutions Use: To remove toxic substances normally excreted by the kidney.

Tarun K. Mandal, Ph.D.

PARENTERALS

*Poisoning *Kidney Failure *Renal Transplants Commercially available solutions contain: *Dextrose *Peptides

Tarun K. Mandal, Ph.D.

PARENTERALS

Solutions are made to be hypertonic (with Dextrose) to plasma to avoid absorption of water from the dialysis solutions into the circulation. Peritoneal cavity acts as a semipermeable membrane.

Tarun K. Mandal, Ph.D.

PARENTERALS

BIOLOGICALS: -vaccines -toxins -toxoids -antitoxins -immune serums -blood derivatives -diagnostic aids
Tarun K. Mandal, Ph.D.

PARENTERALS

Storage: Refrigerator at 2 deg C to 8 deg C, avoid freezing These preparation should meet the std. of the bureau of biologies of the FDA.

Tarun K. Mandal, Ph.D.

PARENTERALS

IMMUNITY:
Power of the body to resist and overcome infection.

NATURAL OR NATIVE IMMUNITY:


Individuals resistance to a particular toxic agent because of race, endocrine balance, etc.

ACQUIRED IMMUNITY:
Specific immunity that may be acquired (Active or Passive)
Tarun K. Mandal, Ph.D.

PARENTERALS

ACTIVE IMMUNITY: *Naturally acquired active immunity--occurs in response to an infection *Artificially acquired active immunity-- response to a specific vaccine or toxoid PASSIVE IMMUNITY: Introduce already formed antibodies into body to combat a specific antigen
Tarun K. Mandal, Ph.D.

PARENTERALS

VACCINES: Administered primarily for prophylactic action for the development of active acquired immunity. TOXOIDS: Toxins modified and detoxified by moderate heat and chemical treatment Example: Diphtheria, Tetanus
Tarun K. Mandal, Ph.D.

PARENTERALS

ANTITOXINS: Prepared from blood of animal immunized by repeated injections of bacterial toxins

Tarun K. Mandal, Ph.D.

PARENTERALS

ANTISERUMS:

Prepared in same manner as antitoxins except that viruses or bacteria injected to produce antibodies. Produce passive immunity human immune serums and globulins. Serums containing specific antibodies obtained from blood of humans who have had the disease or have been immunized against it with a specific biologic product.

Tarun K. Mandal, Ph.D.