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Theoretical Principles and Practical Skills of Aseptic Manipulations

Maulita Cut Nuria, M.Sc., Apt

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Definitions Background Laminar Air Flow Hoods Aseptic Technique

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Enteral Of or relating to the intestines (e.g. by mouth, orally, per tube) Parenteral Introduced other than by way of the intestines ( (e.g. i t intravenous, i t intramuscular, l subcutaneous) b t ) Aseptic Technique The technique for manipulations of compounded sterile products and parenteral preparations that prevents contamination
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Definitions (cont.)
Contamination any effect or action that has a negative impact on a product's integrity making it unfit for use Chemical composition pH Sterility (e.g. microorganism contamination) Pyrogenicity y g y Biological or therapeutic potency Physical appearance Particulate matter (e.g. dust, glass or precipitation)

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Background g
Sources of p product contamination People (most common) Touch contamination Generation of particulates from shedding cells or hair Supply air Heating Ventilation and Air Conditioning C ( (HVAC) C) Infiltration Particles from adjacent spaces Internal generation Walls, floors, ceilings, packaging, equipment
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Laminar Air Flow Hoods

The underlying principle of a laminar air flow hood is that a constant flow of HEPA filtered air at a rate of approximately 90 linear feet per minute physically sweeps the work area and prevents the entry of contaminated air The hood workspace is used to prevent the contamination of compounded sterile products. HEPA filter - High Efficiency Particulate Air filter removes 99 99.97% 97% of all air particles 0.3 0 3 mm or larger

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Laminar Air Flow Hoods (cont.)

HEPA Filter in Frame

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Laminar Air Flow Hoods (cont.)

Horizontal Flow ( (Laminar Flow Hood) ) Air blows towards worker Vertical Flow (Biological Safety Cabinet or Chemotherapy Hood) Air blows from top down to maintain sterility and protect the worker

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Laminar Air Flow Hoods (cont.)

The HEPA filter is located in the fragile mesh between thin metal strips at the back of the hood behind the HEPA filter screen Nothing should be permitted to come in contact with the HEPA filter NO cleaning solution NO aspirate i t from f syringes i NO glass from ampules NO fluids fluids, even if sterile DO NOT touch HEPA filter

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Laminar Air Flow Hoods (cont.)

Only products essential to product preparation should be placed in the laminar flow hood to minimize the potential for contamination

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Laminar Air Flow Hoods (cont.)

Eating, g, drinking, g, and smoking g is always y prohibited Talking or coughing should be directed away f from the th hood h dt to minimize i i i air i fl flow t turbulence b l A mask covering mouth and nose must be worn while working in the hood The use of a laminar flow hood alone without the observance of aseptic technique, cannot insure product d t sterility t ilit

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Autoclaves Safe and Effective Use

Principles p of Steam Sterilization Sterilization: the complete p destruction of all forms of microbial life, including bacterial spores Steam under pressure reaches high temperatures
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Principles of Steam Sterilization (cont.)

The killing killing power power of steam is due to its latent heat of vaporization 80 calories: 1 L water to boiling g 540 calories: 1 L boiling water to steam

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Action of Steam

Steam contacts a cooler surface, surface condenses, causing a huge decrease in volume and setting g up p a negative g pressure that draws more steam Condensation occurs as long g as there is a temperature differential Action of steam ensures: Surface heating, penetration, and protein coagulation
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Standard Temperature and Pressure

250 F F (121 C) C) and 15 p.s.i. Berapa atm ?

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How Long to Autoclave

Dry yg goods @ 121 C

Time ( (min) ) 15 30 30

Glassware, empty, inverted Instruments, wrapped Utensils, wrapped

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Special Situations
Media boils out of bottles differs among media Very dense or closed dry objects beaker full of microfuge tubes with aluminum foil covering

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