laf dan autoclave_fix

Theoretical Principles and Practical Skills of Aseptic Manipulations

Maulita Cut Nuria, M.Sc., Apt

Page 1

Definitions

Background

Laminar Air Flow Hoods

Aseptic Technique

Free Powerpoint TemplatesPage 2

Definitions

• 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. intravenous, intramuscular, subcutaneous)

• Aseptic Technique

– The technique for manipulations of compoundedsterile products and parenteral preparations thatprevents contamination

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Definitions (cont.)

• Contamination – any effect or action that has a negativeimpact on a product's integrity making it unfit for use

– Chemical composition

– pH

– Sterility (e.g. microorganism contamination)

– Pyrogenicity

– Biological or therapeutic potency

– Physical appearance

– Particulate matter (e.g. dust, glass or precipitation)

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Page 5

Background

Sources of product contamination

• People (most common)

– Touch contamination

– Generation of particulates from shedding cells or hair

• Supply air

– Heating Ventilation and Air Conditioning (HVAC)

• Infiltration

– Particles from adjacent spaces

• Internal generation

– Walls, floors, ceilings, packaging, equipment

Laminar Air Flow Hoods

• The underlying principle of a laminar air flowhood is that a constant flow of HEPA filtered airat a rate of approximately 90 linear feet perminute physically sweeps the work area andprevents the entry of contaminated air

• The hood workspace is used to prevent thecontamination of compounded sterile products.

• HEPA filter - High Efficiency Particulate Air filterremoves 99.97% of all air particles 0.3 mm orlarger

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Laminar Air Flow Hoods (cont.)

HEPA Filter in Frame

Page 7

Laminar Air Flow Hoods (cont.)

• Horizontal Flow (Laminar Flow Hood)

– Air blows towards worker

• Vertical Flow (Biological Safety Cabinet orChemotherapy Hood)

– Air blows from top down to maintain sterility andprotect the worker

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Laminar Air Flow Hoods (cont.)

• The HEPA filter is located in the fragile mesh betweenthin metal strips at the back of the hood behind theHEPA filter screen

• Nothing should be permitted to come in contact with theHEPA filter

– NO cleaning solution

– NO aspirate from syringes

– NO glass from ampules

– NO fluids, even if sterile

– DO NOT touch HEPA filter

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Laminar Air Flow Hoods (cont.)

• Only products essential to product preparationshould be placed in the laminar flow hood tominimize the potential for contamination

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No Paper, Pens, Calculators, or Labels in the Hood

Laminar Air Flow Hoods (cont.)

• Eating, drinking, and smoking is alwaysprohibited

• Talking or coughing should be directed awayfrom the hood to minimize air flow turbulence

• A mask covering mouth and nose must be wornwhile working in the hood

• The use of a laminar flow hood alone without theobservance of aseptic technique, cannot insureproduct sterility

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AutoclavesSafe and Effective Use

Principles of Steam Sterilization

Sterilization: the complete destruction of allforms of microbial life, including bacterial spores

Steam under pressure reaches hightemperatures

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Principles of Steam Sterilization (cont.)

The “killing power” of steam is due to its latentheat of vaporization

– 80 calories: 1 L water to boilingg

– 540 calories: 1 L boiling water to steam

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Action of Steam

Steam contacts a cooler surface,condenses, causing a huge decrease involume and setting up a negativepressure that draws more steam

Condensation occurs as longg as there isa temperature differential

Action of steam ensures: Surfaceheating, penetration, and proteincoagulation

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Standard Temperature and Pressure

250 °F (121 °C) and 15 p.s.i.

Berapa atm ?

How Long to Autoclave

Dry goods @ 121 °C

Glassware, empty, inverted

Instruments, wrapped

Utensils, wrapped

Time (min)

15

30

30

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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 aluminumfoil covering

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TERIMAKASIH

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