Sterilization () refers to any process that removes, kills, or deactivates all forms of life (particularly microorganisms such as fungi, bacteria, spores, and unicellular eukaryotic organisms) and other biological agents (such as prions or viruses) present in fluid or on a specific surface or object. Sterilization can be achieved through various means, including heat, chemicals, irradiation, high pressure, and filtration. Sterilization is distinct from disinfection, sanitization, and pasteurization, in that those methods reduce rather than eliminate all forms of life and biological agents present. After sterilization, fluid or an object is referred to as being sterile or aseptic.
Applications
Foods One of the first steps toward modernized sterilization was made by Nicolas Appert, who discovered that application of heat over a suitable period of time slowed the decay of foods and various liquids, preserving them for safe consumption for a longer time than was typical. In the early 1900's the use of baby bottle sterilization became an important tool in the battle of infant mortality. Canning of foods is an extension of the same principle and has helped to reduce food borne illness ("food poisoning"). Other methods of sterilizing foods include ultra-high temperature processing (which uses a shorter duration of heating), food irradiation, and high pressure (pascalization).
In the context of food, sterility typically refers to commercial sterility, defined as "the absence of microorganisms capable of growing in the food at normal non-refrigerated conditions at which the food is likely to be held during distribution and storage" according to the Codex Alimentarius. (see also 21 CFR 113, a definition focused on thermal processing)
Medicine and surgery
In general, surgical instruments and medications that enter an already aseptic part of the body (such as the bloodstream, or penetrating the skin) must be sterile. Examples of such instruments include scalpels, hypodermic needles, and artificial pacemakers. This is also essential in the manufacture of parenteral pharmaceuticals.
Preparation of injectable medications and intravenous solutions for fluid replacement therapy requires not only sterility but also well-designed containers to prevent entry of adventitious agents after initial product sterilization.
The sterilization of surgical instruments by boiling, dry heat, and flame treatment was introduced around 1883 by Octave Roche Simon Terrillon. The general introduction of steam sterilization into surgery occurred in 1886 by Ernst von Bergmann and Curt Schimmelbusch.Paul Diepgen, Heinz Goerke: Aschoff/Diepgen/Goerke: Kurze Übersichtstabelle zur Geschichte der Medizin. 7., neubearbeitete Auflage. Springer, Berlin/Göttingen/Heidelberg 1960, S. 50–51. In 1890 the use of an Arnold Steam Cooker for sterilization of medical instruments was published and demonstrated by Dr. Herbert E. Smith. Within a year modifications were made creating the Arnold Sterilizer as laboratories and physicians documented the success of steam sterilization.Most medical and surgical devices used in healthcare facilities are made of materials that are able to undergo steam sterilization. However, since 1950, there has been an increase in medical devices and instruments made of materials (e.g., plastics) that require low-temperature sterilization. Ethylene oxide gas has been used since the 1950s for heat- and moisture-sensitive medical devices. Within the past 15 years, a number of new, low-temperature sterilization systems (e.g., vaporized hydrogen peroxide, peracetic acid immersion, ozone) have been developed and are being used to sterilize medical devices.
Spacecraft There are strict international rules to protect the contamination of Solar System bodies from biological material from Earth. Standards vary depending on both the type of mission and its destination; the more likely a planet is considered to be habitable, the stricter the requirements are.
Many components of instruments used on spacecraft cannot withstand very high temperatures, so techniques not requiring excessive temperatures are used as tolerated, including heating to at least , chemical sterilization, oxidization, ultraviolet, and irradiation.
Quantification Sterilization aims to reduce or eliminate microorganisms and other potential pathogens present in a material. In laboratory and industrial practice, steam sterilization is commonly used for culture media, except when treating heat-labile components. Microbial death by heat generally follows first-order kinetics, which can be expressed as:
where is the number of viable microorganisms, is the sterilization time, and is the specific death rate constant. Integration yields:
where is the initial number of microorganisms.
The degree of sterilization is often quantified using the decimal reduction time, or D-value, defined as the time required to reduce by one logarithmic unit (i.e to ) or when initial population is reduced by one-tenth (). The surviving population after time t can then be expressed as:
The D-value is a function of sterilization conditions and varies with the type of microorganism, temperature, water activity, pH, etc.
As in any first-order reaction, the reaction rate increases with temperature due to an increase in the rate constant . For thermal destruction of microorganisms, the specific death rate remains constant at a constant temperature. The dependence of on temperature follows the Arrhenius relationship:
where is a constant known as Arrhenius factor, is the activation energy, is the gas constant, and is absolute temperature. Now, combining the first-order death kinetics with the Arrhenius equation results in the following expression for heat sterilization of a pure culture at constant temperature:
or,
Theoretically, the likelihood of the survival of an individual microorganism is never zero. To compensate for this, the overkill method is often used. Using the overkill method, sterilization is performed by sterilizing for longer than is required to kill the bioburden present on or in the item being sterilized. This provides a sterility assurance level (SAL) equal to the probability of a non-sterile unit.
For high-risk applications, such as medical devices and injections, a sterility assurance level of at least 10−6 is required by the United States of America Food and Drug Administration (FDA).
Heat
Steam
Steam sterilization, also known as moist heat sterilization, uses heated saturated steam under pressure to inactivate or kill microorganisms via denaturation of macromolecules, primarily proteins. This method is a faster process than dry heat sterilization. Steam sterilization is performed using an autoclave, sometimes called a converter or steam sterilizer. The object or liquid is placed in the autoclave chamber, which is then sealed and heated using pressurized steam to a temperature set point for a defined period of time. Steam sterilization cycles can be categorized as either pre-vacuum or gravity displacement. Gravity displacement cycles rely on the lower density of the injected steam to force cooler, denser air out of the chamber drain.Steam Sterilization | Disinfection & Sterilization Guidelines | Guidelines Library | Infection Control | CDC In comparison, pre-vacuum cycles create a vacuum in the chamber to remove cool dry air prior to injecting saturated steam, resulting in faster heating and shorter cycle times. Typical steam sterilization cycles are between 3 and 30 minutes at at , but adjustments may be made depending on the bioburden of the article being sterilized, its resistance (D-value) to steam sterilization, the article's heat tolerance, and the required sterility assurance level. Following the completion of a cycle, liquids in a pressurized autoclave must be cooled slowly to avoid boiling over when the pressure is released. This may be achieved by gradually depressurizing the sterilization chamber and allowing liquids to evaporate under a negative pressure, while cooling the contents.
Proper autoclave treatment will inactivate all resistant bacterial spores in addition to fungi, bacteria, and viruses, but is not expected to eliminate all prions, which vary in their heat resistance. For prion elimination, various recommendations state for 60 minutes or for at least 18 minutes. The 263K scrapie prion is inactivated relatively quickly by such sterilization procedures; however, other strains of scrapie and strains of Creutzfeldt-Jakob disease (CKD) and bovine spongiform encephalopathy (BSE) are more resistant. Using mice as test animals, one experiment showed that heating BSE positive brain tissue at for 18 minutes resulted in only a 2.5 log decrease in prion infectivity.
Most autoclaves have meters and charts that record or display information, particularly temperature and pressure as a function of time. The information is checked to ensure that the conditions required for sterilization have been met. Indicator tape is often placed on the packages of products prior to autoclaving, and some packaging incorporates indicators. The indicator changes color when exposed to steam, providing a visual confirmation.
Biological indicators can also be used to independently confirm autoclave performance. Simple biological indicator devices are commercially available, based on microbial spores. Most contain spores of the heat-resistant microbe Geobacillus stearothermophilus (formerly Bacillus stearothermophilus), which is extremely resistant to steam sterilization. Biological indicators may take the form of glass vials of spores and liquid media, or as spores on strips of paper inside glassine envelopes. These indicators are placed in locations where it is difficult for steam to reach to verify that steam is penetrating that area.
For autoclaving, cleaning is critical. Extraneous biological matter or grime may shield organisms from steam penetration. Proper cleaning can be achieved through physical scrubbing, sonication, ultrasound, or pulsed air.
Pressure cooking and canning is analogous to autoclaving, and when performed correctly renders food sterile.
To sterilize waste materials that are chiefly composed of liquid, a purpose-built effluent decontamination system can be utilized. These devices can function using a variety of sterilants, although using heat via steam is most common.
Dry
Dry heat was the first method of sterilization and is a longer process than moist heat sterilization. The destruction of microorganisms through the use of dry heat is a gradual phenomenon. With longer exposure to lethal temperatures, the number of killed microorganisms increases. Forced ventilation of hot air can be used to increase the rate at which heat is transferred to an organism and reduce the temperature and amount of time needed to achieve sterility. At higher temperatures, shorter exposure times are required to kill organisms. This can reduce heat-induced damage to food products. The standard setting for a hot air oven is at least two hours at . A rapid method heats air to for 6 minutes for unwrapped objects and 12 minutes for wrapped objects. Dry heat has the advantage that it can be used on powders and other heat-stable items that are adversely affected by steam (e.g., it does not cause rusting of steel objects).
Flaming