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Guideline for Disinfection and Sterilization in Healthcare Facilities, 2008

Guideline for Disinfection and Sterilization in Healthcare Facilities, 2008 [PDF - 948 KB]

Formaldehyde

Overview.. Formaldehyde is used as a disinfectant and sterilant in both its liquid and gaseous states. Liquid formaldehyde will be considered briefly in this section, and the gaseous form is reviewed elsewhere 570. Formaldehyde is sold and used principally as a water-based solution called formalin, which is 37% formaldehyde by weight. The aqueous solution is a bactericide, tuberculocide, fungicide, virucide and sporicide 72, 82, 571-573. OSHA indicated that formaldehyde should be handled in the workplace as a potential carcinogen and set an employee exposure standard for formaldehyde that limits an 8-hour time-weighted average exposure concentration of 0.75 ppm 574, 575. The standard includes a second permissible exposure limit in the form of a short-term exposure limit (STEL) of 2 ppm that is the maximum exposure allowed during a 15-minute period 576. Ingestion of formaldehyde can be fatal, and long-term exposure to low levels in the air or on the skin can cause asthma-like respiratory problems and skin irritation, such as dermatitis and itching. For these reasons, employees should have limited direct contact with formaldehyde, and these considerations limit its role in sterilization and disinfection processes. Key provisions of the OSHA standard that protects workers from exposure to formaldehyde appear in Title 29 of the Code of Federal Regulations (CFR) Part 1910.1048 (and equivalent regulations in states with OSHA-approved state plans) 577

Mode of Action. Formaldehyde inactivates microorganisms by alkylating the amino and sulfhydral groups of proteins and ring nitrogen atoms of purine bases 376.

Microbicidal Activity. Varying concentrations of aqueous formaldehyde solutions destroy a wide range of microorganisms. Inactivation of poliovirus in 10 minutes required an 8% concentration of formalin, but all other viruses tested were inactivated with 2% formalin 72. Four percent formaldehyde is a tuberculocidal agent, inactivating 104 M. tuberculosis in 2 minutes 82, and 2.5% formaldehyde inactivated about 107 Salmonella Typhi in 10 minutes in the presence of organic matter 572. The sporicidal action of formaldehyde was slower than that of glutaraldehyde in comparative tests with 4% aqueous formaldehyde and 2% glutaraldehyde against the spores of B. anthracis 82. The formaldehyde solution required 2 hours of contact to achieve an inactivation factor of 104, whereas glutaraldehyde required only 15 minutes.

Uses. Although formaldehyde-alcohol is a chemical sterilant and formaldehyde is a high-level disinfectant, the health-care uses of formaldehyde are limited by its irritating fumes and its pungent odor even at very low levels (<1 ppm). For these reasons and others—such as its role as a suspected human carcinogen linked to nasal cancer and lung cancer 578, this germicide is excluded from Table 1. When it is used, , direct exposure to employees generally is limited; however, excessive exposures to formaldehyde have been documented for employees of renal transplant units 574, 579, and students in a gross anatomy laboratory 580. Formaldehyde is used in the health-care setting to prepare viral vaccines (e.g., poliovirus and influenza); as an embalming agent; and to preserve anatomic specimens; and historically has been used to sterilize surgical instruments, especially when mixed with ethanol. A 1997 survey found that formaldehyde was used for reprocessing hemodialyzers by 34% of U.S. hemodialysis centers—a 60% decrease from 1983 249, 581. If used at room temperature, a concentration of 4% with a minimum exposure of 24 hours is required to disinfect disposable hemodialyzers reused on the same patient 582, 583. Aqueous formaldehyde solutions (1%–2%) also have been used to disinfect the internal fluid pathways of dialysis machines 583. To minimize a potential health hazard to dialysis patients, the dialysis equipment must be thoroughly rinsed and tested for residual formaldehyde before use.

Paraformaldehyde, a solid polymer of formaldehyde, can be vaporized by heat for the gaseous decontamination of laminar flow biologic safety cabinets when maintenance work or filter changes require access to the sealed portion of the cabinet.

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Glutaraldehyde

Overview. Glutaraldehyde is a saturated dialdehyde that has gained wide acceptance as a high-level disinfectant and chemical sterilant 107. Aqueous solutions of glutaraldehyde are acidic and generally in this state are not sporicidal. Only when the solution is "activated" (made alkaline) by use of alkalinating agents to pH 7.5–8.5 does the solution become sporicidal. Once activated, these solutions have a shelf-life of minimally 14 days because of the polymerization of the glutaraldehyde molecules at alkaline pH levels. This polymerization blocks the active sites (aldehyde groups) of the glutaraldehyde molecules that are responsible for its biocidal activity.

Novel glutaraldehyde formulations (e.g., glutaraldehyde-phenol-sodium phenate, potentiated acid glutaraldehyde, stabilized alkaline glutaraldehyde) produced in the past 30 years have overcome the problem of rapid loss of activity (e.g., use-life 28–30 days) while generally maintaining excellent microbicidal activity 584-588. However, antimicrobial activity depends not only on age but also on use conditions, such as dilution and organic stress. Manufacturers' literature for these preparations suggests the neutral or alkaline glutaraldehydes possess microbicidal and anticorrosion properties superior to those of acid glutaraldehydes, and a few published reports substantiate these claims 542, 589, 590. However, two studies found no difference in the microbicidal activity of alkaline and acid glutaraldehydes 73, 591. The use of glutaraldehyde-based solutions in health-care facilities is widespread because of their advantages, including excellent biocidal properties; activity in the presence of organic matter (20% bovine serum); and noncorrosive action to endoscopic equipment, thermometers, rubber, or plastic equipment (Tables 4 and 5).

Mode of Action.The biocidal activity of glutaraldehyde results from its alkylation of sulfhydryl, hydroxyl, carboxyl, and amino groups of microorganisms, which alters RNA, DNA, and protein synthesis. The mechanism of action of glutaraldehydes are reviewed extensively elsewhere 592, 593.

Microbicidal Activity. The in vitro inactivation of microorganisms by glutaraldehydes has been extensively investigated and reviewed 592, 593. Several investigators showed that >2% aqueous solutions of glutaraldehyde, buffered to pH 7.5–8.5 with sodium bicarbonate effectively killed vegetative bacteria in <2 minutes; M. tuberculosis, fungi, and viruses in <10 minutes; and spores of Bacillus and Clostridium species in 3 hours 542, 592-597. Spores of C. difficile are more rapidly killed by 2% glutaraldehyde than are spores of other species of Clostridium and Bacillus 79, 265, 266. Microorganisms with substantial resistance to glutaraldehyde have been reported, including some mycobacteria (M. chelonae, Mycobacterium avium-intracellulare, M. xenopi) 598-601, Methylobacterium mesophilicum 602, Trichosporon, fungal ascospores (e.g., Microascus cinereus, Cheatomium globosum), and Cryptosporidium 271, 603. M. chelonae persisted in a 0.2% glutaraldehyde solution used to store porcine prosthetic heart valves 604.

Two percent alkaline glutaraldehyde solution inactivated 105 M. tuberculosis cells on the surface of penicylinders within 5 minutes at 18ºC 589. However, subsequent studies82 questioned the mycobactericidal prowess of glutaraldehydes. Two percent alkaline glutaraldehyde has slow action (20 to >30 minutes) against M. tuberculosis and compares unfavorably with alcohols, formaldehydes, iodine, and phenol 82. Suspensions of M. avium, M. intracellulare, and M. gordonae were more resistant to inactivation by a 2% alkaline glutaraldehyde (estimated time to complete inactivation: ~60 minutes) than were virulent M. tuberculosis (estimated time to complete inactivation ~25 minutes) 605. The rate of kill was directly proportional to the temperature, and a standardized suspension of M. tuberculosis could not be sterilized within 10 minutes 84. An FDA-cleared chemical sterilant containing 2.5% glutaraldehyde uses increased temperature (35ºC) to reduce the time required to achieve high-level disinfection (5 minutes) 85, 606, but its use is limited to automatic endoscope reprocessors equipped with a heater. In another study employing membrane filters for measurement of mycobactericidal activity of 2% alkaline glutaraldehyde, complete inactivation was achieved within 20 minutes at 20ºC when the test inoculum was 106 M. tuberculosis per membrane 81. Several investigators 55, 57, 73, 76, 80, 81, 84, 605 have demonstrated that glutaraldehyde solutions inactivate 2.4 to >5.0 log10 of M. tuberculosis in 10 minutes (including multidrug-resistant M. tuberculosis) and 4.0–6.4 log10 of M. tuberculosis in 20 minutes. On the basis of these data and other studies, 20 minutes at room temperature is considered the minimum exposure time needed to reliably kill Mycobacteria and other vegetative bacteria with >2% glutaraldehyde 17, 19, 27, 57, 83, 94, 108, 111, 117-121, 607.

Glutaraldehyde is commonly diluted during use, and studies showed a glutaraldehyde concentration decline after a few days of use in an automatic endoscope washer 608, 609. The decline occurs because instruments are not thoroughly dried and water is carried in with the instrument, which increases the solution's volume and dilutes its effective concentration 610. This emphasizes the need to ensure that semicritical equipment is disinfected with an acceptable concentration of glutaraldehyde. Data suggest that 1.0%–1.5% glutaraldehyde is the minimum effective concentration for >2% glutaraldehyde solutions when used as a high-level disinfectant 76, 589, 590, 609. Chemical test strips or liquid chemical monitors 610, 611 are available for determining whether an effective concentration of glutaraldehyde is present despite repeated use and dilution. The frequency of testing should be based on how frequently the solutions are used (e.g., used daily, test daily; used weekly, test before use; used 30 times per day, test each 10th use), but the strips should not be used to extend the use life beyond the expiration date. Data suggest the chemicals in the test strip deteriorate with time 612 and a manufacturer's expiration date should be placed on the bottles. The bottle of test strips should be dated when opened and used for the period of time indicated on the bottle (e.g., 120 days). The results of test strip monitoring should be documented. The glutaraldehyde test kits have been preliminarily evaluated for accuracy and range 612 but the reliability has been questioned 613. To ensure the presence of minimum effective concentration of the high-level disinfectant, manufacturers of some chemical test strips recommend the use of quality-control procedures to ensure the strips perform properly. If the manufacturer of the chemical test strip recommends a quality-control procedure, users should comply with the manufacturer's recommendations. The concentration should be considered unacceptable or unsafe when the test indicates a dilution below the product's minimum effective concentration (MEC) (generally to <1.0%–1.5% glutaraldehyde) by the indicator not changing color.

A 2.0% glutaraldehyde–7.05% phenol–1.20% sodium phenate product that contained 0.125% glutaraldehyde–0.44% phenol–0.075% sodium phenate when diluted 1:16 is not recommended as a high-level disinfectant because it lacks bactericidal activity in the presence of organic matter and lacks tuberculocidal, fungicidal, virucidal, and sporicidal activity 49, 55, 56, 71, 73-79, 614. In December 1991, EPA issued an order to stop the sale of all batches of this product because of efficacy data showing the product is not effective against spores and possibly other microorganisms or inanimate objects as claimed on the label 615. FDA has cleared a glutaraldehyde–phenol/phenate concentrate as a high-level disinfectant that contains 1.12% glutaraldehyde with 1.93% phenol/phenate at its use concentration. Other FDA cleared glutaraldehyde sterilants that contain 2.4%–3.4% glutaraldehyde are used undiluted 606.

Uses. Glutaraldehyde is used most commonly as a high-level disinfectant for medical equipment such as endoscopes 69, 107, 504, spirometry tubing, dialyzers 616, transducers, anesthesia and respiratory therapy equipment 617, hemodialysis proportioning and dialysate delivery systems 249, 618, and reuse of laparoscopic disposable plastic trocars 619. Glutaraldehyde is noncorrosive to metal and does not damage lensed instruments, rubber. or plastics. Glutaraldehyde should not be used for cleaning noncritical surfaces because it is too toxic and expensive.

Colitis believed caused by glutaraldehyde exposure from residual disinfecting solution in endoscope solution channels has been reported and is preventable by careful endoscope rinsing 318, 620-630. One study found that residual glutaraldehyde levels were higher and more variable after manual disinfection (<0.2 mg/L to 159.5 mg/L) than after automatic disinfection (0.2–6.3 mg/L)631. Similarly, keratopathy and corneal decompensation were caused by ophthalmic instruments that were inadequately rinsed after soaking in 2% glutaraldehyde 632, 633.

Healthcare personnel can be exposed to elevated levels of glutaraldehyde vapor when equipment is processed in poorly ventilated rooms, when spills occur, when glutaraldehyde solutions are activated or changed, 634, or when open immersion baths are used. Acute or chronic exposure can result in skin irritation or dermatitis, mucous membrane irritation (eye, nose, mouth), or pulmonary symptoms 318, 635-639. Epistaxis, allergic contact dermatitis, asthma, and rhinitis also have been reported in healthcare workers exposed to glutaraldehyde 636, 640-647.

Glutaraldehyde exposure should be monitored to ensure a safe work environment. Testing can be done by four techniques: a silica gel tube/gas chromatography with a flame ionization detector, dinitrophenylhydrazine (DNPH)-impregnated filter cassette/high-performance liquid chromatography (HPLC) with an ultraviolet (UV) detector, a passive badge/HPLC, or a handheld glutaraldehyde air monitor 648. The silica gel tube and the DNPH-impregnated cassette are suitable for monitoring the 0.05 ppm ceiling limit. The passive badge, with a 0.02 ppm limit of detection, is considered marginal at the Americal Council of Governmental Industrial Hygienists (ACGIH) ceiling level. The ceiling level is considered too close to the glutaraldehyde meter's 0.03 ppm limit of detection to provide confidence in the readings 648. ACGIH does not require a specific monitoring schedule for glutaraldehyde; however, a monitoring schedule is needed to ensure the level is less than the ceiling limit. For example, monitoring should be done initially to determine glutaraldehyde levels, after procedural or equipment changes, and in response to worker complaints 649. In the absence of an OSHA permissible exposure limit, if the glutaraldehyde level is higher than the ACGIH ceiling limit of 0.05 ppm, corrective action and repeat monitoring would be prudent 649.

Engineering and work-practice controls that can be used to resolve these problems include ducted exhaust hoods, air systems that provide 7–15 air exchanges per hour, ductless fume hoods with absorbents for the glutaraldehyde vapor, tight-fitting lids on immersion baths, personal protection (e.g., nitrile or butyl rubber gloves but not natural latex gloves, goggles) to minimize skin or mucous membrane contact, and automated endoscope processors 7, 650. If engineering controls fail to maintain levels below the ceiling limit, institutions can consider the use of respirators (e.g., a half-face respirator with organic vapor cartridge 640 or a type "C" supplied air respirator with a full facepiece operated in a positive pressure mode) 651. In general, engineering controls are preferred over work-practice and administrative controls because they do not require active participation by the health-care worker. Even though enforcement of the OSHA ceiling limit was suspended in 1993 by the U.S. Court of Appeals 577, limiting employee exposure to 0.05 ppm (according to ACGIH) is prudent because, at this level, glutaraldehyde can irritate the eyes, throat, and nose 318, 577, 639, 652. If glutaraldehyde disposal through the sanitary sewer system is restricted, sodium bisulfate can be used to neutralize the glutaraldehyde and make it safe for disposal.

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Hydrogen Peroxide

Overview. The literature contains several accounts of the properties, germicidal effectiveness, and potential uses for stabilized hydrogen peroxide in the health-care setting. Published reports ascribe good germicidal activity to hydrogen peroxide and attest to its bactericidal, virucidal, sporicidal, and fungicidal properties 653-655. (Tables 4 and 5) The FDA website lists cleared liquid chemical sterilants and high-level disinfectants containing hydrogen peroxide and their cleared contact conditions.

Mode of Action.Hydrogen peroxide works by producing destructive hydroxyl free radicals that can attack membrane lipids, DNA, and other essential cell components. Catalase, produced by aerobic organisms and facultative anaerobes that possess cytochrome systems, can protect cells from metabolically produced hydrogen peroxide by degrading hydrogen peroxide to water and oxygen. This defense is overwhelmed by the concentrations used for disinfection 653, 654.

Microbicidal Activity. Hydrogen peroxide is active against a wide range of microorganisms, including bacteria, yeasts, fungi, viruses, and spores 78, 654. A 0.5% accelerated hydrogen peroxide demonstrated bactericidal and virucidal activity in 1 minute and mycobactericidal and fungicidal activity in 5 minutes 656. Bactericidal effectiveness and stability of hydrogen peroxide in urine has been demonstrated against a variety of health-care–associated pathogens; organisms with high cellular catalase activity (e.g., S. aureus, S. marcescens, and Proteus mirabilis) required 30–60 minutes of exposure to 0.6% hydrogen peroxide for a 108 reduction in cell counts, whereas organisms with lower catalase activity (e.g., E. coli, Streptococcus species, and Pseudomonas species) required only 15 minutes' exposure 657. In an investigation of 3%, 10%, and 15% hydrogen peroxide for reducing spacecraft bacterial populations, a complete kill of 106 spores (i.e., Bacillus species) occurred with a 10% concentration and a 60-minute exposure time. A 3% concentration for 150 minutes killed 106 spores in six of seven exposure trials 658. A 10% hydrogen peroxide solution resulted in a 103 decrease in B. atrophaeus spores, and a >105 decrease when tested against 13 other pathogens in 30 minutes at 20ºC 659, 660. A 3.0% hydrogen peroxide solution was ineffective against VRE after 3 and 10 minutes exposure times 661 and caused only a 2-log10 reduction in the number of Acanthamoeba cysts in approximately 2 hours 662. A 7% stabilized hydrogen peroxide proved to be sporicidal (6 hours of exposure), mycobactericidal (20 minutes), fungicidal (5 minutes) at full strength, virucidal (5 minutes) and bactericidal (3 minutes) at a 1:16 dilution when a quantitative carrier test was used 655. The 7% solution of hydrogen peroxide, tested after 14 days of stress (in the form of germ-loaded carriers and respiratory therapy equipment), was sporicidal (>7 log10 reduction in 6 hours), mycobactericidal (>6.5 log10 reduction in 25 minutes), fungicidal (>5 log10 reduction in 20 minutes), bactericidal (>6 log10 reduction in 5 minutes) and virucidal (5 log10 reduction in 5 minutes) 663. Synergistic sporicidal effects were observed when spores were exposed to a combination of hydrogen peroxide (5.9%–23.6%) and peracetic acid 664. Other studies demonstrated the antiviral activity of hydrogen peroxide against rhinovirus 665. The time required for inactivating three serotypes of rhinovirus using a 3% hydrogen peroxide solution was 6–8 minutes; this time increased with decreasing concentrations (18-20 minutes at 1.5%, 50–60 minutes at 0.75%).

Concentrations of hydrogen peroxide from 6% to 25% show promise as chemical sterilants. The product marketed as a sterilant is a premixed, ready-to-use chemical that contains 7.5% hydrogen peroxide and 0.85% phosphoric acid (to maintain a low pH) 69. The mycobactericidal activity of 7.5% hydrogen peroxide has been corroborated in a study showing the inactivation of >105 multidrug-resistant M. tuberculosis after a 10-minute exposure 666. Thirty minutes were required for >99.9% inactivation of poliovirus and HAV 667. Three percent and 6% hydrogen peroxide were unable to inactivate HAV in 1 minute in a carrier test 58. When the effectiveness of 7.5% hydrogen peroxide at 10 minutes was compared with 2% alkaline glutaraldehyde at 20 minutes in manual disinfection of endoscopes, no significant difference in germicidal activity was observed 668. ). No complaints were received from the nursing or medical staff regarding odor or toxicity. In one study, 6% hydrogen peroxide (unused product was 7.5%) was more effective in the high-level disinfection of flexible endoscopes than was the 2% glutaraldehyde solution 456. A new, rapid-acting 13.4% hydrogen peroxide formulation (that is not yet FDA-cleared) has demonstrated sporicidal, mycobactericidal, fungicidal, and virucidal efficacy. Manufacturer data demonstrate that this solution sterilizes in 30 minutes and provides high-level disinfection in 5 minutes 669. This product has not been used long enough to evaluate material compatibility to endoscopes and other semicritical devices, and further assessment by instrument manufacturers is needed.

Under normal conditions, hydrogen peroxide is extremely stable when properly stored (e.g., in dark containers). The decomposition or loss of potency in small containers is less than 2% per year at ambient temperatures 670.

Uses. Commercially available 3% hydrogen peroxide is a stable and effective disinfectant when used on inanimate surfaces. It has been used in concentrations from 3% to 6% for disinfecting soft contact lenses (e.g., 3% for 2–3 hrs) 653, 671, 672, tonometer biprisms 513, ventilators 673, fabrics 397, and endoscopes 456. Hydrogen peroxide was effective in spot-disinfecting fabrics in patients' rooms 397. Corneal damage from a hydrogen peroxide-soaked tonometer tip that was not properly rinsed has been reported 674. Hydrogen peroxide also has been instilled into urinary drainage bags in an attempt to eliminate the bag as a source of bladder bacteriuria and environmental contamination 675. Although the instillation of hydrogen peroxide into the bag reduced microbial contamination of the bag, this procedure did not reduce the incidence of catheter-associated bacteriuria 675.

A chemical irritation resembling pseudomembranous colitis caused by either 3% hydrogen peroxide or a 2% glutaraldehyde has been reported 621. An epidemic of pseudomembrane-like enteritis and colitis in seven patients in a gastrointestinal endoscopy unit also has been associated with inadequate rinsing of 3% hydrogen peroxide from the endoscope 676.

As with other chemical sterilants, dilution of the hydrogen peroxide must be monitored by regularly testing the minimum effective concentration (i.e., 7.5%–6.0%). Compatibility testing by Olympus America of the 7.5% hydrogen peroxide found both cosmetic changes (e.g., discoloration of black anodized metal finishes) 69 and functional changes with the tested endoscopes (Olympus, written communication, October 15, 1999).

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