General Microbiology (BCH2232) Project
Name / Student ID: Alvinder Kaur (PC090728605) Fabian Mok (PC090726528) Ngai Kok Leong (PC090727319) Lee Jia Yin (PC090727720) Hii Khang Jiet (PC080725452) Validation of Sterilization Process (Gas and Chemical Sterilization) 20th August 2010 3rd October 2010.

Experiment 2: Date of Project: Date of Submission:

contained in a fluid. The formaldehyde is widely use in pathology and surgery department. preventive maintenance. viruses. The mixture of steam and formaldehyde at temperature of 80. or for contact with broken skin. Liquid sterility and high disinfectants typically include oxidizing agents such as Hydrogen Peroxide. where the treat of infection exists. Validation on the sterilization methods must be conducted. calibration. and many plastics which would be damage if high-pressure steam of dry-heat steam sterilization is apply. Sterilization of microbiological material. The product achieved is called sterile product and the method employed to attain sterility is termed sterilization. spore forms that is present on a surface.Chemical sterilization also known cold sterilization is an alternative to high-pressure steam or dry-heat sterilization. For sterilization process to be effective. Validation of sterilization method is a very reliable way to secure the effectiveness of pharmaceutical products. Chemical sterilization and Gas sterilization are the few method of sterilization of pharmaceutical product that is recognizes by British Pharmacopoeia (2010). Glutaraldehyde and formaldehyde are the most common disinfectants for chemical sterilization used in practical sterility for some instruments. bacteria. mucosal surface or internal organ.10% v/v) of very reactive chemical and oxidizing agents such as hydrogen peroxide and ozone. such as laparoscopes which cannot be heated. fiber optics. Peracetic Acid and Aldehydes such as Glutaraldehyde and more recently o-phthalaldehyde.Sterilization is a term referring to any process that eliminates or kills all forms of life. . cycle review and approval and annual reviews are activities that make up validation of sterilization methods. electronics. soiled dressing and other contaminated items is necessary to minimize the health hazard associated with these articles. or in a compound such as biological culture media.. including transmissible agents such as fungi. 60. Sterility is the complete absence of viable microorganism from a product. routine bioburden monitoring. Chemical sterilization is used to the heat sensitive materials such as biological materials. in medication. Low temperature Chemical sterilizers function by exposing the articles to be sterilized to high concentrations (typically 5 . 65. 55 or 50°C will penetrate and kill any microorganisms. In the case of sterilization processes. Sterilization is an essential stage in the processing of any product destined for parental administration. Validation of sterilization methods has proven to be vital and most useful in insuring safety of the product outcome.

eyes and respiratory tract. the usage of ethylene oxide is far more common than the usage of formaldehyde. hydroxyl and carbonyl groups on proteins and amino groups of nucleic acids. . conjunctiva and nasal mucosa. The disadvantages for both solutions can cause skin irritation. as are those protected from the gas by inclusion in crystalline or dried organic deposits. Organisms are more resistant to ethylene oxide in a dried state. medical.The advantages of glutaraldehydes and formaldehyde solutions are not readily inactivated by organic materials and used for item that will not tolerate heat sterilization such as laparoscopes and some heat sensitive materials. and to lesser degree glutaraldehydes are irritating to skin. therefore. amino. it’s potentially mutagenic and carcinogenic level and acute toxicity including skin irritation. Ethylene oxide treatment is also considered a good alternative to radiation sterilization in the production of medical devices. therefore a minimum level of 30%-70% humidity in the immediate product environment is wanted. chemically reactive gases like ethylene oxide and formaldehyde possess broad spectrum biocidal activity in many industries . diagnostic and electrical equipment as well as the surface sterilization of powders.These include the sterilization of re-usable surgical instruments.Both glutaraldehydes and formaldehyde require special handling and leave a residue on treated instruments. these two techniques differ in the degree of sterility assurance as heat methods so they are usually reserved for heat sensitive items. The mechanism of antimicrobial action of ethylene oxide and formaldehyde is assumed to be through the alkylation of sulphudryl.75oC . The vapors from formaldehyde (classified a potential carcinogen). Concentration ranges (given as weight of gas per unit chamber volume) are usually in order of 800-1200mg-1 for ethylene oxide with operating temperatures of 45oC-63oC while 5-100mg-1 for formaldehyde with operating temperatures of 70oC. On a regular basis. The disadvantages of Ethylene oxide and formaldehyde gases is. A major disadvantage of formaldehyde is low penetrating power and this limits the packaging materials that can be employed to principally paper and cotton fabric. In Gas sterilization. However. rinsing with sterile water is essential if the item must be kept sterile.

reliability and consistency of the sterilization method. Accuracy describes the closeness of mean test result obtain from the sterilization method. therefore Limit of Detection (LOD) parameter is important to produce the lowest number of micro-organisms that the methods are able to detect in a given sample. For a new method that is being validated. Consequently.Result from validation can be used to judge the quality. This is done before the validation of the actual method. The robustness of the sterilization technique should remain unaffected when being subjected towards an operational and environmental variable. Precision of the sterilization method can be judge by repeating the method under same condition. Validation on the qualitative test for the presence or absence of microorganism can be done to prove the efficacy of the sterilization methods . There are few fundamental parameter of validation that requires being access to prove the reliability of the sterilization method. Accuracy of the sterilization method can be determine by running the alternative method analysis and then is compare with the pharmacopoeia method. Both methods should give an equivalence results and this conclude that the alternative route is acceptable. Mean value of the accuracy should be within 15% of its actual value. Sterilization cannot be governed by complete absence of microorganism. Both the method should give a degree of equivalence.The basic definition of a validation is to formulate a documented proved that a method being used is able to give a reliable results and the consistency is being achieved. This is to validate the suitability and applicability of the new method. This indicates the reliability of the sterilization methods. Other parameter under observation is the linearity where the results obtained from the alternative methods should be proportional towards the concentration of micro-organisms from the sample being used. A validation test is done in to an alternative route to prove that it is acceptable. the value obtained should fall into a agreed range. is it crucial to conceive and elucidate the methods to achieve a certain results which are being concerned. This is governed by the variance. standard deviation and coefficient of variation. The primary validation is done on a summary of new alternative methods which suppose to be validated. A sterilization method is done to kill microorganism to produce sterile product. it consists of two levels. This validation is then matching towards the pharmacopoeia methods. .

Biological indicator is common being used for medical devices. The carrier used should be shield by any chance of contamination or degradation at the same time allowing the contact between the sterilizing agent and the micro-organism. The microorganism used as the indicator for the gaseous sterilization is added directly towards the gas being sterilized. Hence. The micro-organism being used employs a population of spores that is placed on an inert carrier. it could also be administered into the gas which mimics the one which is in concern to be sterilized. Validation of the Chemical and Gaseous Sterilization are conducted to confirm Chemical and Gas Sterilization process is evaluated and under control.Minimum Bactericidal Concentration(MBC) and the extinction time of the microorganism. isolators. they are selected to test the sterilization procedure of Chemical sterilization. Hence. The resistance of microorganism is determine by the Minimum Inhibitory Concentration(MIC) . This microorganism is chosen based on its defined and stable resistance towards Glutaraldehyde and formaldehyde. Sterilization of Ethylene Oxide a Gas sterilizer is being validated with the spores of Bacillus subtilis. By using a sequence of reliable Biological.The Process of Validation in Chemical Sterilization and Gaseous Sterilization for Pharmaceuticals products. This micro-organism is chosen based on its defined and stable resistance towards ethylene oxide. chambers. Chemical and Mechanical Indicators at regular intervals. Sterilization of Glutaraldehyde and Formaldehyde as Chemical Sterilizer is being validated with the spores of Bacillus licheniformis. . they are selected to test the sterilization procedures. However. Besides that. Chemical and Gaseous sterilization is mainly used with the aim of discharging the preliminary treatment of bioburden which further accommodating an adequate safety margin. This biological indicator is packed in a way that the sterilizing agent is readily penetrated into it. A standardized preparation of chosen micro-organism is used to assay the efficiency of a sterilization process that is conducted. etc. the sterilization procedures are able to be monitored. it is crucial that the gas product produce no inhibiting effect towards the micro-organism used. This is done by exposing the components of concern towards a chemical sterility which present in gaseous phase for gaseous sterilization and in liquid phase for chemical sterilization.

Other method of validation of Chemical and Gas sterilization include Mechanical indicator. dirt. moisture. This includes heat sensitive tape. A sterilization process should always be considered to compromise between achieving good antimicrobial activity and maintaining product stability.formaldehyde in solution in chemical sterilization and ethylene oxide solution in Gas sterilization meets the manufacturer's recommendation for a minimum effective concentration (MEC) needed to effectively kill organisms. Sterilization processes must be revalidated at least annually in the absence of any changedriven revalidation. Some acceptable approaches include the use of single runs rather than 3 consecutive runs are sufficient in the absence of recurring problems. temperature and pressure for that specific sterilization procedure. labels or glass vials containing pellets. Sterilization control and sterilization assurances are comprise an attempt to achieve as far as .Other method of Validation of Chemical and Gas sterilization include the use Chemical indicator. computer printouts that evaluate the sterilizer’s function by providing an apparent record of the time. It is carryout outside the sterilizer such as observing gauges. microorganism. animal and insects. Chemical monitors that indicate if a glutaraldehyde . It therefore must be validated against a suitable test organism and its efficacy is continuously monitored during use.the revalidation of the worst case loading pattern . in a separate enclosed area with limited excess that is used to store sterile and clean patient care supplies. Solution should be tested prior to immersing into medical devices into the solution to ensure that the solution is at its Minimum concentration (MEC). Chemical indicator are used for equipment control as a way to find out whether or not the solution is doing its job or is at the minimum concentration (MEC) needed to effectively kill organisms. This storage area is best located next to or connected to where sterilization occurs. The revalidation can be a subset of the original validation work. Mechanical monitoring may be the first indicator to show that something has gone awry. revalidation of each type of cycle . The entire sterile item should be stored in an area and manner whereby the packs or containers will be protected from dust. the selection of a worst case load pattern for revalidation . This validation is crucial to show something has gone inappropriate.

cn/v29240/usp29nf24s0_c1035.com/.pharmacopeia.cn/v29240/usp29nf24s0_c1211. Reference : http://findarticles. however it is an additional check and continued compliance with the test does give confidence as to the efficacy of a sterilization or aseptic process..com/p/articles/mi_m0BPC/is_1_29/ai_n8708450/ http://www.microtestlabs.org/ip/instrum/inm12..html .html#usp29nf24s0_c1211 www. The sterility test on its own provide no guarantee.engenderhealth.html#usp29nf24s0_c1035 http://www.pharmacopeia./steam-eo-chemical-sterilization.possible the continuous monitoring of a particular sterilization process.html www.

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