A brief description
Ozone has a very high energy, very unstable, at room temperature, normal pressure, the structure of the ozone is easy to change, and will soon decompose itself into oxygen and a single oxygen atom. The latter has a very strong oxidizing effect, the half-life of ozone is 22~25min. the raw material for generating ozone is mainly air and electricity. Ozone sterilization mechanism: (1) the role of the cell membrane, resulting in an increase in the permeability of the cell membrane, intracellular material outflow, so that the cell is inactive; (2) so that the cell activity is necessary for the enzyme inactivation; (3) the destruction of intracellular genetic material or to make it lose its function. Ozone kills viruses by directly destroying ribonucleic acid or deoxyribonucleic acid. While killing bacteria, mold microorganisms is the first role of ozone and cell membranes, so that the composition of the cell membrane received damage, resulting in metabolic disorders and inhibit their growth, ozone continues to penetrate the destruction of intramembranous tissues, until death. Different clean rooms can be based on its volume, the choice of appropriate ozone generator, the host set in the HVAC system for the supply and return of air ducts or purification of air-conditioning box at the back end of the intermediate filter in a suitable location, the power control system is placed in the machine room. Disinfection, close the new air inlet (or close about 90% of the new air inlet valve) and the return air to the external discharge valve, is the entire clean room air being disinfected through the purification system ducts to form an internal cycle. Daily power on 1 ~ 1.5h for air sterilization; alternative chemicals fumigation disinfection and sterilization of equipment surfaces need to be powered on 2 ~ 2.5h. 100,000 levels of clean area, ozone dosage concentration of 10MG/M3. ozone attenuation rate of 0.375. According to the size of the space dosage calculation: ozone generator output = volume * 10/0.375 according to the calculated ozone generator output, the basic Can reach 100,000 levels of clean area clean standards.
Two, the purpose
Preparation of the production process is mainly completed in the clean area, clean area with the effect of ozone sterilization directly affects the quality of the product, in order to confirm the reliability of the ozone sterilization, ozone sterilization instead of formaldehyde fumigation sterilization, to reduce the damage to the health of the staff, specially formulated for the validation of this verification program to verify. Validation process should be strictly in accordance with the contents of this program, if for special reasons need to change, should fill out the verification program change application and approval letter, reported to the verification team for approval.
Three, the principle of the process
The ozone gas will be led to the main air supply duct, the use of HVAC systems in the purification of the fan generated by the pressure of the air source, diffusion to the control of the entire clean area.
Four, validation plan
(1), clean area air disinfection method principle:
1.1 is the use of ozone generator to produce ozone, through the HVAC system unit of positive pressure air supply section and air supply pipe into the clean area, and then back to the unit through the return ducts, composed of a closed air circulation system, to control the disinfection process, the circulation time or airtight time, the disinfection process. Thus, a certain concentration of ozone gas in the specified time to kill microorganisms in the clean area.
1.2 In order to ensure that the disinfection effect, should ensure that air disinfection, HVAC system operation or disinfectant gas cycle, the outside world, fresh air shall not enter the HVAC system, in order to prevent the disinfectant gas concentration drop, not up to the required disinfection concentration.
(2), air sterilization mechanism and characteristics:
2.1 Ozone sterilization mechanism and characteristics:
2.1.1 Ozone at room temperature, atmospheric pressure, the molecular structure is not stable, and will soon decompose itself into oxygen and individual oxygen atoms, which has a strong activity, the latter has a very strong oxidation of bacteria, can be oxidized to break down the bacteria inside the oxidation of the enzymes essential for grapes, thus destroying the cell membrane. The latter has a strong oxidizing effect on bacteria and can oxidize the enzymes necessary for the oxidation of grapes inside the bacteria, thus destroying its cell membrane and killing it. The excess oxygen atoms will recombine themselves to become universal oxygen molecules, there is no harmful residue, so it is called non-polluting disinfectant;
2.1.2 Ozone is a broad-spectrum bactericidal agent that can kill bacterial propagules and spores, viruses, fungi, etc., and can destroy botulinum toxin. However, it is corrosive, ozone is a strong oxidizing agent, can damage a variety of items, the higher the concentration of items the more serious the damage, so in the use of ozone disinfection, should be easy to be oxidized or accelerated aging in the clean area of the items (such as plastics, rubber), products and so on for protection.
2.1.3 Ozone fumigation disinfection, high relative humidity is good, low is poor, almost no sterilization of dry bacteria.
2.1.4 Ozone concentration in the air up to 0.01-0.02mg / L when you can smell, the concentration reaches 1mg / L, can cause accelerated respiration, change, chest tightness and other symptoms, in the 2.5-5mg / L, can cause accelerated pulse rate, fatigue, headache, stay for 1 hour can occur emphysema, or even death, so the ozone disinfection, it is forbidden to have a person with a clean area in the operation.
2.1.5 Ozone stability is extremely poor, room temperature can be decomposed into oxygen, ozone generator stops generating, HVAC system continues to run ventilation 30-60 minutes, so that the ozone is completely decomposed, the personnel can enter the clean area to operate.
2.1.6 "Disinfection Technical Specification" stipulates that, for airborne bacteria, ozone sterilization concentration of 2-4PPM, for the object surface of the settlement of bacteria, ozone sterilization concentration of 10-15PPM, the decline rate of ozone 1H is about 62.25%. Should be based on the volume of space such as disinfection (V), the concentration of microorganisms required to kill (C) and the decline rate of ozone 1H to determine the amount of ozone generator. Formula: W (ozone generation) = (CV) / (1-62.25%) mg / h.
(3), the frequency of disinfection:
3.1 100,000 clean area air disinfection must be carried out in the following cases:
3.1.1 daily disinfection: clean room and HVAC system daily disinfection is only carried out using ozone disinfection, the purpose of the elimination of Airborne bacteria;
3.1.2 Regular monthly disinfection: The monthly disinfection of the cleanroom and HVAC system is carried out by alternating formaldehyde and ozone to prevent the production of drug-resistant strains of bacteria, with the aim of destroying the environmental planktonic and settled bacteria;
3.1.3 Environmental monitoring: When the environmental microbiological test results in the clean area exceed the standard, regular monthly disinfection will be carried out according to the following conditions;
3.1.4 Production shutdown: shut down production in the clean area for more than a week, before production according to the monthly periodic disinfection.
(4), disinfection equipment and disinfectants:
4.1 Alcohol lamp, 36% formaldehyde solution;
4.2 Ozone generator, ozone;
(5), disinfection methods:
5.1 Disinfection of the preparatory work before:
5.1.1 The clean room should be carried out in the full range of hygiene and cleanliness, including plant
5.1.2 The clean room should be free of intermediates, semi-finished products, raw and auxiliary materials exposed storage and no operators;
5.1.3 With clean rags jiaojiao purified water to wipe the equipment, the outer surface of the walls, with clean mop jiaojiao drinking water to wipe the ground to keep the clean area wet;
5.1.4 Close the workshop all the strong exhaust port, open the clean area
5.1.4 Close all the strong exhaust vents in the workshop, and open the inner door of the transfer window in the clean area;
5.1.5 Close the fresh air outlet valve of the HVAC system and fully open the total return air valve.
5.1.6 Ozone and formaldehyde disinfection before the preparation of the same, but the use of formaldehyde, should be in direct contact with the product of equipment, containers and other internal surfaces should be protected to prevent the formaldehyde collection attached to their surfaces, while the ozone can not be considered.
5.2 Ozone disinfection method:
5.2.1 After the normal operation of the HVAC system, turn on the ozone generator power supply, the power indicator is on, the voltmeter shows that the voltage is 220V;
5.3.1 The ozone generator button screwed to the "manual" position, start the ozone generator (A, B), the ozone generator (A, B), the ozone generator (A, B), the ozone generator (A, B) and the ozone generator (A, B). Generator (A, B) disinfection button, the ammeter shows the working current 3.6A, while the indicator light, the ozone generator began to work, record the start of disinfection time;
5.3.2 Disinfection time: ozone disinfection time of 1-1.5 hours a day (to eliminate the airborne bacteria), the regular monthly disinfection time of 2-2.5 hours (to eliminate environmental bacteria). -2.5 hours (to eliminate the environment of planktonic bacteria and precipitating bacteria).
5.3.3 Daily disinfection or monthly periodic disinfection at the end of the specified disinfection time, close the ozone generator start button, turn off the main power supply, ozone generator stop working.
5.3.4 After the end of ozone disinfection, the clean area requires personnel to operate, the HVAC system should continue to run for 30min, and then open the process exhaust fan and the new air valve to supplement the fresh air, the unit will continue to run for 30min after the ozone in the clean area is completely decomposed, the personnel can enter the clean area.
5.3.5 After the end of ozone disinfection, if there is no need for personnel to operate in the clean area (such as disinfection at the end of the day after production), you can directly turn off the HVAC system fan, the next day, the HVAC system operation to reach the self-cleaning time (40min), the personnel can be directed to enter the clean area operation.
V. Validation program
5.1 Daily ozone sterilization
5.1.1 Sterilization cycle: 3 days.
5.1.2 Sterilization time
5.1.2.1 1 hour
5.1.2.2 1.5 hours
5.1.2.3 2 hours
5.1.3 Post-sterilization testing (each time consecutively to do the testing of the three cycles)
5.1.3.1 Inspection indicators
5.1 .3.1.1.1 Detection of settled bacteria
5.1.3.1.2 Bio-indicator for bacterial challenge test, bio-indicator strains of Bacillus subtilis is selected, the use of the determination of the initial number of bacteria to be determined before the use of the determination should be not less than 10. Before sterilization, the surface dish containing the bioindicator is placed in the central ground in the room to be tested, open the surface dish before sterilization, after the end of sterilization, the recovery of bio-indicator into the thioglycolate liquid medium, incubated at 37 ℃ for 3 days.
5.1.3.2 Accreditation standards:
5.1.3.2.1 Sedimentation bacteria: 100 level, sedimentation bacteria ≤ 1 per dish, 10,000 level, sedimentation bacteria ≤ 3 per dish, 100,000 level, sedimentation bacteria ≤ 10 per dish
5.1.3.2.2 Bio-indicators: should be sterile growth
5.1.4 Sedimentation bacteria, bio-indicator test Records are shown in the validation implementation report
5.2 Great sterilization
5.2.1 Sterilization cycle: half a month.
5.2.2 Sterilization time
5.2.2.1 2 hours
5.2.2.2 2.5 hours
5.2.2.2.3 3 hours
5.2.3 Post-sterilization testing (three consecutive cycles of testing at each time)
5.2.3.1 Inspection indicators
5.2 .3.1.1 Detection of settled bacteria
5.2.3.1.2 Bio-indicator for bacterial challenge test
Bio-indicator strains of Bacillus subtilis are selected, and the initial bacterial count should be determined before using the assay and should be not less than 10. Before sterilization, the surface dish containing the bio-indicator is placed on the central floor in the room to be tested, and the surface dish is opened before sterilization, and after sterilization, the bio-indicator is recovered and put into the thioglycolate liquid medium, and incubated for 3 days at 37 degrees Celsius
5.2.3.2 Accreditation standards:
5.2.3.2.1 Sedimentation bacteria: 100 level, sedimentation bacteria ≤ 1 per dish, 1 Class 10,000, Sedimentation bacteria ≤ 3/dish, Class 100,000, Sedimentation bacteria ≤ 10/dish
5.2.3.2.2.2 Bio-indicator: should be free of bacterial growth
5.2.4 Sedimentation bacteria, bio-indicator testing records are shown in the validation implementation report
5.3 Preparation of the revalidation program cycle:
When there is a change in the sterilization time, the sterilizing equipment or Sedimentation bacteria regular monitoring of the data have a negative trend, should be in accordance with the methods used in the previous validation of ozone sterilization re-validation, or according to the actual situation of repairing the validation program and then re-validation, in order to confirm the sterilization effect.
5.4 Validation results of the total assessment:
5.4.1 Quality Assurance Department is responsible for the collection of the verification, test results reported to the validation team. Validation team is responsible for the validation of the results of the review, to make a validation conclusion, issue certificates, and the validation of the results of the review fill in the "validation of the results of the integrity of the total evaluation record sheet", which includes:
5.4.1.1 Validation of the results of the omission?
5.4.1.2 validation implementation process of the validation program has not been modified, the reasons for modification, basis, whether approved?
5.4.1.3 Is the verification record complete?
5.4.1.4 Test verification results in accordance with the standard requirements? Is the deviation and the explanation of the deviation reasonable? Is there a need for further supplementary testing?
5.4.1.5 Is the deviation and the description of the deviation reasonable? Is additional testing required?
5.4.2 General evaluation and countersigning of validation results:
5.4.2.1 General evaluation:
5.4.2.2 Countersigning
6 Evaluation of validation program and final approval:
6.1 Evaluation of validation program:
6.2 Final approval of the validation program:
6.6 Validation of the original records
6. p>
Sedimentation bacteria detection record (daily sterilization)
Sterilization date Sterilization time Sedimentation bacteria Detection date Detector
2010-10-14 1 hour
1.5 hours
2 hours 15 2010-10-14 Deng XX
2010-10-15 10 2010-10-15 Liang XX
2010-10-16 8 2010-10-16 Lin XX
Comprehensive Evaluation
The sterilizer basically meets the requirements
Confirmed
QA: Luo XX
Date: 2010-10-17 Verification Team: Yang XX
Date: 2010-10-17
Sedimentation bacteria detection record (large sterilization)
Sterilization date Sterilization time Sedimentation bacteria Detection date Detection person
2010-10-18 2 hours
2.5 hours
3 hours 8 2010-10-18 Yang XX
2010-10-19 5 2010-10-19 Zhang XX
2010-10-20 3 2010-10-20 XX Zhang
Comprehensive Evaluation
The sterilizer basically meets the requirements of the 10,000 level
Confirmation
QA: XX Zhu
Date: 2010-10-21 Verification Team: XX Zhu
Date: 2010-10-21
Bio-indicator test record (large sterilization)
Sterilization date Colony count Detection date Testing person
2010-10-22
5 2010-10-22
Deng XX
2010-10-23
4 2010-10- 23
Leung XX
2010-10-24
3 2010-10-24
Lin XX
Comprehensive Evaluation
The sterilizer basically complies with the requirements
Confirmation
QC: Luo XX
Date: 2010-10-25 Verification team ：Yang XX
Date: 2010-10-25
Verification Report
Item Result Analysis and Evaluation Conclusion
Sedimentation bacteria check The ozone generator installed in the air supply port of the fan was turned on, and the samples were taken respectively after sterilization for 30min, 45min, 60min and 90min, and the sediments were all in accordance with the requirements of GMP.
Competent
Overall Evaluation According to the verification test, our infusion workshop decided to use sampling sterilization 30 min disinfection, disinfection effect has been verified to meet the expected standards.
Report Reviewer: XX Lin Date: 2010-11-4
Verification Summary
Ozone sterilization has long been studied by scientists, the national industry has done to affirm and recommend the application of ozone sterilization, but the application of ozone sterilization in the pharmaceutical production environment is only in recent years is gradually being widely used.
Characteristics of ozone disinfection
Ozone disinfection and sterilization methods compared with conventional disinfection and sterilization methods have the following characteristics:
(1) high efficiency: ozone disinfection does not require any other auxiliary materials and additives. Disinfection when the ozone generator produces a certain amount of ozone, in a relatively sealed environment, diffusion uniformity, inclusiveness, permeability, overcoming the problem of ultraviolet sterilization of the existence of disinfection of the dead zone, to achieve a full range of fast, efficient disinfection sterilization purposes. In addition, because of its sterilization broad-spectrum, not only can kill bacterial propagules, germ cells, viruses, fungi and protozoa spores and other microorganisms, but also destroy botulinum and toxins and rickettsiae, etc., but also has a strong mold, fishy, smelly and other odor removal function.
(2) high cleanliness: ozone in the environment can be naturally decomposed into oxygen, which is ozone as a disinfectant and sterilizer unique advantages. Ozone generated using the oxygen in the air, disinfection and oxidation process, the excess oxygen atoms in 30 minutes and then combined into molecular oxygen, there is no residual material, to solve the disinfectant disinfection, the residue of the secondary pollution problem, while eliminating the end of disinfection and cleaning again.
(3) Convenience: ozone sterilization equipment is generally installed in the indoor or central air conditioning system, air purification system, or sterilization equipment. According to the sterilization of the required concentration and time, automatically set the timing control of ozone sterilization equipment, easy to operate and use. And formaldehyde, epoxyacetic acid fumigation disinfection for a long time, and ozone sterilization can be turned on every day at regular intervals to use.
(4) economic: through the ozone sterilization sterilization in many pharmaceutical industry GMP applications, as well as the use of medical and health units and the operation of the comparison, ozone disinfection method compared with other methods have great economic and social benefits.
After practical application and summarize the ozone in the pharmaceutical industry is widely used in the following areas:
1, clean area disinfection and sterilization
China's Ministry of Health issued the "toxic technical specifications", the ozone sterilizing effect of the use of the scope of use and use of the method have clear provisions. Affirmed that ozone is a broad-spectrum bactericidal agent, can kill bacterial propagules and germ cells, viruses, fungi, etc., can destroy botulinum toxin. Ozone sterilizes faster than chlorine.
In the pharmaceutical factory clean area is large, almost all have a central air-conditioning purification system to complete the purification and disinfection of each clean area. The traditional disinfection method is to use formaldehyde fumigation, as mentioned above, formaldehyde fumigation has many disadvantages. Ozone sterilization to replace the most simple and easy to implement a good method. The method is extremely simple, according to the size of the clean plant space to choose a standardized ozone generator. Then the ozone generator directly into the air-conditioning purification system in the air duct, known as built-in ozone generator. Ozone with the airflow of the air duct into the clean area, the clean area for space sterilization sterilization, the remaining ozone inhaled into the air outlet, taken away by the central air conditioning. The ozone generator can also be placed in the air-conditioning unit, the ozone into the central air-conditioning duct, and then be sent to the clean room, called external ozone generator. External ozone generators are easy to install and service, but the manufacturing cost is a little higher. Both methods have the same disinfection effect. According to the Ministry of Health disinfection specifications, the ozone concentration of air disinfection is 5ppm, but in fact the clean area disinfection is not only the disinfection of the air, in fact, also includes equipment, appliances and other object surface disinfection, so the general design concentration of 10ppm, equivalent to 20mg of ozone content per cubic meter of air. Every day before going to work two hours of power on, when going to work off, you can ensure that a day in the clean area of the planktonic bacteria and settling bacteria to meet the requirements of GMP.
2, space sterilization
For some pharmaceutical production processes when there is no central air conditioning or do not need to set up a central air conditioning, but the room needs to be individually sterilized. The method is based on the size of the room space to select a certain concentration of ozone generator, installed directly in the room. According to the need to set the disinfection time, the end of disinfection will automatically shut down, so it is very convenient to use.
For rooms with cleanliness requirements, the machine is equipped with a filter, so that the ozone generator has a self-cleaning function to achieve the purpose of disinfection and cleanliness.
In short, ozone sterilization to the present has been widely used in the national pharmaceutical industry. Ozone generator from the product to the application of technology have been greatly improved and development. In the selection of the important consideration of their own actual situation and suitable characteristics, but also to consider its half-decay and decline, as well as the relationship between input and economic benefits. In order to ozone in the pharmaceutical industry to get better application.
The above, only from a few aspects of the ozone in the production of clean areas, you can see that the ozone in the field is still a great deal of work. GMP verification for the development of ozone technology provides a broad opportunity, we should seize this opportunity, so that it can make a breakthrough. At the same time, due to the successful application of ozone sterilization process in the pharmaceutical production process, but also for the GMP verification provides a new type of powerful weapon, more conducive to the pharmaceutical production enterprises as soon as possible through the GMP verification.