Application of sterilization equipment when producing sterile drugs
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Sterilization involves the sterilization of plants, equipment, containers, sanitary ware, work clothing, raw and auxiliary materials, finished products, packaging materials, instruments, and the like. Sterile drugs, especially injections, corneal wounds or surgical eye drops, etc., are drugs that directly enter the human blood circulation system to produce curative effects, so there are special requirements for their quality, which must comply with the Pharmacopoeia sterility test. Claim. To this end, in the production process, efforts should be made to eliminate microbial contamination and kill microorganisms in the drug, which is the purpose of sterilization.
The methods of sterilization are basically divided into two categories: physical methods (dry heat sterilization, moist heat sterilization, radiation sterilization, filtration sterilization) and chemical methods (gas sterilization, liquid sterilization). Several commonly used sterilization methods are described below.
* Dry heat sterilization method and equipment
1. Dry heat sterilization Dry heat can be used for the sterilization of articles that can withstand higher temperatures but are not suitable for being penetrated by steam or easily damaged by damp heat. At the same time, dry heat is also one of the methods used in the pharmaceutical industry to remove pyrogens. Since dry heat has a much lower killing effect on microorganisms than saturated steam at the same temperature, dry heat sterilization requires higher temperatures or longer sterilization times.
Non-aqueous substances or extremely viscous liquids such as glycerin, liquid paraffin, oil, etc. which are ineffective by moist heat sterilization can be sterilized by this method. When sterilized, the oil, ointment base or powder should be laid into a thin layer of no more than 0.6 cm to allow better penetration of heat.
When evaluating the relative sterilization ability of dry heat sterilization, a lethality unit similar to the Fo value and a biological indicator can generally be used. The Fo value differs from the Fo value in the verification of the moist heat sterilization system: the Fo value for evaluating the relative sterilization ability of the dry heat sterilization process is an equivalent time when the time and temperature conditions are changed to 170 ° C. At the same time, the Z value is set to 20 ° C or FH value. The BP 1993 edition stipulates that only the dry heat sterilization system with sterilization as the end goal must ensure that the small FH value of the zui is greater than 170 ° C for 60 min. For systems sterilized by dry heat survival probability method, the verification results should be able to prove that the sterilization process ensures that the killing effect on heat-resistant microorganisms reaches the recognized low survival probability, and the probability of not being killed is generally 10-6. The probability of residual microbial contamination of a sterilized item after sterilization is one in a million. For the verification results of the dry heat overkill sterilization system, it should be proved that the sterilization process ensures that the microorganisms which are highly tolerant to dry heat produce more than 12 log decrements, and the survival probability of microorganisms below 10~12 is obtained. That is, the possibility of microbial contamination remaining after the sterilization of the sterilized article is less than one in a million, and the conventional evaluation of the amount of the bacteria is not required in the verification of the overkill sterilization process.
Second, the main equipment for dry heat sterilization The main equipment for dry heat sterilization is dry heat sterilization cabinet and tunnel sterilization system. Dry heat sterilization equipment generally consists of the following important parts:
1. Heater - It is the main component of dry heat sterilization equipment, which has a great impact on the sterilization effect. Heater failure is one of the main reasons for the decline in sterilization effect of dry heat sterilization equipment. It is mainly manifested by a decrease in heating rate; affecting heat distribution; and generating dust.
The cause of the heater failure is mainly due to the long-term use of the heater or the poor air quality through the heater. Therefore, the current monitor should be configured in the heating system of the sterilizer, and the fault should be discovered in time.
2, high-efficiency filter - in the dry heat sterilization equipment, the air should pass through the high-efficiency filter; remove the dust generated in the internal air circulation system requires high-efficiency filter to prevent the pollution of the exhaust air can also be installed with high-efficiency filter . Therefore, the high-efficiency filter must meet the requirements of the air volume for the dry heat sterilization process and can withstand the corresponding wind pressure.
3, buffer plate, damper, damper or air baffle - buffer plate or air baffle is used to control the air flow of the dry heat sterilizer, more installed near the inlet or exhaust duct or heater, Used for positive pressure control. The damper can be installed near the high-efficiency filter in the exhaust system to prevent backflow wind pollution. The positive pressure can be controlled by controlling the exhaust air volume, and the air flow regulator can also be used to control the air inlet and exhaust air volume to maintain a positive pressure. In general, the pressure in the chamber of the dry heat sterilizer is slightly higher than the adjacent non-sterile area and slightly lower than the adjacent aseptic area.
4, the fan - the airflow cycle in the dry heat sterilizer has a great impact, the wind volume of the fan should be measurable, and can be adjusted, if necessary, suppliers can be required to increase this requirement to their equipment standards, because the fan air volume The measurement indication value can provide a basis for checking the fan status during the use of the device.
5, conveyor belt (only for continuous method) - the speed of the conveyor belt, in the continuous transmission of dry heat sterilization system is a very important factor in the sterilization effect. Conveyor speed controllers are currently available in most dry heat continuous sterilization systems, but there are fewer speed recording devices. Since the conveyor speed determines the amount of heat that the material receives when it passes through the sterilizer, its speed record is one of the basic data to prove that the sterilizer is operating under control. It is also necessary to save the conveyor belt for each sterilization process in the production process. Speed ​​record. At the same time, the SOP of the dry heat sterilizer should also clearly specify the operating speed range of the conveyor belts of various sterilization processes, which should be verified by verification.
6, running chain control system - dry heat sterilizer chain control system is equipped with: door interlock control system; pressure sensor; temperature sensing, control, stop chain operation of the chain control device, etc., to ensure that under any circumstances The temperature is below the design requirements to prevent the sterilized item from passing through the sterilizer below the sterilization temperature.
7, temperature controller and recorder - in the dry heat sterilization system, temperature detection, sensing, control, recording system is the basis of the entire sterilization process control, its control system must be able to ensure sterilization in the sterilizer chamber The temperature can be maintained within the set sterilization temperature range, and the recording system must record the temperature detection and temperature readings of the sensing system accurately.
Section 2 Hygrothermal Sterilization Method and Equipment
1. The moist heat sterilization method is a method of sterilizing in saturated steam or boiling water or circulating steam. Since the latent heat of the steam is large and the penetrating power is strong, the protein is easily denatured or solidified, so the sterilization efficiency is higher than the dry heat sterilization method. This is a widely used sterilization method in the production of injections. It has the advantages of reliable sterilization, convenient operation, easy control and economy. However, it does not apply to the sterilization of drugs that are unstable to heat and humidity.
Factors affecting moist heat sterilization are:
1, the type and number of bacteria: different bacteria, the same developmental stage of the same bacteria have different resistance to heat. The smaller the number of bacteria, the shorter the sterilization time. For example, even if the bacteria are all killed in the injection, the bacterial corpses in the injection are too much, which may cause clinical adverse reactions.
2, the nature of the drug and sterilization time: Generally speaking, the higher the sterilization temperature, the shorter the sterilization time, but considering the stability of the drug, the sterilization temperature should be appropriately reduced or shortened under the premise of effective sterilization. Sterilization time. For example, vitamin C injection was sterilized by circulating steam for 15 minutes, and Nacl infusion was sterilized at 115 ° C for 30 minutes.
3. Properties of steam: steam has saturated steam, wet saturated steam and superheated steam. The saturated steam has a high heat content and a large heat penetration force, so the sterilization effect is high. Wet saturated steam has moisture, low heat content, poor penetrating power, and low sterilization efficiency. The superheated steam temperature is higher than the saturated steam, but the penetration is poor and the sterilization efficiency is low.
4, the nature of the medium: the preparation contains nutrients, such as sugar, protein, etc., to enhance the heat resistance of bacteria. The viability of bacteria is also affected by the pH of the medium. Generally, the neutral environment heat resistance is large, the alkaline is second, and the acidity is not conducive to bacterial development.
Second, the main equipment of the moist heat sterilization method The main equipment of the moist heat sterilization method is the sterilization kettle. There are many types of sterilization kettles, and the performance difference is also great, but the basic structure is similar, and the material used is a strong alloy. At present, the square autoclave used by many domestic enterprises can seal the pressure, and has a vent, a safety valve, a pressure and a temperature indicating device. The jacketed sterilizer is equipped with a tracked cart and is divided into several compartments. The top of the sterilizer is equipped with two pressure gauges, one for indicating the pressure inside the steam jacket and the other for indicating the pressure inside the cabinet. The middle of the two pressure gauges is a thermometer. An exhaust valve should also be installed above the sterilizer to remove non-condensable gases when heating steam is introduced. In recent years, some pharmaceutical companies have begun to adopt steam sterilizers with vacuum pumping capacity and Fo control. At present, there are two main types of imported sterilizers used in China: a rotary sterilizer with steam as a sterilization medium. For example, Austria's HDRR009.20/IS, another spray sterilizer with superheated water as medium, such as FOW5-500 of FEDEGARI, Italy. The main advantage of the imported sterilizer is that the batch volume is large, the accuracy and precision of the temperature control system are good, and the heat is relatively uniform during the sterilization process.
The basic sterilization procedure for the sterilizer is:
Bottling → heating, steam replacement air → sterilization → exhaust steam → preheated water cooling → unloading bottles.
Third, the precautions for the use of hot press sterilizer:
1. The air in the sterilizer must be drained. If air is present in the sterilizer, the pressure indicated on the table is the total pressure of both steam and air rather than pure steam pressure. As a result, although the pressure reached a predetermined level, the temperature did not reach. If the gauge pressure is inconsistent with the thermometer indication, there may be that the air is not exhausted, or the pressure gauge may be out of order, or it may be unsaturated steam. The cause should be solved.
2. The sterilization time must be counted from when all the liquids actually reach the required temperature. Although the warm-up time (heating time) during damp heat sterilization can be shorter than the dry heat time, it is important to measure and shorten the warm-up time. Sex, it is no less than dry heat sterilization. For example, 250ml-500ml glass bottle infusion, preheating time is generally 15~30 minutes, and many can reach 45 minutes.
3. After the sterilization is completed, stop the heating. Generally, the pressure indicated by the pressure gauge should be gradually reduced to zero before the steam in the pot can be released. After the pressure in the pot is equal to the atmospheric pressure, the sterilization pot is slightly opened until 10~15. After a minute, turn it all on again. This avoids the pressure difference between the inside and the outside being too large to cause the article to rush out of the pan and cause the glass bottle to burst. This must be taken care of to avoid work accidents. In order to shorten the sterilization cycle, there is also a spray of cooling water to the closed container containing the solution in the sterilizer to accelerate the cooling. The cooling effect at the water temperature of 18 ° C and the spray size of 80 μm is large, and the excessively cold spray will cause the bottle to blast. For materials that require drying but are not easily damaged after sterilization, the steam in the sterilizer is released immediately after sterilization to facilitate drying.
4. Circulating steam sterilization method and boiling sterilization method The steam sterilization method is steam sterilization in an unsealed container, and the pressure is equal to atmospheric pressure, that is, steam sterilization at 100 °C. At present, China's pharmaceutical factories produce injections, especially 1~2ml of injections and varieties that are not resistant to high temperatures. Basically, this method is adopted. The boiling sterilization method is to put ampoules or other items into water and boil them for sterilization. Circulating steam sterilization and boiling sterilization, generally 100 ° C for 30 minutes to 60 minutes. This method does not guarantee the killing of all spores, such as spores of anaerobic bacteria such as tetanus, so contamination should be avoided as much as possible during the preparation process. The steam sterilizer can be opened by a hot press sterilizer for sterilization. Steamer can also be used for sterilization in rural or field conditions. If the boiling point of water is reduced in the plateau, 1-2% sodium carbonate and 5% phenol are often added to the water to increase the boiling point of water. Prolong the sterilization time to solve the problem. Generally, for every 300 meters increase, the sterilization time is extended by 20%. This method has a large sterilization effect on proliferating microorganisms, but it cannot guarantee the killing of spores.
5. Low-temperature intermittent sterilization method: the method to be sterilized is heated at 60-80 ° C for 1 hour, and the bacterial propagules are killed, and then placed at room temperature or in an incubator for 24 hours, wherein the spores develop into breeding. Body, then sterilized a second time. Operate continuously three times or more until all spores are eliminated. This method is applicable to preparations or medicines that must be sterilized by heat but are not resistant to higher temperatures. The disadvantage is that the time is long and the effect of eliminating spores is not complete enough. For the preparations or medicines sterilized by this method, in addition to the antibacterial property itself, an appropriate amount of bacteriostatic agent should be added to increase the sterilization efficiency.
The above several sterilization methods are several sterilization methods of moist heat sterilization. The factory now adopts hot pressing sterilization method and boiling sterilization method. For example, the oral liquid workshop adopts boiling sterilization for 40 minutes to the liquid. The bottle and cap are sterilized at 121 ° C for 20 minutes or at 105 ° C for 30 minutes. If the oral solution is sterilized by zui, the potted drug is sterilized at 105 ° C for 30 minutes within the specified time.
Section III Other sterilization methods and disinfection methods
1. Radiation Sterilization Method A method of gamma ray sterilization by radioisotope (60C0 or 137Cs). The ray can directly ionize the molecules of the organic compound, generate free radicals that destroy normal metabolism, and cause decomposition of molecular compounds in the microorganism. Radiation sterilization is characterized by not increasing the temperature of the sterilized product, and having strong penetrability. It is suitable for sterilization of heat-resistant drugs, sterilization of medical devices, polymer materials, packaging materials, and the like. The sterilization dose is generally 2.5 x 104 GY (Gray). This law is contained in the British Pharmacopoeia (1998), the 13th edition of the Japanese Pharmacopoeia. Some companies in China also use 60C0 to sterilize certain Chinese medicines and medical devices. Radiation sterilization, high preparation cost, may reduce the efficacy of certain drugs, produce toxic substances or heat-generating substances, and should also pay attention to safety protection measures when using radiation sterilization.
Second, the ultraviolet sterilization method refers to a method of killing microorganisms by ultraviolet irradiation. The ultraviolet wavelength generally used for sterilization is 200 to 300 nm, and the wavelength at which the sterilization force is strong is 253.7 nm. Ultraviolet light acts on nucleic acid proteins to cause denaturation, and at the same time, the air is exposed to ultraviolet rays to generate trace amounts of ozone, thereby co-sterilizing. Ultraviolet light travels in a straight line and can be reflected by different surfaces. The penetration is weak, but it is easier to penetrate clean air and pure water. Therefore, the method is applicable to the sterilization of the sterilized, sterile room air and water on the surface of the irradiated object; it is not suitable for the sterilization of the liquid medicine, the deep sterilization of the solid substance; the ordinary glass absorbs the ultraviolet light, so it is installed in the glass container. The drugs in this medicine cannot be sterilized by this method. Ultraviolet rays are exposed to the human body for a long time, and conjunctivitis, erythema and skin burning may occur. Therefore, the ultraviolet light is usually turned on for 1 to 2 hours before the person enters the room, and the talent enters the clean room after being closed. If it is necessary to sterilize ultraviolet rays after the person has entered, the human skin and eyes should have effective protective measures. Generally, a 30-watt UV lamp can be installed in a space of 6 to 15 m3, preferably 2.5 to 3 m from the ground.
Pay attention to the following problems when sterilizing with ultraviolet radiation:
1. The bactericidal power of ultraviolet light decreases with the increase of the use time. When the normal use time reaches 70% of the rated time, the ultraviolet lamp should be replaced to ensure the sterilization effect. The average life of domestic UV lamps is generally 2000h.
2, the bactericidal effect of ultraviolet light varies with the strain, and the amount of mold killing is 40 to 50 times larger than that of the bacillus.
3. Ultraviolet radiation is usually designed on the basis of a relative humidity of 60%. When the indoor humidity increases, the exposure should be increased accordingly.
4, the effect of ultraviolet sterilization is related to the length of the irradiation, which requires verification to determine the irradiation time.
5. The installation form and height of the UV irradiation lamp should be determined according to the actual situation and with reference to the instructions for use.
3. Ethylene Oxide Sterilization Ethylene Oxide Sterilization is a method of sterilizing by using ethylene oxide gas. It is a traditional sterilization method, which can be applied to sterilization of work clothes sterilization, medicines that are not resistant to heat sterilization, medical instruments, facilities, equipment, and the like. The ethylene oxide sterilization system mainly has the following four important factors that restrict each other to affect the sterilization effect:
1, temperature;
2, humidity;
3. Gas concentration;
4. Sterilization time.
Because ethylene oxide is a flammable and explosive substance, open flame can cause combustion, and at the same time, it may cause explosion due to gas decomposition. Safety concerns should be paid attention to in ethylene oxide sterilization.
Ethylene oxide has a very strong penetrating and diffusing ability, and its sterilization mechanism is mainly strong oxidation, so it has a wide spectrum of sterilization and strong sterilization ability, and has strong killing effect on microbial propagules and spores. . Although ethylene oxide has irritating odor and toxicity at high concentrations, it is moderately toxic and generally volatilizes quickly and does not leave residual poison on the inner surface of the system.
The sterilization mechanism is mainly the substitution property of its alkyl group, the hydrogen atoms on the -cooH, -NH2, -OH, and SH groups in the bacterial protein molecule are replaced by the hydroxyethyl group of ethylene oxide to make the cells The metabolism of cells produces irreversible damage.
CH2-CH2+R-SH→RS-CH2-CH2OHo
R-SH represents a thiol-containing protein molecule.
The procedure of sterilizing with ethylene oxide gas is generally to place the sterilized article in a sterilizer, to remove air under reduced pressure, to preheat, and to use a mixed gas for ethylene oxide gas (generally, ethylene oxide gas contains 10- 20%, carbon dioxide containing 80~90%), input mixed gas under reduced pressure, maintain a certain concentration, temperature and humidity, after a certain period of time, vacuum to remove ethylene oxide gas, and then into the sterile air replacement ring Oxygen ethane gas until it is replaced.
4. Filtration sterilization The filtration sterilization method is a method of removing living or dead microorganisms by filtration, and is a mechanical sterilization method. This machine is called a sterilization filter. It is mainly used for the sterilization of heat-labile drug solutions, gases and water. The filter for sterilization is required to effectively remove microorganisms from the solution, and the solution smoothly passes through the filter, and the filtrate does not fall into any unnecessary substances, the filter is easy to clean, and the operation is simple.
The reproductive bacteria are rarely less than 1 μm, and the spore size is 0.5 μm or less. Therefore, for filters filtered by sieving, such as cellulose ester membrane filters, the pore size must be small enough to prevent bacteria and spores from entering the pores and having a pore size of about 0.2 μm. The pore size of the filter that is retained in the pores or electrostatically acts may be slightly larger than the particles to be filtered, but if the pressure is too large or fluctuating, the particles may be squeezed. Usually, the method of measuring the pore diameter is carried out by filtering with a suspension of B. prodigilsus having a size of about 0.7 μm, and the filtrate is subjected to a culture experiment to observe the presence or absence of growth of the bacterium. The commonly used sterilization filter has a microporous membrane device, a folded cellulose filter rod, and generally has a pore size of 0.22 μm. The filtration method should be combined with aseptic technique and the finished product must be sterilised to ensure its sterilization quality. The filter membrane is preferably sterilized by CH2-CH2. O
V. Ozone disinfection method The principle of ozone (O3) disinfection is: ozone has unstable molecular structure at normal temperature and pressure, and quickly decomposes into oxygen (O2) and single oxygen atom (O). The latter has strong activity. It has a strong oxidation effect on bacteria. Ozone oxidizes and decomposes the enzymes necessary for oxidizing glucose inside the bacteria, thereby destroying its cell membrane and killing it. The excess oxygen atoms recombine themselves into universal oxygen molecules (O2) without any harmful residues, so they are called non-polluting disinfectants. It not only has a strong killing ability for various bacteria (including hepatitis virus, Escherichia coli, Pseudomonas aeruginosa and bacteria), but also is effective for mold.
The raw materials for the production of ozone are mainly electric energy and air, which are generally obtained by a high-frequency ozone generator (electronic sterilizer). When disinfecting, the ozone generator can be placed directly in the middle of the room.
The ozone generator technical indicators to be confirmed and corrected in the verification of ozone disinfection effect are: ozone production, ozone concentration and time timer, and the disinfection time is determined by verifying the number of bacteria.
6. Formaldehyde disinfection method 1. Calculate the volume of the room and weigh out the formaldehyde at a ratio of 10g/m3.
2, pour formaldehyde into the formaldehyde generator or heating plate or beaker, and put the humidification water, if necessary, also add potassium permanganate (2 ~ 3g / m3), and then heat (formaldehyde generator steam heating, heating plate Or the beaker is heated with hot water therein to evaporate it into a gas.
3. Sterilization process: air conditioner stops running → start formaldehyde gas generator or heat formaldehyde in heating plate → let formaldehyde gas spread for about 30 minutes → start air conditioner to let formaldehyde gas circulate for about 30 minutes → stop air conditioner, room fumigation , time is not less than 8 hours → room exhaust, replaced with fresh air for about 2 hours → resume normal operation.
When the relative humidity is above 65% and the temperature is between 24 and 40 °C, the disinfection effect of formaldehyde gas is good.
The amount of gas generated by formaldehyde sterilization, fumigation time, and ventilation time should be determined after verification. It has also been reported that peracetic acid is used for indoor sterilization in the workshop, and its efficacy is two and a half times larger than that of the same concentration of formaldehyde. It is non-irritating to the mucous membrane and has no effect on wood, metal products and medical devices.
7. Disinfectant disinfection Cleanroom wall, ceiling, doors and windows, machinery and equipment, instruments, consoles, cars, tables, chairs, etc., as well as human hands (gloves) should be cleaned regularly with disinfectant during environmental verification and daily production. Spraying, common disinfectants are isopropanol (75%), ethanol (75%), glutaraldehyde (1%), and clean (0.1 to 0.2%). It is also possible to use 0.5 to 3% of stone carbonate or 2 to 5% of cresol.
The disinfectant for the sterile room must be in a laminar flow table and filtered with a 0.22 μm filter before it can be used.
Section 4 Sterilization Management Points
First, the selection of filtration equipment and treatment methods should meet the process requirements, and the microporous membrane should be tested for integrity before and after use to ensure its sterilization effect.
Second, the liquid medicine starts from preparation to sterilization, and the interval must not exceed the time specified in the process.
Third, sterilization should have temperature, pressure and heating time, constant temperature time, quantity and temperature and pressure curve or temperature curve of the whole process.
4. Sterilize according to the batch number, and do not mix batches. Intermediate products before and after sterilization should have reliable anti-mixing measures.
5. The sterilizing equipment must be verified before use. After the process is changed or overhauled, it must be re-verified to ensure the sterilization effect.
Sixth, the sterilizer should be selected as a double-twisted sterilizer. The temperature uniformity and sterilization effect should be verified before use, and the FO value should be controlled above 8.0.
7. The packaging materials, equipment and other items that are in direct contact with the drug shall be stipulated from the time of sterilization to the time of use.
8. When a batch of products is sterilized using multiple sterilizers, the sub-lot number should be added.
Section V Steam Sterilization Verification
Steam autoclave verification, the general verification steps are as follows:
1. Formulation of the verification plan;
2. Approval of the verification plan;
3. Preparation of instruments and documents;
Fourth, pre-confirmation;
5. Installation confirmation - electrical installation, steam installation, cold support but water installation;
Sixth, operation confirmation - single machine or system test.
VII. Performance Confirmation - Thermal Distribution Test, Thermal Penetration Test, Biological Indicator Verification Test, and Challenge Test of Microbial Spores.
Eight, verification report - data analysis, verification results, evaluation and recommendations.
9. Approval conclusions.
How to carry out steam autoclave verification, the former State Administration of Medicine approved GMP, GSP committee prepared "Guidelines for the verification of pharmaceutical production" (published in April 1996) P263 Appendix IV "Example of Sterilization Validation" for reference.