ASEPTIC TECHNIQUES/BEHAVIOUR

ASEPTIC TECHNIQUES/BEHAVIOUR

 

A clean room is a room in which the concentration of airborne particles is controlled, and which is constructed and used in a manner to minimize the introduction, generation, and retention of particles inside the room and in which other relevant parameters, e.g. temperature, humidity, and pressure, are controlled as necessary. ”

The basic function of a clean room is to protect the drug product/substance from contamination. Air born contamination from outside air is an ordinary problem associated with clean rooms.

The biggest source of contamination in the clean room is personnel. Clean rooms should maintain with minimum number of the personnel. Adherence to basic aseptic technique is a continues requirement for operators in an aseptic processing operation.

ASEPTIC TECHNIQUES

1.	Contact Sterile Materials only with sterile instruments: Sterile instruments (e.g: forceps) are should always be used in the handling of sterilized materials. These instruments should be replaced as necessary throughout the operation.   After initial gowning, sterile gloves should be regularly sanitized to minimize the risk of contamination. Personnel should not directly contact sterile products, containers, closures or critical surfaces.
2.	Moving slowly and deliberately: Rapid movements can create unacceptable turbulence in the critical zone. Such movements disrupt the sterile field, presenting a challenge beyond intended clean room design and control parameters. The principle of slow, careful movement should be followed throughout the clean room.
3.	Keeping the entire body out of the path of laminar air: Laminar air flow design is used to protect sterile equipment surfaces, container closures and the product. Personnel should not disrupt the path of laminar flow air in the aseptic processing zone.
4.	Approaching a necessary manipulation in a manner that does not compromise sterility of the product : In order to maintain sterility of nearby sterile materials, a proper aseptic manipulation should be approached from the side and not above the product (in vertical laminar flow operations) Also speaking  when in direct proximity to an aseptic processing line is not an acceptable practice.

 

ASEPTIC BEHAVIOUR

1.	Only qualified persons are allowed to enter the clean room.
2.	The personnel who entering in the aseptic processing area should be gowned properly. An aseptic processing area gown should provide a barrier between the body and exposed sterile materials and prevent contamination from particles generated by and microorganisms shed from, the body. Gowns needs to be sterile and non shedding, and should cover the skin and hair. If an element of gown found to be torn or defective, it should be changed immediately.
3.	Gloves should be applied in such a way to prevent contamination of the glove surface. The most widely recommended method is to grip the gloves at the wrist and slide the gloves onto the hand, without touching the surfaces of the glove that will later come in contact with items in the clean room. Gloves should be applied last, immediately before entering the clean room.
4.	It is recommended that personnel entering the clean room do not open the door with their gloves to prevent particles from the door from contaminating them.
5.	Only clean room compatible materials are allowed in clean rooms.
6.	Personnel should attempt to avoid coughing or sneezing as much as possible, but if it cannot be avoided, you should turn your head and try to direct it away from the item you are working with as much as possible. Because the face mask is an imperfect seal, particles can easily get around the mask when you cough or sneeze. Masks are often replaced after sneezing or coughing.
7.	The primary source of particulate contamination in the clean room is users and the rates at which users shed particles correlate to the level of physical activity or motion by the individual. Keep movements to a minimum and avoid activities like pacing, extraneous walking, or horseplay.
8.	The number of personnel working in the clean room to be minimized  at any given time.
9.	Person should always be correctly positioned in relation to the production process. For instance it is bad practice to lean over the product because this would allow the particles from clean room garments to fall on to and around the product.
10.	Speaking should be avoided when working close proximity to a product.
11.	Avoid touching surfaces within the clean room
12.	Hands should be held away from the clean room garment to reduce the risk of contaminating the gloves.
13.	Clean room gloves should be cleaned on regular basis to reduce the risk of transferring contaminants by touch.
14.	Cloths used in clean rooms should be used only once or a predetermined number of times. After use a clean room cloth should be discarded.

Can Leptospira species penetrate sterilizing-grade filters?

CAN LEPTOSPIRA SPECIES PENETRATE STERILIZING-GRADE FILTERS?   (Leptospiral challenges for sterile filtration)

 

Leptospira  are Gram-negative aerobic spirochetes that are flexible, highly motile, and spiral-shaped with internal flagella. The bacteria measure 1μm in diameter and 10-20 μm in length. Leptospira  are obligate aerobes that use oxygen as the electron receptor and long-chain fatty acids as a major source of energy. While some of the Leptospira are harmless fresh-water saprophytes, other species are pathogenic and can cause leptosporosis, a significant disease in humans and animals.Leptospira  can occupy diverse environments and habitats; these bacteria are found throughout the world, except in Antarctica. Leptospiral contamination can occur in products sterilized by filtration. Heat sterilization of water eliminates this possibility. 

FDA is aware of a recent report  of Leptospira licerasiae contamination in cell cultures. There is no indication that this bacterium ultimately contaminated either the finished drug substance or drug product. This bacterium has been found to pass through 0.1 µm pore size rated sterilizing-grade membrane filters. While this specific species was the identified contaminant in this case, other Leptospira species also are capable of passing through 0.1 µm pore size rated filters. Compendial microbiological test methods typically used in association with upstream biotechnology and pharmaceutical production are not capable of detecting this type of bacteria.

Based on current information, Leptospira  contamination does not appear to occur frequently, and purification steps that follow cell culture in a typical biotechnology operation would be expected to prevent carryover to the finished drug substance. Testing of bulk drug substances produced in the reported cases did not detect Leptospira  spp., and no evidence of deleterious effects on in-process product were observed in the known case study. However, these types of bacteria can potentially:

• penetrate sterilizing-grade membrane filters
• be present in the manufacturing site environment
• impact in-process production (e.g., production yields, impurity levels, process performance)
• go undetected due to the limitations of current compendial bioburden tests in detecting this microbial genus

As a general principle, manufacturers should use sound risk management and be aware of unusual microbiota reported in the literature that may impact their manufacturing processes (e.g., cell culture biotechnology, conventional sterile drug manufacturing).

Manufacturers should assess their operations, be aware of potential risks, and apply appropriate risk management based on an understanding of possible or emerging contamination risks. As appropriate, preventive measures should be implemented during the product and process lifecycle.

 

Process Validation - EMA & US Guidance Comparison

Process Validation  - EMA & US Guidance Comparison

 

Similarities
Incorporates product life cycle, QRM and efficient quality system practices (ICH Q8, Q9 & Q10).
Emphasis on continued process verification through analysis of pre and post release data to provide confidence of an ongoing valid process.
Acknowledgement and provision of scope to emerging processing technologies, such as PAT, to assist the validation effort.
Enhanced detail to provide understanding of regulator expectation on what constitutes an appropriate validation effort.
EMA	US
Definition “documented evidence that the process, operated within established parameters, can perform effectively and reproducibly to produce a medicinal product meeting its predetermined specifications and quality attributes.”	Definition “the collection and evaluation of data, from the process design stage throughout production, which establishes scientific evidence that a process is capable of consistently delivering quality product.”
The EMA draft guideline states “a minimum of three consecutive batches”, with justification to be provided (there are some exceptions to this statement).	The US FDA guidance states that the number of batches must be sufficient to provide statistical confidence of the process. It is a subtle, but important distinction in the approaches.
The US FDA guidance emphases documenting the development phase as part of PV.	The EMA draft encourages the use of the product development activities, but is less prescriptive on requirements.
The EMA guideline specifically allows the use of CPV to replace traditional validation efforts.	US FDA approach does not place high emphasis on CPV, and requires all three stages of process validation to be fully addressed, regardless of whether contemporary or traditional methods are utilised.
The US FDA guidance considers equipment and process design, as well as equipment qualification as part of the overall process validation effort.	The EMA guideline sees process as independent from equipment and facility. Currently, the EMA still relies on Annex 15 of the GMP guide for instruction on equipment qualification.

 

Process Validation definitions

 

Process Validation definitions

 

According to US FDA  

In 1978 

 “A validation manufacturing process is one which has been proved to do what it purports or is represented to do. The proof of validation is obtained through the collection and evaluation of data, preferably, beginning from the process development phase and continuing the production phase. Validation necessarily includes process qualification (the qualification of materials, equipment, system, building, personnel), but it also includes the control on the entire process for repeated batches or runs”. 

In 1987

“Process validation is establishing documented evidence which provides a high degree of assurance that a specific process (such as the manufacture of pharmaceutical dosage forms) will consistently produce a product meeting its Pre determined specifications and quality characteristics”.

In 2011

“Process Validation is defined as the collection and  evaluation of data, from the process design stage throughout production, which establishes scientific evidence that a process is capable of consistently delivering quality products”.

 According to EMEA

In March 2012

“Process validation can be defined as documented evidence that the process, operated within established parameters, can perform effectively and reproducibly to produce a medical product meeting its predetermined specifications and quality attributes.”

Continuous process verification (PCV) has been introduced to cover an alternative approach to process validation based on a continuous monitoring of manufacturing performance. This approach is based on the knowledge from product and process development studies and / or previous  manufacturing experience. CPV may be applicable to both a traditional and enhanced approach to pharmaceutical development. It may use extensive in-line, on-line or at-line monitoring and / or controls to evaluate process performance. Process validation should confirm that the control strategy is sufficient to support the process design and quality of the product. The validation should cover all

manufactured strengths and all manufacturing sites used for production of the marketed product.

Process Validations Interview Questions And Answers

                                                              

Process Validation  - Interview Questions & Answers

 

Sr.No.	Interview Questions & Answers
1.	What is process validation?   EMA Definition “documented evidence that the process, operated within established parameters, can perform effectively and reproducibly to produce a medicinal product meeting its predetermined specifications and quality attributes.”                                                                                                                                        USFDA Definition “The collection and evaluation of data, from the process design stage throughout production, which establishes scientific evidence that a process is capable of consistently delivering quality  product.”
2.	Which is the latest guidance document for process validation published by USFDA?   Process Validation: General Principles and Practices, (published on Jan.2011)
3.	According to regulatory guidelines (USFDA), what are the stages of process validation?   Process validation involves a series of activities taking place over the lifecycle of the product and process. There are three stages for process validation activities.   Stage 1 – Process Design: The commercial manufacturing process is defined during this stage based on knowledge gained through development and scale-up activities.   Stage 2 – Process Qualification: During this stage, the process design is evaluated to determine if the process is capable of reproducible commercial manufacturing.   Stage 3 – Continued Process Verification: Ongoing assurance is gained during routine production that the process remains in a state of control.
4.	How many batches to be considered for process validation?   The EMA draft guideline states “a minimum of three consecutive batches”, with justification to be provided (there are some exceptions to this statement).   The US FDA guidance states that the number of batches must be sufficient to provide statistical confidence of the process. It is a subtle, but important distinction in the approaches.
5.	Explain the strategy for industrial process validation of solid dosage forms?   ·       The use of different lots of raw materials should be included. i.e., active drug substance and major excipients. ·       Batches should be run in succession and on different days and shifts (the latter condition, if appropriate). ·       Batches should be manufactured in the equipment and facilities designated for eventual commercial production. ·       Critical process variables should be set within their operating ranges and should not exceed their upper and lower control limits during process operation. Output responses should be well within finished product specifications. ·       Failure to meet the requirements of the Validation protocol with respect to process input and output control should be subjected to process requalification
6.	What is Validation Protocol?   A written plan of actions stating how process validation will be conducted; it will specify who will conduct the various tasks and define testing parameters; sampling plans, testing methods and specifications; will specify product characteristics, and equipment to be used. It must specify the minimum number of batches to be used for validation studies; it must specify the acceptance criteria and who will sign/approve! Disapprove the conclusions derived from such a scientific study.
7.	What should be the content of process validation protocol? 1. General information 2. Objective 3. Background/Pre validation Activities, Summary of development and tech transfer (from R&D or another Site) activities to justify in-process testing and controls; any Previous validations. 4. List of equipment and their qualification status 5. Facilities qualification 6. Process flow charts 7. Manufacturing procedure narrative 8. List of critical processing parameters and critical excipients 9. Sampling, tests and specifications 10. Acceptance criteria
8.	In- process validation studies what should be the blend sample size? 1x – 3x dosage unit range on case to case basis. As per USFDA guidance, sampling size can be increased from 1x –10x with adequate scientific justification.
9.	According to USFDA guidance how many sampling points should be considered for collecting blend samples? At least 10 sampling locations to be considered to represent potential areas of poor blending.   In tumbling blenders (ex: V-blenders, double cones, or drum mixers),samples should be selected from at least two depths along the axis of blender.   At least 20 locations are recommended to adequately validate connective blenders (ex: ribbon blender)  What will be the reason of within location variance of blend data? Inadequacy of blend mix, sampling error or agglomeration
10.	What is the difference between EMA & US guideline on process validation?   EMA US Definition “documented evidence that the process, operated within established parameters, can perform effectively and reproducibly to produce a medicinal  product meeting its predetermined specifications  and quality attributes.”   Definition “the collection and evaluation of data, from the process design stage throughout production, which establishes scientific evidence that a process is capable of consistently delivering quality product.” The EMA draft guideline states “a minimum of three consecutive batches”, with justification to be provided (there are some exceptions to this statement). The US FDA guidance states that the number of batches must be sufficient to provide statistical confidence of the process. It is a subtle, but important distinction in the approaches. The EMA draft encourages the use of the product development activities, but is less prescriptive on requirements. The US FDA guidance emphases documenting the development phase as part of PV. The EMA guideline specifically allows the use of CPV to replace traditional validation efforts. US FDA approach does not place high emphasis on CPV, and requires all three stages of process validation to be fully addressed, regardless of whether contemporary or traditional methods are utilised. The US FDA guidance considers equipment and process design, as well as equipment qualification as part of the overall process validation effort. The EMA guideline sees process as independent from equipment and facility. Currently, the EMA still relies on Annex 15 of the GMP guide for instruction on equipment qualification.
11.	Why hopper challenge study is performing during process validation?   To evaluate effect of vibrations during compression on blend uniformity, hopper study shall be carried out.
12.	What are the critical process variables in coating?   Pan RPM, inlet & exhaust temperature, spray rate, gun distance and air pressure.
13.	Why blending is a critical parameter in tablet manufacturing?    Less blending will result in non-uniform distribution of drug and poor flow whereas more blending will result in de-mixing leading to non-uniform distribution of drug and increase in disintegration time.
14.	What are the critical parameters to be checked during dry mixing?  Mixing time and mixing speed
15.	What are the critical parameters to be checked during binder preparation and addition?  Amount of binder solution and mixing time
16.	What the major variables in tablet compression? Speed of machine, and hopper level are the major variables.
17.	What is the revalidation criteria for process validation? 1. Change in formulation, procedure or quality of pharmaceutical ingredients. 2. Change of equipment, addition of new equipment and major breakdowns/maintenance, which affect the performance of equipment. 3. Major change of process, parameters. 4. Change in manufacturing site. 5. On appearance of negative quality trends. 6. On appearance of new findings based on current knowledge. 7. Batch size change
18.	What are the benefits of process validation?   ·       Consistent through output. ·       Reduction in rejections and reworks. ·       Reduction in utility cost. ·       Avoidance of capital expenditures. ·       Fewer complaints about process related failure. ·       Reduced testing process and finished goods. ·       More rapid and accurate investigations into process deviation. ·       More rapid and reliable start-up of new equipment. ·       Easier scale-up from development work. ·       Easier maintenance of equipment. ·       Improve employee awareness of processes.
19.	What are the common variables in the manufacturing of tablets?   · Particle size of the drug substance · Bulk density of drug substance/excipients · Powder load in granulator · Amount & concentration of binder · Mixer speed & mixing timings · Granulation moisture content · Milling conditions · Lubricant blending times · Tablet hardness · Coating solution spray rate
20.	What is the action plan if a test failure observed during process validation?   Any test during process validation shall investigate to determine the case of failure. Where the case of failure is not obvious, it may useful to us an investigation procedure to ensure that all the possible areas of potential failure are covered. Once the case of the process validation failure has been identified, the failure shall classified into the following categories.   Type I: where the failure can be attributed to an occurrence which is not intrinsic to the process for example, an equipment failure raw material that it can be agreed to complete the validation exercise substituting another batch for the one that failed. This investigation and the subsequent action shall be included in the validation report.   Type II: where the failure may be attribute failure or where the investigation is inconclusive than the validation exercise has failed. In this case the validation terms decide and justify the course of action to be taken, recording its justification and recommendations.   This decision shall consider: ·       Re-testing - if investigation of the analytical results supports the decision. ·       Introduction a change in operation parameters, process steps. ·       Changing the process equipment or the procedure for using the equipment. ·       Suspension of the process validation exercise until further technical evaluation and/or development has been carried out. ·       Changing the sampling regime. ·       Review of historical data. ·       Change of the process validation acceptance criteria. ·       Change to an analytical procedure.