An Essentiality In The Pharmacy

Tarun Virmani

The development of a drug product is a lengthy process involving drug discovery, laboratory testing, animal studies, clinical trials and regulatory registration.

To further enhance the effectiveness and safety of the drug product after approval, many regulatory agencies such as the United States Food and Drug Administration (FDA) also require that the drug product be tested for its identity, strength, quality, purity and stability before it can be released for use. For this reason, pharmaceutical validation and process controls are important in spite of the problems that may be encountered¹. Process controls include raw materials inspection, in-process controls and targets for final product. The purpose is to monitor the on-line and off-line performance of the manufacturing process and then validate it. Even after the manufacturing process is validated, current good manufacturing practice also requires that a well written procedure for process controls is established to monitor its performance². This paper provides an overview of pharmaceutical validation and process controls in drug development. The validation concept can be applied to new drugs, new dosage forms and generic drug development.

Essentials of Pharmaceutical Validation

Validation is an integral part of quality assurance; it involves the systematic study of systems, facilities and processes aimed at determining whether they perform their intended functions adequately and consistently as specified. A validated process is one which has been demonstrated to provide a high degree of assurance that uniform batches will be produced that meet the required specifications and has therefore been formally approved. Validation in itself does not improve processes but confirms that the processes have been properly developed and are under control³. Adequate validation is beneficial to the manufacturer in many ways:

• It deepens the understanding of processes; decreases the risk of preventing problems and thus assures the smooth running of the process.
• It decreases the risk of defect costs.

• It decreases the risk of regulatory noncompliance.
• A fully validated process may require less in-process controls and end product testing.

Validation should thus be considered in the following situations:

• Totally new process
• New equipment
• Process and equipment which have been altered to suit changing priorities and
• Process where the end-product test is poor and an unreliable indicator of product quality.

When any new manufacturing formula or method of preparation is adopted, steps should be taken to demonstrate its suitability for routine processing. The defined process should be shown to yield a product consistent with the required quality. In this phase, the extent to which deviations from chosen parameters can influence product quality should also be evaluated. When certain processes or products have been validated during the development stage, it is not always necessary to revalidate the whole process or product if similar equipment is used or similar products have been produced, provided that the final product conforms to the in-process controls and final product specification. There should be a clear distinction between in-process control and validation. In production, tests are performed each time on a batch to batch basis using specifications and methods devised during the development phase. The objective is to monitor the process continuously⁴.

Major Phases in Validation

The activities relating to validation studies may be classified into three:

Phase 1:

This is the Pre-validation Qualification Phase which covers all activities relating to product research and development, formulation pilot batch studies, scale-up studies, transfer of technology to commercial scale batches, establishing stability conditions and storage, and handling of in-process and finished dosage forms, equipment qualification, installation qualification, master production document, operational qualification and process capacity.

Phase 2:

This is the Process Validation Phase. It is designed to verify that all established limits of the critical process parameter are valid and that satisfactory products can be produced even under the worst conditions.

Phase 3:

Known as the Validation Maintenance Phase, it requires frequent review of all process related documents, including validation of audit reports, to assure that there have been no changes, deviations, failures and modifications to the production process and that all standard operating procedures (SOPs), including change control procedures, have been followed. At this stage, the validation team comprising of individuals representing all major departments also assures that there have been no changes/deviations that should have resulted in requalification and revalidation. A careful design and validation of systems and process controls can establish a high degree of confidence that all lots or batches produced will meet their intended specifications. It is assumed that throughout manufacturing and control, operations are conducted in accordance with the principle of good manufacturing practice (GMP) both in general and in specific reference to sterile product manufacture⁵. The validation steps recommended in GMP guidelines can be summarized as follows:

• As a pre-requisite, all studies should be conducted in accordance with a detailed, pre-established protocol or series of protocols, which in turn is subject to formal – change control procedures.

• Both the personnel conducting the studies and those running the process being studied should be appropriately trained and qualified and be suitable and competent to perform the task assigned to them.

• All data generated during the course of studies should be formally reviewed and certified as evaluated against pre-determined criteria.

• Suitable testing facilities, equipment, instruments and methodology should be available.

• Suitable clean room facilities should be available in both the ‘local’ and background environment. There should be assurance that the clean room environment as specified is secured through initial commissioning (qualification) and subsequently through the implementation of a programme of re-testing – in-process equipment should be properly installed, qualified and maintained.

• When appropriate attention has been paid to the above, the process, if aseptic, may be validated by means of “process simulation” studies.

• The process should be revalidated at intervals and

• Comprehensive documentation should be available to define support and record the overall validation process⁶.

Process Validation

Process validation is the means of ensuring and providing documentary evidence that processes (within their specified design parameters) are capable of repeatedly and reliably producing a finished product of the required quality⁵. It would normally be expected that process validation be completed prior to the release of the finished product for sale (prospective validation). Where this is not possible, it may be necessary to validate processes during routine production (concurrent validation). Processes, which have been in use for some time without any significant changes, may also be validated according to an approved protocol (retrospective validation) ^6-13.

Pre-requisites for Process Validation

Before process validation can be started, manufacturing equipment and control instruments as well as the formulation must be qualified. The information on a pharmaceutical product should be studied in detail and qualified at the development stage, i.e., before an application for marketing authorization is submitted. This involves studies on the compatibility of active ingredients and recipients, and of final drug product and packaging materials, stability studies, etc. Other aspects of manufacture must be validated including critical services (water, air, nitrogen, power supply, etc.) and supporting operations such as equipment cleaning and sanitation of premises. Proper
training and motivation of personnel are prerequisites to successful validation^14-16.

Process validation decision

The following model may be useful in determining whether or not a process should be validated:

Figure 1 - Process validation decision tree
The model shown describes a decision tree that a manufacturer can follow when deciding on whether a process needs to be validated. The process under consideration in this model is the simplest possible - many processes may be large and/or a complex set of sub-processes.
Each process should have a specification describing both the process parameters and the output desired. The manufacturer should consider whether the output can be verified by subsequent monitoring or measurement (A). If the answer is positive, then the consideration should be made as to whether or not verification alone is sufficient to eliminate unacceptable risk and is a cost effective solution (B). If yes, the output should be verified and the process should be appropriately controlled (C). If the output of the process is not verifiable then the decision should be to validate the process (D); alternatively, it may become apparent that the product or process should be redesigned to reduce variation and improve the product or process (E). Also, a change in a manufacturing process may result in the need for process validation even though the process formerly only required verification and control. The risk or cost may also be reduced by redesigning the product or process to a point where simple verification is an acceptable decision (E).

The Pharmaceutical Process Equipment

The key idea of validation is to provide a high level of documented evidence that the equipment and the process conform to a written standard. The level (or depth) is dictated by the complexity of the system or equipment. The validation package must provide the necessary information and test procedures required to provide that the system and process meet specified requirements. Validation of pharmaceutical process equipment involves the following:

• Installation Qualification

This ensures that all major processing and packaging equipment, and ancillary systems are in conformity with installation specification, equipment manuals schematics and engineering drawing. It verifies that the equipment has been installed in accordance with manufacturers recommendation in a proper manner and placed in an environment suitable for its intended purpose.

• Operational Qualification

This is done to provide a high degree of assurance that the equipment functions as intended. Operational qualification should be conducted in two stages:

•  
  1. Component Operational Qualification, of which calibration can be considered a large part.

•  
  1. System Operational Qualification to determine if the entire system operates as an integrated whole.

• Process Performance Qualification

This verifies that the system is repeatable and is consistently producing a quality product. These exercises assure, through appropriate performance lists and related documentation, that equipment, ancillary systems and sub-systems have been commissioned correctly. The end results are that all future operations will be reliable and within prescribed operational limits. At various stages in a validation exercise there are needs for protocols, documentation, procedures, specifications and acceptance criteria for test results. All these need to be reviewed, checked and authorized. It would be expected that representatives from the professional disciplines, e.g., engineering, research and development, manufacturing, quality control and quality assurance are actively involved in these undertakings with the final authorization given by a validation team or the quality assurance representative¹⁷.

Conclusion:

It is necessary, before approval of a new drug, that an accurate and reliable assessment for its effectiveness and safety for the intended indication and target patient population is demonstrated. Pharmaceutical validation which includes assay validation, cleaning validation, equipment validation as well as the overall process validation is crucial in stability analysis, animal studies and early phases of clinical development such as bioavailability/bioequivalence studies. After the drug is approved, pharmaceutical validation and process control are necessary to ensure that the drug product will meet/set pharmaceutical standards for identity, strength, quality, purity, stability, evaluation safety and efficacy. In general, pharmaceutical validation and process control provide a certain assurance of batch uniformity and integrity of the product manufactured.

References:

1. Sharp JR. The Problems of Process Validation. Pharm J 1986; 1:43-5.
2. Chow S. Pharmaceutical Validation and Process Controls in Drug Development. Drug Inf J 1997; 31: 1195-201.
3. Committee on Specifications for Pharmaceutical Preparations. Good Manufacturing Practices for Pharmaceutical Products. WHO Technical Report Series no. 82. Geneva: World Health Organization, 1992, pp 14-79.
4. South African Guide to Good Manufacturing Practice. Pretoria: Medicines Control Council, 1996. http://www.pharmanet.co.za/mcc /inpectorate/ins-71998.htm .
5. Guide to Inspections of Oral Solid Dosage Forms Pre/Post Approval Issued for Development and Validation. WashingtonDC: US Food and Drug Administration, 1994.
6. Therapeutics Products Programme. Process Validation: Aseptic Processes for Pharmaceuticals. http://www.hc-sc.gc.ca/hpbdgps/ therapeutic; downloaded March 30, 2001.
7. Rosendale DM. Process Equipment 1990. www.vectorcorporation.com/download/val_in terphex.
8. Cleaning Validation in Active Pharmaceutical Ingredient Manufacturing Plants. Brussels: Active Pharmaceutical Ingredients Committee. http://www.apic.cefic.org/pub4cleaningval/1999pdf; downloaded September 1999.
9. Guide to Inspections Validation of Cleaning Processes. WashingtonDC: US Food and Drug Administration. http://www.fda.gov/ora/inspect _ref/igs/valid.html.
10. Cleaning Validation Guidelines. Ottawa, Canada: Health Products and Food Branch Inspectorate, Health Canada, May 2000, p 11.
11. Harder SW. The Validation of Cleaning Procedures. Pharm Technol 1984; 8(5): 29-34.
12. Jenkins KM, Vanderwielen AJ. Cleaning Validation: An Overall Perspective. Pharm Technol 1994; 18(4): 60-74.
13. United States Pharmacopoeia and the National Formulary XXIII, 18th ed,. Rockville, MD: The United States Pharmacopoeia Convention Inc., 1995, pp 1982 – 1984.
14. Chapman GM, Amer G, Boyce C, Brower G, Green C, Hall WE, Harpaz D, Mullendore B. Proposed Validation Standard VS1: Non-aseptic Pharmaceutical Processes. J Val Technol 2000; 6:502-20.
15. LeBlane DA. Establishing scientifically justified acceptance criteria for cleaning validation of finished drug product. Pharm Technol 1998; 23(10): 136-48.
16. WHO Expert Committee on Specifications for Pharmaceutical Preparations, 34th Report. WHO Technical Report Series no. 863, Annex 6, Geneva: WHO, 1966, pp 80-96.
17. Good Manufacturing Practices for Pharmaceutical Products, WHO/Pharm./93.562/Annex: Guidelines on Validation of Manufacturing Process. Geneva: WHO.

Sterilization Process Validation

Sterilization Process Validation
Title: Sterilization Process Validation
Manual Number: 039
Prepared by: Date: Supersedes:
Checked by: Date: Date Issued:
Approved by: Date: Review Date:
5
4 Responsibilities
All sterile manufacturing sites or its contractors are responsible for ensuring that
sterilization processes used to produce items are properly validated.
5 Guideline
Validation of processes used to sterilize drug products and equipment are the most critical
validation activities undertaken. Common elements in the validation of any sterilization
process include:
􀂇 Sterilization Cycle Development
􀂇 Biological and Physical Measurement Controls
􀂇 Empty Chamber Studies
􀂇 Loaded Chamber Studies
􀂇 Routine Use/Ongoing Monitoring
􀂇 Validation Maintenance/Change Control/Revalidation
The sterilization method chosen depends on the application. The following methods are
typically available:
Method Typical application
Steam
sterilization
For the sterilization of fluids in ampoules, vials etc, or the
sterilization of processing equipment, reactors, preparation
tanks, solution delivery piping, etc. In general, sterilization
through the application of saturated steam under pressure is the
preferred method of sterilization. The principles apply to SIP
processes as well.
Sterilization by
filtration
Used for those products that cannot be sterilized due to the
heat sensitivity of the product or where heat labile packaging
is chosen since it provides a distinct patient benefit. Not the
preferred sterilization method.
Dry heat
sterilization and
Depyrogenation
Used to sterilize/depyrogenate containers (ampoules, vials,
etc.), pharmaceutical raw materials and processing equipment.
The use of dry heat has little application for the sterilization of
pharmaceutical drug products.
Radiation
sterilization
To sterilize packaging equipment, consumables, garments etc.
that are difficult to sterilize using steam or other methods.
Radiation sterilization is mostly used for medical devices.
5.1 Steam Sterilization
Steam Sterilization is the most common type of sterilization employed in the
pharmaceutical manufacturing environment. The principles of steam sterilization
are applicable to processes conducted within autoclaves as well as sterilizationin-
place (SIP) processes.
For the sterilization of fluids in e.g. vials and ampoules, a fluids load autoclave
cycle is used. The steam (or superheated water) is used as a heat transfer medium
to heat the contents of the vials/ampoules. The moisture required for sterilization
is derived from the contents in the vial/ampoule.
9
minute of the exposure stage.
The difference between the control probe, recording chart probe and independent
sensor (usually a thermocouple) during the exposure stage should not exceed + 1.0􀂃C.
5.1.1.2 Loaded Chamber Temperature Distribution
This study must be included to demonstrate that the equipment loading
patterns do not significantly change the chamber temperature distribution
within the chamber. Typically thermocouples are distributed throughout the
chamber (not in contact with load items) as for the Empty Chamber Temperature
Distribution study and cycles may be run using both the maximum and minimum
loads.
Loaded Chamber Temperature Distribution studies should meet an acceptance
criteria for the temperature to be within + 1􀂃C of the mean loaded chamber
temperature after one minute of the exposure stage.
The difference between the control probe, recording chart probe and independent
sensor during the exposure stage should not exceed + 1.0􀂃C.
5.1.1.3 Heat Penetration Studies
Loaded chamber heat penetration studies must be performed to demonstrate
that the pre-required time at temperature criteria are met for the loads being
validated. The heat penetration locations to be monitored are assessed
using both temperature probes and BIs. The thermocouples and the BIs should
be placed at the same locations wherever possible. Special emphasis is placed on
those locations identified during cycle development studies as being difficult for
steam to penetrate/difficult to heat (cold spot determination). Such locations
typically include the interior of hoses, filter housings, large objects, filling
apparatus and items with multiple layers of protective wrapping.
The placement of temperature probes and BIs within the load must not enhance the
penetration of steam into the load item. Where the load includes multiple items of
the same configuration in the load (e.g. bags of stoppers), BIs should be placed in a
second item adjacent to that containing the thermocouple.
This is to prevent the presence of the thermocouple from enhancing the penetration of
steam to the BI.Where items in the load are unique, the BIs must be placed in the load
item near the probe and precautions taken to prevent enhanced steam penetration.
Heat penetration cycles performed, as part of an initial validation exercise
must be repeated several times, e.g. three times,to demonstrate consistency.
These studies should be performed using established load patterns, though a
minimum and maximum load may be used to represent each particular load
pattern for qualification purposes. The purpose of the heat penetration study is to
document that the load items (including the cold spot) receives the minimum
required pre-determined time at temperature/F0.
16
Fans or blowers are generally used to help assure uniform distribution of heat
throughout the chamber. Fan speed should be determined during validation and
thereafter periodically monitored to assure operation within the acceptable range.
Typical test during the validation of a dry heat oven include:
5.4.1.1 Empty Chamber Temperature Distribution
Temperature probes (thermocouples) are distributed throughout the empty
chamber (in free space) and a temperature profile is produced. A uniform
temperature profile is expected. A probe should be located next to the controlling
sensor. Note the come-up time to temperature and cool-down times, as these
should be consistent in an empty chamber.
5.4.1.2 Loaded Chamber Temperature Distribution and Heat Penetration
In dry heat applications loaded chamber mapping and heat penetration
studies must be performed. Typically, these studies may be performed at the
same time. Thermocouples must be distributed throughout the chamber (in
free space) for heat distribution information. Thermocouples must also be
placed inside of the container, equipment or component being treated for the
heat penetration information. The penetration thermocouples need to make
contact with the surface of the item. This is because, due to the mass of the item,
the time to reach sterilization/depyrogenation temperature can substantially lag
behind the temperature of the surrounding air.
Biological indicators or endotoxin-spiked vials should be located adjacent to the
penetration thermocouples. Heat penetration studies, conducted as part of the
initial validation, should be repeated several times (e.g. three times) to
demonstrate consistency.
Product containers should have container mapping performed similar to that
described for steam sterilization. Load patterns are important as air is a poor
conductor of heat and the distribution of mass can greatly affect heating
characteristics. Assigning a temperature profile requirement is more problematic
as the profile is load specific. The temperature profile for a specific load should
how good reproducibility
5.4.2 Dry Heat Tunnels
Validation of dry-heat tunnels is demonstrated by both temperature measurements
and inactivation of bacterial endotoxins (depyrogenation).
Similar studies to dry-heat ovens are performed, i.e., empty tunnel temperature
distribution and loaded tunnel endotoxin challenge. The temperature variation
within the sterilization/depyrogenation zone may be greater than seen in an oven.
Higher temperatures are usually selected - ranging from 270􀂃􀂃 - 350􀂃C, due to the
shorter exposure time required and the greater temperature variation. Points to consider
for physical measurements include:
Belt Speed - determines the exposure time
Temperature - determines the time required for inactivation
19
5.6 Validation Maintenance/Change Control/Revalidation
Validation of sterilization processes is not a one-time exercise. Validation
maintenance is a phrase that describes a number of activities that support the
ongoing validated state of a process. In the case of sterilization processes, routine
bioburden monitoring, preventive maintenance, calibration, cycle review and
approval and annual reviews are activities that make up validation maintenance.
Bioburden monitoring must take place for every batch of aseptically
produced drug product. The preventive maintenance program should provide
clear instructions on reporting any unusual observations or equipment
breakdowns/mechanical failures so that an evaluation can be made on impact to
the validated state. Cycle review and approval should be in accordance with
detailed SOPs that have a direct traceability to the qualification and validation
activities, set point parameters and acceptance ranges.
Chamber vacuum leak testing, air detector device performance (if so equipped),
Bowie-Dick type testing for steam penetration, thermometric testing for small
loads, and other types of testing or review that may be required by local
regulatory authority expectation or requirement should be understood and
implemented as appropriate. For example, steam quality testing is a requirement
of the UK authorities. These tests are also mentioned in the PDA Technical
Monograph #1 and FDA􀂶s Guideline on Drug Products Produced By Aseptic
Processing.
The change control program should ensure that technical experts and the Quality
Assurance Function assess any planned changes to the equipment, process, loads,
procedures or documentation as to whether the qualified or validated state may be
impacted by the change. Any additional work necessary to demonstrate the
ongoing validated state should be reviewed and approved as part of the change
control process. Sterilization processes are critical processes. Be vigilant in
assuring that any changes or repairs are fully assessed for potential impact
via the change control process. Perform a periodic assessment of the
potential cumulative effect of changes that individually may not be
significant enough to prompt revalidation but, taken together, indicate a
need to re qualify or revalidate.
Sterilization processes must be revalidated at least annually in the absence of
any change-driven revalidation. The revalidation can be a subset of the original
validation work. Some acceptable approaches include:
􀂇 Single runs rather than 3 consecutive runs are sufficient in the absence of
recurring problems
􀂇 The selection of a worst case load pattern for revalidation
􀂇 The revalidation of each type of cycle (e.g. cycle for stoppers, cycle for
filling parts, cycle for products, etc.) but not the revalidation of each cycle
loading pattern.
􀂇 The revalidation of the worst case loading pattern and one selected
loading pattern, with the rotation of the remaining loading patterns at
subsequent annual revalidations.
􀂇 The revalidation of the empty chamber, maximum and minimum loading

WATER PURIFIED SYSTEMS VALIDATION

Abstract. Work methodology and professionals involved in pharmaceutical industry in Brazil has been changed in
the later years. At long Current Good Manufacturing Practices (cGMP) have been used by industrial processes,
especially in the multinational industries ones that use the main branch procedures. On the other hand, smaller
companies use own cGMPs demanding great effort from their employees. Anvisa´s RDC 134, reedit by RDC 210,
is the document that formalizes the so called cGMPs of World Health Organization (WHO) approved in 1994, as
well as obeying Mercosul standards. Since RDC was implemented, professionals of pharmaceutical industries have
been working to answer all the requirements, especially process validation. Pharmaceutical industry, as well as the
cosmetic, veterinarian and food ones answer with the qualification policies that are used to validate processes to the
sources of pressure and stimulus which are represents by the market and govern health agencies. Validation is a
typical procedure that refers mostly to Good Manufacturing Practices, whose main objective is to prevent
medicines to offer risks to the consumer, to the environment and to the operator. Water is essential to the
production of medicine and health products. In most cases it is added to the product during the manufacturing
process. Since it is suitable to the human consume, being drinkable doesn’t assure that it also can be used in
machines, industrial facilities, medicine, production, food products, cosmetics or any kind of chemical or
pharmaceutical products. In the present paper, a water purification system was studied to guarantee the
requirements of United States Pharmacopoea XXIII and XXIV. Filtration system, deionizer unit and an ultraviolet
(UV) light equipment, are part of the system. The monitorated parameters were Conductivity, Oxidizable
Substances, pH, Carbon Dioxide and Bacteriological Purity. The system was under control and it was validated
based on the results obtained.1. Introduction
In the last years, mainly at the end of 90´s, the validation approach in pharmaceutical industries has been
discussed due to its real importance within a productive process in relation to the products quality attributes, like
purity, safety and effectiveness, that constitutes the base of the thought and working of professionals involved
to the process validation (Santos, 2001). The U.S. Food and Drug Administration (FDA) in 1987 in his most
recently proposed named Guidelines on General Principles of Process Validation, has offered a definition to
process validation, after a series of incidents involved cross-contamination problems by some pharmaceutical
manufacturing establishments:
“Process validation is establishing documented evidence which provides a high degree of assurance that a
specific process will consistently produce a product meeting its predetermined specifications and quality
characteristics”.
Therefore, validation is legible through the documentation establishment, the guarantee of that manufacturing
process assures the product quality with homogeneity. The validation concept applies to the end product, and all
the previous stages are called qualification. So, the validation consists of a series of qualifications that involves
tests, systems verifications and also critical process parameters, that are its regulating keys, and can vary within
an acceptance limit. The different qualifications form than, the stages of validation: Design Qualification,
Installation Qualification, Operation Qualification and Performance Qualification (Jönck, 2002a). Each one must
follow its own schedule and protocol.
The water, in a pharmaceutical industry, is the most important raw material for the accomplishment of any
process. Therefore, it is essential to not only assure that water, but also installations and procedures are
validated. (Veneranda, 2004). So, it is necessary to choose a well-designed water system, by using a combination
of methods that allows reaching the quality levels of water to a determinate application, by optimizing its
particular capacity of remove contaminants.
The objective of this work consists in to validate the Steviafarma Industrial S/A company water purification
system, according to the specified requisites in United States Pharmacopeia (USP) XXIII and XXIV, adopting the
concurrent validation and the Statistical Control Process. The software Statistica 6.0 was the tool work used for
the data statistical treatment.
2.1. Equipaments
The water purified system is composed by a pre-filtration unit, with a 1 mm cellulose filter (Cunolatina)
followed by an activated carbon bed (Cunolatina). The first one is responsible to remove non-dissolved ions and
the second one to remove chlorine compounds and low molecular weight organic compounds, present in the feed
water, to the deionizer column protection. After the carbon bed, comes the deionizer unit, represented by a mixed
bed ion exchange column (Permution), with the objective of to removal the dissolved ions. The purified water than
passes through the ultraviolet equipment with a 254 nm wave length (Germetec), with germicide action for its
sterilization.
One of the most important variables in the process of purified water for pharmaceutical apllications is the
quality and the credibility of the water supplying system. In the present case, the water supplying is obtained
from SANEPAR – Paraná State Sanitation Company. The conductivity data were measured in a portable
equipment (Alpax) and pH data were obtained by using a Quimis pHmeter.
2.2. Methodology
The validation methodology applied to this system was the concurrent one, which was realized during the
routines operations. The installation qualification was only applied to UV equipment, because it was obtained
with the others equipment suppliers, documents certifying their own technical and functional specifications.
To UV equipment, an installation verification were made. Also, the technical specifications were checked, in
accordance with the design requirements, supplier instructions and supported documentation. The operational
qualification, had the purpose of to verify that the equipment operates as intended in all expected operational
range, according to their functional specifications. The project qualification was not carried through, because it is
more specific to new equipments or systems. According to Jönck (2002b), this stage is an activity that, through
critical process analysis, objectives to certify that the technical documentation incorporates the concepts of the
User Requirements Specifications (URS). Therefore, as the case was an installed system, the validation effort
started with the installation of the qualification.
The performance qualification consisted in to evaluate the system performance during the water production in
attempting to USP requirements. This stage was done in two phases. During the first phase, the system was kept
under normal operating levels during extensive and frequent samplings for four months in order to profile the
system. The second phase is the phase 1 continuation, but with lower frequency of sampling than phase 1, being
also carried out in 4 months. Generally each phase of testing should take 4-6 months to complete or longer, if
necessary, as proposed by Johnson (1993). The author also describes that a full validation of a system could
require as long as a year because of the operating problems, equipment failures, and maintenance errors, which
should be expected to occur during the validation period.
The system sampling procedure sampling consisted, first, in opening the water register and, after that the
feeding valve. Water was circulated through out the system during 15 minutes and, all the valves were sanified
with a 70% alcohol solution.
Chlorine Technique
For free chlorine evaluation, it was placed 1 mL of orto-toluidine in an erlenmeyer, and added 100 ml of the
water sampling. After the mixture, the color was compared immediately with permanent chlorine standards
(Pregnolatto e Pregnolatto, 1985).
Hardness Techinique
50 mL of water were tranfered to a 250 mL erlenmeyer and added 1 mL of ammonia buffer solution and a small
quantity (0,05 g) of Eriocromo Black T indicator. After that, the solution were titulated with EDTA 0,01 M until
purple color turn blue (Pregnolatto e Pregnolatto, 1985).
Viable Bacteria Count (Multiple Tube Fermentation Technique)
§ It were used 12 tubes series containing 10 mL of lauryl tryptose broth. The test was realized in triple. 1 mL of
water was added to 10 mL of lauryl tryptose in the first tube series and the dilutions 1:100 and 1:1000 were
prepared from the dilution 1:10. The three tubes left were the controls.
§ After the dilution has finished, they were taken to stoven during 48 hours at 30-350C.
§ If in a tube gas were present, it were considered positive for grown of microorganisms. The results were
expressed in MPN/mL according to a specific table (Farmacopéia Brazileira, 1988).
Yeasts Count (Pour Plate Method)
§ The plates were prepared in duplicata. 2 mL of sampling was separated, and in each one 1,0 mL were used.
§ After the Sabouraud Dextrose Agar culture medium be cold between the temperature of 44 to 48ºC, 20 mL was
added to the plates.
§ The plates under were rested for 10 to 15 minutes until the solidification. After that, the plates were taken to
stoven to 22-25°C for 48 - 72 hours, with a control (Farmacopéia Brasileira, 1988).
Heterotrophic Bacteria Count (Pour Plate Method)
The applied technique for yeasts determination was the same to bacteria. The only differences were in the
culture medium used: Plate Count Agar (PCA) and in the time and temperature of incubation: 30-35°C during 24
to 48 hours (Farmacopéia Brasileira, 1988).
Oxidizable Substances Determination
To 100 mL of purified water, 10 mL of 2 N H2SO4 was added and after that, the solution was heated until
boiling. Then, 0,1 mL of potassium permanganate 0,1 N were mixed to the solution, marking 10 minutes of boil
(USP XXIII, 1995).
Carbon Dioxide Determination
To 25 mL of purified water was added 25 mL of calcium hidroxide SR. The mixture had to remain clear (USP
XXIII, 1995).

Selecting and Validating Aqueous Detergents

Malcolm C. McLaughlin
May 1999
Today, the terms "critical" or "precision" cleaning are used interchangeably to refer to any cleaning process in which residue can cause a failure in the function of the surface being cleaned. In that regard, there is perhaps no more demanding application for cleaning than pharmaceutical manufacturing, where solids and liquids come into contact with plastic, glass, and metal piping and processing equipment, and where cross-contamination can be costly, in terms of both lost product and risk to human and animal health. Many leading drug companies, as well as firms that manufacture medical devices, are finding that aqueous cleaners provide the scrupulous cleaning required for manufacturing healthcare related products. Examples:
Capsules and tablets. Some pharmaceutical ingredients resist going into solution, making tablet presses and dies difficult to clean. Even stubborn, sustained-release product residues come clean quickly with appropriate aqueous cleaners.
Suspensions. Aqueous cleaners also eliminate intensive scrubbing and human contact in cleaning large stainless steel tanks as much as 2000 gal. used in manufacturing liquid suspensions.
Intermediates. Aqueous cleaners are suitable for cleaning glass-lined chemical reactors used in processing pharmaceutical intermediates such as powders, fillers, binding agents, and other chemicals. Aqueous cleaners are synthetic detergent cleaning agents used in a water solution. The chemical and mechanical action involved includes a number of processes: Solubilization increases the solubility of a substance in a particular medium.
Wetting lowers surface and interfacial tensions so that the cleaner penetrates small spaces while getting under the soil to lift it from the substrate.
Emulsification creates an oil/water mixture by coating oil droplets with surfactant to keep them from recombining and migrating to the surface of a cleaning bath.
Deflocculation prevents agglomeration by breaking soil into fine particles and dispersing them through the cleaning medium.
Sequestration is the reaction with ions such as calcium, magnesium, or heavy metals to prevent the formation of insoluble byproducts (such as soap scum).
Saponification is the alkaline hydrolysis of fat by the reaction of fatty acids with alkalies to form water-soluble soaps. The following reviews the principal components of aqueous critical cleaners, considerations for their selection, and methods of pharmaceutical validation of their use.
Components of Aqueous Cleaners
Water, the "universal solvent," is an important basic component of aqueous cleaners because it dissolves many types of soils. Water--which may be municipal tap water, well water, deionized, or distilled water, depending on the cleaning application--also functions as a carrying medium for detergent compounds. Water is a polar solvent that is good at dissolving a wide range of polar soils.
Water has a unique "V" shaped structure with two hydrogen atoms at the top of the "V" and an oxygen at the bottom. This directional shape towards the base of the "V" is called a dipole moment. Polar molecules such as water have a dipole moment. This dipole moment is important because it allows stable solutions of other dissolved polar-soil molecules to become arranged in more thermodynamically stable alternating positive and negative ends of molecules.
In addition to having a desirable dipole moment that helps dissolve other polar soils, water is capable of bonding to soils by a mechanism called hydrogen bonding. The ability of water to undergo hydrogen bonding is relatively unique among solvents. This capability gives water significant additional ability to dissolve soils as compared with other solvents. But while water is capable of dissolving many inorganic and some organic contaminants, not all soils dissolve readily in water. For this reason, aqueous detergent cleaners are complex mixtures specifically formulated to create greater chemical and mechanical cleaning action with ingredients that enhance the ability of water to hold soils. Typically these include surface-active agents (surfactants) and builders, which react with dissolved metal ions in the water to help stop them from interfering with cleaning.
It is critical that the detergent be able to clean effectively and to rinse away without leaving interfering residues and a scientifically formulated detergent will also typically include nondepositing rinse-aids. And since corrosion is the enemy of high-quality metal parts, corrosion inhibitors are often used in aqueous-cleaning formulations.
Detergent Selection
The major variables in cleaning using aqueous methods include the following:
Cleaner. The cleaner or detergent used should be matched to the desired cleaning method, and the surface and types of soils being cleaned. For instance, a low-foaming detergent should be used for spray or machine cleaning; a good anti-redeposition detergent for soak and ultrasonic cleaning; and a high emulsifying and wetting detergent for manual cleaning. The detergent, temperature, and degree of agitation should be strong enough to remove the soil to the desired level of cleanliness without harming the substrate being cleaned.
Agitation. Agitation can be none (as in soaking), or be performed manually (with a cloth, sponge, brush), ultrasonically, via flow-through clean-in-place (for pipes, tanks, and tubes), spray cleaning (clean-in-place spray balls), and high-pressure spray cleaning. In general, the more agitation, the more effective the cleaning on bulk soils. Cleaning can often be enhanced by pre-soaking, particularly if soils are dried or baked onto the part to be cleaned. It is always desirable, whenever possible, to clean prior to soils becoming dried or baked onto surfaces.
Temperature. In general, higher-temperature cleaning solutions result in better cleaning. In practice, there is typically an optimum temperature for a given combination of cleaning variables. Many soak, manual, and ultrasonic cleaning methods work best, for example, at 50-55¡C. Many spray washing techniques work best at 60-70ºC. Waxy or oily soils are more easily cleaned at somewhat higher temperatures. Particulate soils tend to be more easily cleaned at slightly lower temperatures.
Cleaning time. As a rule, the longer the cleaning time, the more thorough the cleaning. Many cleaning mechanisms-such as emulsifying, dissolving, suspending, and penetrating-are time-dependent. Up to the point where cleaning has been completed, the longer they're employed, the more cleaning is accomplished. Cleaning time can be accelerated by increased agitation, the use of more aggressive detergents, and by increasing temperature. If you cannot increase agitation, detergent, or temperature, then you must be prepared to use longer cleaning times to achieve the desired cleanliness. (There are some instances when long cleaning times may promote substrate corrosion, weakening, or swelling.)
Type of rinse. A thorough rinse will remove soils which have been cleaned from the surface and any residue from the detergent itself. Whatever contaminants are present in the rinse water can also be present after rinsing. Therefore, in pharmaceutical process equipment and medical device cleaning, high-purity rinse water is required.
Drying method. Drying can affect residues and corrosion since impurities from rinse water can be deposited during evaporation. Water, particularly high purity rinse water, can be corrosive to metal substrates during heated and air drying. The use of physical water removal drying techniques can help minimize such corrosion. It is also important to keep in mind that a cleaner's pH value can have a direct effect on cleaning effectiveness and each detergent formulation has maximum effectiveness at a specific pH value. An acidic cleaner would be effective for removing insoluble metal salts such as those used in time release coatings. Alkaline (basic) cleaners can be formulated to remove organic soils. Enzyme cleaners can be highly effective on protein soils. Alkaline cleaners work best when the soil can be hydrolyzed (typically natural oils and fats, fingerprints, natural greases, some types of pharmaceuticals, and protein residues). The cleaning process should be enclosed to avoid exposure hazards. Most cleaners are alkaline in nature since hydrolysis--and the chelation and dispersing of soils-typically occurs most effectively at alkaline pH levels. However, the higher the pH the more corrosive the cleaner.
Other Factors
It is important to select detergents for testing that are manufactured with appropriate quality-control procedures, with lot-number tracking and certificates of analysis from the manufacturer. These certificates document each lot of detergent to assure consistency and quality control from lot to lot in order to control for potential cleaning failure due to inconsistencies in manufacturing or unannounced formulation changes. It is desirable to choose a detergent from a manufacturer that maintains quality control on their raw materials and in turn keeps retained samples of each lot of detergent that is used to be able to respond to concerns about a particular batch. The detergent should be widely available and economical to use (for optimum economy, a concentrated detergent is typically used at 1:100 to 2:100 dilutions). The detergent concentrate should be diluted according to the manufacturer's instructions; typically, warm (about 50 degrees C) or hot (about 60 degrees C) water is used. Ambient temperature water may be acceptable, especially with presoaking. For difficult soils, very hot water should be used (>65 degrees C), and the recommended detergent concentration doubled.
Validation
In pharmaceutical manufacturing of human and animal health products, acceptable levels of residues must be carefully calculated. This is accomplished using a variation of the "Lilly Calculation," which involves determining a maximum dosage of the ingredient, the number of doses per volume of process equipment, and the surface area of that equipment: Maximum Residue= (maximum dose X number of doses in equipment) /surface area of equipment (Maximum dose is the highest amount to which a person can effectably be exposed--e.g., three logs below the LD50 of the compound.)
In practice, analytical methods can detect well below calculated maximum residues. The acceptable detection limits for a detergent are usually set well below, since the pharmaceutically active ingredients of a medicine will usually require much higher levels of cleanliness.
If 10 L of rinse water were used to extract the 100 L tank, then a rinse water residue of 10 ppm (100 mg/10 L) would correspond to 7 mg/square centimeter (100 mg/14,000 square centimeter). This is way below the typical calculated numbers.
There are a number of ways to analyze such residue based on the brand of detergent, including anionic surfactant analysis, direct UV/Visible determination, high performance liquid chromatography (HPLC), atomic absorption (AA) or inorganic residues, and liquid chromatography (LC), phosphate detection, protease enzyme detection, and total organic carbon (TOC). Detergent suppliers should be able to provide information on residue detection.
When rinsing with deionized water, conductivity has been used to detect conductive salts. Standard solutions of known dilution should be made up to determine the detection limits. There are two basic types of tests:
Rinse water test. Test a parameter of the rinsewater before and after rinsing the surface. No significant change in the parameter measured indicates no detected residue.
Wipe or swab test. Wipe a known area of the surface you wish to test with a clean swab or filter paper that is soaked with extraction solution such as 50/50 isopropanol/high purity water. Digest or extract the swab or filter paper and analyze for residue. Use glass fiber filter paper or swab for extraction methods. Do not use isopropanol mixtures for UV or TOC detection methods.
By choosing an appropriate cleaning method, detergent, and approach to residue detection, aqueous cleaners can be effectively used for cleaning pharmaceutical process equipment and medical devices.

Tutorial On HEPA Filtration

By: Jason Kelly
March 2007


HEPA filters are used in cleanrooms in many different industries, including semiconductor, pharmaceutical medical devices, nuclear, and biotechnology. The main function of a HEPA filter is to provide clean air to the cleanroom. The HEPA filter is constructed with many pleated layers of filter media paper; this design prevents particles from freely passing through the filter as they become trapped and stick onto the filter fibers (Figure 1). There are four mechanisms at work: capture by straining, impaction, interception, and diffusion (Figure 2). Straining/sieving is defined as when a particle is too large and becomes trapped between two filter fibers. Impaction is when a particle of relatively greater mass is unable to follow the curved streamline around the fiber and, as a result of momentum, travels in a straight line into the filter fiber and sticks. Interception occurs when a section of a particle “runs into” a filter fiber. Diffusion capture occurs when particles leave the streamline due to random collisions with the surrounding fluid molecules and strike the fibers, where they again stick.
LEAK TESTING
ISO 14644-2 outlines the frequency of cleanroom validation according to cleanroom classification. Part of this validation includes leak testing of the HEPA filter. ISO 14644-3 outlines the testing procedure to follow.
EQUIPMENT REQUIRED
Aerosol Generator: used to produce an aerosol upstream of the HEPA filter. A stable test aerosol has particles that have the following distribution:

• More than 20% by mass of particles less than 0.5 µm
• More than 50% by mass of particles less than 0.7 µm
• More than 75% by mass of particles less than 1.0 µm

An annual output test should be carried out to verify that the aerosol meets this distribution.
Photometer: used to measure the upstream aerosol concentration and downstream penetration of the HEPA filter by the aerosol; it should be calibrated at least annually.
Particle Counter: used to measure the concentrations of particles of different sizes downstream of the HEPA filter; it should be calibrated at least annually.

Figure 1: Filter media magnification x500

Figure 2: Filter mechanisms at work

AEROSOL GENERATOR OUTPUT TEST
The aerosol generator output test, carried out annually by the supplier, verifies that the aerosol generator is capable of producing a stable distribution. The photometer verifies that the sensor/optics and flow rates are within tolerances. Some new digital photometers now on the market have the ability to verify on “power up” the condition of the sensor/optics and flow rates as part of a self-diagnostics program and, therefore, improves the reliability of the unit continually instead of waiting annually for the calibration and hoping it passes. Photometer operators can be satisfied that the photometer is free from contamination around the sensor/optics, the flow path is unobstructed, the unit is performing correctly, and that the results are accurate.
TEST METHOD
A linear photometer may be used to measure upstream concentrations provided it is calibrated.
This upstream concentration may be used as the 100% reference, enabling percentage penetration downstream to be measured directly. The linear photometer may be used to test filters to 0.01%.
TEST REQUIREMENTS
Each filter requires an upstream challenge of 20–100 µg/L for best results. One must find the area of the filter and the airflow through the filter before calculating the throughput of the filter. The product of throughput and concentration are required to give the aerosol output required. Alternatively, the aerosol can be adjusted to reference the photometer at 100%.
TEST PROCEDURE
Disperse the test aerosol upstream of the HEPA filter to produce a uniform challenge concentration in the region of 20–100 µg/L. Using the photometer, measure the upstream challenge concentration.
Maintain this concentration throughout the test. Adjust the aerosol generator such that the challenge concentration at the upstream filter face is at a level such that the photometer can be set and maintained at a stable reading of 100%. The photometer is then set at 100%.
Using the same photometer, scan the entirety of the downstream face and perimeter inclusive of the sealing device with the sampling probe. Hold the probe approx 25 mm away from the area tested and pass over the entire area in slightly overlapping strokes, at a rate of 5 cm/sec. Record the location of any steady, repeatable reading of the photometer that exceeds 0.01% for grades A through D in the relevant class of environmental cleanliness. Refer to ISO 14644-3 section B.6.2.5 for a more detailed procedure.
GUIDELINES
Care should be taken when generating the upstream aerosol as too much aerosol will over-concentrate the filter and filter replacement will be necessary; use too little aerosol and there may be insufficient aerosol to effectively scan the filter sufficiently. Discuss these scenarios with to reach an agreement regarding the upstream concentration.
Once a leak has been detected, repair of the filter will be necessary, following the manufacturer’s procedure regarding the type of repair media and guidelines on resealing the filter and performing tests after the repair. After successful testing, a particle counter positioned under the filter can evaluate the condition of the filter and can be used as a back up to the photometer.
References

• ISO 14644-3
• ISO 14644-2
• IEST-RP-CC034
• IEST-RP-CC007

Jason Kelly is the Director of OptiCal Sciences,Ireland Ltd.and has been working in the semiconductor and pharmaceutical/medical device cleanroom industry for the last ten years.He can be reached at www.optical-sciences.ie.