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).