Supporting Clean Areas & Clean Areas Separation in Aseptic Product Manufacturing – USFDA

Supporting Clean Areas

Supporting clean areas can have various classifications and functions.  Many support areas function as zones in which nonsterile components, formulated products, in-process materials, equipment, and container/closures are prepared, held, or transferred.  These environments are soundly designed when they minimize the level of particle contaminants in the final product and control the microbiological content (bioburden) of articles and components that are subsequently sterilized.  

The nature of the activities conducted in a supporting clean area determines its classification.  FDA recommends that the area immediately adjacent to the aseptic processing line meet, at a minimum, Class 10,000 (ISO 7) standards (see Table 1) under dynamic conditions. 

Manufacturers can also classify this area as Class 1,000 (ISO 6) or maintain the entire aseptic filling room at Class 100 (ISO 5).  An area classified at a Class 100,000 (ISO 8) air cleanliness level is appropriate for less critical activities (e.g., equipment cleaning).

                   C.        Clean Area Separation 

An essential part of contamination prevention is the adequate separation of areas of operation.  To maintain air quality, it is important to achieve a proper airflow from areas of higher cleanliness to adjacent less clean areas.  It is vital for rooms of higher air cleanliness to have a substantial positive pressure differential relative to adjacent rooms of lower air cleanliness.  For example, a positive pressure differential of at least 10-15 Pascals (Pa)[1] should be maintained between adjacent rooms of differing classification (with doors closed).  When doors are open, outward airflow should be sufficient to minimize ingress of contamination, and it is critical that the time a door can remain ajar be strictly controlled (Ref. 4).  

In some cases, the aseptic processing room and adjacent cleanrooms have the same classification.  Maintaining a pressure differential (with doors closed) between the aseptic processing room and these adjacent rooms can provide beneficial separation.  In any facility designed with an unclassified room adjacent to the aseptic processing room, a substantial overpressure (e.g., at least 12.5 Pa) from the aseptic processing room should be maintained at all times to prevent contamination.  If this pressure differential drops below the minimum limit, it is important that the environmental quality of the aseptic processing room be restored and confirmed.

The Agency recommends that pressure differentials between cleanrooms be monitored continuously throughout each shift and frequently recorded.  All alarms should be documented and deviations from established limits should be investigated.

Air change rate is another important cleanroom design parameter.  For Class 100,000 (ISO 8) supporting rooms, airflow sufficient to achieve at least 20 air changes per hour is typically acceptable.  Significantly higher air change rates are normally needed for Class 10,000 and Class 100 areas.

A suitable facility monitoring system will rapidly detect atypical changes that can compromise the facility’s environment.  An effective system facilitates restoration of operating conditions to established, qualified levels before reaching action levels.  For example, pressure differential specifications should enable prompt detection (i.e., alarms) of an emerging low pressure problem to preclude ingress of unclassified air into a classified room.


[1] Equal to 0.04-0.06 inches of water gauge.  

Sterile Product Building, Facility and Clean Area Classification Guideline By USFDA

Sterile Product Building, Facility and  Clean Area Classification Guideline By USFDA has clearly mentioned what conditions to be followed to maintain the Sterile Conditions in Sterile Product Manufacturing site.

21 CFR 211.42(b) states, in part, that “The flow of components, drug product containers, closures, labeling, in-process materials, and drug products through the building or buildings shall be designed to prevent contamination.”

21 CFR 211.42(c) states, in part, that “Operations shall be performed within specifically defined areas of adequate size.  There shall be separate or defined areas or such other control systems for the firm’s operations as are necessary to prevent contamination or mixups during the course of the following procedures: * * * (10)  Aseptic processing, which includes as appropriate:  (i)  Floors, walls, and ceilings of smooth, hard surfaces that are easily cleanable; (ii) Temperature and humidity controls;  (iii)  An air supply filtered through high-efficiency particulate air filters under positive pressure, regardless of whether flow is laminar or nonlaminar; (iv) A system for monitoring environmental conditions;   (v) A system for cleaning and disinfecting the room and equipment to produce aseptic conditions; (vi) A system for maintaining any equipment used to control the aseptic conditions.”

21 CFR 211.46(b) states that “Equipment for adequate control over air pressure, micro-organisms, dust, humidity, and temperature shall be provided when appropriate for the manufacture, processing, packing, or holding of a drug product.”   21 CFR 211.46(c) states, in part, that “Air filtration systems, including prefilters and particulate matter air filters, shall be used when appropriate on air supplies to production areas * * *.”

21 CFR 211.63 states that “Equipment used in the manufacture, processing, packing, or holding of a drug product shall be of appropriate design, adequate size, and suitably located to facilitate operations for its intended use and for its cleaning and maintenance.”   21 CFR 211.65(a) states that “Equipment shall be constructed so that surfaces that contact components, inprocess materials, or drug products shall not be reactive, additive, or absorptive so as to alter the safety, identity, strength, quality, or purity of the drug product beyond the official or other established requirements.” 21 CFR 211.67(a) states that “Equipment and utensils shall be cleaned, maintained, and sanitized at appropriate intervals to prevent malfunctions or contamination that would alter the safety, identity, strength, quality, or purity of the drug product beyond the official or other established requirements.” 21 CFR 211.113(b) states that “Appropriate written procedures, designed to prevent microbiological contamination of drug products purporting to be sterile, shall be established and followed.  Such procedures shall include validation of any sterilization process.”

As provided for in the regulations, separate or defined areas of operation in an aseptic processing facility should be appropriately controlled to attain different degrees of air quality depending on the nature of the operation.  Design of a given area involves satisfying microbiological and particle criteria as defined by the equipment, components, and products exposed, as well as the operational activities conducted in the area. 

Clean area control parameters should be supported by microbiological and particle data obtained during qualification studies.  Initial cleanroom qualification includes, in part, an assessment of air quality under as-built, static conditions.  It is important for area qualification and classification to place most emphasis on data generated under dynamic conditions (i.e., with personnel present, equipment in place, and operations ongoing).  An adequate aseptic processing facility monitoring program also will assess conformance with specified clean area classifications under dynamic conditions on a routine basis.

The following table summarizes clean area air classifications and recommended action levels of microbiological quality (Ref. 1).

TABLE 1- Air Classifications

 Clean Area Classification (0.5 um particles/ft3)ISO  Designationb> 0.5 µm particles/m3Microbiological Active Air Action Levelsc (cfu/m3 )Microbiological Settling Plates Action Levelsc,d (diam. 90mm; cfu/4 hours)
10053,5201e1e
1000635,20073
10,0007352,000105
100,00083,520,00010050

a- All classifications based on data measured in the vicinity of exposed materials/articles during periods of activity.  b- ISO 14644-1 designations provide uniform particle concentration values for cleanrooms in multiple industries.  An ISO 5 particle concentration is equal to Class 100 and approximately equals EU Grade A. c- Values represent recommended levels of environmental quality.  You may find it appropriate to establish alternate microbiological action levels due to the nature of the operation or method of analysis.

  • The additional use of settling plates is optional.
  • Samples from Class 100 (ISO 5) environments should normally yield no microbiological contaminants.

Two clean areas are of particular importance to sterile drug product quality: the critical area and the supporting clean areas associated with it.

   A.        Critical Area – Class 100 (ISO 5)

A critical area is one in which the sterilized drug product, containers, and closures are exposed to environmental conditions that must be designed to maintain product sterility (§ 211.42(c)(10)).  Activities conducted in such areas include manipulations (e.g., aseptic connections, sterile ingredient additions) of sterile materials prior to and during filling and closing operations.  

This area is critical because an exposed product is vulnerable to contamination and will not be subsequently sterilized in its immediate container.  To maintain product sterility, it is essential that the environment in which aseptic operations (e.g., equipment setup, filling) are conducted be controlled and maintained at an appropriate quality.  One aspect of environmental quality is the particle content of the air.  Particles are significant because they can enter a product as an extraneous contaminant, and can also contaminate it biologically by acting as a vehicle for microorganisms (Ref. 2).  Appropriately designed air handling systems minimize particle content of a critical area.  

Air in the immediate proximity of exposed sterilized containers/closures and filling/closing operations would be of appropriate particle quality when it has a per-cubic-meter particle count of no more than 3520 in a size range of 0.5 µm and larger when counted at representative locations normally not more than 1 foot away from the work site, within the airflow, and during filling/closing operations.  This level of air cleanliness is also known as Class 100 (ISO 5).   

We recommend that measurements to confirm air cleanliness in critical areas be taken at sites where there is most potential risk to the exposed sterilized product, containers, and closures.  The particle counting probe should be placed in an orientation demonstrated to obtain a meaningful sample.  Regular monitoring should be performed during each production shift.  We recommend conducting nonviable particle monitoring with a remote counting system.  These systems are capable of collecting more comprehensive data and are generally less invasive than portable particle counters.   See Section X.E. for additional guidance on particle monitoring.

Some operations can generate high levels of product (e.g., powder) particles that, by their nature, do not pose a risk of product contamination.  It may not, in these cases, be feasible to measure air quality within the one-foot distance and still differentiate background levels of particles from air contaminants.  In these instances, air can be sampled in a manner that, to the extent possible, characterizes the true level of extrinsic particle contamination to which the product is exposed.  Initial qualification of the area under dynamic conditions without the actual filling function provides some baseline information on the non-product particle generation of the operation. 

HEPA-filtered[1] air should be supplied in critical areas at a velocity sufficient to sweep particles away from the filling/closing area and maintain unidirectional airflow during operations.  The velocity parameters established for each processing line should be justified and appropriate to maintain unidirectional airflow and air quality under dynamic conditions within the critical area (Ref. 3).[2]  

Proper design and control prevents turbulence and stagnant air in the critical area.  Once relevant parameters are established, it is crucial that airflow patterns be evaluated for turbulence or eddy currents that can act as a channel or reservoir for air contaminants (e.g., from an adjoining lower classified area).  In situair pattern analysis should be conducted at the critical area to demonstrate unidirectional airflow and sweeping action over and away from the product under dynamic conditions.  The studies should be well documented with written conclusions, and include evaluation of the impact of aseptic manipulations (e.g., interventions) and equipment design.  Videotape or other recording mechanisms have been found to be useful aides in assessing airflow initially as well as facilitating evaluation of subsequent equipment configuration changes.  It is important to note that even successfully qualified systems can be compromised by poor operational, maintenance, or personnel practices.

Air monitoring samples of critical areas should normally yield no microbiological contaminants.  We recommend affording appropriate investigative attention to contamination occurrences in this environment.


[1] High Efficiency Particulate Air filter

[2] A velocity of 0.45 meters/second (90 feet per minute) has generally been established, with a range of plus or minus 20 percent around the setpoint.  Higher velocities may be appropriate in operations generating high levels of particulates.

Reference :-

Sterile Drug Products  Produced by Aseptic Processing — Current Good Manufacturing Practice (USFDA)

TEST FOR SPECIFIED MICRO-ORGANISMS (Ph. Eur. method 2.6.13.)

  1. TEST FOR SPECIFIED MICRO-ORGANISMS (Ph. Eur. method 2.6.13.) INTRODUCTION The tests described hereafter will allow determination of the absence or limited occurrence of specified micro-organisms that may be detected under the conditions described. The tests are designed primarily to determine whether a substance or preparation complies with an established specification for microbiological quality. When used for such purposes, follow the instructions given below, including the number of samples to be taken, and interpret the results as stated below. Alternative microbiological procedures, including automated methods, may be used, provided that their equivalence to the Pharmacopoeia method has been demonstrated.
  2. GENERAL PROCEDURES The preparation of samples is carried out as described in general chapter 2.6.12. If the product to be examined has antimicrobial activity, this is insofar as possible removed or neutralised as described in general chapter 2.6.12. If surface-active substances are used for sample preparation, their absence of toxicity for micro-organisms and their compatibility with inactivators used must be demonstrated as described in general chapter 2.6.12.
  3. GROWTH-PROMOTING AND INHIBITORY PROPERTIES OF THE MEDIA, SUITABILITY OF THE TEST AND NEGATIVE CONTROLS The ability of the test to detect micro-organisms in the presence of the product to be tested must be established. Suitability must be confirmed if a change in testing performance, or the product, which may affect the outcome of the test is introduced. 3-1 PREPARATION OF TEST STRAINS Use standardised stable suspensions of test strains or prepare them as stated below. Seed lot culture maintenance techniques (seed-lot systems) are used so that the viable micro-organisms used for inoculation are not more than 5 passages removed from the original master seed-lot. 3-1-1 Aerobic micro-organisms  Grow each of the bacterial test strains separately in casein soya bean digest broth or on casein soya bean digest agar at 30-35 °C for 18-24 h. Grow the test strain for Candida albicans separately on Sabouraud-dextrose agar or in Sabouraud-dextrose broth at 20-25 °C for 2-3 days.  — Staphylococcus aureus such as ATCC 6538, NCIMB 9518, CIP 4.83 or NBRC 13276;  — Pseudomonas aeruginosa such as ATCC 9027, NCIMB 8626, CIP 82.118 or NBRC 13275;  — Escherichia coli such as ATCC 8739, NCIMB 8545, CIP 53.126 or NBRC 3972;  — Salmonella enterica subsp. enterica serovar Typhimurium, such as ATCC 14028 or, as an alternative, Salmonella enterica subsp. enterica serovar Abony such as NBRC 100797, NCTC 6017 or CIP 80.39;  — Candida albicans such as ATCC 10231, NCPF 3179, IP 48.72 or NBRC 1594. Use buffered sodium chloride-peptone solution pH 7.0 or phosphate buffer solution pH 7.2 to make test suspensions. Use the suspensions within 2 h or within 24 h if stored at 2-8 °C. 3-1-2 Clostridia  Use Clostridium sporogenes such as ATCC 11437 (NBRC 14293, NCIMB 12343, CIP 100651) or ATCC 19404 (NCTC 532 or CIP 79.03) or NBRC 14293. Grow the clostridial test strain under anaerobic conditions in reinforced medium for clostridia at 30-35 °C for 24-48 h. As an alternative to preparing and then diluting down a fresh suspension of vegetative cells of Cl. sporogenes, a stable spore suspension is used for test inoculation. The stable spore suspension may be maintained at 2-8 °C for a validated period. 3-2 NEGATIVE CONTROL To verify testing conditions, a negative control is performed using the chosen diluent in place of the test preparation. There must be no growth of micro- organisms. A negative control is also performed when testing the products as described in section
  4. A failed negative control requires an investigation. 3-3 GROWTH PROMOTION AND INHIBITORY PROPERTIES OF THE MEDIA Test each batch of ready-prepared medium and each batch of medium prepared either from dehydrated medium or from ingredients. Verify suitable properties of relevant media as described in Table 2.6.13.-1
 Test for growth promoting properties, liquid media  

 Inoculate a portion of the appropriate medium with a small number (not more than  100 CFU) of the appropriate micro-organism. Incubate at the specified temperature  for not more than the shortest period of time specified in the test. Clearly visible  growth of the micro-organism comparable to that previously obtained with a  previously tested and approved batch of medium occurs. 


 Test for growth promoting properties, solid media  

 Perform the surface-spread method, inoculating each plate with a small number  (not more than 100 CFU) of the appropriate micro-organism. Incubate at the  specified temperature for not more than the shortest period of time specified in the  test. Growth of the micro-organism comparable to that previously obtained with a  previously tested and approved batch of medium occurs. 


 Test for inhibitory properties, liquid or solid media  

 Inoculate the appropriate medium with at least 100 CFU of the appropriate micro- organism. Incubate at the specified temperature for not less than the longest  period of time specified in the test. No growth of the test micro-organism occurs. 


 Test for indicative properties  

 Perform the surface-spread method, inoculating each plate with a small number  (not more than 100 CFU) of the appropriate micro-organism. Incubate at the  specified temperature for a period of time within the range specified in the test.  Colonies are comparable in appearance and indication reactions to those  previously obtained with a previously tested and approved batch of medium. 

 3-4 SUITABILITY OF THE TEST METHOD

 For each product to be tested, perform the sample preparation as described in the  relevant paragraph in section 4. Add each test strain at the time of mixing, in the  prescribed growth medium. Inoculate the test strains individually. Use a number of  micro-organisms equivalent to not more than 100 CFU in the inoculated test  preparation. 

 Perform the test as described in the relevant paragraph in section 4 using the  shortest incubation period prescribed. 

 The specified micro-organisms must be detected with the indication reactions as  described in section 4.  

 Any antimicrobial activity of the product necessitates a modification of the test  procedure (see 4-5-3 of general chapter 2.6.12). 

 If for a given product the antimicrobial activity with respect to a micro-organism for  which testing is prescribed cannot be neutralised, then it is to be assumed that the  inhibited micro-organism will not be present in the product. 

 4 TESTING OF PRODUCTS

 4-1 BILE-TOLERANT GRAM-NEGATIVE BACTERIA


 4-1-1 Sample preparation and pre-incubation  Prepare a sample using a 1 in  10 dilution of not less than 1 g of the product to be examined as described in  general chapter 2.6.12, but using casein soya bean digest broth as the chosen  diluent, mix and incubate at 20-25 °C for a time sufficient to resuscitate the  bacteria but not sufficient to encourage multiplication of the organisms (usually 2 h  but not more than 5 h). 


 4-1-2 Test for absence  Unless otherwise prescribed, use the volume  corresponding to 1 g of the product, as prepared in 4-1-1, to inoculate  enterobacteria enrichment broth-Mossel. Incubate at 30-35 °C for 24-48 h.  Subculture on plates of violet red bile glucose agar. Incubate at 30-35 °C for 18-24  h. 

 The product complies with the test if there is no growth of colonies. 

 4-1-3 Quantitative test 


 4131Selection and subculture --- . Inoculate suitable quantities of  enterobacteria enrichment broth-Mossel with the preparation as described under 4- 1-1 and/or dilutions of it containing respectively 0.1 g, 0.01 g and 0.001 g (or 0.1  mL, 0.01 mL and 0.001 mL) of the product to be examined. Incubate at 30-35 °C  for 24-48 h. Subculture each of the cultures on a plate of violet red bile glucose  agar. Incubate at 30-35 °C for 18-24 h. 


 4132Interpretation --- . Growth of colonies constitutes a positive result. Note the  smallest quantity of the product that gives a positive result and the largest quantity  that gives a negative result. Determine from Table 2.6.13.-2 the probable number of  bacteria. 
 4-2 ESCHERICHIA COLI


 4-2-1 Sample preparation and pre-incubation  Prepare a sample using a 1 in  10 dilution of not less than 1 g of the product to be examined as described in  general chapter 2.6.12, and use 10 mL or the quantity corresponding to 1 g or 1  mL to inoculate a suitable amount (determined as described under 3-4) of casein  soya bean digest broth, mix and incubate at 30-35 °C for 18-24 h. 


 4-2-2 Selection and subculture  Shake the container, transfer 1 mL of casein  soya bean digest broth to 100 mL of MacConkey broth and incubate at 42-44 °C  for 24-48 h. Subculture on a plate of MacConkey agar at 30-35 °C for 18-72 h. 


 4-2-3 Interpretation  Growth of colonies indicates the possible presence of E.  coli. This is confirmed by identification tests. 

 The product complies with the test if no colonies are present or if the identification  tests are negative. 

 4-3 SALMONELLA


 4-3-1 Sample preparation and pre-incubation  Prepare the product to be  examined as described in general chapter 2.6.12, and use the quantity  corresponding to not less than 10 g or 10 mL to inoculate a suitable amount  (determined as described under 3-4) of casein soya bean digest broth, mix and  incubate at 30-35 °C for 18-24 h. 


 4-3-2 Selection and subculture  Transfer 0.1 mL of casein soya bean digest  broth to 10 mL of Rappaport Vassiliadis Salmonella enrichment broth and incubate  at 30-35 °C for 18-24 h. Subculture on plates of xylose, lysine, deoxycholate agar.  Incubate at 30-35 °C for 18-48 h. 


 4-3-3 Interpretation  The possible presence of Salmonella is indicated by the  growth of well-developed, red colonies, with or without black centres. This is  confirmed by identification tests.  

 The product complies with the test if colonies of the types described are not  present or if the confirmatory identification tests are negative. 

 4-4 PSEUDOMONAS AERUGINOSA


 4-4-1 Sample preparation and pre-incubation  Prepare a sample using a 1 in  10 dilution of not less than 1 g of the product to be examined as described in  general chapter 2.6.12, and use 10 mL or the quantity corresponding to 1 g or 1  mL to inoculate a suitable amount (determined as described under 3-4) of casein  soya bean digest broth and mix. When testing transdermal patches, filter the  volume of sample corresponding to 1 patch of the preparation described under 4-5- 1 in general chapter 2.6.12 through a sterile filter membrane and place in 100 mL  of casein soya bean digest broth. Incubate at 30-35 °C for 18-24 h. 


 4-4-2 Selection and subculture  Subculture on a plate of cetrimide agar and  incubate at 30-35 °C for 18-72 h. 


 4-4-3 Interpretation  Growth of colonies indicates the possible presence of P.  aeruginosa. This is confirmed by identification tests. 

 The product complies with the test if colonies are not present or if the confirmatory  identification tests are negative. 

 4-5 STAPHYLOCOCCUS AUREUS


 4-5-1 Sample preparation and pre-incubation  Prepare a sample using a 1 in  10 dilution of not less than 1 g of the product to be examined as described in  general chapter 2.6.12, and use 10 mL or the quantity corresponding to 1 g or 1  mL to inoculate a suitable amount (determined as described under 3-4) of casein  soya bean digest broth and mix. When testing transdermal patches, filter the  volume of sample corresponding to 1 patch of the preparation described under 4-5- 1 in general chapter 2.6.12 through a sterile filter membrane and place in 100 mL  of casein soya bean digest broth. Incubate at 30-35 °C for 18-24 h. 


 4-5-2 Selection and subculture  Subculture on a plate of mannitol salt agar and  incubate at 30-35 °C for 18-72 h. 


 4-5-3 Interpretation  The possible presence of S. aureus is indicated by the  growth of yellow/white colonies surrounded by a yellow zone. This is confirmed by  identification tests.  

 The product complies with the test if colonies of the types described are not  present or if the confirmatory identification tests are negative. 

 4-6 CLOSTRIDIA


 4-6-1 Sample preparation and heat treatment  Prepare a sample using a 1 in  10 dilution (with a minimum total volume of 20 mL) of not less than 2 g or 2 mL of  the product to be examined as described in general chapter 2.6.12. Divide the  sample into 2 portions of at least 10 mL. Heat 1 portion at 80 °C for 10 min and  cool rapidly. Do not heat the other portion. 


 4-6-2 Selection and subculture  Use 10 mL or the quantity corresponding to 1  g or 1 mL of the product to be examined of both portions to inoculate suitable  amounts (determined as described under 3-4) of reinforced medium for clostridia.  Incubate under anaerobic conditions at 30-35 °C for 48 h. After incubation, make  subcultures from each container on Columbia agar and incubate under anaerobic  conditions at 30-35 °C for 48-72 h. 


 4-6-3 Interpretation  The occurrence of anaerobic growth of rods (with or without  endospores) giving a negative catalase reaction indicates the presence of  clostridia. This is confirmed by identification tests. 

 The product complies with the test if colonies of the types described are not  present or if the confirmatory identification tests are negative. 

 4-7 CANDIDA ALBICANS


 4-7-1 Sample preparation and pre-incubation  Prepare the product to be  examined as described in general chapter 2.6.12, and use 10 mL or the quantity  corresponding to not less than 1 g or 1 mL to inoculate 100 mL of Sabouraud- dextrose broth and mix. Incubate at 30-35 °C for 3-5 days. 


 4-7-2 Selection and subculture  Subculture on a plate of Sabouraud-dextrose  agar and incubate at 30-35 °C for 24-48 h. 


 4-7-3 Interpretation  Growth of white colonies may indicate the presence of C.  albicans. This is confirmed by identification tests. 

 The product complies with the test if such colonies are not present or if the  confirmatory identification tests are negative. 


 The following section is given for information.  

 5 RECOMMENDED SOLUTIONS AND CULTURE MEDIA

 The following solutions and culture media have been found to be satisfactory for the  purposes for which they are prescribed in the test for microbial contamination in  the Pharmacopoeia. Other media may be used provided that their suitability can be  demonstrated. 


 Stock buffer solution  Place 34 g of potassium dihydrogen phosphate in a 1000  mL volumetric flask, dissolve in 500 mL of purified water, adjust to pH 7.2 ± 0.2  with sodium hydroxide, dilute to 1000.0 mL with purified water and mix. Dispense  into containers and sterilise. Store at 2-8 °C. 

Reference :-

  1. TEST FOR SPECIFIED MICRO-ORGANISMS (Ph. Eur. method 2.6.13.)

SOP For Operation, Maintenance And Calibration Of Ultrasonic Water Bath

The purpose of this Standard Operating Procedure (SOP) is to lay down the procedure for Operation, maintenance and Calibration of Ultrasonic Water Bath.

  • PROCEDURE:
    1. PRELIMINARY CHECK:
      1. Fill the Water bath with water up to height of minimum 7cm from bottom of tank. Whenever the Ultrasonic Water Bath is required with specific temperature then heater to be kept “ON”.
      1. Ensure that there is no leakage of water from water bath.
      1. Do not put any heavy objects on the bottom of the tank as it can damage the transducers. Always use tray supplied with ultrasonic bath.
      1. Ensure that liquid does not splash on the controls and switches.
  • Basic Operation :Connect the Ultrasonic Water Bath to the main power supply with the connecting plug the cleaner into grounded outlet after power on the temperature displays the actual environment temperature time and temperature default as the last setting ones.( 3 minutes default time for initial use temperature display 0°C)Press + Button adjustable for timer working time increase 1 minute hold the key the time raised by 10 minutes continuous in the contrary.Press – Button adjustable for timer working time reduce 1 minute hold the key the reduced by 10 minutes continuous the cleaning stops when the counts down 00:00.Press Button for heating key for temperature setting with the range 0°C-80°C.Press + Button adjustable for heating a time temperature will rise 1°C holding the key temperature will rise continuously with 10°C in the contrary.Press – Button temperature reduce 1°C holding the key temperature reduced continuously with 10°C usually the best results are with 40°C-60°C.When setting the temperature if the setting temperature exceed the environment temperature Press button heating key the heating working and indicator light up below the environment temperature heating can’t start and the indicator light turn off.Heating won’t shut off automatically when the environment temperature below the setting press Button heating key in to stop heating (heating can’t stop if you don’t press Button heating key).After time and temperature setting press Button heating key en press any other keys Degas,Soft, Normal Machine start to work indicator light up if need to stop heating or ultrasonic press corresponding key again then corresponding indicator lights turn off.Machine stop working after 8hours working time (power saving mode) in this made machine restored to the standby state once you press any key.Three ultrasonic working mode:Degas mode: Press Degas Button intermittent operation of ultrasonic power for rapid removal of entrained air from liquids ultrasonic work 6 seconds and stop 2 seconds same cycle to be proceed & press Degas key again to stop it.Normal mode: Press Normal Button to start the normal faction strong ultrasonic power with large current press again to stop it.Soft mode: Press Soft Button to start soft function continuous slight variation of ultrasonic frequency to eliminate hot spots dead zones and standing waves press again to stop it. During working you will hear the “sizzling“ voice that means the cleaner running properly.
  • MAINTENANCE  : Ensure system is off and disconnect the power cable from the main socket before cleaning.Clean the water bath and replace the water on daily basis or as and when the water gets turbid.
  • CALIBRATION PROTOCOL:

            Calibration Frequency: Yearly

  • Carry out the Calibration of temperature indicator controller with sensor and time by an approved external party.

Sterile Drug Products Produced by Aseptic Processing – USFDA Guideline for Buildings & Facilities

21 CFR 211.42(b) states, in part, that “The flow of components, drug product containers, closures, labeling, in-process materials, and drug products through the building or buildings shall be designed to prevent contamination.
21 CFR 211.42(c) states, in part, that “Operations shall be performed within specifically defined areas of adequate size. There shall be separate or defined areas or such other control systems for the firm’s operations as are necessary to prevent contamination or mixups during the course of the following procedures: * * *
(10) Aseptic processing, which includes as appropriate:

(i) Floors, walls, and ceilings of smooth, hard surfaces that are easily cleanable;

(ii) Temperature and humidity controls;

(iii) An air supply filtered through high-efficiency particulate air filters under positive pressure, regardless of whether flow is laminar or nonlaminar;

(iv) A system for monitoring environmental conditions;

(v) A system for cleaning and disinfecting the room and equipment to produce aseptic conditions;

(vi) A system for maintaining any equipment used to control the aseptic conditions.”

21 CFR 211.46(b) states that “Equipment for adequate control over air pressure, micro-organisms, dust, humidity, and temperature shall be provided when appropriate for the manufacture, processing, packing, or holding of a drug product.”
21 CFR 211.46(c) states, in part, that “Air filtration systems, including prefilters and particulate matter air filters, shall be used when appropriate on air supplies to production areas * * *.”
21 CFR 211.63 states that “Equipment used in the manufacture, processing, packing, or holding of a drug product shall be of appropriate design, adequate size, and suitably located to facilitate operations for its intended use and for its cleaning and maintenance.”
21 CFR 211.65(a) states that “Equipment shall be constructed so that surfaces that contact components, inprocess materials, or drug products shall not be reactive, additive, or absorptive so as to alter the safety, identity, strength, quality, or purity of the drug product beyond the official or other established requirements.”
21 CFR 211.67(a) states that “Equipment and utensils shall be cleaned, maintained, and sanitized at appropriate intervals to prevent malfunctions or contamination that would alter the safety, identity, strength, quality, or purity of the drug product beyond the official or other established requirements.”
21 CFR 211.113(b) states that “Appropriate written procedures, designed to prevent microbiological contamination of drug products purporting to be sterile, shall be established and followed. Such procedures shall include validation of any sterilization process.”

EU Guidelines For Computerized System Part- 2

Project Phase

  1. Validation
    4.1 The validation documentation and reports should cover the relevant steps of the life cycle. Manufacturers should be able to justify their standards, protocols, acceptance criteria, procedures and records based on their risk assessment.

4.2 Validation documentation should include change control records (if applicable) and reports on any deviations observed during the validation process.
4.3 An up to date listing of all relevant systems and their GMP functionality (inventory)
should be available.
For critical systems an up to date system description detailing the physical and logical
arrangements, data flows and interfaces with other systems or processes, any hardware and software pre-requisites, and security measures should be available.
4.4 User Requirements Specifications should describe the required functions of the
computerised system and be based on documented risk assessment and GMP impact. User requirements should be traceable throughout the life-cycle.
4.5 The regulated user should take all reasonable steps, to ensure that the system has been developed in accordance with an appropriate quality management system. The supplier should be assessed appropriately.
4.6 For the validation of bespoke or customised computerised systems there should be a process in place that ensures the formal assessment and reporting of quality and performance measures for all the life-cycle stages of the system.
4.7 Evidence of appropriate test methods and test scenarios should be demonstrated.
Particularly, system (process) parameter limits, data limits and error handling should be considered. Automated testing tools and test environments should have documented assessments for their adequacy.
4.8 If data are transferred to another data format or system, validation should include checks that data are not altered in value and/or meaning during this migration process.

EU Guidelines For Computerized System Part -1

Principle
This annex applies to all forms of computerised systems used as part of a GMP regulated activities. A computerised system is a set of software and hardware components which together fulfill certain functionalities.
The application should be validated; IT infrastructure should be qualified.
Where a computerised system replaces a manual operation, there should be no resultant decrease in product quality, process control or quality assurance. There should be no increase in the overall risk of the process.


General

  1. Risk Management
    Risk management should be applied throughout the lifecycle of the computerised system taking into account patient safety, data integrity and product quality. As part of a risk management system, decisions on the extent of validation and data integrity controls should be based on a justified and documented risk assessment of the computerised system.
  2. Personnel
    There should be close cooperation between all relevant personnel such as Process Owner, System Owner, Qualified Persons and IT. All personnel should have appropriate qualifications, level of access and defined responsibilities to carry out their assigned duties.
  3. Suppliers and Service Providers
    3.1 When third parties (e.g. suppliers, service providers) are used e.g. to provide, install, configure, integrate, validate, maintain (e.g. via remote access), modify or retain a computerised system or related service or for data processing, formal agreements must exist between the manufacturer and any third parties, and these agreements should include clear statements of the responsibilities of the third party. IT-departments should be considered analogous.
    3.2 The competence and reliability of a supplier are key factors when selecting a product or service provider. The need for an audit should be based on a risk assessment.
    3.3 Documentation supplied with commercial off-the-shelf products should be reviewed by regulated users to check that user requirements are fulfilled.
    3.4 Quality system and audit information relating to suppliers or developers of software and implemented systems should be made available to inspectors on request.

Manufacture of Liquids, Creams and Ointments

Principle
Liquids, creams and ointments may be particularly susceptible to microbial and other
contamination during manufacture. Therefore special measures must be taken to prevent any contamination.


Premises and equipment

  1. The use of closed systems for processing and transfer is recommended in order to protect the product from contamination. Production areas where the products or open clean containers are exposed should normally be effectively ventilated with filtered air.
  2. Tanks, containers, pipework and pumps should be designed and installed so that they may be readily cleaned and if necessary sanitised. In particular, equipment design should include a minimum of dead-legs or sites where residues can accumulate and promote microbial proliferation.
  3. The use of glass apparatus should be avoided wherever possible. High quality stainless steel is often the material of choice for parts coming into contact with product.
    Production
  4. The chemical and microbiological quality of water used in production should be specified and monitored. Care should be taken in the maintenance of water systems in order to avoid the risk of microbial proliferation. After any chemical sanitisation of the water systems, a validated flushing procedure should be followed to ensure that the sanitising agent has been effectively removed.
  5. The quality of materials received in bulk tankers should be checked before they are
    transferred to bulk storage tanks.
  6. Care should be taken when transferring materials via pipelines to ensure that they are delivered to their correct destination.
  7. Materials likely to shed fibres or other contaminants, like cardboard or wooden pallets, should not enter the areas where products or clean containers are exposed.
  8. Care should be taken to maintain the homogeneity of mixtures, suspensions, etc. during filling. Mixing and filling processes should be validated. Special care should be taken at the beginning of a filling process, after stoppages and at the end of the process to ensure that homogeneity is maintained.
  9. When the finished product is not immediately packaged, the maximum period of storage and the storage conditions should be specified and adhered to.

Sampling on Starting and Packaging Materials

Principle
Sampling is an important operation in which only a small fraction of a batch is taken. Valid conclusions on the whole cannot be based on tests which have been carried out on non representative samples. Correct sampling is thus an essential part of a system of Quality Assurance.
Note
Sampling is dealt with in Chapter 6 of the Guide, items 6.11. to 6.14. This annex gives
additional guidance on the sampling of starting and packaging materials.


Personnel

  1. Personnel who take samples should receive initial and on-going regular training in the
    disciplines relevant to correct sampling. This training should include:
    — sampling plans,
    — written sampling procedures,
    — the techniques and equipment for sampling,
    — the risks of cross-contamination,
    — the precautions to be taken with regard to unstable and/or sterile substances,
    — the importance of considering the visual appearance of materials, containers and labels,
    — the importance of recording any unexpected or unusual circumstances.
    Starting materials
  2. The identity of a complete batch of starting materials can normally only be ensured if individual samples are taken from all the containers and an identity test performed on each sample. It is permissible to sample only a proportion of the containers where a validated procedure has been established to ensure that no single container of starting material has been incorrectly labelled.
  3. This validation should take account of at least the following aspects:
    — the nature and status of the manufacturer and of the supplier and their
    understanding of the GMP requirements of the Pharmaceutical Industry;
    — the Quality Assurance system of the manufacturer of the starting material;

— the manufacturing conditions under which the starting material is produced and
controlled;
— the nature of the starting material and the medicinal products in which it will be
used.
Under such a system, it is possible that a validated procedure exempting identity testing of
each incoming container of starting material could be accepted for:
— starting materials coming from a single product manufacturer or plant;
— starting materials coming directly from a manufacturer or in the manufacturer’s
sealed container where there is a history of reliability and regular audits of the
manufacturer’s Quality Assurance system are conducted by the purchaser (the
manufacturer of the medicinal product) or by an officially accredited body.
It is improbable that a procedure could be satisfactorily validated for:
— starting materials supplied by intermediaries such as brokers where the source of
manufacture is unknown or not audited;
— starting materials for use in parenteral products.

  1. The quality of a batch of starting materials may be assessed by taking and testing a
    representative sample. The samples taken for identity testing could be used for this purpose. The number of samples taken for the preparation of a representative sample should be determined statistically and specified in a sampling plan. The number of individual samples which may be blended to form a composite sample should also be defined, taking into account the nature of the material, knowledge of the supplier and the homogeneity of the composite sample.
    Packaging material
  2. The sampling plan for packaging materials should take account of at least the following:
    the quantity received, the quality required, the nature of the material (e.g. primary
    packaging materials and/or printed packaging materials), the production methods, and what is known of the Quality Assurance system of the packaging materials manufacturer based on audits. The number of samples taken should be determined statistically and specified in a sampling plan.

EU Guideline For Outsourced Activities

EU Guidelines For Outsourced Activities Any activity covered by the GMP Guide that is outsourced should be appropriately defined, agreed and controlled in order to avoid misunderstandings which could result in a product or operation of unsatisfactory quality. There must be a written Contract between the Contract Giver and the Contract Acceptor which clearly establishes the duties of each party. The Quality Management System of the Contract Giver must clearly state the way that the Qualified Person certifying each batch of product for release exercises his full responsibility

1 There should be a written Contract covering the outsourced activities, the
products or operations to which they are related, and any technical arrangements
made in connection with it.
2 All arrangements for the outsourced activities including any proposed changes
in technical or other arrangements should be in accordance with regulations in force,
and the Marketing Authorisation for the product concerned, where applicable.
3 Where the marketing authorization holder and the manufacturer are not the
same, appropriate arrangements should be in place, taking into account the principles described in this chapter.


The Contract Giver


4 The pharmaceutical quality system of the Contract Giver should include the
control and review of any outsourced activities. The Contract Giver is ultimately
responsible to ensure processes are in place to assure the control of outsourced
activities. These processes should incorporate quality risk management principles and notably include:
5 Prior to outsourcing activities, the Contract Giver is responsible for assessing
the legality, suitability and the competence of the Contract Acceptor to carry out
successfully the outsourced activities. The Contract Giver is also responsible for
ensuring by means of the Contract that the principles and guidelines of GMP as
interpreted in this Guide are followed.
6 The Contract Giver should provide the Contract Acceptor with all the
information and knowledge necessary to carry out the contracted operations correctly in accordance with regulations in force, and the Marketing Authorisation for the product concerned. The Contract Giver should ensure that the Contract Acceptor is fully aware of any problems associated with the product or the work which might
pose a hazard to his premises, equipment, personnel, other materials or other products.
7 The Contract Giver should monitor and review the performance of the
Contract Acceptor and the identification and implementation of any needed
improvement.
8 The Contract Giver should be responsible for reviewing and assessing the
records and the results related to the outsourced activities. He should also ensure,
either by himself, or based on the confirmation of the Contract Acceptor’s Qualified
Person, that all products and materials delivered to him by the Contract Acceptor have been processed in accordance with GMP and the marketing authorisation.


The Contract Acceptor
9 The Contract Acceptor must be able to carry out satisfactorily the work
ordered by the Contract Giver such as having adequate premises, equipment,
knowledge, experience, and competent personnel.
10 The Contract Acceptor should ensure that all products, materials and
knowledge delivered to him are suitable for their intended purpose.
11 The Contract Acceptor should not subcontract to a third party any of the work
entrusted to him under the Contract without the Contract Giver’s prior evaluation and
approval of the arrangements. Arrangements made between the Contract Acceptor
and any third party should ensure that information and knowledge, including those
from assessments of the suitability of the third party, are made available in the same
way as between the original Contract Giver and Contract Acceptor.
12 The Contract Acceptor should not make unauthorized changes, outside the
terms of the Contract, which may adversely affect the quality of the outsourced
activities for the Contract Giver.
13 The Contract Acceptor should understand that outsourced activities, including
contract analysis, may be subject to inspection by the competent authorities.


The Contract
14 A Contract should be drawn up between the Contract Giver and the Contract
Acceptor which specifies their respective responsibilities and communication
processes relating to the outsourced activities. Technical aspects of the Contract
should be drawn up by competent persons suitably knowledgeable in related
outsourced activities and Good Manufacturing Practice. All arrangements for
outsourced activities must be in accordance with regulations in force and the
Marketing Authorisation for the product concerned and agreed by both parties.
15 The Contract should describe clearly who undertakes each step of the
outsourced activity, e.g. knowledge management, technology transfer, supply chain,
subcontracting, quality and purchasing of materials, testing and releasing materials, undertaking production and quality controls (including in-process controls, sampling
and analysis).
16 All records related to the outsourced activities, e.g. manufacturing, analytical
and distribution records, and reference samples, should be kept by, or be available to,
the Contract Giver. Any records relevant to assessing the quality of a product in the
event of complaints or a suspected defect or to investigating in the case of a suspected falsified product must be accessible and specified in the relevant procedures of the Contract Giver.
17 The Contract should permit the Contract Giver to audit outsourced activities,
performed by the Contract Acceptor or his mutually agreed subcontractors.

Reference:-

  • EU Guideline For Outsourced Activities – EUDraLex Volume -4 Chapter 7