Cleanroom, Classification, Design and

A brief review on ISO 14644 Standard on Cleanroom Design and testings. With a look at PIC/S and WHO standards

1. Cleanrooms Slide 1 of 68 November 2014 Tehran University of Medical Sciences School of Pharmacy Cleanrooms Classification, Design and Testing Ahmadreza Barazesh Under the supervision of Dr. Vatanara

2. References ISO 14644 Part 1: Classification of air cleanliness Part 2: Continued compliance with Part 3: Metrology and test methods Part 4: Design, construction and start- up WHO Technical Report Series, No. 902, 2002 Annex 6: Good manufacturing practices for sterile pharmaceutical products WHO Technical Report Series, No. 961, 2011 Annex 5: WHO guidelines on good manufacturing practices for heating, ventilation and air-conditioning systems for non-sterile pharmaceutical dosage forms Annex 6: WHO good manufacturing practices for sterile pharmaceutical products PIC/S GMP Guide (Part I: Basic Requirements For Medicinal Products) PIC/S Guide To Good Manufacturing Practice For Medicinal Products - Annexes Disclaimer : The TUMS logo included, neither indicates that the lecturer is an official lecturer of TUMS nor the content is approved by TUMS. It is just indicative of the department in which the lecture was prepared and presented.

3. Scope What will be covered during this presentation: A Brief review on cleanroom classification Introduction of Design Concepts and Considerations based on ISO 14644 Series Standards, PIC/S and WHO Guidelines. Testing Methods and Procedures According to

4. Introduction Cleanrooms provide for the control of airborne contamination to levels appropriate for accomplishing contamination-sensitive activities. Aerospace, Microelectronics, Pharmaceuticals, Medical devices, Healthcare (Hospitals) Food.

5. Definitions Cleanroom: room in which: The concentration of airborne particles is controlled , Constructed and used in a manner to minimize the introduction , generation , and retention of particles inside the room, Other parameters ( temperature , humidity , and pressure) are controlled Installation: cleanroom or one or more clean zones, together with all associated structures, air-treatment systems, services, and utilities. Classification: level of airborne particulate cleanliness, represents maximum allowable concentrations (in particles per cubic metre of air) for considered sizes of particles

6. Definitions Particle: Solid or liquid object which, for purposes of classification of air cleanliness, falls within a threshold size in the range from 0.1 to 5µm Occupancy states As-built: installation is complete, all services functioning, no production equipment, materials, or personnel present At-rest: no personnel present Operational: the installation is functioning in the specified manner , specified number of personnel present and working

7. Classification The particulate cleanliness of air shall be defined in one or more of three occupancy states, viz. “as-built”, “at-rest”, or “operational” The maximum permitted concentration of particles, C n , for each considered particle size, D, In which, N is the ISO classification number, which shall not exceed a value of 9. (ISO Class 1 to 9)

8. Classification

9. Classification Graphical representation of ISO-class concentration limits for selected ISO classes

10. Classification PIC/ S Guide To GMP For Medicinal Products Annex 1

11. Classification WHO Technical Report Series, No. 902, 2002 Annex 6

12. Classification WHO Technical Report Series, No. 902, 2002 Annex 6 This comparison is defined based on at-rest limitations.

13. Classification Class maximum particles/m 3 FED STD 209E equivalent EU GMP Classification ≥0.1 µm ≥0.2 µm ≥0.3 µm ≥0.5 µm ≥1 µm ≥5 µm ISO 1 10 2.37 1.02 0.35 0.083 0.0029 ISO 2 100 23.7 10.2 3.5 0.83 0.029 ISO 3 1,000 237 102 35 8.3 0.29 Class 1 ISO 4 10,000 2,370 1,020 352 83 2.9 Class 10 ISO 5 100,000 23,700 10,200 3,520 832 29 Class 100 Grade A and Grade B ISO 6 1.0×10 6 237,000 102,000 35,200 8,320 293 Class 1,000 ISO 7 1.0×10 7 2.37×10 6 1,020,000 352,000 83,200 2,930 Class 10,000 Grade C ISO 8 1.0×10 8 2.37×10 7 1.02×10 7 3,520,000 832,000 29,300 Class 100,000 Grade D ISO 9 1.0×10 9 2.37×10 8 1.02×10 8 35,200,000 8,320,000 293,000 Room air

14. Classification PIC/ S Guide To GMP For Medicinal Products Annex 1

15. Classification: Designation The designation of airborne particulate cleanliness for clean rooms and clean zones shall include: the classification number, expressed as “ISO Class N”; the occupancy state the considered particle size(s), and the concentration(s), 0,1µm through 5 µm. Example designation: ISO Class 4; operational state; considered sizes: 0,2µm (2 370 particles/m 3 ), 1 µm (83 particles/m 3 )

16. Airborne particle physical control: Filtration (HEPA) Dilution (Higher Airchange Rate) Isolation HEPA class retention (total) retention (local) E10 > 85% --- E11 > 95% --- E12 > 99.5% --- H13 > 99.95% > 99.75% H14 > 99.995% > 99.975% U15 > 99.9995% > 99.9975% U16 > 99.99995% > 99.99975% U17 > 99.999995% > 99.9999%

17. Classification: PIC/S Grade A: The local zone for high risk operations: F illing zone, open ampoules and vials, making aseptic connections. P rovided by a LAF work station with a homogeneous air speed in a range of 0.36 – 0.54 m/s (guidance value ) A unidirectional air flow and lower velocities may be used in closed isolators and glove boxes. Grade B: For aseptic preparation and filling, this is the background environment for the grade A zone. Grade C and D: Clean areas for carrying out less critical stages in the manufacture of sterile products.

18. PIC/S General Paragraphs Clean Areas: Entry through airlocks for personnel and/or for equipment and materials . Supplied with air has passed through filters of an appropriate efficiency. The various operations of component preparation , product preparation and filling  in separate areas within the clean area Manufacturing operations: P roduct is terminally sterilized, C onducted aseptically at some or all stages . In order to meet “in operation” conditions, areas should be designed to reach certain air-cleanliness levels in the “at rest” occupancy state.

19. PIC/S General Paragraphs “In operation” classification may be demonstrated during normal operations , simulated operations or during media fills (worst - case) Clean rooms and clean air devices should be routinely monitored Monitoring locations based on risk analysis and the results of classification Grade A : full duration of critical processing Grade A: Such a frequency and sample size that all interventions, transient events is captured and alarms triggered Grade B : The same as grade A; the sample frequency may be decreased . Grade C and D : in accordance with the principles of quality risk management .

20. PIC/S General Paragraphs Terminally Sterilized Products Preparation of components and most products should be done in at least a grade D environment Where the product is at a high or unusual risk of microbial contamination  G rade C Filling of products for terminal sterilization  Grade C Where the product is at unusual risk of contamination from the environment, filling  G rade A with Grade C background . Preparation and filling of ointments, creams, suspensions and emulsions should  grade C before terminal sterilization

21. PIC/S General Paragraphs Aseptic Preparation Components after washing  G rade D Handling of sterile starting materials, unless subjected to sterilization or filtration  G rade A with Grade B background. Otherwise  Grade C Handling and filling of aseptically prepared products  Grade A T ransfer of partially closed containers, as used in freeze drying,  either in a Grade A environment with grade B background or in sealed transfer trays in a grade B environment

22. PIC/S Paragraphs on Premises A ll exposed surfaces should be smooth, impervious and unbroken To reduce accumulation of dust and to facilitate cleaning there should be no uncleanable recesses and a minimum of projecting ledges, shelves, cupboards and equipment. False ceilings should be sealed. Sinks and drains should be prohibited in grade A/B areas Changing rooms should be designed as airlocks, The final stage of the changing room should, in the at-rest state, be the same grade as the area into which it leads.

23. PIC/S Paragraphs on Premises Both airlock doors should not be opened simultaneously; interlocking system or a visual and/or audible warning system should be operated. A filtered air supply should maintain a positive pressure and an air flow relative to surrounding areas of a lower grade. a pressure differential of 10-15 pascals It should be demonstrated that air-flow patterns do not present a contamination risk. A warning system should be provided to indicate failure in the air supply.

24. Planning and Design A project plan shall be developed to define the requirements of the products, the processes and the scope of the installation . A process equipment list shall be compiled, and shall include the critical requirements for each piece of process equipment . Diversity factors shall be defined, considering peak and average demand for each utility and environmental control system . A contamination control concept shall be developed for each zone of an installation.

25. Design: Control and segregation concepts For economic, technical and operational reasons, clean zones are often enclosed or surrounded by further zones of lower cleanliness. T he zones with the highest cleanliness demands is reduced to the minimum size. Movement of material and personnel between adjacent clean zones gives rise to the risk of contamination transfer,  management of material and personnel flow

26. Design: Control and segregation concepts

27. Design: Personnel flow and Material flow Personnel flows considered: Manufacturing personnel Maintenance personnel Quality control personnel Material flows considered: Raw materials Finished goods Waste Product (In-process, Intermediate & Final) Equipment Clean and dirty components Portable equipment Product containers

28. Design: Personnel flow and Material flow

29. Design: Personnel flow and Material flow Desirable Layout Less Desirable Layout

30. Design: Air Flow Patterns Air flow patterns: Cleanroom airflow patterns can be categorized as either unidirectional or non- unidirectional (or mixed) Unidirectional airflow ISO Class 5 and cleaner may be either vertical or horizontal airflow rely upon a final filtered air supply and return inlets are nearly opposite air supplies to maintain the airstream straight

31. Design: Air Flow Patterns non -unidirectional airflow cleanrooms Air flow outlets located in multiple positions. Filter outlets may be distributed at equal intervals or grouped over the core process. The final filter location may be remote, (avoid contamination ingress between filters and cleanroom) Return air locations in non-unidirectional airflows are not as critical Distribute the returns to minimize dead zones within the cleanroom

32. Disturbance of unidirectional airflow

33. Contamination Control Concepts

34. In order to protect cleanrooms from contamination from adjacent less clean spaces Displacement concept (low pressure differential, high airflow ) by means of a low turbulent "displacement" airflow , >0,2 m /s Pressure differential concept (high pressure differential, low airflow ) The pressure differential in the range of 5 - 20 Pa, to allow doors to be opened and to avoid unintended turbulence. Physical barrier concept U se of an impervious barrier to prevent contamination transfer to a clean zone from a less clean zone. Design: Segregation Concepts

35. Design: Layout of an installation General Considerations: Size: of cleanroom should be minimum . if a large space is required, it should be divided, with or without physical barriers . Workstation siting and organization: critical workstations away from, major traffic pathways. less clean operations site downstream of cleaner operations. Ancillary areas and adjacent cleanrooms: Pressure or flow differentials, Access and communication arrangements ( such as airlocks , speech panels and intercoms cross-contamination from less clean zones does not compromise the cleaner zones.

36. Design: Layout of an installation General Considerations (contd.) Utility services and ancillary equipment General: Utility services should be designed and installed such that the cleanroom is not compromised by contamination. exposed piping, tubing and cable runs should be minimized , Vacuum-cleaning equipment Sprinkler systems Communication systems: to reduce personnel movement Glazing: Avoid heat loss and solar gain, non-opening double glaze

37. Design: Layout of an installation Access: General : The number of openings should be minimized. Normal (non-emergency) access should be through airlocks for both personnel and material. Airlocks : In order to maintain pressure differential and integrity of during entry and exit, airlocks or transfer hatches (pass- throughs ) are normally required . Emergency exits : Emergency exits should be provided with means to show that they have been opened.

38. Design: Layout of an installation Access (contd.) Changing rooms: Have three functional zones: Entry : access from ancillary. appropriate for removal , storage, disposal and/or redonnning of garments not permitted within the cleanroom; T ransition zone : where garments or personal equipment dedicated to the cleanroom are stored , donned or removed. I nspection /access zone: where inspection of the completed gowning is accomplished and provides access to cleanroom. The three functional zones may be separated by a physical barrier (e.g. a stepover bench or airlock )

39. Changing Rooms

40. Design: Layout of an installation The following requirements should be defined: number of people passing through the gowning procedure the gowning procedure (i.e. what garments are to be taken off and put on) the frequency of garment replacement . Consideration should be given to the following provisions: Storage and disposal of garments; Storage before use and disposal of consumable items Storage of personal items; Hand-washing and drying or other decontamination processes; Display or posting of gowning sequence, with clear instructions; full-length mirrors to check effective fit.

41. Design: Construction and materials The materials used should be selected to meet the requirements of the installation, and should take into account the following : the cleanliness class; effects of abrasion and impact; cleaning and disinfection methods and frequencies; chemical/microbiological attack and corrosion . Surface cleanliness and cleanability of materials of construction Fittings in airlocks: Minimum horizontal surfaces

42. Design: Construction and materials Ceilings : Ceilings should be sealed , penetration points should be kept minimum. Walls: Materials and surface finishes should meet all general requirements. Particular consideration to impact and abrasion. ( r ubbing strips, protective bars) Cover strips or seals between panels should be smooth, with rounded edges Use double glazing, with airtight seal, which can enable flush mounting Doors should present as few horizontal surfaces as possible , thresholds avoided . Consider use of push plates, automatic openings, or appropriate door-swing Floors: Floors or floor coverings should be non-porous, slip-resistant, abrasion-resistant, conductive if necessary.

43. Design: Construction and materials

44. Design: Control of air Cleanliness Air filtration systems Air filtration systems including filter elements , mounting frames , housings , gaskets , sealants and clamping systems should be selected to suit both the cleanliness and using condition. Three basic stages of air filtration are recommended: prefiltering of the outside air to ensure adequate quality of air supply secondary filtering in the air conditioning plant to protect the final filters; final filtering before cleanroom supply . “Sacrificial " filters or temporary filters: considered to protect the air cleanliness of air-handling systems during construction and commissioning.

45. HVAC Systems

46. HVAC Systems

47. HVAC Systems

48. Tests Methods Cleanroom tests: Required Tests : An airborne particle count test shall be carried out in order to classify an installation Optional Tests: Airborne particle count for ultrafine and/or Micro-particles Airflow test Air pressure difference tests Installed filter system leakage test Air flow direction tests and visualization Temperature, Humidity and Electrostatic tests Particle deposition tests Recovery tests Containment leak tests

49. Tests Methods Airborne particle count for classification and test measurement: M easurement of airborne particle concentrations with size 0.1 - 5 μm . A discrete-particle-counting, light-scattering instrument is used to determine the concentration of airborne particles. Prior to testing, verify that all aspects of the cleanroom and functioning in accordance with specifications. Airflow rate or velocity tests; Pressure difference test; Containment leakage test; Filter leakage test.

50. Airborne particle count for classification and test measurement: Sampling Minimum number of sampling point locations: Sampling locations evenly distributed, at the height of the work activity. Sample a sufficient volume of air that a minimum of 20 particles would be detected if the particle concentration for the largest considered particle size were at the class limit for the designated ISO class. The volume sampled at each location shall be at least 2 litres , with a minimum sampling time at each location of 1 min. Compute the overall mean of the averages, standard deviation, and 95% upper confidence limit from the average particle concentrations for all locations. Tests Methods

51. Tests Methods Airborne particle count for ultrafine particles Smaller than 0,1μm DPC, with a capability for accurate particle size definition up to at least 1μm . Condensation nucleus counter ( CNC ) S mall sampling flow & long sampling tube  diffusion loss. Airborne particle count for macro-particles Larger than 5 μm . There are two general categories of macroparticle measurement methods . collection by filtration or inertial effects, followed by microscopic measurement in situ measurement of the concentration and size of macroparticles with a time-of-flight particle counter or DPC

52. Tests Methods T wo general categories of macroparticle measurement methods: collection by filtration or inertial effects, followed by microscopic measurement of the number and size, or measurement of the mass of collected particles: filter collection and microscopic measurement will report macroparticles using particle size based upon the agreed diameter; cascade impactor collection and microscopic measurement will report macroparticles using particle size based upon the microscopist's choice of reported particle diameter ; cascade impactor collection and weight measurement will report macroparticles using particle size based upon an aerodynamic diameter; I n situ measurement of the concentration and size of macroparticles with a time-of-flight particle counter or a DPC: DPC measurement of particle size based upon an equivalent optical diameter; T ime -of-flight particle size measurement based upon an aerodynamic diameter.

53. Tests Methods Airflow Test T o measure airflow velocity and uniformity , and supply airflow rate Measurement of velocity distribution is necessary in unidirectional airflow cleanrooms, and supply airflow rate in non-unidirectional cleanrooms . S upply airflow rate ( air volume supplied to the clean installation per unit of time) can also be used to determine the air changes . Airflow rate is measured either downstream of final filters or in air supply ducts; both methods rely upon measurement of velocity of air passing through a known area .

54. Tests Methods: Air Flow Test Procedure for unidirectional airflo w installation test Supply airflow velocity M easured at approximately 150-300 mm from the filter face. N umber of measuring points should be the square root of 10 times of area in m 2 but no less than 4. At least 1 point for each filter outlet Uniformity of velocity within the cleanroom measured at approximately 150-300 mm from the filter face and the subdivision into grid cells should be defined Supply airflow rate measured by filter face velocity The results of the airflow velocity test can be used to calculate the total supply airflow rate. Supply airflow rate in air ducts by volumetric flowmeters (orifice meters, Venturi meters and anemometers)

55. Tests Methods: Air Flow Test Procedure for non-unidirectional airflow installation test Air volume supply rate and air-change rate are the most important parameters . Supply airflow rate measured at the inlet Because of local turbulence, use of a flowhood that captures all of the air issuing from each supply diffuser is recommended . Supply airflow rate calculated from filter face velocity Evaluation of the supply airflow rate without a flowhood may be done with an anemometer downstream of each final filter. The supply airflow rate is determined from the airflow velocity multiplied by the area of exit.

56. Test Methods Air pressure difference test Verify the capability of the complete installation to maintain the specified pressure difference between separate spaces With all doors closed, the pressure difference between the cleanroom and any surrounding should be measured and recorded . The following should be considered: installation of permanent measuring points; take measurements near the middle of the cleanroom away from any supply air inlets or return air outlet.

57. Test Methods Installed filter system leakage test To confirm that the filter system is properly installed and that leaks have not developed I ntroducing an aerosol challenge upstream of the filters and scanning immediately downstream of the filters and support frame or by sampling in a downstream duct . A pplied to cleanrooms in “as-built” or in “at-rest” occupational states, and when commissioning new cleanrooms, or existing installations require re-testing, or after the final filters have been replaced Detection of leakage by Scanning / Stationary remeasuring

58. Test Methods: Filter Leakage Test

59. Test Methods Apparatus and materials for installed filter system leakage tests Aerosol photometer Discrete-particle counter (DPC ) Suitable pneumatic or thermal aerosol generator(s ) Suitable aerosol dilution system . Suitable aerosol source substances

60. Test Methods Airflow direction test and visualization To confirm that the airflow direction and its uniformity conform to the design and performance specifications can be performed by the following four methods: Tracer thread method ; silk threads, single nylon fibers, flags or thin film tapes and effective lighting Tracer injection method ; tracer particles illuminated by high intensity light sources (DI Water , alcohol/glycol) Airflow visualization method by image processing techniques ; (Quantitative) Airflow visualization method by the measurement of velocity distribution.

61. Test Methods: Air Flow Visualization

62. Test Methods Temperature test Capability to maintain the air temperature level within the control Measured at a minimum of one location for each temperature-controlled zone. Measurement time should be at least 5 min with one value recorded at least every minute. Comprehensive temperature test: At least 1 h after the air-conditioning system has been operated The number of measuring locations should be at least two. Probe should be positioned at work-level height and at a distance of no less than 300 mm from the ceiling, walls, or floor of the installation

63. Test Methods Humidity test Capability to maintain the air humidity level Expressed as relative humidity or dew point The sensor should be located at least at one location for each humidity control zone, and sufficient time should be allowed for the sensor to stabilize. The measurement time should be at least 5 min.

64. Test Methods Particle deposition test Sizing and counting particles that can be deposited from the air onto product or work surfaces in the installation. Particles are collected on witness plates with surface characteristics similar to those of the at-risk surface Are sized and counted using optical microscopes, electron microscopes, or surface scanning apparatus. The witness plate should be placed in the same plane as the at-risk surface. And at the same electrical potential as the test surface.

65. Test Methods Recovery test Ability of the installation to eliminate airborne particles. Only important and recommended for non-unidirectional airflow systems This test is not recommended for ISO Classes 8 and 9. 100:1 recovery time is defined as the time required for decreasing the initial concentration by a factor of 0,01

66. Test Methods Containment leak test Determine if there is intrusion of contaminated air into the clean zones from non-controlled areas Particle concentration outside should be greater than the cleanroom concentration by a factor of 10 3. If the concentration is less, generate an aerosol. To check for leakage through construction joints, cracks or service conduits, scan inside the enclosure at a distance of not more than 5 cm from the joint, at a scan rate of approximately 5 cm/s.