Particle Counting. - Theory - Guidelines - Monitoring

- Theory - Guidelines - Monitoring 1

Agenda Introduction to Cleanroom Classification of Air Cleanliness: ISO 14644 Guidance for the Manufacture of Sterile Medicinal Products EU GMP, Annex 1 Classification In-Process Monitoring Best Practices for FMS Solutions Particle Counter Theory and Calibration: IS0 21501-4 -------- On-line TOC Analyzer EP&USP Harmonization for Liquid Particle 2

Agenda Introduction to Cleanroom Classification of Air Cleanliness: ISO 14644 Guidance for the Manufacture of Sterile Medicinal Products EU GMP, Annex 1 Classification In-Process Monitoring Best Practices for FMS Solutions Particle Counter Theory and Calibration: IS0 21501-4 -------- On-line TOC Analyzer EP&USP Harmonization for Liquid Particle 3

Why do we care about Particle Counts? Particles as contamination negatively affect a process or product Change chemical composition Affect stability Affect purity Affect safety Affect reliability Contamination affects the yield of a process Increases cost to produce Increases cost to maintain or service Increases cost of ownership [COO] Decreases ROI of capital equipment purchase 4

Why do we care about Particle Counts? Pharmaceutical, Biotechnology Particles in injection could cause occlusion of blood vessels Red Blood cells are about 5 µm Capillary (5 to 10 µm) Large veins (10 to 50 µm) Viables in injection can trigger infection Possibility of reaction to foreign substances (RES/allergic reaction) Medical Devices Poor adhesion of medicated coating in stent creates embolisms Medical and photographic films Contamination prevents complete image recovery 5

Why monitor for particles? Greatest concern is for viable microorganisms Technology is not available today to measure viable counts in real time Requires incubation time Total non-viable particle counts used as a surrogate Non-viable counts Includes all types of airborne material Solid particles Fibers Microorganisms Skin flakes Non-viable particle count monitoring is a critical component of the total Environmental Monitoring program Non-viable particle counting offers potential for real-time response 6

Relative Sizes of Particulate Matter 100 90 80 70 60 50 40 30 20 10 0 Human hair: 50-100 µm Visible: 35-50 µm Skin Flake: about 20 µm Bacteria: 1 to 8 µm 7

Relative Sizes of Particles Table salt Bacteria Pencil cells 8

Particles Found in a Cleanroom Aluminosilicate 9

Guidance Suggested Monitoring 10 9 8 7 Event driven 6 5 4 3 2 Bacterium 2-8 µm 5 µm Baseline 1 0 0.5 µm Virus: 0.001 µm 10

How an airborne particle counter works Detector Particle Light Trap Laser Diode Mirror Principle: Light Scattering 11

Three Common Methods of Sampling Manually with a handheld or portable particle counter Sequentially with a manifold/scanner and particle counter Continuously with portable or remote particle counters 3400 Series Cleanroom classification Environmental monitoring Short-term online sampling HHPC Handheld particle counter Used for troubleshooting 6000 & 7000 Series for production monitoring Grade A & B areas 12

Manual Monitoring Advantages: - Traditional method of sampling -Simple - Flexible - Low Initial Cost Disadvantages: - Inconsistent in Time - Inconsistent in Position of Probe - Intrusive to Work Process - Usually is not continuous - Production worker often not informed of high counts - High Labor cost (labor intensive) 13

Sequential Monitoring Advantages: - Consistently samples fixed positions - Samples every position one or more times per hour - Can operate 24 hours per day - Frequent sampling of each point - Consistent sampling position - Statistical data for historical purposes - Consistency of data allows SPC analysis - Can be part of an FMS system - Single counter to calibrate A2400 Disadvantages: - Samples at fixed locations - Can miss events - Particle loss in tubing - Requires auxiliary pump(s) to maintain constant flow through all sample tubes R2400 14

Tubing Transport Loss 15

Advantages: - Able to detect fast, random events - Uses dedicated counter at each sample point - Avoids effects of long tubing runs - Consistent sample points - All points monitored simultaneously - Quick operator feedback - Build statistical database - Relate particle events to process activity opportunity to improve process Disadvantages: Continuous Monitoring - Requires good plan of monitoring strategy Sample points must be chosen at the beginning Decisions must be made about alarms and reports - More expensive to implement More instruments Utility support: data cables, vacuum lines - Calibration needed for each counter - Data must be processed or summarized 6000 7000 16

Flexibility Cost Labor Calibration Quality 100 90 80 70 60 50 40 30 20 10 0 Flexibility Initial Cost Daily Labor Calibration Quality of data Manual Sequential Continuous 17

Cleanroom and Clean Zone Classification Classification is the formal process of qualifying the environment by the number of particles using a standard method (ISO 14644) Performed on a regular basis but not frequently Grade A areas: Six months, ISO 4.8 Grade B areas: Six months, ISO 5 Grade C, D areas: Annually, ISO 7/8 Standards define minimum number of points Based on area of cleanroom or clean zone Standards define minimum amount of air to be sampled Minimum volumes for statistically valid samples (typically 1 minute at 1 cfm) Grade A requires minimum 1 m 3 (Annex 1) Classification is a rigid protocol 18

Environmental Monitoring Determine readiness of room to carry out designated task Performed whenever relevant activity will occur daily work Typically performed with portable counters Number of sample points defined by risk assessment Area of cleanroom or clean zone Activity to be performed Risk to product Frequency and volume of sample points defined by risk assessment Frequent enough to show control Frequent enough to manage risk of product contamination Inferred guidance, not rigid by regulation 19

Aseptic Process Monitoring Governed by EMEA and FDA Guidance Augments ISO 14644 requirements for critical (Grade A) areas Recommends continuous monitoring Number of sample points based on risk assessment Not rigidly defined by guidance Location of points based on risk assessment Limited guidance on sampling location Sample volumes defined by type of equipment Published guidance, not rigid company specific SOPs 20

Regulations and Guidance for the Manufacture of Sterile Medicinal Products ISO 14644 and EU GMP Annex 1 21

Agenda Introduction to Cleanroom Classification of Air Cleanliness: ISO 14644 Guidance for the Manufacture of Sterile Medicinal Products EU GMP, Annex 1 Classification In-Process Monitoring Best Practices for FMS Solutions Particle Counter Theory and Calibration: IS0 21501-4 -------- On-line TOC Analyzer EP&USP Harmonization for Liquid Particle 22

Classification Standards for Airborne Particles General Air Monitoring Standards 1999 ISO 14644-1 Classification of air cleanliness ISO 14644-2 Specifications for testing and monitoring to prove continued compliance with ISO 14644-1 ISO 14644-3 Guidance on instrumentation to be used for testing for compliance with ISO 14644-1 ISO 14644 23

ISO 14644 General Standard Covers All Industries Electronics Semiconductor Flat Panel Circuit Board Optical MEMS/Nanomachines Life Sciences Pharmaceutical Biotechnology Medical Devices Hospitals/Pharmacies Aerospace Launch Vehicles Satellites Commercial/Military Aircraft Laboratories Analytical Laboratories Universities Other Nuclear Photographic, X-ray films Automobile Painting Laboratory Aerospace Other Life Sciences Electronics 24

Classification Standard ISO 14644-1 Defines cleanroom classes in a zone Establishes minimum sampling volumes Purpose: Gather a sample volume with theoretically at least 20 particles for a statistically valid sample Establishes minimum number of points to classify area, based on statistical criteria Gather from a valid number of locations for a representative sample of the total air volume 25

Classification: ISO 14644-1 A sample size with at least 20 theoretical particles, or Minimum sample volume 2.0 liter Minimum sample time 1 minute Minimum number of locations whichever is greatest 1 with at least 3 samples total 26

Classification Limits ISO 14644-1 Class Number of Particles per Cubic Meter by Micrometer Size 0.1 μm 0.2 μm 0.3 μm 0.5 μm 1 μm 5 μm ISO 1 10 2 ISO 2 100 24 10 4 ISO 3 1,000 237 102 35 8 ISO 4 10,000 2,370 1,020 352 83 ISO 5 100,000 23,700 10,200 3,520 832 29 ISO 6 1,000,000 237,000 102,000 35,200 8,320 293 ISO 7 352,000 83,200 2,930 ISO 8 3,520,000 832,000 29,300 ISO 9 35,200,000 8,320,000 293,000 27

What volume do I need to collect? ISO 14644 requires sample with 20 theoretical particles ISO 14644 allows 3520 particles (0.5um) in 1m 3 (1000 liters) Volume = 20 particles 3520 particles X 1000 liters Volume = 5.7 liters Greater than 2.0 liters so minimum sample volume is satisfied 28

ISO 14644-1 Minimum Sample Time at 1 CFM Time required (in minutes) at 1 cfm (28.3 lpm) flow rate with 1-minute limit imposed 0.1 um 0.2 um 0.3 um 0.5 um 1 um 5 um ISO Class 1 70.64 353.20 ISO Class 2 7.06 29.43 70.64 176.60 ISO Class 3 1.00 2.98 6.93 20.18 88.30 ISO Class 4 1.00 1.00 1.00 2.01 8.51 ISO Class 5 1.00 1.00 1.00 1.00 1.00 24.36 ISO Class 6 1.00 1.00 1.00 1.00 1.00 2.41 ISO Class 7 1.00 1.00 1.00 ISO Class 8 1.00 1.00 1.00 ISO Class 9 1.00 1.00 1.00 29

Continued Compliance: ISO 14644-2 Schedule of Tests to Demonstrate Continuing Compliance Test Parameter Class Maximum Time Interval Test Procedure Particle Count Test All classes ISO 5 24 6 Months months > ISO 5 12 Months ISO 14644-1 Annex A Important Air Pressure note: If Particle counts and Differential ISO 14644-1 All Classes 12 Months Pressure Difference are monitored continuously (for Annex B5 example, by a Facility Monitoring System) then ISO 14644-1 the Airflow Maximum All Time Classes Interval 12 for Months re-certification Annex B4 can be extended 30

Example: ISO 14644-1 Classification Vial Washing Station 5 m Lyo 1 8 m Lyo 2 Calculations for Number of Points: Area of clean zone = 80 m² Take the SQRT (80) = 8.94 Rounding up to next integer = 9 sample positions 5 m Lyo 3 4 m 31

Example: ISO 14644-1 Classification Vial Washing Station 1 2 3 4 5 6 Lyo 1 7 Lyo 2 8 Calculations for Number of Points: Area of clean zone = 80 m² Take the SQRT (80) = 8.94 Rounding up to next integer = 9 sample positions 9 Lyo 3 32

Example ISO 14644-1 Calculations 1 2 3 4 5 6 7 8 9 10 Vial Washing System Freeze Dryer 1 Need to adjust for equipment in room. Under ISO 14644-1, if you sample at 10 or more positions, you can avoid the added calculation of the UCL (Upper Confidence Limit). Calculation of the UCL is only mandated when the number of positions used is between 2 and 9. Best to sample near potential problem spots which are near entrances and exits and near operator positions. Freeze Dryer 2 Freeze Dryer 3 33

Example ISO 14644-1 Calculations Vial Washing System 1 2 9 8 10 Freeze Dryer 1 3 4 5 6 7 11 Need to adjust for equipment in room. 12 Freeze Dryer 2 Under ISO 14644-1, if you sample at 10 or more positions, you can avoid the added calculation of the UCL (Upper Confidence Limit). Calculation of the UCL is only mandated when the number of positions used is between 2 and 9. Best to sample near potential problem spots which are near entrances and exits and near operator positions. 13 14 Freeze Dryer 3 34

Example ISO 14644-1 Calculations 1. Average sample data values at each position 2. Normalize the average to number of particles per cubic meter 3. Compare normalized value to the target class limit; normalized value at each and every sample point must be less than the limit for the given size and target room classification If the number of points sampled is more than 1 but less than 10, then the UCL factor must be applied: a) Calculate the standard deviation b) Use Student s T-factor from tables c) Calculate UCL d) Compare to classification limit; UCL must not exceed the applicable limit 35

Probable Revisions to ISO 14644-1, -2 1. Frequency of re-certification Now: If zone is ISO Class 5, every 6 months If zone is ISO Class 6, every 12 months Proposed: For all zones, every 12 months 36

Probable Revisions to ISO 14644-1, -2 2. Eliminate Student s T test for sample plans with 1 to 9 sample positions 3. Method of determining number of sample positions Replace with stated number of minimum sample positions as a look-up chart Based on 95% confidence levels not on SQRT of area May mean a small increase in the number of sample points 37

B.4.1.1 Derive the minimum number of sampling locations NL from table 3. Table 3 shows the number of sample locations related to the area of each cleanroom or clean zone to be classified and provides at least 95% confidence that at least 90 % of all locations do not exceed the class limits. Different levels of confidence and verification can be specified and agreed upon by the customer and supplier. 38

Room Size (ft 2 ) Room Size (m 2 ) ISO 14644-1: 1999 ISO 14644-1: 2007 (95,90) FS209E Unidirectional (all classes) FS209E Turbulent Class 1000 FS209E Turbulent Class 10K FS209E Turbulent Class 100K 21.5 2 2 1 2 2 2 2 43.1 4 2 2 2 2 2 2 64.6 6 3 3 3 3 2 2 86.1 8 3 4 4 3 2 2 107.6 10 4 5 5 4 2 2 129.2 12 4 6 6 5 2 2 150.7 14 4 7 7 5 2 2 172.2 16 4 8 7 6 2 2 193.8 18 5 9 8 7 2 2 279.9 26 6 10 12 9 3 2 301.4 28 6 11 13 10 4 2 344.4 32 6 12 14 11 4 2 366.0 34 6 13 15 12 4 2 387.5 36 6 14 16 13 4 2 409.0 38 7 15 17 13 5 2 559.7 52 8 16 23 18 6 2 581.3 54 8 17 24 19 6 2 624.3 58 8 18 25 20 7 2 796.5 74 9 19 32 26 8 3 839.6 78 9 20 34 27 9 3 1033.3 96 10 21 42 33 11 4 1248.6 116 11 22 50 40 13 4 1485.4 138 12 23 60 47 15 5 1894.4 176 14 24 76 60 19 6 2152.8 200 15 25 87 69 22 7 3229.2 300 18 26 130 103 33 11 5382.0 500 23 27 216 171 54 18 10763.9 1000 32 28 431 341 108 35 21527.8 2000 45 29 862 681 216 69 53819.6 5000 71 29 2153 1702 539 171 107639.1 10000 100 29 4306 3404 1077 341 39

Probable Revisions to ISO 14644-1, -2 4. Remove possibility to classify at 5 micron for ISO Class 5 Limit number of 29 removed Note (b) Due to sampling difficulties affecting collection of large particles in low concentrations, specification of sampling by DPC is inappropriate for classification at the indicated class. 5. Indicate that single digit limits for ISO Class 1 and 2 create challenges to timely execution 40

Agenda Introduction to Cleanroom Classification of Air Cleanliness: ISO 14644 Guidance for the Manufacture of Sterile Medicinal Products EU GMP, Annex 1 Classification In-Process Monitoring Best Practices for FMS Solutions Particle Counter Theory and Calibration: IS0 21501-4 -------- On-line TOC Analyzer EP&USP Harmonization for Liquid Particle 41

Guidance for Airborne Particles Pharmaceutical Industry Compliance Guidance FDA cgmp Guidance for Industry Sterile Drug Products Produced by Aseptic Processing EU GMP Annex I (EMEA) Manufacture of Sterile Medicinal Products EU GMP Annex 1 is more rigid than FDA cgmp Guidance 42

EU GMP Annex 1-2003 From EU Annex 1 The guidance has been reviewed in the light of the international standard EN/ISO 14644-1 and amended in the interests of harmonisation but taking into account specific concerns unique to the production of sterile medicinal products. Emphasizes continuous monitoring for Grade A and recommends for Grade B areas A continuous measurement system should be used for monitoring the concentration of particles in the Grade A zone, and is recommended for the surrounding Grade B areas. Introduces concept of measuring one cubic meter for routine testing (Meaning: classification, not in-process monitoring) 43

Revisions EU GMP Annex 1 2009 5μ particle counts particularly significant when several are seen consecutively and indicates up to 20/m 3 for Grade A and 29/m 3 for Grade B (now equivalent to ISO 5) Accepts that particle counting may not be possible at the point of fill due to the generation of particles or droplets from the product itself Clearly differentiates classification and monitoring The sample sizes taken for monitoring purposes using automated systems will usually be a function of the sampling rate of the system used. It is not necessary for the sample volume to be the same as that used for formal classification of clean rooms and clean air devices. - 1minute sampling time for continuous monitoring - 1m3 for classification 44

EU GMP Annex 1 Revised: 1 March 2009 Limits at 5 microns for Grade A 1 per cubic meter 20 per cubic meter At Rest In Operation Grade Maximum permitted number of particles/m 3 equal to or above 0.5 µm 5 µm 0.5 µm 5 µm A 3 500 520 20 1 3 500 520 20 1 B 3 500 520 29 1 350 352 000 2 000 900 C 350 352 000 2 000 900 3 500 520 000 20 29 000 D 35000 520 000 20 29 000 not defined not defined 45

Intuitive User Interface! Wizard to conduct pass/fail test for ISO, EU-GMP, FS and BS standards Improved area and sample location management Easy sampling recipe management through Group menu Multiple user level log in for data security 46

Intuitive User Interface! Test wizard for ISO, EU-GMP, FS, and BS standard compliance Wizard to conduct pass/fail test for ISO, EU- GMP Annex I, FS 209E and BS5295 standards No expertise in standards required. Few clicks to compliance The wizard guides an operator step by step to sample data, process data and product reports 47

Intuitive User Interface! Vial Filling Room 1 2 3 4 5 6 7 9 8 11 10 Freeze Dryer 1 12 Freeze Dryer 2 Better area/location management 13 14 Freeze Dryer 3 Multiple locations with unique location ID and repeatable location name can be created A group of locations can be assigned to a larger cleanroom space called an area Location setting can be copied 48

Intuitive User Interface! Better area/location management 3400 # 1 3400 # 2 3400 # 3 49

Intuitive User Interface! Location #1 Location #2 Group = Sampling Recipe Group is a collection of pre-set sampling parameters and alarm limits Better group management Area #1 Location #4 Location #3 Location #N Individual Location setting Group is a collection of desired sampling parameters New group can be created or existing group can be loaded A location can be added to or removed from a group Settings button allows pre-selecting sampling parameters such as sampling time, delay, hold time, count and environmental alarm, etc. 50

Intuitive User Interface! 21 CFR Part 11 compliance: unique user log in Multiple level of user accounts Basic (no log in) Operator Administrator No limits to number of log in accounts An administrator can create operator level user account and assign selective access rights and group access 51

Met One 3445 3445: 100 LPM Flow Rate Higher flow rate Same packaging Same accessories and support documents Same battery functionality Wireless capability maintained Meets ISO 21501-4 Resolution Sensitivity 52

Met One 3445 Rapid classification and routine sampling of Pharmaceutical areas Battery-powered yet still light-weight Smooth Stainless Steel enclosure is designed with the needs to maintain ultra-clean or aseptic areas Color touchscreen provides a clear window into cleanroom contamination Supported by software for stand-alone use and for continuous monitoring requirements PortAll 2.4 UVO EnVigil Lite OPC Server 53

Met One 3445 revised 3400 Datasheet 54

Met One 3445 revised 3400 Datasheet 55

Met One 3445 - Specifications 56

PortAll Software Organize, archive and report particle count data Download Utility for Airborne Particle Counters Two Versions Generic Industries Life Science Demo version provided with all portable and handheld counters 30-day operation Can shift or upgrade License type Generate Spreadsheets and Graphs of Data Generate reports based on Standards/Guidances ISO 14644 FS209E EU GMP Annex 1 57

PortAll 2.4 Life Sciences 21 CFR Part 11 Support Lockout for multiple attempts Auto Logoff for Inactivity Password Expiration Unique User Accounts Satisfy Corporate standards Length of User Name Length of Password Audit Trail of User actions 58

What s new in PortAll 2.4 Paperless Secure Data Transfer in a 21CFR Part 11 Environment USB Memory Stick data are now transferred in a binary format Supports 21CFR Part11 for data security and integrity Report Generation Just Got Easier Report wizard now fully supports EU GMP Annex 1 reporting Secure, Paperless Compliance Reports Export and archive electronic reports in a secure PDF format No Need to Bring Laptops into the Cleanroom Leverage your existing company network to transfer data Ethernet, wireless and serial communications networks fully supported 59

PortAll 2.4 What s new Support for USB Memory Stick transfer Binary format cannot be edited Only recent Raptor versions of 3400 60

PortAll 2.4 What s new Crystal Reports engine Standard reports Custom by user Custom by Systems Group New! EU GMP Annex 1 report 61

PortAll 2.4 What s new Compliance reports stored in a locked PDF format Suitable for archive in LIMS or other data management system Fully supports 21 CFR 11 62

PortAll 2.4 New Data Sheet 63

Agenda Introduction to Cleanroom Classification of Air Cleanliness: ISO 14644 Guidance for the Manufacture of Sterile Medicinal Products EU GMP, Annex 1 Classification In-Process Monitoring Best Practices for FMS Solutions Particle Counter Theory and Calibration: IS0 21501-4 -------- On-line TOC Analyzer EP&USP Harmonization for Liquid Particle 64

EU Annex 1 Summary: Monitoring Section 12: The sample sizes taken for monitoring purposes using automated systems will usually be a function of the sampling rate of the system used. It is not necessary for the sample volume to be the same as that used for formal classification of clean rooms and clean air devices. It is not necessary to sample 1m 3 during verification or monitoring Particle counters used for monitoring may have the same or different flow rate from those used for classification. Note: Revision of EU Annex 1 2003 completed, effective date of 01 March 2009 65

EU Annex 1 Summary: Monitoring Section 8 Clean rooms and clean air devices should be routinely monitored in operation and the monitoring locations based on a formal risk analysis study and the results obtained during the classification of rooms and/or clean devices Note: Revision of EU Annex 1 2003 completed, effective date of 01 March 2009 66

EU Annex 1 Summary: Monitoring Section 9 The Grade A zone should be monitored at such a frequency and with suitable sample size that all interventions, transient events and any system deterioration would be captured and alarms triggered if alert limits are exceeded. = continuous!!! Note: Revision of EU Annex 1 2003 completed, effective date of 01 March 2009 67

Monitoring Positions: Risk-based Approach Vial Sterilizing Tunnel 1 4 5 Lyo 1 7 2 3 Monitoring must follow the workflow, covering areas where product is exposed Annex 1 Where open vials exit de-pyrogenation human interaction (1) Where vials are filled (2,3) Surrounding Grade B background (4) Where the vials are partially stoppered (5) Loading area in front of lyophilizers must be Grade A if product is not fully stoppered (6,7) 6 Lyo 2 Lyo 3 68

Use Monitoring System for Classification Reduce the number of manually monitored classification points SAVE TIME! Add a wireless portable for room classification 69

Agenda Introduction to Cleanroom Classification of Air Cleanliness: ISO 14644 Guidance for the Manufacture of Sterile Medicinal Products EU GMP, Annex 1 Classification In-Process Monitoring Best Practices for FMS Solutions Particle Counter Theory and Calibration: IS0 21501-4 -------- On-line TOC Analyzer EP&USP Harmonization for Liquid Particle 70

Placement of Sample Probes Annex 1: No guidance FDA: Sample near to exposed product Generally near work height and exposed product If liquid sterile fill, guidance is to sample air approaching the product within 12 (30 cm) of exposed Sample near to points of intervention by operators Examples: Descrambler table Filling needles Stoppering process Less than 1 foot (12 inches or 30 cm) 71

Sampling Probes and Mounting Unidirectional air bathing the exposed product during manufacturing Isokinetic sample probe for particle monitoring (not positioned directly over exposed product) Exposed product or vials/ampoules 72

Where to Monitor? 73

Positioning the Probes Turntable Probe shown with Cap in place 74

Positioning the Probes Filling Line <305mm 75

Positioning the Probes Stoppering Station <305mm 76

Monitoring Powder Fill Section 9: For Grade A zones, particle monitoring should be undertaken for the full duration of critical processing, including equipment assembly, except where justified by contaminants in the process that would damage the particle counter or present a hazard, e. g. live organisms and radiological hazards. In such cases monitoring during routine equipment set up operations should be undertaken prior to exposure to the risk. Monitoring during simulated operations should also be performed. Note: Revision of EU Annex 1 2003 completed, effective date of 01 March 2009 77

Interference from Product (Powder Filling) Unidirectional air bathing the exposed product during manufacturing Higher sample probe for monitoring during production Sample probe to demonstrate air quality before filling process Exposed product or vials/ampoules 78

Recommendations Powder Fill Line Inlet HEPA Filter Outfeed from tunnel = Position of monitoring during filling = Positions of monitoring at rest 79

Continuous Monitoring An FMS System can monitor: Airborne particulates, Room pressures and air flow Room temperature and humidity Other parameters such as door interlocks, etc System configurations available: Multiple Pumps: Multiple sampling positions, greatest flexibility/redundancy, but higher install cost Central Vacuum: Multiple sampling positions, good flexibility/redundancy and medium install cost Integrated Pumps: Multiple sampling positions, good flexibility/redundancy and lowest install cost 80

System Overview Multiple Pumps 230VAC Host RS485, ethernet Pump Controller With Flow sensor Pump Controller With Flow sensor Vacuum Pump Vacuum Pump 24Vdc PSU Ceiling void Computer with FMS Software Wall Plate Clean room Portable APC Remote APC in Enclosure WiFi capable 81

System Overview Central Vacuum 24 VDC Host RS485, ethernet Vacuum Ring Main Pump Control Host PC Smart Socket Controller, Flow sense and vacuum control valve Smart Socket Controller, Flow sense and vacuum control valve Central Vacuum Pumps Main /Standby Plant room Computer with FMS Software Clean room Wall Plate Portable APC Remote APC in Enclosure WiFi capable 82

230VAC System Overview Integrated Pumps Host RS485, ethernet Host PC 24VDC 6A PSU 24VDC 6A PSU 24VDC 1 A PSU Ceiling void Computer with FMS Software Clean room R2315 Portable APC Remote APC with integrated pump WiFi capable 83

Networking Ethernet FMS System Server Node QA Network Client View Node Production Area Prep area Filling Vial capping Address 0 Address 0 Address 0 Engineering Network Client View Node Address 1 Address 2 Address 3 Smart Sockets Connected via Local Ethernet or company wide Intranet services 84

Maintaining Compliance Early detection of out of compliance Walking window technique Previous results per cubic foot Newest results per cubic foot x x 0 0 0 0 0 0 0 0 0 0 0 1 0 1 x x Sum over last 36 samples is 2 ALARM raised for 5.0 µm particle counts 36 rolling samples 85

Software Software is designed to be compliant with GAMP, Title 21CFR Part 11 and EU-GMP Annex 1 86

Human Machine Interface Plan views of Facilities animated to show alarm conditons from Autocad files. Password Access to all panels and system functions ensure security Menu screens can be configured in local language 87

Real Time Trending Trends provide multiple traces, multiple Y axes, real time and historic switching with alarm limits Cursor provides analysis features to read actual point values, min,max etc 88

Historic Trending Calendar date entry enables user to select start and stop date required Cursor allows trace values to be read, traces and alarm levels may be switched on/off. Up to six signature boxes for sign off. 89

Alarm Handling Alarms can be manually or automatically acknowledged and are time and date stamped on generation, acknowledgement and clearing down. 90

Alarm Reconciliation Alarm reconciliation provides users with detailed information concerning alarm transitions and in compliance and out of compliance operation 91

6000/7000 Series Designed to meet ISO 21501 requirements. Built for sterile processing environments with VHP-resistant optics, automatic flow control valve and NEMA-rate enclosure (7000) Built-in user notification of measurement alarms, flow problems, and sensor condition Multiple communication and mechanical installation options External alarm and environmental sensor capabilities reduce monitoring system complexity and costs Built-in memory and Hach OPC Server provide complete data security during network/computer problems Featuring Long Life Laser technology 92

Introduction to 6000 Series Multi-color status indicator LED Detail diagnostics by status indicator LED 0.2 µm minimum sensitivity Long Life Laser Models 6002, 6003, 6015 Standard 2 channel, optional 4 channel Built-in flow sensor option. Multiple I/O option Remote status indicator light option RH-Temp probe option Relative Humidity-Temperature probe connection Service port for instrument set-up Also used for remote indicator light stack 93

Introduction to 6000 Series Improved Sensitivity Flexible Communications Multiple Installation Options Enhanced Instrument Diagnostics Long Life Laser Diode Worldwide service and support 94

Improved Sensitivity and Long Life Laser Identify smaller particles reliably and accurately that could potentially improve product quality and yield with a sensitivity range of 0.2 to 5.0 μm 0.5 to 10.0 μm The Long Life Laser with improved MTTF reduces overall cost of ownership Design for Electronics application OR Life Science application (if sensor installed outside cleanroom) 95

Flexible Communications Save FMS integration costs, repair and inventory related costs with multiple communications Serial I/O (Pulse, RS-232, RS-485 with Modbus RTU or FX protocol) Ethernet Analog Wireless 5-Pin Phoenix connector and RJ-45 Ethernet I/O 10-Pin Phoenix connector Pulse, Serial RS232 Serial RS485 Modbus (RTU or FX) Analog 96

Installation Options - Connections Reduces re-installation time during routine calibration and maintenance with DIP switch option Easy Installation with multiple connection options Side vacuum connection Bottom vacuum connection DIP switch for RS485 instrument address setting Makes installation and removal easier Optional side vacuum connection Bottom vacuum connection 97

Installation Options - Mounting Various mounting options offer shorter installation and removal time DIN Rail (Standard) Mounting Option Utility Terminal Box Mounting Option Wall Plate Mounting Option 98

7000 Series 99

Introduction to 7000 Series ISO 21501 compliant design Multiple I/O option Built-in enhanced instrument status and user notification indicator Long Life Laser VHP tolerant flow path and optics Wall plate, umbilical cords and tubes with quick connect fittings for ease of installation and removal Up to 1000 sampling data storage 0.3 µm sensitivity, 0.1 or 1.0 cfm flow rate Built-in flow sensor option Remote status indicator light option Standard 2 channel, optional 4 channel Sealed enclosure with sealed connectors RH-Temp probe option Free set-up utility program Optional Relative Humidity- Temperature probe connection Multi-color status indicator LED Optional remote indicator light port Service port for instrument set-up Also can be used for optional remote display unit (future enhancement) 100

Flexible Communication Options Save FMS integration costs, repair and inventory related costs with multiple communications Serial I/O (RS-485 with Modbus RTU or FX protocol) Ethernet Analog Wireless RJ 45 sealed connector for Ethernet, RS485 serial or Analog I/O Makes integration to any FMS easier 101

Ease of Installation Features Easy installation with AC or DC input power options 100~230 VAC 50/60 Hz house power 24 VDC local power network Reduces re-installation time during routine calibration and maintenance with DIP switch feature for serial units Quick-connect type sealed vacuum connection Quick connect type vacuum connector For ease of installation DIP switch for setting address of RS485 serial I/O instrument Makes installation and removal easier AC or DC input power option Makes installation easier 102

Ease of Installation Features Wall plate for terminating end-user cables and tubing, reduces disruption to connectivity during routine calibration Umbilical with quick connect sealed connectors protect counter from wash down and make reinstallation easy Quick-connect type sealed vacuum connection makes reinstallation during routine calibration easy Wall Plate - Front Makes installation and removal of counter easier User terminates wiring, tubing at the rear end of the wall plate Makes installation easier Umbilical cords and tubing supplied to connect the counter with wall plate For ease of installation and subsequent removal for routine calibration and maintenance. Ensure sealed connections 103

Installation Features - Standard Accessories Reduces initial installation time Minimizes removal and re-installation time during routine calibration and maintenance Provides wash-down tolerance with sealed umbilical and tubing Item Number Description 1 7000 Counter 2 Mounting Plate 3, 4 Isokinetic Probe (0.1 or 1.0 cfm) 5 Wall Plate 6 Communication Cable with Sealed RJ- 45 Connectors 7 Power Cable AC or DC 8 Vacuum Tubing with Sealed Quick Connectors 104

Enhanced Instrument Diagnostics Enhanced diagnostics reduce trouble shooting time and downtime Built in or optional remote multi-color status indicator LED Instrument can be diagnosed for flow, sensor, communication failures, and count alarm or count alert through built-in or external light stack Steady Green = Normal (set by software or internal) Flashing Red = Count Alarm (set by software or internal) Flashing Yellow = Count Alert (set by software only) One short flash, one long flash Blue = Flow Fail (set by software or internal) Solid Blue = Sensor Fail (set by software or internal) Flashing Blue (2 Hz) = Communication Failure (internally set based on communication timeout) 105

Enhanced Instrument Diagnostics Enhanced instrument status indication reduce troubleshooting time and downtime Built in or optional remote multi-color status indicator LED Instrument can be diagnosed for flow, sensor, communication failures, and count alarm or count alert through built-in or external light indicator Steady Green = Normal (set by software or internal) Flashing Red = Count Alarm (set by software or internal) Flashing Yellow = Count Alert (set by software only) One short flash, one long flash Blue = Flow Fail (set by software or internal) Solid Blue = Sensor Fail (set by software or internal) Flashing Blue (2 Hz) = Communication Failure (internally set based on communication timeout) 106

107 Met One 6000/7000 Setup Utility Program Reduce set-up time with the Met One 6000/7000 setup utility program Local Setup LAN Setup Data Display Reduce trouble shooting time Run Met One 6000/7000 series remote counters without FMS software Obtain instrument information such as model, serial number and built in flow sensor

Local Setup Screen Capture 108

LAN Setup Screen Capture 109

Data Display Screen Capture 110

Environmental Sensors Measurement of Temp/RH, Differential Pressure, Air Flow and Temperature of Fridges, Freezers and Incubators Up to 1 km and 1000 channels Differential Pressure Sensor Temp/RH Sensor 111

Environmental Sensors DP Panels provide central location for signal connection for Temp/RH, PT100s, 4-20mA, DP tubes and digital I/O ( alarms etc.) 112

Alarm Indication Must provide alarm feedback e.g. via traffic light stack via alarm message display via volt free contact to BMS via network via pager/sms text 113

114 Alarm Indication Delivered where it is needed Red/Green lamps at each point of fill Alarm Message displays in area Computer screen in clean area

Alarm Message Display Alarm Indicators and Message Displays can be distributed around the facility 115

Alarm Message Display Alerts staff within facility that monitored parameters are out of limits Four line LCD display shows local parameters currently in alarm Built - in Sounder announces arrival of new alarm conditions Local Mute Button, silences sounder without leaving the room 116

Scalable Software Solutions For small FMS systems using only Met One 6000/7000 (best at < 20 counters), Pharmagraph envigil-lite provides an easy-to-implement solution Large FMS systems requiring: more than 10 20 sensors, alarm messaging, remote client PCs, custom screens & maps, consolidated reports, batch features, etc. are best supported with custom integrated systems such as Hach UltraVision Online, Pharmagraph envigil FMS, Wonderware, Intellution, etc Hach s OPC Server enables flexibility for ALL software solutions 117

Low-cost, Simple envigil Lite 2 for Aseptic Production Monitoring Ethernet RS-485 118

Large, Customized, Batch-Driven UVO Systems for Aseptic Production Monitoring Client # 1 Fill Line A Client # 2 Fill Line B Ethernet RS-485 Server/client with database Fill Line A Ethernet or RS-485 Grade C & D monitoring Fill Line B 119

OPC Server for Aseptic Production Met One OPC Server Customer Software Real-time data & control Sensor Control & OPC Server Sensor network Sensors controlled via MODBUS / TCP (SCADA, UVO, etc.) Historical electronic data Database Data from sensors configured for online operation Paper data & PDF files Crystal Reports Data from sensors configured for offline portable operation Translator that converts data from all Met One sensors into the language of the customers preferred system. 120

Pharmaceutical FMS Installations Many hundreds since 1987 Every major pharmaceutical company on every continent 121

Summary Review your whole particle monitoring program relative to evolving regulations and standards Review the amount of manual, paper-based monitoring Use automation as an opportunity to improve compliance while simultaneously gaining efficiency Involve all internal stake holders Work with flexible suppliers using open architectures to enable maximum integration and preserve options for future expansion 122

Table: Sampling Strategies based on ISO 14644 and EU GMP Annex 1 Grade Classification Regular EM Program Process Monitoring A 1. Frequency = 6 months 2. Minimum number of sample positions: ISO 14644-1 = SQRT area 3. Minimum sample volume: Annex 1 = 1 cubic meter 1. Frequency = a/r (daily????) 2. a/r; sometimes based on Minimum number of sample positions: ISO 14644-1 = SQRT area 3. Minimum sample volume: a/r 4. best practice: (3) 1-minute counts 1. Continuous during process 2. Key high risk positions 3. No minimum volume defined B 1. Frequency = 6 months 2. Minimum number of sample positions: ISO 14644-1 = SQRT area 3. Minimum sample volume: Annex 1 = based on ISO 14644 minimum 4. Min volume (5µm) = 24.36 cubic feet (0.69 m3) 1. Frequency = a/r (daily????) 2. a/r; sometimes based on Minimum number of sample positions: ISO 14644-1 = SQRT area 3. Minimum sample volume: a/r 4. best practice: (3) 1-minute counts 1. Frequent during process; continuous may be preferred 2. Key elevated risk positions 3. No minimum volume defined a/r = as required 123

Table: Sampling Strategies based on ISO 14644 and EU GMP Annex 1 Grade Classification Regular EM Program Process Monitoring C 1. Frequency = 12 months 2. Minimum number of sample positions: ISO 14644-1 = SQRT area 3. Minimum sample volume: (5 µm) Annex 1 > ISO 14644-1 = 0.24 cubic foot 1. Frequency = a/r (weekly????) 2. a/r; sometimes based on Minimum number of sample positions: ISO 14644-1 = SQRT area 3. Minimum sample volume: a/r 4.best practice: (3) 1-minute counts 1. no requirement 2. Low risk 3. No minimum volume defined D 1. Frequency = 12 months 2. Minimum number of sample positions: ISO 14644-1 = SQRT area 3. Minimum sample volume: (5 µm) Annex 1 > ISO 14644-1 = 0.024 cubic foot 1. Frequency = a/r (monthly????) 2. a/r; sometimes based on Minimum number of sample positions: ISO 14644-1 = SQRT area 3. Minimum sample volume: a/r 4.best practice: (3) 1-minute counts 1. no requirement 2. Low risk 3. No minimum volume defined a/r = as required 124

Regulations and Guidance for the Manufacture of Sterile Medicinal Products ISO 14644 and EU GMP Annex 1 125

Particle Count Theory and Calibration (ISO 21501) 126

Agenda Introduction to Cleanroom Classification of Air Cleanliness: ISO 14644 Guidance for the Manufacture of Sterile Medicinal Products EU GMP, Annex 1 Classification In-Process Monitoring Best Practices for FMS Solutions Particle Counter Theory and Calibration: IS0 21501-4 -------- On-line TOC Analyzer EP&USP Harmonization for Liquid Particle 127

Optical Particle Sensor Configuration 128

Sensor View Volume Inlet Tube The View Volume is The area where the air stream and the laser beam intersect Laser beam viewed in cross section Requires: Even illumination Precise alignment 129

Particle Sizing The larger the particle, the larger the corresponding output pulse from the sensor. 130

Counting Electronics +V Threshold Circuit This circuitry is duplicated from one channel up to six channels depending on the model counter Analog Signal in +V CH3 CH2 Digital signals to counting circuitry +V CH. 3 CH. 2 CH. 1 CH1 131

Air Particle Counter Calibration ISO 21501 and 14644 - the link to GMP Regulators inspect to EU GMP, which calls up ISO14644 Next revision ISO14644 will refer to ISO21501-4 ISO 21501-4 states Instruments that conform to this part of ISO 21501 are used for the classification of air cleanliness in cleanrooms and associated controlled environments in accordance with ISO 14644-1 132

What is ISO 21501? ISO 21501 is a new family of standards describing the instruments and calibration requirements for determining particle size distribution using light interaction methods for both liquid and airborne particle counters. ISO 21501 represents the culmination of work by instrumentation manufacturers and industry users and comes at a critical time for the life science industry with the increasing trend for real-time air particle monitoring in cleanrooms using light scattering air particle counters. 133

What is ISO 21501 replacing? Previous calibration methods guidelines: ASTM F 328-98(2003) Standard Practice for Calibration of an Airborne Particle Counter Using Monodisperse Spherical Particles (withdrawn May 2007). IEST-RP-CC014.1 Calibration and Characterization of Optical Airborne Particle Counters (providing actual methods to perform the calibration). JIS B 9921:1997 - Japanese standard which comprehensively deals with OPC design performance, most notably in the area of counting efficiency. 134

ISO 21501 Additional Tests Before ISO 21501-4 Size calibration Size resolution False count rate Sampling time ISO 21501-4 Size calibration Verification of size setting Counting efficiency Size resolution False count rate Concentration limit Sampling flow rate Sampling time Sampling volume 135

Parameter ISO 21501 Requirements 21501-2 LS Liquid 21501-3 LE Liquid 21501-4 Airborne Size calibration +/- 2.5% +/- 2.5% +/- 2.5% Verification of size setting +/- 15% Cal curve (minimum = 3 points) +/- 10% Reference material +/- 10% Cal curve (minimum = 3 points) 50% Counting efficiency 50 +/- 30% N/A 50 +/- 20% 100% Counting efficiency 100 +/- 30% 1.5 to 3X minimum size 100 +/- 20% Reference material < 2X minimum size 100 +/- 10% 1.5 to 2X minimum size Size resolution 10% 10% 15% False count rate Particles per liter (minimum size) ISO 21501 Requirements N/A Comments Standard uncertainty of the mean size Median voltage Use 10% (As-received) Close to the minimum detectable size. Concentration < 25% of concentration limit. Use 1.5 to 2X minimum size Manufacturer's recommended particle size. Use area under the curve. >1.5X CC sets markers at 5% medium value (counts) Particles per cubic Poisson distribution with a 95% confidence limit meter (minimum size) On screen calculator Maximum particle concentration Particles per cubic centimeter Particles per cubic centimeter Particles per cubic 10% Coincidence loss (calculated) meter (minimum size) On screen calculator Sample flow rate MFR standard uncertainty MFR standard uncertainty +/- 5% Volumetric flow rate If unit has flow rate control system Include flow rate uncertainty (all) Sample time +/- 1% +/- 1% +/- 1% On screen calculator Sample volume +/- 5% +/- 5% N/A On screen calculator Response rate N/A N/A 0.5% 10 min at concentration limit (minimum size), Sample (<60 sec) then filter (10 sec), then sample (< 60 sec) 136

Calibrating Air Counters with PSL Spheres f Mean size Std Deviation size NIST-traceable standard particles have normal distribution 137

Particle Count Distribution Number of Particles millivolts (Particle Size ) 138

Resolution/Counting Efficiency (Accuracy) N Particle size 0.3 µm 0.5 µm Good resolution showing separation of channels 139

Resolution/Counting Efficiency (Accuracy) N Particle size 0.3 µm 0.5 µm Poor resolution leads to poor counting accuracy: undercounting in smaller sized channels 140

Counting Efficiency ISO 21501 At the counter minimum specified size Using PSL that is 1.5 2 times the minimum specified size Counting efficiency must be 50% ±20% Counting efficiency must be 100% ±10% AND Channel 1 Channel 1 The counting efficiency for PSL particles of the minimum measurable particle size value marked in the specifications shall be within the range from 30% to 70% AND PSL particles of 1.5 2 times minimum measurable particle size shall be 90% to 110% 141

Illumination Uniformity Affects Resolution Laser beam in cross section Particle = Ideal: Even illumination in the view volume provides the same pulse height regardless of where the particle passes through the beam = = Not ideal: Uneven illumination causes pulse height variance. High illumination intensity leads to high intensity peak, and low intensity illumination leads to low peak height 142

Effects of Flow Rate on Pulse Height The amplitude of the sensor output is a function of the particle s residence time in the view volume. 28.3 lpm flow rate At 28.3 LPM, residence time yields a strong response from 0.5µ particle 50 lpm flow rate As flow rate is increased, residence time decreases and response decreases 100 lpm flow rate At very high flow rates, performance can be challenging as the particles pass through the view volume very quickly and calibration is based on a weaker signal 143

Sensor View Volume Optics and Flow Small inlet/outlet Nozzle Good optical uniformity Low flow turbulence Large inlet/outlet Nozzle Poor optical uniformity Higher flow turbulence 144

145 Design Considerations Pump Type Image Advantages Disadvantages Roots (Hach Met One Patent) Blower High flow Low power Lightweight Best vacuum High flow Low power Lightweight Poor vacuum (requires larger inlet nozzle) Carbon Vane High flow High power Heavyweight Maintenance

Hach Ultra and ISO 21501 Designed Compliant Instruments: 3400 Series 6000 Series 7000 Series 146

HACH Calibration Software Core Cal 3 Ability to provide a premium ISO 21501 calibration to the customer resulting in increased service opportunity. Same procedures & software for all HACH distributors Equivalent to a Factory calibration 147

Core Cal 3 Features Continued Automatic counter detection Real-time graphing capability 148

149 Professional Documentation Professional Abobe PDF certificates Custom logo capable Incorporated into Hach calibration procedures

Competitor s 100 lpm PHA Curve (0.5 um) Poor optical performance leads to very poor resolution Poor signal-to-noise leads to poor differentiation between noise and real particle counts This particle counter does not meet the JIS 9921 or ISO 21501 standard for counting efficiency 150

Ultra High Flow Considerations (100 lpm) Impact on immediate environment and airflow patterns in critical areas Isokinetic probe must be matched to particle counter flow rate High exhaust rate of sampled air may disturb room air flow Disturbance of airflow patterns in restricted areas due to high rate of sampled air Modest air supply of most LAF cabinets may not be sufficient to support high flow rates without substantial disturbance Recommend smoke studies to evaluate potential disturbance of controlled air flow Accuracy of size resolution and counting efficiency Pulsations in high flow pumps cause inconsistent air flow within the counter resulting in poor size resolution performance Long term performance higher maintenance requirements 151

Sampling Probes and Mounting High-Flow Unidirectional air bathing the exposed product during manufacturing Air from outside Grade A zone REMEMBER: There is no requirement to sample 1m 3 of air when monitoring 152

Effect of Exhaust Air: Issues in using High-Flow No exhaust air flow REMEMBER: There is no requirement to sample 1m 3 of air when monitoring Exhaust air flow creates turbulence and worse 153

Met One 3400 Roots Pump Operating at 100 lpm (0.5 μm) Clearly identified peak, excellent resolution Exceeds ISO21501 calibration standard 154

155 ISO 21501 Calibration Standard Improved compliance Removes ambiguity by providing a single internationally recognized standard method for calibration Harmonization between ISO and GMP guidance Improved unit-to-unit reproducibility All particle counters are referenced to a gold standard Improved counting accuracy Reduces false alarms in Grade A areas All current Met One particle counters from Hach may be calibrated using ISO 21501 at your facility

Portfolio 156

Total Organic Carbon 157

Why? Regulation TOC tousp<643>, EP 2.2.44 Conductivity to USP<645>, EP 2.2.38 Suitable for WFI and PW Water contamination Biofilm 158

TOC increase and Biofilm 159

Correlation between endotoxines and TOC 160

On-line vs. conventional lab testing 161

Anatel Theory 162

USP <643> & EP 2.2.44 Requirements Attribute USP <643> EP 2.2.44 Technology to be used share the objective of completely oxidizing the organic molecules in an aliquot of sample water to carbon dioxide have in common the objective of completely oxidising the organic molecules in the sample water to produce carbon dioxide Apparatus on-line or off-line using a calibrated instrument Use a calibrated instrument on-line or offline has demonstrated acceptable system suitability shown to have acceptable system suitability. 163

164 USP <643> & EP 2.2.44 Requirements Attribute USP <643> EP 2.2.44 Limit of detection 0.05 mg of carbon per liter 0.05 mg of carbon per litre Blank and water (r w ) TOC Water high purity water as defined under <661> containers (conductivity measured just prior to dispensing, of not more than 0.15 µs/cm) TOC level of not more than 0.25 mg per liter Reagent Water not greater than 1.0 µs.cm -1. not greater than 0.1 mg/l.

165 USP <643> & EP 2.2.44 Requirements Attribute USP <643> EP 2.2.44 System Suitability frequency Standard Solution (r s ) System Suitability Solution (r ss ) Response Efficiency periodically demonstrated USP Sucrose USP 1,4-benzoquinone RE = 100[(r ss r w )/(r s r w ] not less than 85% and not more than 115% System suitability SOPs should include error trapping for systematic errors. at suitable intervals Reagent grade sucrose Reagent grade 1,4-benzoquinone RE = 100[(r ss r w )/(r s r w ] not less than 85% and not more than 115% System suitability SOPs should include error trapping for systematic errors. Limit response r s r w limit response (not necessarily 500 ppb) r s r w limit response (not necessarily 500 ppb)

USP <643> & EP 2.2.44 Requirements Attribute USP <643> EP 2.2.44 On-line locations reflect the quality of the water used. (changed from earlier draft specifying quality of the water at the use points ) representative of the water used. 166

Anatel Theory 167

Anatel Theory of Operation The organic compounds in each sample are completely oxidized to CO 2 in the presence of UV light and a TiO 2 catalyst Conductivity of the water sample is measured before and after oxidation The difference in conductivity is due to the conductive species produced from the dissolved CO 2 in water, and is the basis for calculating the amount of organic carbon in the sample The end point of the oxidation process is determined precisely and dynamically by a sophisticated set of algorithms Inorganic species that contribute conductivity are constant, and are compensated for in the background measurement 168

Principle of operation Organics oxidized to CO 2 Conductivity δ conductivity UV on Time 169

Product Portfolio Positioning Anatel TOC Pharmaceuticals Electronics PAT700 A1000XP A643a A1000 170

171 Anatel Performance Criteria Low detection limit of 1.0 ppb TOC and conductivity in a single sensor Wide sample temperature range Complete, rapid oxidation Multiple sensor network for data acquisition and control Analog, digital and serial data outputs Convenient system suitability Multi-point user calibration Easy validation check Confirm conductivity meter accuracy and cell constant Measure grab samples (for model A643 & PAT700)

A643 overview On Line TOC analyzer Available in 2 versions (Portable or Stationary) Calibration, Suitability & Validation tests pre-programmed on the transmetter Completed oxydation of the sample Grab sample feature 172

PAT 700 173

Using On-line TOC analyzers to Meet USP <643> TOC and <645> Conductivity Process piping Isolation valve Cell Constant can be verified without removing from the line Water system does not have to be re-sanitized Drain 174

PAT 700 Key features - Video On-line TOC analysis with complete sample oxidation Science-based, risk managing instrument designed to meet all requirements of USP, EP and JP OASIS TM Onboard, Automated Standards Introduction System Integral color touch screen display Multiple inputs/outputs including separate analogs for TOC, temperature and conductivity Dual UV lamps with UV Detect technology for improved reliability and diagnostics IP 56 stainless-steel enclosure improves protection from water and particulates 175

PAT700 TOC Analyzer with OASIS TM Onboard, Automated Standards Introduction System 176

PAT700 TOC Analyzer with OASIS TM Reduces operator intervention inserting/changing out bottles and entering data All bottles for a test installed at once and used in sequence -Reduces risk Information about each standard stored in RFID (Radio Frequency Identification) tag on the bottle (concentration, C of A value, lot number, expiration date ) -Information can be written to the bottle by the analyzer Analyzer tracks amount of standard used and number of reps Voids bottle following test - avoids reuse of bottles Sensor serial number on excursion bottles 177

OASIS TM Technology - RFID RFID - Radio Frequency Identification Store all information concerning standards 178

Bottle Mode Excursion Mode Empty sample bottle with RFID tag can be loaded for excursion sampling Analyzer automatically fills bottle with sample from water system under User configured TOC value USP conductivity 35 or 36 error conditions Manually Information on sample programmed to RFID tag on the bottle Date, time, serial number, previous TOC, previous conductivity and previous temperature Validation bottle can be run automatically to verify analyzer Selection of excursion with or without validation sample Reinforces PAT capability through process analysis and validation 179

UV Detect TM Diagnostics Direct measurement of UV source performance Diagnosis of UV when TOC readings are skeptical Fast, real-time lamp feedback On-line: Verified after every TOC analysis Off-line: Can be conducted in diagnostic mode after lamp replacement Provides confidence for on-line water release Can be used for rouge monitoring Supports PAT initiative through reduced risk More reliable than firmware or simple hours-of-operation counters Reduces cost-of-ownership 180

Integral Color touch screen Integral color touch screen display for ease of use and access to information, configuration and analyzer operation 181

182 PAT700 Portable or Stationary Optional handle sold separately Customer installed Recommended to be used with quick connect version of analyzer

reflect the quality of the water used Water treatment plant Purified Water Tank Points Of Use WFI treatment plant WFI Tank Points Of Use 183

184 Cleaning in Place Validation Typical Process: Rinse vessel Grab samples Perform laboratory with TOC HPLC Drawback Labor intensive Material intensive Slow process Vessel being cleaned unavailable.

185 Cleaning in Place Validation TOC & Conductivity Conductivity/TOC Process: Rinse vessel Monitor conductivity When conductivity reach a low level, start TOC Valid CIP after TOC reach low value Advantage PAT Quick answer Lab independent

186

Cleaning in Place Validation TOC & Conductivity Pre-rinse 1 & 2 : Low or ambient temperature. Aim is to remove majority of product residues. Wash 1: Use detergent (sodium hydroxide based). Aim is to clean the vessel. Increase of temperature in order to boos this process Post Rinse 1: Aim is to remove caustic residues Wash 2: Use detergent (citric acid). Aim is to remove caustic residues Post rinse 2: Aim is to remove acidic residues. High temperature Final Rinse: Ambient temperature, for accurate measurement. 187

Cleaning in Place Validation TOC & Conductivity Conductivity TOC Time Final Rinse Start TOC measure Stop CIP 188

Lab move to online TOC 189

Total Organic Carbon 190

Liquid 191

Effects of Particulate Contamination Focus has been on injectable liquids Possibility to occlude (block) capillaries and arteries Red Blood cells are about 5 µm Capillary (5 to 10 µm) Large veins (10 to 50 µm) Threat of microbial infection Possibility of reaction to foreign substances (RES/allergic reaction) 192

193 Global Regulations:Particles in Liquids Primary method Optical Particle Counter [OPC] Light Obscuration Counter Secondary method Optical microscope Subjective Labor intensive Requires more time to process samples

194 Volume Definitions: SVI, LVI SVI ex., Antibiotics, Insulin, Cytotoxic drug Small Volume Injectable Less than or egal to100 ml Also known as SVP [Small Volume Parenteral] LVI ex., NaCl solution ( IV bag ) Large Volume Injectable More than 100 ml Also known as LVP [Large Volume Parenteral]

Global Regulations: Particles in Liquids USP 32-NF 26 <1> Injections USP 32-NF 26 <788> Particulate Matter in Injections USP 32-NF 26 <789> Particulate Matter in Ophthalmic Solutions 195

Global Regulations: Particles in Liquids EP 6 (6.3) Parenteral Preparations EP 6 (6.3) - (2.9.19) Particulate Contamination: Sub-visible Particles 196

197 Harmonization of USP 788 with EP Current version USP 32 May 2009 Current version EP6 <788> Particulate Matter in Injection

Harmonization of USP 788 Almost a verbatim copy of EP 2.9.19 Change in calibration management At least (4) sample aliquots to be taken USP 788 has previously required only (3) At least 25 ml pooled sample for SVI<25mL USP 788 has previously required only 20 ml Environment test changed Test at 10 microns only; no 25 micron limit now Exactly (5) sample draws of (5) ml At 10 microns, must be no more than 25 particles in 25 ml 198

Comparison of Compendial Requirements for Liquid Particles in Injections Feature USP 31-NF 26 (USP30-33(2) IRA) EP JP/KP Number of 4 4 3 samples Size of aliquot 5 ml 5 ml 5 ml or portion Discard #1 Yes Yes Yes SVI: 10 um 6000 per container 6000 per container 6000 per container counts SVI: 25 um 600 per container 600 per container 600 per container counts LVI: 10 um 25 per ml 25 per ml 25 per ml counts LVI: 25 um 3 per ml 3 per ml 3 per ml counts Environment 25 per 25 ml 25 per 25 ml 5 per 10 ml @ 10 um Environment @ 25 um No test No test 2 per 10 ml 199

Comparison of Compendial Requirements for Liquid Particles in Injections Container Type USP 29 <788> Volume <25 ml SVI/SVP Table 1 SVI 25 ml <= Volume <100 ml SVI/SVP Table 1 SVI Volume = 100 ml SVI/SVP Table 1 SVI 100 ml > Volume LVI/LVP Table 1 LVI EP 2.9.19 USP 31- NF 26 Test 1.B Test 1.B Counts per container Test 1.B Test 1.B Counts per container Test 1.B Test 1.B Counts per container Test 1.A Test 1.A Counts per ml 200

Ophtalmic products USP <789> Title: Particulate Matter in Ophthalmic Solutions Based on methods and standardization tests of USP <788> Increases particulate matter limits for ophthalmic solutions 50 or fewer particles of 10 micron or larger 5 or fewer particles of 25 micron or larger Excludes these formulations from particulate testing Gels Suspensions Emulsions Medical devices EP By microscope only, and only for eye-drops: No more than 20 particles 25 microns and larger No more than 2 particles 50 microns and larger. No particles greater than 90 microns. [Refer to General section, 1163, Eye Preparations]. 201

Calibration & System Suitability USP <788> 31 and earlier Instrument Standardization IST - Sample Volume accuracy - Sample Flow Rate - Calibration (Moving Windows) - Sensor Resolution - Accuracy with USP Particle Count RS Do be performed every 6 months 202

Calibration & System Suitability What is the USP Particle Count RS? 2 bottles of blank water 2 bottles containing 10 and 15µm particles, with a well know 10µm particles concentration and a well know ratio of counts at 10µm to the counts at 15µm 203

Calibration & System Suitability 204

Calibration & System Suitability USP <788> 32 Harmonized with EP & JP - Calibration with Particle Sphere Latex (sizes between 10 and 25µm) - System Suitability with USP Particle Count RS No timeframe commonly every year 205

Calibration & System Suitability Hach recommendation Calibration + IST every 6 months (Sample Volume accuracy; Sample Flow Rate; Calibration; Sensor Resolution; Particle Counting Accuracy USP Particle Count RS) On a regular basis (1 to 3 months), system suitability with USP Particle Count RS. 206

PharmSpec 2.2 Software Added USB to connect 9705 to computer Supports 21 CFR Part 11 compliance Familiar environment minimizes learning curve and training time Network backup and archiving Designed for Windows 2000 and XP Create and control user-defined tests 207

PharmSpec 2.2 Software Supports Compendial standards from USP, EP, JP and KP for Injectables and Ophthalmics 208

PharmSpec 2.2 Software Support Standardization tests: -Moving Windows -Sensor resolution -Volume Accuracy -USP Counting accuracy -Flow rate -Electronic resolution 209

PharmSpec 2.2 Software Permits users to comply with FDA 21 CFR Part 11 - Electronic Records, Electronic Signatures 210

Update for PharmSpec http://www.hachultra.com/index.php/hachultra/tools/downloads/pharmspec_2_0_downloads Customer can download compendial update at no charge from hachultra.com website 211

Product Portfolio Positioning 8103-1990 9703-1997 9705-2006 212

9705 Features: Sides Handles Easily moved Sensor Cleanout Quick access for removing blockages Thermal printer Delivers data immediately Counts Averages Standard Deviation Software for computer PHARMSPEC Include all EP & USP procedure test User procedure builder IQ/OQ doc 213

Liquid 214