Validation Guide Sartopore® 2 Cartridges, MaxiCaps...

Validation Guide Sartopore® 2 Cartridges, MaxiCaps®, MidiCaps® and Capsules

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1. Introduction 51.1 cGMP Quality Assurance

from Sartorius 61.2 Quality Assurance 61.3 Prevention of Contamination 61.4 Complete Traceability 61.5 Sartorius Drug Master File –

DMF 5967 61.6 Quality Management System 6

2. Technical Specifications 72.1 Type and Part Number Overview 72.1.1 Standard Cartridges 72.1.2 Mini Cartridges 82.1.3 MaxiCaps® 92.1.4 MidiCaps® 102.1.5 Capsules 150|300 112.2 Filter Material 122.3 Mechanism of Filtration 122.4 Pore Size Combinations 122.5 Materials of Construction 122.6 Fiber Release 122.7 Test Methods for the Quality

Assurance of Sartorius Sartopore® 2 Filter Cartridges and Capsules 13

2.8 Dimensions 142.8.1 Standard Cartridges 142.8.2 Mini Cartridges 142.8.3 MaxiCaps® 152.8.4 Total Height MidiCaps® 162.8.5 Capsules 150|300 172.9.1 Cartridges 182.9.2 MaxiCaps® 182.9.3 MidiCaps® 182.9.4 Capsules 150|300 182.10 Maximum Back Pressure

for Cartridges, MaxiCaps®, MidiCaps® and Capsules 150|300 18

2.11 Wetting the Filters for Integrity Testing 18

2.12 Rinse-Up Required for Particle Removal 18

2.13 Sterilization 182.14 Integrity Test Limits 192.14.1 Standard Cartridges 192.14.2 Mini Cartridges, MaxiCaps®,

MidiCaps® and Capsules 150|300 19

3. Flow Rates 203.1 Sartopore® 2 with 0.45 µm Final

Membrane 213.2 Sartopore® 2 with 0.2 µm Final

Membrane 243.3 Sartopore® 2 with 0.1 µm Final

Membrane 28

4. Chemical Stability – Sartopore® 2 33

5. Integrity Test Limits 355.1 Basis for the Determination of

Integrity Test Values 355.2 Bacteria Retention Test 365.3 Diffusion Test Limits for

Sartopore® 2 Filter Elements, 0.45 µm 37

5.3.1 Standard Cartridges and MaxiCaps® (10"|250 mm) 0.45 µm 37

5.3.2 Mini Cartridges and MidiCaps® 0.45 µm (0.2 m2|2.2 ft2) 39

5.4 Diffusion Test Limits for Sartopore® 2 Filter Elements, 0.2 µm 41


5.4.2 Standard Cartridges (5"|157 mm) 0.2 µm 43

5.4.3 Mini Cartridges and MidiCaps® (0.2 m2|2.2 ft2) 0.2 µm 45



5.4.6 Capsules 300 (0.03 m2|0.32 ft2) 0.2 µm 51

5.4.7 Capsules 150 (0.015 m2|0.16 ft2) 0.2 µm 53

5.5 Diffusion Test Limits for Sartopore® 2 Filter Elements, 0.1 µm 55


5.5.2 Standard Cartridges (5"|157 mm) 0.1 µm 57




5.5.6 Capsules 300 (0.03 m2|0.32 ft2) 0.1 µm 65

5.5.7 Capsules 150 (0.015 m2|0.16 ft2) 0.1 µm 67

5.6 Manual Determination of Maximal Allowable Pressure Drop 69

5.6.1 Sartopore® 2 Filter Elements with 0.45 µm Final Membrane 70

5.6.2 Sartopore 2 Filter Elements with 0.2 µm Final Membrane 71

5.6.3 Sartopore® 2 Filter Elements with 0.1 µm Final Membrane 72

6. Mycoplasma Retention 736.1 Introduction 736.2 Purpose 736.3 Test Methods 736.3.1 Qualification of the

Monitoring Filters 736.3.2 Test Set-up 746.4 Test Procedure 756.5 Test Results

Sartopore® 2 (10"|250 mm) 0.1 µm 756.6 Discussion 75

7. Thermal Stability 767.1 Steam Sterilization of Cartridges 767.1.1 Effects on Water Flow Rates 777.1.2 Effects on Diffusion Values 797.1.3 Effects on Bubble Point Values 817.1.4 Bacteria Challenge Test Values 83

Table of Contents

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7.2 Autoclaving of MaxiCaps®, MidiCaps® and Capsules 84

7.2.1 Effects on Water Flow Rates 857.2.2 Effects on Diffusion Values 877.2.3 Effects on Bubble Point Values 897.2.4 Bacteria Challenge Test Values 91

8. Testing According to USP 928.1 Particle Content of the Filtrate 938.1.1 Sartopore® 2 Filter Elements

with 0.2 µm Final Membrane 948.1.2 Sartopore® 2 Filter Elements

with 0.1 µm Final Membrane 968.2 Determination of Oxidizable

Substances of the Filtrate 988.2.1 Standard Cartridges 988.2.2 Mini Cartridges 998.2.3 MaxiCaps® 998.2.4 MidiCaps® 1008.2.5 Capsules 1018.3 Determination of pH Values and

Conductivity of the Filtrate 1028.3.1 Standard Cartridges 1038.3.2 Mini Cartridges 1058.3.3 MaxiCaps® 1068.3.4 MidiCaps® 1078.3.5 Capsules 1088.4 Determination of Chloride, Sulfate

and Ammonia in the Filtrate 1108.4.1 Determination of Chloride 1108.4.2 Determination of Sulfate 1148.4.3 Determination of Ammonia 1188.5 Biocompatibility 122

9. Endotoxin Testing 126

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Pharmaceutical products, such as injectable and infusion solutions or those which come in contact with open wounds, must conform to exactly defined quality standards. The desired quality of the final product can only be obtained when the entire production process is adequately safeguarded against contamination. Final product quality meeting the standards of the respective pharmacopeias can be achieved by using membrane filter technology at critical points where particles or microbes could contaminate a product or must be separated from it. Heat-stable final products can be sterilized practically and effectively by autoclaving. This process, however, does not remove particles or dead micro organisms which may release pyrogens.

Therefore, a prior membrane filtration run is required by cGMP regulations (Current Good Manufac-turing Practice of the US Food and Drug Administration) to ensure that particles and microbes are removed. Solutions containing heat-labile products, such as anti biotics, can be cold sterilized by membrane filtration immediately before aseptic filling. Microbe retentive filtration (bacteria retentive according to the European Pharmacopeia 6) or sterile filtration (sterilization by filtration in conformance with the current USP), respectively, is an important process step in the manufacture of sterile pharmaceutical products. When sterilizing filters are used in the manufacture of pharmaceu-ticals, the aseptic process must be validated, taking all aspects of the product and the production process into consi deration.

Sartopore® 2, pleated membrane filter elements with a heteroge-neous membrane, reliably fulfills the product-specific requirements which have to be imposed on a sterilizing grade filter. Validation is indispensable for guaranteeing the safety of pharmaceuticals, and is a logical supplement and significant part of the cGMP regulations which have been in force for quite some time. Guidelines for validation are given in the US Code of Federal Regulations Title 21 and the current USP. In addition, guidelines have been established jointly by the Committee for Laboratories and Official Drug Product Inspection Services and the Department of Industrial Pharmacists of the Feder-ation Internationale Pharmaceu-tique (F.I.P.), which is the European counterpart of the FDA. The term validation is defined by the F.I.P. guidelines as follows: ”Validation, as used in these guidelines, comprises the systematic testing of essential production steps and equipment in the R & D and production departments, including testing and inspection of pharmaceutical products with the goal of ensuring that the finished products can be manufactured reliably and repro-ducibly and in the desired quality in keeping with the established production and quality control procedures“.

We have compiled this validation guide so users of Sartopore® 2 Filter Cartridges, MaxiCaps®, MidiCaps® and Capsules can plan, implement and document their own validation procedures.

1. Introduction

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1.3 Prevention of ContaminationSartopore® 2 Filter Cartridges and MaxiCaps® are sealed in protective plastics bags in a controlled production area. During production Sartopore® 2 Filter Cartridges and MaxiCaps® are dried to reliably prevent microbial growth, and thus rule out the possibility of pyrogen synthesis during shipping and storage.

Sartopore® 2 MidiCaps® and Capsules sizes 4 and 5 are sealed in steam permeable protective plastic bags in a controlled production area. Following this step they are heat treated with steam to reliably prevent microbial growth, and thus rule out the possibility of pyrogen synthesis during shipping and storage.

1.4 Complete TraceabilityThe pore size, type and lot number are printed on the label of the protective plastic bag and on the label of the box in which the Cartridge or Capsule is packed. In addition, these specifications are imprinted on the outer cage of each Filter Cartridge or on the housing of each Capsule. The traceable lot number allows convenient retrieval of all data complied on the materials used, production steps and QC tests.

1.1 cGMP Quality Assurance from SartoriusConsistent high quality of Sartorius Membrane Filters, Capsules (ready-to-connect filtration units) and Filter Cartridges is assured by careful selection of the raw materials, well-planned and validated production technologies and an exceptionally efficient Quality Assurance Department, all of which results in high batch-to-batch reproducibility. The test procedures used are based both on external standard methods, such as the USP, EP and ASTM, and on in-house methods which are the result of Sartorius’ experience over the past 60 years.

1.2 Quality AssuranceFor quality assurance, all materials are selected carefully in accordance with current regulations, such as the FDA CFR’s, cGMP’s in-house guidelines and the specifications of our Research and Development Depart ment including the terms of delivery and acceptance of our Purchasing Department. Documen-tation begins with the inspection of the incoming raw materials including in-process materials, molded parts and sealing materials, etc. for manufacture. Adherence to cGMP requirements (clean-room conditions, gowning and employee hygiene, etc.) which are monitored by documented in-process controls, ensures optimal quality control in standard operating procedures for production. Finished Sartorius Capsules and Filter Cartridges undergo final product quality control. This involves 100 % non-de-structive testing of each individual product and other individual tests carried out on a representative number of samples. A lot is not released until all in-process and final quality control data are available.

1.5 Sartorius Drug Master File – DMF 5967In compliance with Title 21 Code of Federal Regulations (CFR) 314.420, the required information has been filed in the Sartorius Drug Master File – DMF 5967 under the heading ”Sartopore® 2 Cartridges and Capsules Validation Guide“ (Polyethersulfon)“ at the US Food and Drug Administration (FDA).

1.6 Quality Management SystemSartorius Stedim Biotech implemented Quality Management Systems to assure consistent high quality of Membrane Filters and Filter Cartridges.

Exemplary Quality Systems Certificates:

Quality Management System ISO 9001

Quality Management System ISO 13485

The complete Quality Systems Certificates are continuously updated and can be downloaded on our website:

www.sartorius-stedim.com/ qm-certificates

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2. Technical Specifications

2.1 Type and Part Number Overview

2.1.1 Standard Cartridges Explanation

544Sartopore® 2, heterogeneous double layer Polyethersulfone membrane filter

Pore Size06G 0.45 µm final membrane07H 0.2 µm final membrane58K 0.1 µm final membrane

Height Filtration Area0 5" 0.3 m2| 3.2 ft2

1 10" 0.6 m2| 6.5 ft2

2 20" 1.2 m2|12.9 ft2

3 30" 1.8 m2|19.4 ft2

Adapter21 Double Open end

cartridge with flat gasket

25 S-adapter top, locking bayonet adapter with 226 double O-ring bottom

27 Flat top, double 222 O-ring bottom

Adapter

Pore Size

Height

544 25 07H 1

8 |

Explanation

Adapter

Pore Size

Filtration Area

Units per Package

544 15 07H 9 B

Type of Adapter15 Bayonet adapter with

O-ring18 Plug adapter with

double O-ring


Height Filtration Area7 0.05 m2|0.5 ft2

8 0.1 m2|1.1 ft2

9 0.2 m2|2.2 ft2

Units per PackageB Five pieces

2.1.2 Mini Cartridges

544Sartopore® 2, heterogeneous double layer Polyethersulfone membrane filter

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2.1.3 MaxiCaps® Explanation


Height Filtration Area1 0.6 m2| 6.5 ft2

2 1.2 m2|12.9 ft2

3 1.8 m2|19.4 ft2

ConnectorsS 11⁄2" Tri-Clamp (sanitary)O 1⁄2" single stepped hose

barbF 3⁄4" Tri-Clamp (sanitary)B 3⁄4" to 1" multiple stepped

hose barb

NoteThe first code letter of the connector code represents the inlet, the second code letter represents the outlet connector.

Pore Size

Filtration Area

Connector

544 13 07H 1 00 544Sartopore® 2, heterogeneous double layer Polyethersulfone membrane filter

10 |

2.1.4 MidiCaps® Explanation



8 0.1 m2|1.1 ft2

9 0.2 m2|2.2 ft2

0 0.45 m2|4.8 ft2

ConnectorsS 11⁄2" Tri-Clamp (sanitary)O 1⁄2" single stepped hose

barbF 3⁄4" Tri-Clamp (sanitary)H 1⁄4" multiple stepped hose

barb (only Size 7)

Units per PackageA Four piecesV Two pieces


Pore Size

Filtration Area

Connector

Units per Package

53544 07H 9 00 A 544Sartopore® 2, heterogeneous double layer Polyethersulfone membrane filter

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2.1.5 Capsules 150|300 Explanation

Pore Size06G 0.45 µm (only size 5)07H 0.2 µm final membrane58K 0.1 µm final membrane


5 0.03 m2|0.32 ft2

ConnectorsN 1⁄4" NPT ThreadS ¾" Tri-Clamp (sanitary),

Capsule 150 onlyO 1⁄4" multiple stepped hose

barb with Filling Bell at outlet

Units per PackageB Five pieces


Pore Size

Filtration Area

Connector

Units per Package

13544 07H 4 OO B 544Sartopore® 2, heterogeneous double layer Polyethersulfone membrane filter

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2.2 Filter MaterialHydrophilic highly asymetric; heterogeneous double layer Polyethersulfone membrane filters, with the upstream filter membrane having a larger pore size than the final membrane.

2.3 Mechanism of FiltrationThe retention of particles and microorganisms is achieved by a sieving mechanism through the Polyethersulfone filter membrane. The throughput is enhanced through the use of fractionated filter membrane combinations where the two membranes have different retention ratings

2.4 Pore Size Combinations 0.45 µm + 0.2 µm 0.8 µm + 0.45 µm 0.2 µm + 0.1 µm

2.5 Materials of ConstructionAll materials meet the FDA require-ments as defined in Title 21 Code of Federal Regulations. Biosafety testing, such as the Class VI Plastics Testing as described in the current USP, are also met and exceeded.

Upstream Support Layer: Polypropylene

Filter Membrane: Polyethersulfone, double layer

Downstream Support: Polypropylene

Outer Cage: Polypropylene

Inner Core: Polypropylene

Endcaps: Polypropylene

Capsule Housings: Polypropylene

O-Rings|Gaskets: Silicone; Optional: EPDM or Viton

Filling Bell: Polycarbonate

2.6 Fiber ReleaseSartopore® 2 Filter Cartridges and elements comply with Title 21 Code of Federal Regul ations, Section 211.72 and 210.3 (b) (6) for non-fiber releasing filters.

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2.7 Test Methods for the Quality Assurance of Sartorius Sartopore® 2 Filter Cartridges and Capsules

Bubble Point of the Element

Diffusion Value of the Element

Flow Rate and Throughput of the Membrane

Bacteria Challenge Testing of every Lot of Filter Membrane

Bacteria Challenge Testing

Pyrogen Testing

Flow Rate Testing

Steam Sterilizability

Extractable Substances

Correlation of Diffusion and Bubble Point Values with the HIMA|ASTM Bacteria Challenge Tests

Current USP Class VI Plastics Tests Intracutaneous Test Systemic Injection Test Implantation Test

Particle Release

pH Change of the Filtrate

Conductivity Changes of the Filtrate

Extractable Substances

Water Flow Rates

Temperature and Pressure Resistance

Sterilizability In-Line Steam Sterilization Autoclavability

Evaluation of Integrity Test Values After Long Term Storage

Lot Related Tests 100 % Individual Testing

Routine Testing of Randomly Sampled Filter Cartridges and Capsules

Testing Conducted for the Validation of the Filter Cartridges and Capsules

14 |

‡ 70 mm

Hei

ght

Height Adapter 10" 20" 30" Diameter [mm] [mm] [mm] [mm]

21 250 498 746 70

25 321 568 817 70

27 275 530 786 70

± 3 mm per 10"

Height measurements include adapter and S-top where indicated in the diagram.

2.8 Dimensions

2.8.1 Standard Cartridges

Adapter 15 Adapter 18 Filtration Area Height Height Diameter [m2|ft2] ± 1 mm ± 1 mm ± 1 mm

0.05 |0.5 50 mm 61 mm 56 mm

0.1 |1.1 84 mm 95 mm 56 mm

0.2 |2.2 134 mm 144 mm 56 mm

2.8.2 Mini Cartridges 56 mm

Hei

ght

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Total Height

Filtration Connector Combinations [mm] Area SS SO OO FF BB [m2|ft2]

0.6| 6.5 365 371 377 365 385

1.2|12.9 615 621 627 615 635

1.8|19.4 865 871 877 865 885

2.8.3 MaxiCaps®

Total Diameter (including valves) Filtration All Connector Area Combinations [m2|ft2] [mm]

0.6| 6.5 140.6

1.2|12.9 140.6

1.8|19.4 140.6

Total Diameter (without valves) Filtration All Connector Area Combinations [m2|ft2] [mm]

0.6| 6.5 99.8

1.2|12.9 99.8

1.8|19.4 99.8

Diameter with Valves

Diameter without Valves

Hei

ght


16 |

Hei

ght

Filtration Connector Combinations [mm] Area SS SO OO FF FO FH HH [m2|ft2]

0.45|4.8 330 337 343 – – – –

0.2 |2.2 197 204 210 197 204 – –

0.1 |1.1 148 154 161 148 154 – –

0.05|0.5 115 120 126 115 120 116 120

2.8.4 Total Height MidiCaps®

Total Diameter (including Valves)

Filtration All Connector Area Combinations [m2|ft2] [mm]

0.45|4.8 108.5

0.2 |2.2 108.5

0.1 |1.1 108.5

0.05|0.5 108.5

Total Diameter Housings (without Valves)

Filtration All Connector Area Combinations [m2|ft2] [mm]

0.45|4.8 77

0.2 |2.2 77

0.1 |1.1 77

0.05|0.5 77

Diameter without Valves


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Total Height (without Filling Bell)

Filtration Area Connector Combinations [mm] [m2|ft2] NO SO OO SS

0.015|0.16 96 89 102 87

Total Height (with Filling Bell)

Filtration Area Connector Combinations [mm] [m2|ft2] NO SO OO

0.015|0.16 119 112 125

Total Height (with Filling Bell)

Filtration Area Connector Combinations [mm] [m2|ft2] OO

0.03|0.32 127

Total Height (without Filling Bell)

Filtration Area Connector Combinations [mm] [m2|ft2] OO

0.03|0.32 107

Total Diameter

Filtration Area All Connector Combinations [mm] [m2|ft2] (without Valves)

0.015|0.16 45.7

Total Diameter

Filtration Area All Connector Combinations [mm] [m2|ft2]

0.03|0.32 65

2.8.5 Capsules 150|300

2.8.5.1 Capsules 150

2.8.5.2 Capsules 300

Hei

ght

wit

hout

Fill

ing

Bell

Diameter Capsule Housing with Valves

Hei

ght

wit

h Fi

lling

Bel

l

Diameter Capsule Housing without Valves

Hei

ght

wit

h Fi

lling

Bel

l

Hei

ght

wit

hout

Fill

ing

Bell

Diameter Capsule Housing

18 |18 |

2.11 Wetting the Filters for Integrity TestingFor each filter element, rinse the filters in the direction of flow for 5 minutes with a differential pres sure of 0.3 bar| 4 psi backpressure 0.5 bar|7 psi in order to as sure that the filters have been wetted completely. Generally, filters are wetted with water. In cases where a different wetting medium is used, if the surface tension of the fluid is different from water (>70 dynes/cm), different integrity test values than indicated on the next page may be required.

2.12 Rinse-Up Required for Particle RemovalThe requirements for filtrate quality according to the current USP ”Sterile Water for Injection“ are met without the need for initial flushing of the Filter Cartridge.

2.13 Sterilization Autoclaving of wet Filter Cartridge, MaxiCaps®, MidiCaps® and Capsules up to a maximum temperature of 134 °C, for 30 minutes orIn-line steam sterilization of wetted Cartridges with a maximum of 2.3 bar|34 psi inlet pressure and 2 bar|29 psi outlet pressure (max. Dp= 0.3 bar|5 psi).

NoteMaxiCaps®, MidiCaps® and Capsules cannot be in-line steam sterilized.

2.9 Maximum Allowable Differential PressureThe maximum allowable differential pressure depends on the temper-ature at which the pressure is exerted. Maximum allowable differ-ential pressures in the direction of filtration.

2.9.1 Cartridges

Temperature [°C] 20 80 121 134

Pressure [bar] 5 2 1.5 0.5

Pressure [psi] 72.5 29 22 7

2.9.3 MidiCaps®

Temperature [°C] 20.0 50

Pressure [bar] 5.0 3

Pressure [psi] 72.5 43.5

2.9.2 MaxiCaps®

Temperature [°C] 20 50

Pressure [bar] 4 3

Pressure [psi] 58 43.5

2.9.4 Capsules 150|300


Pressure [bar] 4 2

Pressure [psi] 58 29

2.10 Maximum Back Pressure for Cartridges, MaxiCaps®, MidiCaps® and Capsules 150|300The maximum allowable pressure in reverse of the direction of filtration:

Cartridges

Temperature [°C] 20 100 121 134

Pressure [bar] 2 1 0.5 0.3

Pressure [psi] 29 14.5 7 4

MaxiCaps®, MidiCaps® and Capsules


Pressure [bar] 2 1.5

Pressure [psi] 29 21.8

| 19 | 19

2.14 Integrity Test Limits


Pore Size of the Height Test Pressure Maximum Diffusion Minimum Bubble Point Final Membrane [bar|psi] [mL/min] [bar|psi]

0.45 µm 10" 1.7|25 12 2.2|32

20" 1.7|25 24 2.2|32

30" 1.7|25 36 2.2|32

0.2 µm 5" 2.5|36 10 3.2|46

10" 2.5|36 18 3.2|46

20" 2.5|36 36 3.2|46

30" 2.5|36 54 3.2|46

0.1 µm 5" 4.0|58 14

10" 4.0|58 24

20" 4.0|58 48

30" 4.0|58 72

2.14.2 Mini Cartridges, MaxiCaps®, MidiCaps® and Capsules 150|300

Pore Size of the Filtration Area Test Pressure Maximum Diffusion Minimum Bubble Point Final Membrane [m2|ft2] [bar|psi] [mL/min] [bar|psi]

0.45 µm 0.03 |0.32 1.7|25 2 2.2|32

0.05 |0.5 1.7|25 3 2.2|32

0.1 |1.1 1.7|25 4 2.2|32

0.2 |2.2 1.7|25 6 2.2|32

0.45 |4.8 1.7|25 12 2.2|32

0.6 |6.5 1.7|25 12 2.2|32

1.2 |12.9 1.7|25 24 2.2|32

1.8 |19.4 1.7|25 36 2.2|32

0.2 µm 0.015 |0.16 2.5|36 1 3.2|46

0.03 |0.32 2.5|36 2 3.2|46

0.05 |0.5 2.5|36 4 3.2|46

0.1 |1.1 2.5|36 5 3.2|46

0.2 |2.2 2.5|36 7 3.2|46

0.45 |4.8 2.5|36 14 3.2|46

0.6 |6.5 2.5|36 18 3.2|46

1.2 |12.9 2.5|36 36 3.2|46

1.8 |19.4 2.5|36 54 3.2|46

0.1 µm 0.015 |0.16 4.0|58 1

0.03 |0.32 4.0|58 2

0.05 |0.5 4.0|58 4

0.1 |1.1 4.0|58 6

0.2 |2.2 4.0|58 9

0.45 |4.8 4.0|58 18

20 |

3. Flow Rates

BackgroundTest filter elements are placed into individual Sartorius filter housings (Sartorius housings, Type 340011P25TT112A or 331019P15TT112A). Capsules are directly installed into the piping system, using sanitary flanges. The piping system to and from the filters has an inner diameter of 25 mm|1 inch resp. 15 mm|0.6 inch. The water inlet is opened and the filter housings are completely vented.

The filters are rinsed for approxi-mately 5 minutes at 0.3 bar|4 psi differential pressure to assure complete wetting. The filter elements are then integrity tested to assure that only integral filters are tested. The inlet pressure (Pi) is held constant at 2.5 bar|36 psi.

Through the adjustment of valves on the downstream side of the filter housing, the required differential pressure for the test measurements is established. After achieving a constant differential pressure, the flow rate is recorded from the flow meter and the temperature is noted. The flow meter used in this testing was a Fisher & Porter COPA XM Magnetic Inductive Flow Meter Model D10D1465.

ResultsThe flow rate curves for water through Sartopore® 2 Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® of the various filtration areas versus differential pressure are on the following pages.

NoteThe flow rate is strongly influenced by the viscosity of the medium being filtered. For this reason, all flow rate measurements are taken at 20 °C so that the influence of temperature on viscosity is not a factor.

For flow rate measurements of 20" and 30" filter elements, the flow rates reach a point where the geometry of the piping and the filter housing begin to contribute to the over all differential pressure (resistance to flow). At a flow rate of approx imately 7000 L/h (120 L/min), the filter membrane surface area is no longer the flow limiting factor, but the housing and piping system begin to have increasing effects on differential pressure. For this reason the flow rates are only recorded at limited differential pressures.

Water Stream

Water Stream

Regulating Valve, Inlet

Housing with Test Filter or Capsule

Pressure Gauge Regulating Valve, Outlet

ThermometerFlow Meter

Diffusion and Bubble Point Measurements with the automated integrity test system, Sartocheck® 4 plus

Test Set-up

| 21

3.1 Sartopore® 2 with 0.45 µm Final Membrane

Water Flow Rates for Standard Cartridges and MaxiCaps®

[psi]

7.5

4.5

0

3

Differential Pressure [bar]

Flow Rate [L/h]

0 2500 5000 7500 100000

0.1

0.2

0.3

0.4

0.5

10”6

20” 30”

1.5

Water Flow Rates for Mini Cartridges with 0.05 m2|0.5 ft2, 0.1 m2|1.1 ft2 and 0.2 m2|2.2 ft2 Filtration Area

[psi]

15

7.5

0


Flow Rate [L/h]

0 1000 2000 30000

0.25

0.5

0.75

1.00

0.05 m2|0.5 ft2 0.1 m2|1.1 ft2 0.2 m2|2.2 ft2

22 |

Water Flow Rates for MidiCaps® with 0.45 m2|4.8 ft2 Filtration Area

[psi]

14.5

7.25

0


Flow Rate [L/h]

0 5.0000

0.25

0.5

0.75

1

OO SO SS

1.000 2.000 3.000 4.000


[psi]

14.5

7.25

0


Flow Rate [L/h]

0 3.0000

0.25

0.5

0.75

1

OO SO SSFF

1.000 2.000

| 23


[psi]

14.5

7.25

0


Flow Rate [L/h]

0 1.2000

0.25

0.5

0.75

1

OO SO SSFF

400 800


[psi]

14.5

7.25

0


Flow Rate [L/h]

0 200 8000

0.25

0.5

0.75

1

HH OO FF|SS|SO

400 600

24 |

Water Flow Rate for Sartopore® 2 Capsules 300 with 0.03 m2|0.32 ft2 Filtration Area

[psi]

15

7.5

0


Flow Rate [L/h]

0 100 200 3000

0.25

0.5

0.75

1

SS|SO|OO



[psi]

15

7.5

0


Flow Rate [L/h]

0 2000 4000 6000 8000 10000 12000 140000

0.25

0.5

0.75

1

10” 20” 30”5”

| 25


[psi]

15

7.5

0


Flow Rate [L/h]

0 500 1000 15000

0.25

0.5

0.75

1.00

0.05 m2|0.5 ft2 0.1 m2|1.1 ft2 0.2 m2|2.2 ft2


[psi]

14.5

7.25

0


Flow Rate [L/h]

0 1.000 4.0000

0.25

0.5

0.75

1

OO SO SS

2.000 3.000

26 |


[psi]

14.5

7.25

0


Flow Rate [L/h]

0 500 2.0000

0.25

0.5

0.75

1

OO SO SS

1.000 1.500

FF


[psi]

14.5

7.25

0


Flow Rate [L/h]

0 1.2000

0.25

0.5

0.75

1

FF OO SSSO

400 800

| 27


[psi]

14.5

7.25

0


Flow Rate [L/h]

0 200 8000

0.25

0.5

0.75

1

HH OO|SO|FF SS

400 600


[psi]

30

15

0


Flow Rate [L/h]

0 120 240 3600

0.5

1

1.5

2

SS|SO|OO

28 |28 |



[psi]

15

7.5

0


Flow Rate [L/h]

0 1000 2000 3000 4000 50000

0.25

0.5

0.75

1

10” 20” 30”5”


[psi]

30

15

0


Flow Rate [L/h]

0 60 120 1800

0.5

1

1.5

2

SS|SO|OO

| 29 | 29


[psi]

15

7.5

0


Flow Rate [L/h]

0 100 200 300 400 500 6000

0.25

0.5

0.75

1.00

0.05 m2|0.5 ft2 0.1 m2|1.1 ft2 0.2 m2|2.2 ft2


[psi]

14.5

7.25

0


Flow Rate [L/h]

0 1.5000

0.25

0.5

0.75

1

OO SO SS

500 1.000

30 |


[psi]

14.5

7.25

0


Flow Rate [L/h]

0 6000

0.25

0.5

0.75

1

OO FF SS|SO

200 400


[psi]

14.5

7.25

0


Flow Rate [L/h]

0 100 4000

0.25

0.5

0.75

1

OO SS|SO|FF

200 300

| 31

Water Flow Rate for MidiCaps® with 0.05 m2|0.5 ft2 Filtration Area

[psi]

14.5

7.25

0


Flow Rate [L/h]

0 50 2000

0.25

0.5

0.75

1

SS|SO|OO|FF|HH

100 150



Flow Rate [L/h]

0 50 100 1500

0.5

1

1.5

2

22.5

[psi]

30

15

0

7.5

OO

32 |


[psi]

30

15

0

7.5


Flow [L/h]

0 20 40 60 800

0.5

1

1.5

2

22.5SS|SO|OO

| 33

Compatibility measurement with complete filter element, but different O-ring materials:

Silicone EPDM VitonAcids:

HCL, 30 % – –

HCL, 25 % – –

HNO3, 10 %

HNO3, 65 %

H2SO4, conz. – – – – – –

H2SO4, 25 %

H3PO4, 25 %

Formic acid, conz. – – – –

Formic acid, 25 %

Acetic acid, conz.

Acetic acid, 25 % – –

Trichloracetic acid, 25 % – –

Trichloracetic acid, 10 % – –

Citric acid

Tartaric acid

Lactic acid

Bases:

Ammonia, 10 % – –

Ammonia, 30 % – – – –

NAOH, 1 M

NAOH, 2.5 M – – – –

KOH, 1 M

Solvents:

Acetone – – – – – –

Cyclohexanone – – – – – –

Methylethylketone – – – – – –

Methylisobutyketone – – – –

Diethylether

Methanol, 98 %

Ethanol, 10 %

Ethanol, 98 %

Isopropanol

n-Propanol

n-Amylalcohol

n-Butanol

Glycerol

Etyleneglycol

Methyleneglycol

Dioxane – – – – – –

Tetrahydrofuran – – – – – –

Dimethylsulfoxide – – – – – –

Dimethylformamide – – – – – –

Triethanolamine

4. Chemical Stability – Sartopore® 2

Legend = Compatible = Limited compatibility

depending on concentration, temperature etc.

– – = Not compatible

Test Specifications7 days contact at 20 °C

ImportantCompatibility is influenced by various factors, such as temper-ature, concentration, etc. If necessary, test the compatibility with the solution you wish to filter before performing the actual filtration run.

34 |

Legend = Compatible = Limited compatibility

depending on concentration, temperature etc.

– – = Not compatible

Test Specifications7 days contact at 20 °C

ImportantCompatibility is influenced by various factors, such as temper-ature, concentration, etc. If necessary, test the compatibility with the solution you wish to filter before performing the actual filtration run.

Silicone EPDM VitonMiscellanous:

Aniline – – – – – –

Sodium hypochlorite

Benzylalcohol – – – – – –

Phenol, 10 % – – – – – –

Formalin, 30 %

Hexane

Xylene – – – – – –

Toluene – – – – – –

Benzene – – – – – –

Tetralin – – – – – –

Dekalin – –

Methylenchloride – – – – – –

Chloroform – – – – – –

Carbontetrachloride – – – – – –

Trichloreethylene – – – – – –

Perchloreethylene – – – –

Monochlorbenzol – – – – – –

Methylacetate – – – – – –

Ethylacetate – – – – – –

Amylacetate – –

Propylacetate – –

Terpentine – –

H2O2, 0.3 %

Ammoniumpersulfat, 25 %

Sodiumhypochloride, 5 %

Starch solution

Water

| 35

5. Integrity Test Limits

5.1 Basis for the Determination of Integrity Test ValuesEstablishing a correlation between bacterial retention of a sterilizing grade filter and a non-destructive integrity test is decisive for the reliability of a sterile filtration process.

According to the Health Industry Manufacturers Association (HIMA) Guidelines for ”Microbiological Evaluation of filters for Steril-izing Liquids“ Doc. No. 3, Vol. 4, 1982|ASTM F 838-05 Guideline, and the FDA ”Guideline on Sterile Drug Products Produced by Aseptic Processing“, June 1987, a steril-izing grade Filter Cartridge should produce a sterile effluent when challenged with a minimum concentration of 107 Brevun-dimonas diminuta organisms/cm2 of filter area.

The FDA ”Guidelines on Sterile Drug Products Produced by Aseptic Processing“, June 1987 states:

”After a filtration process is properly validated for a given product, process and filter, it is important to assure that identical filter replacements (membrane or cartridge) used in production runs will perform in the same manner. One way of achieving this is to correlate filter performance data with filter integrity testing data. Normally, integrity testing of the filter is performed after the filter unit is assembled and sterilized prior to use. More importantly, however, such testing should be conducted after the filter is used in order to detect any filter leaks or perfora-tions that may have occurred during filtration.

Test-MethodSeveral hundred Sartopore® 2 Filter Cartridges and Capsules, with 0.1 µm and 0.2 µm pore size membranes, from numerous production lots were tested according to a Bacteria Challenge Test in accordance with the HIMA Document No. 3, Vol. 4 (April 1982) ”Microbiological Evaluation of Filters for Sterilizing Liquids“|ASTM F 838-05 Guideline, and DIN 58356, Part 1.

For Sartopore® 2 Filter Cartridges and Capsules with 0.45 µm final membrane the same methodology was applied using Serratia marce-scens as challenging organism.

Test OrganismBrevundimonas diminuta (0.2 µm) (ATCC 19146)Serratia marcescens (0.45 µm) (ATCC 14756)

NoteFor validation studies of the Sartopore® 2 filter elements, a minimum concentration of 1 ƒ 107 B. diminuta (S. marcescens) per cm2 filtration area for each tested element was used.

Integrity TestThe Sartopore® 2 filter elements were integrity tested by diffusion and bubble point test methods in order to correlate the results of the destructive Bacteria Challenge Test with these non-destructive integrity tests.

The Diffusion Test and the Bubble Point Test are performed utilizing a Sartocheck® 4 Plus automated integrity test unit.

The diffusion values are determined at a test pressure of 2.5 bar|36 psi for 0.2 µm rated filters and 1.7 bar|25 psi for 0.45 µm rated filters. For the determination of the bubble point, air pressure is slowly increased on the upstream side of the filter housing by the Sartocheck® 4 Plus integrity tester.

36 |

Water flow is initiated and the water stream first passes through a sterilizing grade Filter Cartridge. The purpose of this filter is to remove particles and bacteria to assure the Test Filter is only challenged with the bacterial load as described in the HIMA|ASTM Document.

Sterilizing Filter CartridgeSartobran®-P 0.45 + 0.2 µm Membrane Filter 5232507H1-P

The bacterial challenge bioburden that will be introduced to the test Filter Cartridge is controlled by dosing of the bacterial suspension into the water stream with a peristaltic pump. After the bacterial suspension is added to the water stream, the flow is directed through a mixing tube to ensure that proper mixing of the bacterial suspension has occurred. For the control and monitoring of the differential pressure during the Bacteria Challenge Test, pressure gauges and valves have been installed on the upstream and downstream side of the Filter Cartridges. The filtrate that passes through the test filter then flows through the analytical filters. After the completion of the Bacteria Challenge Test, these analytical filters can be examined according to the analytical methods described in the HIMA|ASTM document.

Test ProcedureThe Sartopore® 2 Filter Cartridges or Capsules are installed and wetted as described in the operating instruct ions. The filter system is then sterilized.

The system is then rinsed with water and the test filter is integrity tested with the Sartocheck® 4 plus. The water flow is controlled with the valving of the system and set so that the bacterial suspension can be dosed into the water stream. After the Bacteria Challenge Test, the analytical filters are incubated on agar plates to determine if there was passage of bacteria through the test filter. The analysis of the analytical filters is conducted according to the HIMA|ASTM Method.

5.2 Bacteria Retention Test

Water Stream

Analytical Filters

Water Stream

Regulating Valve

Sterile Prefil-tration

Dosing Pump

Bacteria Suspension

Pressure Gauge

Mixing Tube


Flow Meter


Test Set-up

| 37

5.3 Diffusion Test Limits for Sartopore® 2 Filter Elements, 0.45 µm

5.3.1 Standard Cartridges and MaxiCaps® (10"|250 mm) 0.45 µm

Lot Number Diffusion [mL/min] Bioburden [CFU]* Filtrate Quality

026641 6.1 1.35 ƒ 1011 sterile

018081 6.2 2.45 ƒ 1011 sterile

026841 6.4 3.85 ƒ 1011 sterile

026641 6.4 1.35 ƒ 1011 sterile

026741 6.6 1.35 ƒ 1011 sterile

026741 6.7 4.6 ƒ 1011 sterile

026641 6.7 8.0 ƒ 1010 sterile

026641 6.8 1.35 ƒ 1011 sterile

026841 6.9 2.85 ƒ 1011 sterile

026741 7.0 2.1 ƒ 1011 sterile

026841 7.1 2.85 ƒ 1011 sterile

026741 7.2 1.35 ƒ 1011 sterile

026641 7.2 2.7 ƒ 1011 sterile

017981 7.3 8.0 ƒ 1010 sterile

026641 7.4 1.35 ƒ 1011 sterile

026741 7.5 2.1 ƒ 1011 sterile

026841 7.6 2.1 ƒ 1011 sterile

026741 7.8 2.1 ƒ 1011 sterile

017981 7.9 2.45 ƒ 1011 sterile

026741 8.0 4.6 ƒ 1011 sterile

017681 8.1 1.0 ƒ 1011 sterile

026841 8.2 2.1 ƒ 1011 sterile

026841 8.4 4.45 ƒ 1011 sterile

026841 8.6 4.45 ƒ 1010 sterile

012281 8.8 8.0 ƒ 1010 sterile

026741 9.0 2.1 ƒ 1011 sterile

021381 9.2 4.45 ƒ 1011 sterile

012081 9.4 1.35 ƒ 1011 sterile

026841 9.6 4.45 ƒ 1011 sterile

021341 9.8 4.45 ƒ 1011 sterile

012081 10.2 1.15 ƒ 1011 sterile

021281 10.4 2.7 ƒ 1011 sterile

021381 10.7 2.1 ƒ 1011 sterile

012181 11.2 1.15 ƒ 1011 sterile

021281 11.5 8.0 ƒ 1010 sterile

026841 12.0 4.45 ƒ 1011 sterile

021381 12.2 2.1 ƒ 1011 sterile

012181 12.4 1.35 ƒ 1011 sterile

012281 13.1 8.0 ƒ 1010 sterile

021281 14.3 2.7 ƒ 1011 sterile

026641 15.7 2.6 ƒ 1011 non sterile

021281 18.3 4.45 ƒ 1011 non sterile

026841 19.1 2.1 ƒ 1011 non sterile

* CFU = Colony Forming Units

38 |

ConclusionThe data shows that Filter Cartridges that have diffusion values < 14.3 mL/min always produced a sterile filtrate with 100 % retention of the test organism, Serratia marcessens.

In order to have a high degree of safety when evaluating the test results, and considering that other filter integrity test units or other test methods may be used, a safety margin of 2.3 mL/min has been defined. For a thoroughly water wetted 10" Sartopore® 2 0.45 µm Filter Cartridge, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 1.7 bar|22 psi at 20 °C is:

12 mL/min.

Sartopore® 2 0.45 µm Filter Cartridge of various lengths have the following maximum allowable diffusion values at a test pressure of 1.7 bar|22 psi at 20 °C:

for a 10" Filter Cartridge: 12 mL/min



Bubble Point: $ 2.2 bar|32 psi

NoteThe diffusion and Bubble Point Test results are influenced by the nature of the wetting medium. The diffusion and bubble point values listed in this validation guide are for Sartopore® 2 Filter Cartridges wetted with water at 20 °C. It should be noted, that a variation of the test conditions such as temperature, wetting liquid or type of gas may require a different integrity test limit related to those mentioned above.

If a different test method is selected, for example an integrity test device that measures the values by monitoring the upstream pressure drop, this test method must be verified to the direct methods described above. The upstream pressure drop (pressure hold) test is not only influenced by the diffusion of gas through the wetted filter membranes, but also the upstream volume of the filtration system. Without exact values for the upstream volume of the filtration systems, maximum allowable pressure drop values cannot be calculated for a particular filter system.

Test Cartridge Lot Number

Diffusion [mL/min]

0266

41

Non Sterile

Sterile

0

2

4

6

8

10

12

14

16

18

20

Safety margin

0180

8102

6841

0266

4102

6741

0267

4102

6641

0266

4102

6841

0267

4102

6841

0267

4102

6641

0179

8102

6641

0267

4102

6841

0267

4101

7981

0267

4101

7681

0268

4102

6841

0268

4101

2281

0267

4102

1381

0120

8102

6841

0213

4101

2081

0212

8101

2181

0212

8102

6841

0213

81

0213

81

0121

8101

2281

0212

8102

6641

0212

8102

6841

Standard Cartridges and MaxiCaps® (10"|250 mm) 0.45 µm

| 39

5.3.2 Mini Cartridges and MidiCaps® 0.45 µm (0.2 m2|2.2 ft2)

Lot Number Diffusion [mL/min] Bioburden [CFU] Filtrate Quality

107541 2.0 3.52 ƒ 1011 sterile

107641 2.1 6.25 ƒ 1010 sterile

107641 2.2 6.25 ƒ 1010 sterile

102341 2.3 3.52 ƒ 1011 sterile

107541 2.4 5.5 ƒ 1010 sterile

107541 2.5 3.52 ƒ 1011 sterile

102341 2.5 2.16 ƒ 1011 sterile

107641 2.6 6.25 ƒ 1010 sterile

102341 2.7 2.16 ƒ 1011 sterile

102341 3.1 2.16 ƒ 1011 sterile

107641 3.3 6.25 ƒ 1010 sterile

107541 3.5 3.52 ƒ 1011 sterile

107541 3.7 3.52 ƒ 1011 sterile

107641 4.0 6.25 ƒ 1010 sterile

102341 4.2 2.6 ƒ 1011 sterile

102341 4.3 2.6 ƒ 1011 sterile

102341 4.7 2.6 ƒ 1011 sterile

107641 4.9 6.25 ƒ 1010 sterile

107541 5.3 3.52 ƒ 1011 sterile

107541 5.5 3.52 ƒ 1011 sterile

107641 5.8 6.25 ƒ 1010 sterile

102341 6.0 2.6 ƒ 1011 sterile

102341 6.5 2.6 ƒ 1011 sterile

107641 6.7 6.25 ƒ 1010 sterile

102341 7.3 6.6 ƒ 1011 sterile

107541 7.6 3.52 ƒ 1011 sterile

102341 8.3 2.6 ƒ 1011 non sterile

107541 9.0 3.52 ƒ 1011 non sterile

107641 11.4 6.25 ƒ 1010 non sterile

40 |

ConclusionThe data shows that Mini Cartridge and MidiCaps® and Capsules that have diffusion values # 7.6 mL/min always produced a sterile effluent with 100 % retention of the test organism, Serratia marcessens.

In order to have a high degree of safety when evaluating the test results, and considering that other filter integrity test units or other test methods may be used, a safety margin of 1.6 mL/min has been defined.

For a thoroughly water wetted Sartopore® 2 Mini Cartridge and MidiCap®, with 0.2 m2 filtration area and 0.45 µm pore size (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 1.7 bar|25 psi at 20 °C is:

6 mL/min.

Size 8: 4 mL/minSize 7: 3 mL/minSize 5: 2 mL/min


Lot Number

Diffusion [mL/min]

1075

41

Non Sterile

Sterile

0

2

4

6

8

10

12

Safety margin

1076

4110

7641

1023

4110

7541

1075

4110

2341

1076

4110

2341

1023

4110

7641

1075

4110

7541

1076

4110

2341

1023

4110

2341

1076

4110

7541

1075

4110

7641

1023

4110

2341

1076

4110

2341

1075

4110

2341

1075

4110

7641

Mini Cartridges and MidiCaps® (0.2 m2|2.2 ft2) 0.45 µm

| 41



Note Since most of the filters tested during the validation studies had low diffusion values and produced a sterile filtrate, the following data is a sampling from all filters tested during the validation testing indicating results near the diffusion|sterile filtrate limits.


920841 11.5 1.12 ƒ 1011 sterile

925041 11.6 1.70 ƒ 1011 sterile

920641 11.7 6.12 ƒ 1010 sterile

925041 11.8 1.70 ƒ 1011 sterile

925141 11.9 1.78 ƒ 1011 sterile

925141 12.0 1.78 ƒ 1011 sterile

924941 12.1 1.12 ƒ 1011 sterile

925041 12.3 1.70 ƒ 1011 sterile

925041 12.5 1.78 ƒ 1011 sterile

925141 12.5 1.28 ƒ 1011 sterile

925141 12.5 1.78 ƒ 1011 sterile

920741 12.6 2.63 ƒ 1011 sterile

920641 12.7 7.20 ƒ 1010 sterile

920741 13.1 2.63 ƒ 1011 sterile

925041 13.1 1.66 ƒ 1011 sterile

920741 13.2 2.63 ƒ 1011 sterile

920641 13.4 7.20 ƒ 1010 sterile

920641 13.6 6.12 ƒ 1010 sterile

920641 13.7 7.20 ƒ 1010 sterile

920641 13.9 6.12 ƒ 1010 sterile

920541 13.9 1.66 ƒ 1011 sterile

920641 14.1 7.20 ƒ 1010 sterile

920741 14.4 2.63 ƒ 1011 sterile

920641 14.5 6.12 ƒ 1010 sterile

920641 14.6 2.60 ƒ 1011 sterile

920641 14.9 6.12 ƒ 1010 sterile

920641 14.9 6.12 ƒ 1010 sterile

920641 15.3 7.20 ƒ 1010 sterile

920741 15.7 2.92 ƒ 1011 sterile

917581 16.2 1.78 ƒ 1011 sterile

920741 16.5 2.63 ƒ 1011 sterile

917581 16.8 1.78 ƒ 1011 sterile

917581 17.5 1.78 ƒ 1011 sterile

920741 18.0 1.01 ƒ 1011 sterile

917581 18.9 1.78 ƒ 1011 sterile

924941 19.1 1.12 ƒ 1011 sterile

924941 19.3 1.66 ƒ 1011 sterile

924941 19.4 1.66 ƒ 1011 sterile

920741 19.8 1.01 ƒ 1011 sterile

925141 19.9 1.28 ƒ 1011 sterile

924941 20.8 1.66 ƒ 1011 sterile

917581 22.3 1.78 ƒ 1011 non sterile

925041 24.6 1.66 ƒ 1011 non sterile

924941 25.5 1.66 ƒ 1011 non sterile

920841 27.3 1.12 ƒ 1011 non sterile

* CFU = Colony Forming Units

42 |

ConclusionThe data shows that Filter Cartridges that have diffusion values < 20.8 mL/min always produced a sterile filtrate with 100 % retention of the test organism, Brevundimonas diminuta. In order to have a high degree of safety when evaluating the test results, and considering that other filter integrity test units or other test methods may be used, a safety margin of 2.8 mL/min has been defined. For a thoroughly water wetted 10" Sartopore® 2 0.2 µm Filter Cartridge, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 2.5 bar|36 psi at 20 °C is:

18 mL/min.

Sartopore® 2 0.2 µm Filter Cartridges and MaxiCaps® of various lengths have the following maximum allowable diffusion values at a test pressure of 2.5 bar|36 psi at 20 °C:





NoteThe diffusion and Bubble Point Test results are influenced by the nature of the wetting medium. The diffusion and bubble point values listed in this validation guide are for Sartopore® 2 Filter Cartridges wetted with water at 20 °C. It should be noted, that a variation of the test conditions such as temperature, wetting liquid or type of gas may require a different integrity test limit related to those mentioned above.


Test Cartridge Lot Number

Diffusion [mL/min]

9208

41

Non Sterile

Sterile

0

5

10

15

20

25

30

Safety margin

9250

4192

0641

9250

4192

5141

9251

4192

4941

9250

4192

5041

9251

4192

5141

9207

4192

0641

9207

4192

5041

9207

4192

0641

9206

4192

0641

9206

4192

5041

9206

4192

0741

9206

4192

0641

9206

4192

0641

9206

4192

0741

9175

8192

0741

9175

8192

0741

9175

8192

4941

9249

41

9175

81

9249

4192

0741

9251

4192

4941

9175

8192

5041

9249

4192

0841


| 43

5.4.2 Standard Cartridges (5"|157 mm) 0.2 µm



036002 4.2 1.7 ƒ 1011 sterile

036302 4.5 1.5 ƒ 1010 sterile

036102 5.2 1.7 ƒ 1011 sterile

036302 5.3 1.7 ƒ 1010 sterile

036302 5.4 1.7 ƒ 1010 sterile

036102 5.4 1.1 ƒ 1011 sterile

036302 5.5 1.1 ƒ 1011 sterile

036002 5.6 1.7 ƒ 1010 sterile

036102 5.9 1.7 ƒ 1011 sterile

036102 6.1 1.7 ƒ 1011 sterile

036102 6.3 1.7 ƒ 1011 sterile

036002 6.5 1.4 ƒ 1011 sterile

036002 6.9 1.4 ƒ 1011 sterile

036302 6.9 1.7 ƒ 1010 sterile

036302 7.0 1.7 ƒ 1011 sterile

036302 7.4 1.7 ƒ 1010 sterile

036302 7.8 1.5 ƒ 1010 sterile

036302 8.2 1.7 ƒ 1010 sterile

036102 8.6 1.7 ƒ 1011 sterile

036002 8.9 1.8 ƒ 1010 sterile

036002 9.5 1.4 ƒ 1011 sterile

036102 9.9 1.7 ƒ 1011 sterile

036002 10.5 1.7 ƒ 1011 sterile

036102 10.8 1.4 ƒ 1011 sterile

036002 11.3 1.7 ƒ 1011 sterile

036002 11.6 1.7 ƒ 1010 sterile

036002 12.3 1.5 ƒ 1010 sterile

036102 12.6 1.7 ƒ 1011 non sterile

036002 13.5 1.7 ƒ 1011 non sterile

036002 14.3 1.7 ƒ 1011 non sterile

44 |44 |

ConclusionThe data shows that Filter Cartridges that have diffusion values < 12.3 mL/min always produced a sterile filtrate with 100 % retention of the test organism, Brevundimonas diminuta.

In order to have a high degree of safety when evaluating the test results, and considering that other filter integrity test units or other test methods may be used, a safety margin of 2.3 mL/min has been defined. For a thoroughly water wetted 5" Sartopore® 2 0.2 µm Filter Cartridge, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 2.5 bar|36 psi at 20 °C is:

10 mL/min.

Bubble Point$ 3.2 bar|46 psi

Lot Number

Diffusion [mL/min]

0360

02

Sterile

0

2

4

6

8

10

12

14

16

18

20

Safety margin

0363

0203

6102

0363

0203

6302

0361

0203

6302

0360

0203

6102

0361

0203

6102

0360

0203

6002

0363

0203

6302

0363

0203

6302

0363

0203

6102

0360

0203

6002

0361

0203

6002

0361

0203

6002

0360

0203

6002

0361

0203

6002

0360

02

Non Sterile

Standard Cartridges (5"|157 mm) 0.2 µm

| 45 | 45

5.4.3 Mini Cartridges and MidiCaps® (0.2 m2|2.2 ft2) 0.2 µm


933041 3.7 8.12 ƒ 1010 sterile

933041 3.8 7.28 ƒ 1010 sterile

932941 3.9 7.28 ƒ 1010 sterile

932941 4.0 7.28 ƒ 1010 sterile

933041 4.0 7.28 ƒ 1010 sterile

932941 4.1 7.28 ƒ 1010 sterile

932941 4.3 7.28 ƒ 1010 sterile

927941 4.4 7.14 ƒ 1010 sterile

932941 4.4 8.12 ƒ 1010 sterile

932641 4.5 8.12 ƒ 1010 sterile

932941 4.7 7.28 ƒ 1010 sterile

929441 4.8 2.66 ƒ 1010 sterile

929441 4.9 2.66 ƒ 1010 sterile

929441 5.0 2.60 ƒ 1010 sterile

927941 5.1 7.14 ƒ 1010 sterile

929441 5.4 2.60 ƒ 1010 sterile

930541 5.6 9.10 ƒ 1010 sterile

930541 6.1 9.10 ƒ 1010 sterile

930541 6.4 9.10 ƒ 1010 sterile

932641 6.4 8.12 ƒ 1010 sterile

932641 7.3 8.12 ƒ 1010 sterile

927941 7.4 7.14 ƒ 1010 sterile

932641 7.5 7.28 ƒ 1010 sterile

927941 8.0 7.14 ƒ 1010 sterile

927941 8.5 7.14 ƒ 1010 sterile

930541 8.8 9.10 ƒ 1010 sterile

932941 9.1 8.12 ƒ 1010 non sterile

930541 9.3 9.10 ƒ 1010 non sterile

930541 9.9 9.10 ƒ 1010 non sterile

929441 10.4 2.66 ƒ 1010 non sterile

46 |

ConclusionThe data shows that Mini Cartridges and MidiCaps® that have diffusion values # 8.8 mL/min always produced a sterile effluent with 100 % retention of the test organism, Brevundimonas diminuta.


For a thoroughly water wetted Sartopore® 2 Mini Cartridge and MidiCap®, with 0.2 m2 filtration area, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 2.5 bar|36 psi at 20 °C is:

7 mL/min.

NoteThe diffusion and Bubble Point Test results are influenced by the nature of the wetting medium. The diffusion and bubble point values listed in this validation guide are for Sartopore® 2 Mini Cartridges and Capsules wetted with water at 20 °C. It should be noted, that a variation of the test conditions such as temperature, wetting liquid or type of gas may require a different integrity test limit related to those mentioned above.



Lot Number

Diffusion [mL/min]

9330

41

Sterile

0

2

4

6

8

10

12

Safety margin

9330

4193

2941

9329

4193

3041

9329

4193

2941

9279

4193

2941

9326

4193

2941

9294

4192

9441

9294

4192

7941

9294

4193

0541

9305

4193

0541

9326

4193

2641

9279

4193

2641

9279

4192

7941

9305

4193

2941

9305

4193

0541

9294

41

Non Sterile


| 47



930641 2.7 2.65 ƒ 1010 sterile

930641 2.7 2.65 ƒ 1010 sterile

932541 3.0 6.69 ƒ 1010 sterile

930641 3.1 2.65 ƒ 1010 sterile

932541 3.1 2.59 ƒ 1010 sterile

930641 3.2 2.65 ƒ 1010 sterile

932541 3.3 6.69 ƒ 1010 sterile

932541 3.3 6.69 ƒ 1010 sterile

932541 3.8 2.59 ƒ 1010 sterile

932441 3.9 6.69 ƒ 1010 sterile

932441 4.1 6.69 ƒ 1010 sterile

930641 4.3 2.65 ƒ 1010 sterile

932541 4.4 2.59 ƒ 1010 sterile

932441 4.5 2.59 ƒ 1010 sterile

930641 4.6 2.65 ƒ 1010 sterile

932541 4.7 6.69 ƒ 1010 sterile

930641 5.1 2.65 ƒ 1010 sterile

930641 5.1 2.65 ƒ 1010 sterile

930641 5.1 2.65 ƒ 1010 sterile

932441 5.3 2.59 ƒ 1010 sterile

932541 5.3 2.59 ƒ 1010 sterile

932441 5.4 6.69 ƒ 1010 sterile

932541 5.4 6.69 ƒ 1010 sterile

932441 5.8 2.59 ƒ 1010 sterile

930641 6.1 2.65 ƒ 1010 sterile

932441 6.1 6.69 ƒ 1010 sterile

932541 6.5 2.59 ƒ 1010 sterile

932441 6.7 6.69 ƒ 1010 non sterile

932441 7.3 2.59 ƒ 1010 non sterile

932441 11.4 2.59 ƒ 1010 non sterile

48 |



For a thoroughly water wetted Sartopore® 2 0.2 µm Mini Cartridge and MidiCap, with 0.1 m2 filtration area, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 2.5 bar|36 psi at 20 °C is:

5 mL/min.


Lot Number

Diffusion [mL/min]

9306

41

Non Sterile

Sterile

0

2

4

6

8

10

12

Safety margin

9306

4193

2541

9306

4193

2541

9306

4193

2541

9325

4193

2541

9324

4193

2441

9306

4193

2541

9324

4193

0641

9325

4193

0641

9306

4193

0641

9324

4193

2541

9324

4193

2541

9324

4193

0641

9324

4193

2541

9324

4193

2441

9324

41


| 49



932141 0.7 1.33 ƒ 1010 sterile

930741 1.3 1.33 ƒ 1010 sterile

932041 1.4 1.33 ƒ 1010 sterile

930741 1.5 1.33 ƒ 1010 sterile

930741 1.5 1.33 ƒ 1010 sterile

930741 1.6 1.33 ƒ 1010 sterile

932041 1.6 1.33 ƒ 1010 sterile

930741 1.7 1.33 ƒ 1010 sterile

928141 1.7 1.10 ƒ 1010 sterile

930741 1.8 1.33 ƒ 1010 sterile

930741 1.8 1.33 ƒ 1010 sterile

928141 1.9 1.10 ƒ 1010 sterile

932041 2.0 3.44 ƒ 1010 sterile

932041 2.1 3.44 ƒ 1010 sterile

930741 2.3 1.33 ƒ 1010 sterile

932041 2.6 3.44 ƒ 1010 sterile

932041 2.8 3.44 ƒ 1010 sterile

932041 3.0 1.33 ƒ 1010 sterile

930741 3.2 1.33 ƒ 1010 sterile

932141 3.5 1.33 ƒ 1010 sterile

928141 4.2 1.10 ƒ 1010 sterile

928141 4.2 1.10 ƒ 1010 sterile

932141 4.6 1.33 ƒ 1010 sterile

928141 4.9 1.10 ƒ 1010 sterile

932141 4.9 3.44 ƒ 1010 sterile

932141 5.2 3.44 ƒ 1010 sterile

932141 5.4 3.44 ƒ 1010 non sterile

932141 5.6 3.44 ƒ 1010 non sterile

932141 5.9 1.33 ƒ 1010 non sterile

932141 8.6 3.44 ƒ 1010 non sterile

50 |



For a thoroughly water wetted Sartopore® 2 0.2 µm Mini Cartridge and MidiCap, with 0.05 m2 filtration area, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 2.5 bar|36 psi at 20 °C is:

4 mL/min.


Lot Number

Diffusion [mL/min]

9321

41

Non Sterile

Sterile

0

2

4

6

8

10

Safety margin

9307

4193

2041

9307

4193

0741

9307

4193

2041

9307

4192

8141

9307

4193

0741

9281

4193

2041

9320

4193

0741

9320

4193

2041

9320

4193

0741

9321

4192

8141

9281

4193

2141

9281

4193

2141

9321

4193

2141

9321

4193

2141

9321

41


| 51

5.4.6 Capsules 300 (0.03 m2|0.32 ft2) 0.2 µm

Lot Number Diffusion [mL/min] Filtrate Quality

920881 0.5 sterile

919281 0.6 sterile

920981 0.6 sterile

920881 0.6 sterile

920881 0.7 sterile

919281 0.7 sterile

920981 0.7 sterile

920981 0.7 sterile

920881 0.8 sterile

920981 0.8 sterile

920881 0.8 sterile

920881 0.9 sterile

920981 1.0 sterile

919281 1.1 sterile

920981 1.2 sterile

920881 1.2 sterile

919281 1.3 sterile

919281 1.4 sterile

920981 1.6 sterile

920981 1.7 sterile

920881 1.7 sterile

919281 1.8 sterile

920881 1.8 sterile

919281 2.0 sterile

919281 2.0 sterile

920881 2.1 sterile

920981 2.5 sterile

920981 2.6 sterile

920881 2.7 non sterile



52 |

ConclusionThe data shows that Sartopore® 2 Capsules 300 that have diffusion values # 2.6 mL/min always produced a sterile effluent with 100 % retention of the test organism, Brevundimonas diminuta.


For a thoroughly water wetted Sartopore® 2 Capsule 300 with 0.03 m2 filtration area, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 2.5 bar|36 psi at 20 °C is:

2 mL/min.


Lot Number

Diffusion [mL/min]

9208

81

Non Sterile

Sterile

0

1

2

3

4

5

6

Safety margin

9192

8192

0981

9208

8192

0881

9192

8192

0981

9209

8192

0881

9209

8192

0881

9208

8192

0981

9192

8192

0981

9208

8191

9281

9192

8192

0981

9209

8192

0881

9192

8192

0881

9192

8191

9281

9208

8192

0981

9209

8192

0881

9192

81

9192

81

Capsules 300 (0.03 m2|0.32 ft2) 0.2 µm

| 53

5.4.7 Capsules 150 (0.015 m2|0.16 ft2) 0.2 µm


011581 0.3 sterile

011681 0.3 sterile

011781 0.3 sterile

011781 0.4 sterile

011681 0.4 sterile

011581 0.5 sterile

011581 0.5 sterile

011781 0.5 sterile

011581 0.6 sterile

011681 0.6 sterile

011681 0.7 sterile

011781 0.7 sterile

011581 0.7 sterile

011781 0.8 sterile

011681 0.8 sterile

011681 0.8 sterile

011781 0.8 sterile

011581 0.9 sterile

011781 0.9 sterile

011681 1.0 sterile

011581 1.0 sterile

011781 1.0 sterile

011781 1.2 sterile

011681 1.3 sterile

011581 1.4 sterile

011781 1.5 sterile

011681 1.6 sterile

011781 1.6 sterile




54 |

ConclusionThe data shows that Sartopore® 2 Capsules 150 that have diffusion values # 1.6 mL/min always produced a sterile effluent with 100 % retention of the test organism, Brevundimonas diminuta.



1 mL/min.


Lot Number

Diffusion [mL/min]

0115

81

Non Sterile

Sterile

0

1

2

3

Safety margin

0116

8101

1781

0117

8101

1681

0115

8101

1581

0117

8101

1581

0116

8101

1681

0117

8101

1581

0117

8101

1681

0116

8101

1781

0115

8101

1781

0116

8101

1581

0117

8101

1781

0116

8101

1581

0117

8101

1681

0117

81

0117

81

0115

8101

1681

Capsules 150 (0.015 m2|0.16 ft2) 0.2 µm

| 55



Note

Since most of the Filter Cartridges 10"|250 mm tested during the validation studies had low diffusion values and produced a sterile filtrate, the following data is a sampling from all filters tested during the validation testing indicating results near the diffusion|sterile filtrate limits.


009341 18.2 1.3 ƒ 1011 sterile

009241 18.3 1.3 ƒ 1011 sterile

009241 18.4 6.6 ƒ 1010 sterile

009141 18.6 1.3 ƒ 1011 sterile

009341 18.7 8.6 ƒ 1010 sterile

009141 18.9 1.3 ƒ 1011 sterile

009241 19.1 8.6 ƒ 1010 sterile

009341 19.3 1.3 ƒ 1011 sterile

009241 19.4 1.3 ƒ 1011 sterile

009341 19.5 6.6 ƒ 1010 sterile

009341 19.9 6.6 ƒ 1010 sterile

009341 20.3 6.6 ƒ 1010 sterile

009141 20.5 1.3 ƒ 1011 sterile

009241 20.8 1.3 ƒ 1011 sterile

009241 20.9 9.2 ƒ 1010 sterile

009341 21.3 9.2 ƒ 1010 sterile

009241 21.7 1.3 ƒ 1011 sterile

009141 22.0 6.6 ƒ 1010 sterile

009241 22.3 9.2 ƒ 1010 sterile

009341 22.6 1.3 ƒ 1011 sterile

009341 22.8 1.3 ƒ 1011 sterile

009141 23.1 9.2 ƒ 1010 sterile

009241 23.3 8.6 ƒ 1010 sterile

009341 23.5 8.6 ƒ 1010 sterile

009341 23.6 9.2 ƒ 1010 sterile

009141 23.7 1.3 ƒ 1011 sterile

009241 23.8 6.6 ƒ 1010 sterile

009241 24.0 9.2 ƒ 1010 sterile

009341 24.2 8.6 ƒ 1010 sterile

009141 25.5 1.3 ƒ 1011 sterile

009241 26.7 1.3 ƒ 1011 sterile

009341 28.1 9.2 ƒ 1010 sterile

009141 29.2 8.6 ƒ 1010 sterile

009241 30.1 1.3 ƒ 1011 non sterile

009141 35.2 6.6 ƒ 1010 non sterile

009341 38.6 9.2 ƒ 1010 non sterile

*CFU = Colony Forming Units

56 |

ConclusionThe data shows that Filter Cartridges that have diffusion values < 29.2 mL/min always produced a sterile filtrate with 100 % retention of the test organism, Brevundimonas diminuta. In order to have a high degree of safety when evaluating the test results, and considering that other filter integrity test units or other test methods may be used, a safety margin of 5.2 mL/min has been defined. For a thoroughly water wetted 10" Sartopore® 2 0.1 µm Filter Cartridge, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 4.0 bar|58 psi at 20 °C is:

24 mL/min.

Sartopore® 2 0.1 µm Filter Cartridges and MaxiCaps® of various lengths have the following maximum allowable diffusion values at a test pressure of 4.0 bar|58 psi at 20 °C:




NoteThe Diffusion Test results are influ-enced by the nature of the wetting medium. The Diffusion Test values listed in this validation guide are for Sartopore® 2 filter wetted with water at 20 °C. It should be noted, that a variation of the test condi-tions such as temperature, wetting liquid or type of gas may require a different integrity test limit related to those mentioned above.


Lot Number

Diffusion [mL/min]

0093

41

Sterile

0

5

10

15

20

25

30

35

40

Safety margin

0092

4100

9241

0091

4100

9341

0091

4100

9241

0093

4100

9241

0093

4100

9341

0093

4100

9141

0092

4100

9241

0093

4100

9241

0091

4100

9241

0093

4100

9341

0091

4100

9241

0093

4100

9341

0091

4100

9241

0092

4100

9341

0091

4100

9241

0093

41

0092

4100

9141

0093

41

0091

41

Non Sterile


| 57

5.5.2 Standard Cartridges (5"|157 mm) 0.1 µm



313742 8.1 1.2 ƒ 1010 sterile

313742 8.2 1.2 ƒ 1010 sterile

314442 8.9 2.2 ƒ 1010 sterile

313742 9.2 1.2 ƒ 1010 sterile

314442 9.4 2.2 ƒ 1010 sterile

313742 9.8 1.2 ƒ 1010 sterile

313742 10.3 1.2 ƒ 1010 sterile

314442 10.7 2.2 ƒ 1010 sterile

314442 11.2 2.2 ƒ 1010 sterile

313742 11.8 4.2 ƒ 1010 sterile

314442 12.2 5.0 ƒ 1010 sterile

314442 12.0 5.0 ƒ 1010 sterile

313742 12.8 4.2 ƒ 1010 sterile

313742 13.1 1.2 ƒ 1010 sterile

314642 13.3 3.4 ƒ 1010 sterile

314642 13.5 3.4 ƒ 1010 sterile

314442 13.8 5.0 ƒ 1010 sterile

314442 13.9 5.0 ƒ 1010 sterile

314642 14.1 3.4 ƒ 1010 sterile

314642 14.2 3.4 ƒ 1010 sterile

313742 14.4 4.2 ƒ 1010 sterile

314442 14.6 5.0 ƒ 1010 sterile

314642 14.9 3.4 ƒ 1010 sterile

314642 15.3 3.4 ƒ 1010 sterile

314442 16.0 5.0 ƒ 1010 sterile

314442 16.1 5.0 ƒ 1010 non sterile

314642 16.8 3.4 ƒ 1010 non sterile

314642 17.2 3.4 ƒ 1010 non sterile

58 |

ConclusionThe data shows that Filter Cartridges that have diffusion values < 16.0 mL/min always produced a sterile filtrate with 100 % retention of the test organism, Brevundimonas diminuta.

In order to have a high degree of safety when evaluating the test results, and considering that other filter integrity test units or other test methods may be used, a safety margin of 2 mL/min has been defined. For a thoroughly water wetted 5" Sartopore® 2 0.1 µm Filter Cartridge, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 4.0 bar|58 psi at 20 °C is:

14 mL/min.

Lot Number

Diffusion [mL/min]

3137

42

Sterile

0

2

4

6

8

10

12

14

16

18

20

Safety margin

3137

4231

4442

3137

4231

4442

3137

4231

3742

3144

4231

4442

3137

4231

4442

3144

4231

3742

3137

4231

4642

3146

4231

4442

3144

4231

4642

3146

4231

3742

3144

4231

4642

3146

4231

4442

3144

4231

4642

3146

42

Non Sterile

Standard Cartridges (5"|157 mm) 0.1 µm

| 59



011541 5.4 2.59 ƒ 1010 sterile

011741 5.5 2.1 ƒ 1010 sterile

011441 5.7 2.1 ƒ 1010 sterile

011441 5.8 2.1 ƒ 1010 sterile

011741 6.1 4.8 ƒ 1010 sterile

011541 6.2 4.9 ƒ 1010 sterile

011441 6.4 4.8 ƒ 1010 sterile

011541 6.7 4.9 ƒ 1010 sterile

011741 7.0 2.59 ƒ 1010 sterile

011541 7.2 4.8 ƒ 1010 sterile

011441 7.6 4.8 ƒ 1010 sterile

011741 7.8 4.9 ƒ 1010 sterile

011741 8.1 4.9 ƒ 1010 sterile

011541 8.3 4.9 ƒ 1010 sterile

011441 8.6 4.8 ƒ 1010 sterile

011441 8.8 4.8 ƒ 1010 sterile

011741 9.0 2.1 ƒ 1010 sterile

011541 9.1 2.59 ƒ 1010 sterile

011741 9.3 4.9 ƒ 1010 sterile

011741 9.5 4.9 ƒ 1010 sterile

011541 9.8 4.9 ƒ 1010 sterile

011441 10.2 4.9 ƒ 1010 sterile

011441 10.5 4.8 ƒ 1010 sterile

011441 10.8 4.8 ƒ 1010 sterile

011541 11.3 4.8 ƒ 1010 sterile

011741 11.5 4.8 ƒ 1010 sterile

011741 11.8 4.8 ƒ 1010 sterile

011741 12.1 4.8 ƒ 1010 non sterile

011541 13.6 4.9 ƒ 1010 non sterile

011441 16.1 4.8 ƒ 1010 non sterile

60 |

ConclusionThe data shows that Mini Cartridges and MidiCaps® that have diffusion values # 11.8 mL/min always produced a sterile filtrate with 100 % retention of the test organism, Brevundimonas diminuta.


For a thoroughly water wetted Sartopore® 2 0.1 µm Mini Cartridge with 0.2 m2 filtration area, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 4.0 bar|58 psi at 20 °C is:

9 mL/min.

NoteThe Diffusion Test results are influ-enced by the nature of the wetting medium. The diffusion values listed in this validation guide are for Sartopore® 2 0.1 µm Mini Cartridges and MidiCaps® wetted with water at 20 °C. It should be noted, that a variation of the test conditions such as temperature, wetting liquid or type of gas may require a different integrity test limit related to those mentioned above.


Lot Number

Diffusion [mL/min]

0115

41

Non Sterile

Sterile

0

2

4

6

8

10

12

14

16

18

Safety margin

0117

4101

1441

0114

4101

1741

0115

4101

1441

0115

4101

1741

0115

4101

1441

0117

4101

1741

0115

4101

1441

0114

4101

1741

0115

4101

1741

0117

4101

1541

0114

4101

1441

0114

4101

1541

0117

4101

1741

0117

4101

1541

0114

41


| 61



011641 3.8 2.5 ƒ 1010 sterile

011641 4.0 2.5 ƒ 1010 sterile

011941 4.1 1.05 ƒ 1010 sterile

011841 4.3 3.43 ƒ 1010 sterile

011841 4.5 3.43 ƒ 1010 sterile

011941 4.8 1.05 ƒ 1010 sterile

011941 5.0 1.05 ƒ 1010 sterile

011641 5.2 2.5 ƒ 1010 sterile

011941 5.4 1.05 ƒ 1010 sterile

011841 5.5 3.43 ƒ 1010 sterile

011841 5.7 3.43 ƒ 1010 sterile

011641 5.8 2.5 ƒ 1010 sterile

011641 5.9 2.5 ƒ 1010 sterile

011841 6.0 3.43 ƒ 1010 sterile

011941 6.0 1.05 ƒ 1010 sterile

011841 6.0 3.43 ƒ 1010 sterile

011641 6.1 2.5 ƒ 1010 sterile

011641 6.1 2.5 ƒ 1010 sterile

011841 6.2 3.43 ƒ 1010 sterile

011941 6.3 1.05 ƒ 1010 sterile

011841 6.6 3.43 ƒ 1010 sterile

011841 6.8 3.43 ƒ 1010 sterile

011641 7.0 2.5 ƒ 1010 sterile

011941 7.2 1.05 ƒ 1010 sterile

011941 7.4 1.05 ƒ 1010 sterile

011641 7.6 2.5 ƒ 1010 sterile

011841 7.7 3.43 ƒ 1010 sterile

011941 8.5 3.43 ƒ 1010 non sterile

011641 9.3 1.05 ƒ 1010 non sterile

011941 10.2 2.5 ƒ 1010 non sterile

62 |



For a thoroughly water wetted Sartopore® 2 0.1 µm Mini Cartridge and MidiCap® with 0.1 m2 filtration area, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 4.0 bar|58 psi at 20 °C is:

6 mL/min.

Lot Number

Diffusion [mL/min]

0116

41

Non Sterile

Sterile

0

2

4

6

8

10

12

Safety margin

0116

4101

1941

0118

4101

1841

0119

4101

1941

0116

4101

1941

0118

4101

1841

0116

4101

1641

0118

4101

1941

0118

4101

1641

0116

4101

1841

0119

4101

1841

0118

4101

1641

0119

4101

1941

0116

4101

1841

0118

4101

1641

0119

41


| 63



012041 1.8 1.67 ƒ 1010 sterile

012041 2.0 1.67 ƒ 1010 sterile

012341 2.1 1.0 ƒ 1010 sterile

012141 2.2 1.67 ƒ 1010 sterile

012141 2.3 1.67 ƒ 1010 sterile

012041 2.4 1.67 ƒ 1010 sterile

012141 2.7 1.67 ƒ 1010 sterile

012141 2.9 1.67 ƒ 1010 sterile

012341 3.1 1.0 ƒ 1010 sterile

012141 3.3 1.67 ƒ 1010 sterile

012041 3.5 1.67 ƒ 1010 sterile

012041 3.7 1.67 ƒ 1010 sterile

012341 3.8 1.0 ƒ 1010 sterile

012141 3.9 1.67 ƒ 1010 sterile

012141 4.0 1.67 ƒ 1010 sterile

012341 4.0 1.0 ƒ 1010 sterile

012041 4.0 1.67 ƒ 1010 sterile

012341 4.1 1.0 ƒ 1010 sterile

012141 4.3 1.67 ƒ 1010 sterile

012141 4.3 1.67 ƒ 1010 sterile

012041 4.5 1.67 ƒ 1010 sterile

012341 4.7 1.0 ƒ 1010 sterile

012341 4.8 1.0 ƒ 1010 sterile

012141 4.9 1.67 ƒ 1010 sterile

012041 5.0 1.67 ƒ 1010 sterile

012141 5.1 1.67 ƒ 1010 sterile

012341 5.2 1.0 ƒ 1010 sterile

012041 5.8 1.67 ƒ 1010 non sterile

012141 6.5 1.67 ƒ 1010 non sterile

012341 7.1 1.0 ƒ 1010 non sterile

64 |



For a thoroughly water wetted Sartopore® 2 0.1 µm Mini Cartridge and MidiCaps® with 0.05 m2 filtration area, (keeping in mind this safety factor), the maximum allowable diffusion value at a test pressure of 4.0 bar|58 psi at 20 °C is:

4 mL/min.

Lot Number

Diffusion [mL/min]

0120

41

Non Sterile

Sterile

0

1

2

3

4

5

6

7

8

Safety margin

0120

4101

2341

0121

4101

2141

0120

4101

2141

0121

4101

2341

0121

4101

2041

0120

4101

2341

0121

4101

2141

0123

4101

2041

0123

4101

2141

0121

4101

2041

0123

4101

2341

0121

4101

2041

0121

4101

2341

0120

4101

2141

0123

41


| 65

5.5.6 Capsules 300 (0.03 m2|0.32 ft2) 0.1 µm


920881 0.5 sterile

919281 0.6 sterile

920981 0.6 sterile

920881 0.6 sterile

920881 0.7 sterile

919281 0.7 sterile

920981 0.7 sterile

920981 0.7 sterile

920881 0.8 sterile

920981 0.8 sterile

920881 0.8 sterile

920881 0.9 sterile

920981 1.0 sterile

919281 1.1 sterile

920981 1.2 sterile

920881 1.2 sterile

919281 1.3 sterile

919281 1.4 sterile

920981 1.6 sterile

920981 1.7 sterile

920881 1.7 sterile

919281 1.8 sterile

920881 1.8 sterile

919281 2.0 sterile

919281 2.0 sterile

920881 2.1 sterile

920981 2.5 sterile

920981 2.6 sterile




66 |

ConclusionThe data shows that Sartopore® 2 Capsules 300 that have diffusion values # 2.6 mL/min always produced a sterile filtrate with 100 % retention of the test organism, Brevundimonas diminuta.



2 mL/min.

Lot Number

Diffusion [mL/min]

9208

81

Non Sterile

Sterile

0

1

2

3

4

Safety margin

9192

8192

0981

9208

8192

0881

9192

8192

0981

9209

8192

0881

9209

8192

0881

9208

8192

0981

9192

8192

0981

9208

8191

9281

9192

8192

0981

9209

8192

0881

9192

8192

0881

9192

8191

9281

9208

8192

0981

9209

8192

0881

9192

8191

9281

Capsules 300 (0.03 m2|0.32 ft2) 0.1 µm

| 67

5.5.7 Capsules 150 (0.015 m2|0.16 ft2) 0.1 µm


024881 0.3 sterile

024981 0.3 sterile

025081 0.4 sterile

024981 0.4 sterile

024981 0.4 sterile

024881 0.4 sterile

024881 0.4 sterile

024981 0.5 sterile

024881 0.5 sterile

025081 0.5 sterile

025081 0.6 sterile

024981 0.6 sterile

024881 0.7 sterile

024881 0.7 sterile

024981 0.8 sterile

025081 0.8 sterile

024981 0.9 sterile

024881 0.9 sterile

024981 1.0 sterile

024881 1.0 sterile

025081 1.0 sterile

024981 1.1 sterile

024981 1.2 sterile

024881 1.3 sterile

025081 1.3 sterile

024881 1.4 sterile

024981 1.5 sterile

025081 1.5 sterile




68 |

ConclusionThe data shows that Sartopore® 2 Capsules 150 that have diffusion values # 1.5 mL/min always produced a sterile filtrate with 100 % retention of the test organism, Brevundimonas diminuta.



1 mL/min.

Lot Number

Diffusion [mL/min]

0248

81

Non Sterile

Sterile

0

1

2

3

4

Safety margin

0249

8102

5081

0249

8102

4981

0248

8102

4881

0249

8102

4881

0250

8102

5081

0249

8102

4881

0248

8102

4981

0250

8102

4981

0248

8102

4981

0248

8102

5081

0249

8102

4981

0248

8102

5081

0248

8102

4981

0250

8102

4881

0250

8102

4981

Capsules 150 (0.015 m2|0.16 ft2) 0.1 µm

| 69

NoteWith the use of the Sartorius automated integrity test system, Sartocheck® 4 Plus, the upstream volume is calculated during every integrity test. For the manual determination of the integrity of a filtration system the direct measurement of diffusion or Bubble Point values is recommended.

5.6 Manual Determination of Maximal Allowable Pressure DropSlowly pressurize the filter housing containing the wetted Filter Cartridge. The test pressure which you need to use is dependent upon the pore size of the membrane to be tested. (See integrity test data table). Once the correct pressure is attained, allow for a 5 minute stabilization period and then close the pressure supply. During the 5 minute test period, the pressure drop should not exceed the permis-sible value. This maximum pressure drop is dependent upon a variety of criteria, including the upstream volume of the special filter housing at a constant temperature, and must be calculated according to the general gas equation:

PA–PE= VD·t·PO

V

PA–PE: Pressure drop in mbar after test period (t)

VD: Gas diffusion in mL/min (see table)

t: Test time (min)

PO: Atmospheric pressure (=1000 mbar|14.5 psi)

V: Volume of housing on the inlet side in mL (net volume of housing with installed cartridge + volume of the inlet line to the stop valve + volume of the gas tubing)

70 |

Examples of the maximum allowable pressure drop for Sartopore® 2 filter elements:

5.6.1 Sartopore® 2 Filter Elements with 0.45 µm Final Membrane

MaxiCaps®

Filter Max. Diffusion at 20 °C Upstream Max. allowable Pressure drop Capsule and 1.7 bar|25 psi [mL/min] Volume [mL] at 20 °C and 1.7 bar|25 psi

5441306G1 12 711 84 mbar/5 min (1.22 psi/5 min)

5441306G2 24 1256 96 mbar/5 min (1.39 psi/5 min)

5441306G3 36 1774 101 mbar/5 min (1.46 psi/5 min)

MidiCaps®


5445307H0--SS 12 530 113 mbar/5 min (1.64 psi/5 min)




Cartridges

Filter Max. Diffusion at 20 °C Upstream Max. allowable Pressure drop Housing Filter Cartridge and 1.7 bar|25 psi [mL/min] Volume [mL] at 20 °C and 1.7 bar|25 psi

340011P25TT112A Sartopore® 2 12 1750 34 mbar/5 min 10" 5442506G1 (10") (0.50 psi/5 min)



Mini Cartridges

Filter Max. Diffusion at 20 °C Upstream Max. allowable Pressure drop Housing Filter Cartridge and 1.7 bar|25 psi [mL/min] Volume [mL] at 20 °C and 1.7 bar|25 psi

330019P15TT112A Sartopore® 2 6 275 109 mbar/5 min 5441506G9 (1.58 psi/5 min) 0.2 m2|2.2 ft2



| 71

MaxiCaps®


5441307H1 18 711 126 mbar/5 min (1.83 psi/5 min)

5441307H2 36 1256 143 mbar/5 min (2.07 psi/5 min)

5441307H3 54 1774 152 mbar/5 min (2.2 psi/5 min)

Standard Cartridges

Filter 0.2 µm Max. Diffusion at 20 °C Upstream Max. allowable Pressure drop Housing Filter Cartridge and 2.5 bar|36 psi [mL/min] Volume at 20 °C and 2.5 bar|36 psi

340011P25TT112A Sartopore® 2 18 1750 mL 51 mbar/5 min 10" 5442507H1 (10") (0.73 psi/5 min)



Mini Cartridges

Filter 0.2 µm Max. Diffusion at 20 °C Upstream Max. allowable Pressure drop Housing Filter Cartridge and 2.5 bar|36 psi [mL/min] Volume [mL] at 20 °C and 2.5 bar|36 psi

330019P15TT112A Sartopore® 2 7 275 125 mbar/5 min 5441507H9 (1.81 psi/5 min) 0.2 m2|2.2 ft2



MidiCaps®


5445307H0––SS 14 530 132 mbar/5 min (1.91 psi/5 min)




5.6.2 Sartopore 2 Filter Elements with 0.2 µm Final Membrane

72 |

MaxiCaps®

Filter Max. Diffusion at 20 °C Upstream Max. allowable Pressure drop at Capsule and 4.0 bar|58 psi [mL/min] Volume [mL] 20 °C and 4.0 bar|58 psi

5441358K1 24 711 169 mbar/5 min (2.45 psi/5 min)

5441358K2 48 1256 191 mbar/5 min (2.77 psi/5 min)

5441358K3 72 1774 203 mbar/5 min (2.94 psi/5 min)

NoteWith the use of the Sartorius automated integrity test system, Sartocheck® 4 Plus, the upstream volume is calculated during every integrity test. In order to have a reliable determination of the

Standard Cartridges

Filter 0.1 µm Max. Diffusion at 20 °C Upstream Max. allowable Pressure drop Housing Filter Cartridge and 4 bar|58 psi [mL/min] Volume [mL] at 20 °C and 2.5 bar|36 psi

340011P25TT112A Sartopore® 2 24 1750 68 mbar/5 min 10" 5442558K1 (10") (0.98 psi/5 min)

340012P25TT112A Sartopore® 48 3200 75 mbar/5 min 20" 5442558K2 (20") (1.08 psi/5 min)

340013P25TT112A Sartopore® 72 4700 76 mbar/5 min 30" 5442558K3 (30") (1.10 psi/5 min)

Mini Cartridges

Filter 0.1 µm Max. Diffusion at 20 °C Upstream Max. allowable Pressure drop Housing Filter Cartridge and 4 bar|58 psi [mL/min] Volume [mL] at 20 °C and 2.5 bar|36 psi

330019P15TT112A Sartopore® 9 275 163 mbar/5 min 5441558K9 (2.36 psi/5 min) 0.2 m2|2.2 ft2




MidiCaps®

Filter Max. Diffusion at 20 °C Upstream Max. allowable Pressure drop Capsule and 4.0 bar|58 psi [mL/min] Volume [mL] 20 °C and 4.0 bar|58 psi





integrity of the filtration system without the influence of the upstream volume, it is suggested that the Diffusion Test is used. This test is not dependent on the upstream volume of the filter system.

| 73

6.1 IntroductionBacteria of the mycoplasma group are some of the smallest self-propagating microorganism. Mycoplasma, Acholeplasma as well as Spiroplasma genera belong to this group. They are characterized by the absence of a rigid cell wall and by their parasitic nature. Mycoplasma are found in a variety of polymorphic shapes as coccal cells, filaments or rosettes.

Acholeplasma laidlawii as typical example of mycoplasma can be found commonly as a contami-nation in serum and cell culture media which contain ingredients based on animal plasma.

The cell diameter of mycoplasma is usually found to be between 0.1–0.25 µm.

From filtration standpoint this small small size combined with the deformable properties of their outer shape, due to the lack of a cell wall, exceed current retention capabilities of 0.2 µm rated sterilizing grade membrane filters.

Therefore it is necessary to use membranes with a smaller pore size than 0.2 µm to achieve secure retention of mycoplasma by means of filtration.

As A. laidlawii can be grown under specific conditions to high densities this is a appropriate microorganism to be used for the determination of mycoplasma retention performance of filter elements. Therefore this species of mycoplasma was used for challenging trials of 0.1 µm Sartopore® 2 filters.

6.2 PurposeThe purpose of the performed trials was to investigate about the retention perfomance of Sartopore® 2 0.1 µm Filter Cartridges in 10" format regarding mycoplasma.

6.3 Test Methods

6.3.1 Qualification of the Monitoring FiltersSartobran® P 0.07 µm filter material was qualified for their use as monitoring filters for analyzing the filtrate of the test filters during mycoplasma challenge tests for the passage of any A. laidlawii.

These qualification tests were carried out by testing filter membranes with suspensions of A. laidlawii for evidence of toxicity, capture and retention capabilities.

Filter membrane toxicity was determined by wetting out the membranes with broth base and transferring them to petri-dishes containing mycoplasma growth medium. This medium contains triphenyl tetrazolium chloride (TTC) which promotes specifically the growth of A. laidlawii with a strong red coloration.

Membranes and controll plates were inoculated with A. laidlawii suspension and incubated for 3 days.

ResultA. laidlawii developed well on the membrane with the shiny side up. There was no evidence of toxicity with the counts on the membrane, agar + TTC and agar alone all being similar.

To demonstrate that the retention performance accounts for a broad range of possible concentration of A. laidlawii possibly found in the filtrate the filter membrane retention capability was determined for low and high numbers of A. laidlawii challenges.

To determine the retention capabil-ities for low numbers of A. laidlawii 1 L of broth base was inoculated with 0.02 mL of A. laidlawii suspension containing 100 colony forming units. To confirm the total viability count of the suspension a drop of 0.02 mL was inoculated on a agar plate.

The inoculated broth was filtered at a constant inlet pressure of 2 bar through a 47 mm diameter filter holder equipped with Sartobran® P 0.07 µm flat filters membranes.The filter membrane was placed on TTC agar after filtration and incubated parallel with the control plate.

The same test method was used to determine the retention capability of Sartobran® P 0.07 µm flat membrane using a high titer challenge with a challenge level of 1 ƒ 107 cfu/cm2 filtration area.

ResultsLow titer challenge:After 3 days of incubation 79.5 % of the mycoplasma were recovered. Due to loss of mycoplasma on the pressure vessel walls, in the tubing and through the filter membrane handling the recovery rate was excellent.

High titer challenge:After 4 days of incubation of 99.99 % of A. laidlawii were recovered on the membrane.

ConclusionSartobran® P 0.07 µm membranes show no evidence of toxicity to the development of A. laidlawii colonies and are suitable for the use as analytical filters to determine the amount of mycoplasma CFU´s.

6. Mycoplasma Retention

74 |

6.3.2 Test Set-upSartopore® 2 10" Filter Cartridges from 3 different lots where installed in individual Sartorius filter cartridge housings (Type: 340011P25TT112A) and rinsed according to instructions given in the directions for use leaflet. The filters then where autoclaved at 121 °C for 30 min for steril-ization. Afterwards the filters were tested for integrity by diffusion testing using a Sartocheck® 4 plus automatic integrity tester to assure that only integral filters were tested.

1 liter of activly growing cultures of Acholeplasma laidlawii was added in a pressure vessel to 40 liters of presterilised mycoplasma growth media.

To determine the concentration of colony forming units (mycoplasma) in the challenge solution, agar plates were inoculated with 0.02 mL amounts of a 103 dilution from this suspension to determine the concentration of colony forming units (CFU) in each suspension to be filtered.

The filtrate of the Sartopore® 2 Filter Cartridges was monitored with a Sartobran® P 0.07 µm membrane known to be mycoplasma retentive. This analytical filter was installed in 142 mm diameter stainless steel filter holders and connected to the outlet of the cartridge filter housings. The purpose of this analytical filter is to detect any passage of A. laidlawii.

After filtration the membranes were placed onto agar medium which specifically promotes the growth and development of A. laidlawii colonies accompanied by a strong red colouration after incubation at 36 °C for 7 days. This procedure facilitates colony counting without the need to use membrane staining.

The challenged filters were then intergrity tested for a second time by diffusion test after filtration to assure they maintained their physical integrity during the filtration process.

Nitrogene Pressure Supply

Pressure Vessel

Pressure Gauge

Regulating Valve

Mycoplasma Suspension

Housing with Test Filter

Nitrogene Pressure Supply

Analytical Filters

Test Set-up


| 75

6.4 Test ProcedureThe filter housings containing the filter cartridges were connected with the inlet to the outlet of the pressure vessel containing the suspension of A. laidlawii. The outlet of the filter housing was connected to a “T-piece“ with a sanitary diaphragm valve and to the filter holder containing the analytical filters. The outlet of the filter holder was connected to a tubing that leads the filtrate to a waste container.

A nitrogene pressure source was connected to the inlet of the pressure vessel and light gas pressure was applied. Then the filter housing and the membrane holder were vented. The inlet gas pressure was increased to 2 bar and maintained at constant level till the end of filtration.

After filtration the gas pressure was released from the pressure vessel and connected to the downstream valve. The diaphragm valve was opened to allow the residual rest liquid being passed through the analytical filter using low pressure. This procedure assures that the complete volume of filtrate passed across the analytical filter.

After filtration the filter holder was disassembled and the analytical filters were transferred onto the agar plate and incubated at 36 °C for 7-14 days to detect any passage of A. laidlawii.

6.6 DiscussionThe validation tests of Sartopore® 2 0.1 µm cartridges were performed by an external laboratory specialized in cultivation and handling of Mycoplasma. The retention capability of Sartopore® 2 0.1 µm Filter Cartridges for Mycoplasma was determined using Acholeplasma laidlawii as challenge organism. Identical methodologies and process para meters as described in the HIMA|ASTM regulations for qualification of sterilizing grade filter elements were applied taking into account that no specific regulations for retention qualifi-cation of 0.1 µm filters are existing.

Prior to release flat filter discs of each membrane lot used for production of Sartopore® 2 0.1 µm cartridges are tested for retention of Mycoplasma using a challenge level of 107/cm2 filtration area.

To achieve a challenge level of 107 Acholeplasma laidlawii per cm2 filtration area for qualification of Sartopore® 2 0.1 µm Filter Cartridges with 0.6 m2 filtration area a minimum concentration of 6 ƒ 1010 Mycoplasma in the challenge fluid is required. Challenge tests were performed using 3 Sartopore® 2 0.1 µm 10" Filter Cartridge from 3 different lots.

The results obtained from the validation trials indicate that the typical retention of Sartopore® 2 0.1 µm filter elements for Myco plasma can be stated as 107/cm2 filtration area.

Lot Number Diffusion Value Challenge level Detected colonies [mL/min] [CFU/cm2] [CFU]

009141* 18.7 8.3 ƒ 106 0

004881 17.9 1.9 ƒ 107 0

009341 19.5 1.3 ƒ 108 10

6.5 Test Results Sartopore® 2 (10"|250 mm) 0.1 µm

*Remark: Due to the sensitivity of the Mycoplasma it is extremely difficult to grow the Mycoplasma to the required concentration. Therefore challenge tests were also performed using Mycoplasma concentrations slightly below the required challenge concentration of 107/cm2 filtration area.

76 |76 |

7.1 Steam Sterilization of CartridgesThe materials and construction of the Sartopore® 2 Filter Cartridges allow for exposures to multiple steam sterilization cycles. Since multiple steam sterilization cycles may be required in actual practice, the influences of the thermo-mechanical stresses on the integrity of Sartopore® 2 Filter Cartridges were examined. As a result, recom-mendations and limits for multiple in-line steam sterilization are given.

Test MethodSartopore® 2 Standard Filter Cartridges and Mini Cartridges, with a pore size of 0.2 µm from a number of dif fe rent production lots, were installed into stainless steel filter housing (Sartorius Part Number 340011P25TT112A or 331019P15TT112A) and were in-line sterilized with saturated steam at 2 bar|30 psi for 30 minutes after reaching a steaming temperature of 134°C (measured at the outlet of the housing). Additionally, the differential pressure was held constant and did not exceed 0.3 bar| 4 psi during steam steril-ization. After the steam sterilization cycle, the steam pressure is allowed to drop to atmosphere (in about 3 to 5 minutes) and the system is cooled by filtration with water at a differential pressure of 0.2 to 0.3 bar (3 to 4 psi) for 5 minutes. The in-line steam cycle is then repeated. Before beginning these tests and after 25 in-line steam cycles, the integrity of the Mini Cartridges is verified through diffusion and bubble point testing, as well as the water flow rates. After 25 steam cycles, the filters are tested by the HIMA|ASTM Bacteria Challenge Test to verify that the filters could still produce a sterile effluent.

Important Note for In-line Steam SterilizationAfter the installation and wetting of the Mini Cartridges, the upstream vent valve on the filter housing, all drainage and inlet and outlet valves on the filter housing should be slightly opened and the steam inlet valve should be opened slowly to allow for a slow steam stream coming into the filter system. During the initial phase of pressure increase, the maximum differential pressure should not exceed 0.5 bar|7 psi. As soon as steam is passing through the outlet valve of the housing, the inlet and outlet valves should be manipulated so that the outlet pressure is not more than 2 bar|30 psi. Additionally, the inlet pressure should not be more than 0.2 to 0.3 bar (3 to 4 psi) above the sterilization pressure. After steam sterilization pressures have been achieved, the filters are steamed for 30 minutes under these conditions. After steaming and closing of the steam inlet valve, the upstream and downstream pressures are allowed to drop to atmospheric pressure, the drain valves are closed and the venting valve is opened. If cooling is required to be faster, the system can be rinsed with water at a differential pressure of 0.2 to 0.3 bar (3 to 4 psi). In order to assure that the filters are not chemically attached during steaming, only steam generated with pure water should be used. Water with corrosion reducing agents, which may produce hydrazine or an alkaline steam, should not be used.

In order to demonstrate that the Sartopore® 2 Cartridges have good thermal stability, three cartridges from three different manufacturing lots were tested under the following procedure:

1. The new Filter Cartridges are wetted with water.

2. The filters are then integrity tested by bubble point and diffusion tests. The flow rates were also recorded for these new filters.

3. The filters are in-line steam sterilized.

4. After the first in-line steam sterilization cycle, the filters are integrity tested by bubble point and diffusion tests. The flow rates were also recorded.

5. The filters are then steam sterilized 25 times.

6. After the 25 cycles, the filters are integrity tested by bubble point and diffusion tests. The flow rates were also recorded.

7. The Cartridges are then Bacteria Challenge tested to determine if the steam sterilization cycles had any effect on the bacteria retention properties of the Sartopore® 2 Filter Cartridges.

7. Thermal Stability

| 77 | 77

7.1.1 Effects on Water Flow Rates

Test ProcedureSix Sartopore® 2 Standard Cartridges and six Sartopore® 2 Mini Cartridges from three different lot numbers were installed and wetted in standard filter housings. The flow rate was measured at a differential pressure of 0.5 bar|7 psi. The following table contains the average values for the six Mini Cartridges tested. Flow rate values have been standardized at 20 °C.

Lot Number Flow Rate Prior to Flow Rate Flow Rate Steaming after 1 Steam Cycle after 25 Steam Cycles [L/min] [L/min] [L/min]

920641 50.1 46 42

920741 51 41 40

920841 45 43 41

Flow Rate [l/min]

Prior to Steaming

After 1 Steam Cycle

After 25 Steam Cycles

Lot Number920641 920741 920841

05

1015202530354045505560

Flow Rate [L/min]

7.1.1.1 Standard Cartridges 0.2 µm

78 |78 |

Lot Number Flow Rate Prior to Flow Rate Flow Rate Steaming after 1 Steam Cycle after 25 Steam Cycles [L/min] [L/min] [L/min]

932941 13.6 13.0 12.8

932641 16.0 14.9 14.8

930541 14.8 14.5 13.6

Flow Rate [l/min]

Prior to Steaming

After 1 Steam Cycle


Lot Number932941 932641 930541

0

2

4

6

8

10

12

14

16

18

Flow Rate [L/min]

7.1.1.2 Mini Cartridges 0.2 µm

| 79 | 79

7.1.2 Effects on Diffusion Values

Test ProcedureSix Sartopore® 2 Standard Filter Cartridges and six Sartopore® 2 Mini Cartridges from three different production lots were wetted in standard filter housings. A Diffusion Test utilizing the following parameters was conducted utilizing an automated integrity test system, the Sartocheck® 4 Plus:

Test Pressure: 2.5 bar|36 psiStabilization Time: 5 minutes Test Time: 5 minutes

The following results are the averages for the six Mini Cartridges tested.

Lot Number Diffusion Prior Diffusion Diffusion to Steaming after 1 Steam Cycle after 25 Steam Cycles [mL/min] [mL/min] [mL/min]

920641 16.4 14.2 15.5

920741 15.2 14.1 14.9

920841 14.7 11.4 12.7

Diffusion

Prior to Steaming

After 1 Steam Cycle


Lot Number920641 920741 920841

0

5

10

15

20

Diffusion [mL/min]


80 |

Lot Number Diffusion Prior Diffusion Diffusion to Steaming after 1 Steam Cycle after 25 Steam Cycles [mL/min] [mL/min] [mL/min]

932941 3.9 3.6 3.4

932641 4.9 3.7 3.8

930541 4.0 3.4 3.1

Diffusion [l/min]

Prior to Steaming

After 1 Steam Cycle


Lot Number932941 932641 930541

0

1

2

3

4

5

6

Diffusion [mL/min]


| 81

7.1.3 Effects on Bubble Point Values

Test ProcedureAfter diffusion testing, the same Sartopore® 2 Standard Filter Cartridges and Mini Cartridges are then tested by the bubble point test, utilizing the Sartocheck® 4 Plus automated integrity test system. The following results are the averages for the elements tested.

Bubble Point

Prior to Steaming

After 1 Steam Cycle


Lot Number920641 920741 920841

0

1

2

3

4

5

Bubble Point [bar]


Lot Number Bubble Point Bubble Point Bubble Point prior to Steaming after 1 Steam Cycle after 25 Steam Cycles [bar|psi] [bar|psi] [bar|psi]

920641 3.7|54 3.9|57 4.0|58

920741 3.8|55 3.9|57 3.9|57

920841 3.9|57 4.1|59 4.2|61

82 |

Bubble Point

Prior to Steaming

After 1 Steam Cycle


Lot Number932941 932641 930541

0

1

2

3

4

5

Bubble Point [bar]


Lot Number Bubble Point Bubble Point Bubble Point prior to Steaming after 1 Steam Cycle after 25 Steam Cycles [bar|psi] [bar|psi] [bar|psi]

932941 3.8|55 3.9|57 4.0|58

932641 4.0|58 4.1|59 4.1|59

930541 4.1|59 4.2|61 4.2|61

| 83

Lot Number Bioburden Bacteria Challenge Test [CFU] after 25 Steam Sterilization Cycles

920641 2.6 ƒ 1011 Sterile Filtrate



Lot Number Bioburden Bacteria Challenge Test [CFU] after 25 Steam Sterilization Cycles




7.1.4 Bacteria Challenge Test Values

Test ProcedureWith the Sartopore® 2 Standard Filter Cartridges and Mini Cartridges previously mentioned, a Bacterial Challenge Test was performed. This test was conducted according to the HIMA – Document Number 3, Vol. 4, April 1982, ”The Micro-biological Evaluation of Filters for Sterilizing Liquids“|ASTM F838-05 Guideline. The following bacteria concentrations are averages for the elements tested.

ConclusionThe results indicate that the integrity and bacterial retentive properties of the Sartopore® 2 Standard Filter Cartridges and Mini Cartridges are not effected by 25 In-Line Steam Sterilization Cycles at 134 °C for 30 minutes (12.5 hours total).

NoteThe service life of the filter elements is determined by process conditions, for instance the particle load of the solution being filtered. The service life can also be influ-enced by the steaming conditions. Different qualities of steam and steam process conditions may lead to variations in the service life of the filters when filters are in-line steam sterilized repeatedly. Additional service lifecan be affectedby product residues which are not removed completely from the membrane by flushing.



84 |

7.2 Autoclaving of MaxiCaps®, MidiCaps® and CapsulesThe materials and construction of the Sartopore® 2 MaxiCaps®, MidiCaps® and Capsules allow for exposures to multiple autoclaving cycles. Since multiple autoclaving cycles may be required in actual practice, the influences of the thermo-mechanical stresses on the integrity of Sartopore® 2 MaxiCaps®, MidiCaps® and Capsules were examined. As a result, recom-mendations and limits for multiple autoclaving are given.

Test MethodSartopore® 2 MaxiCaps®, MidiCaps® and Capsules, with a pore size of 0.2 µm from a number of dif fe rent production lots, were installed and were autoclaved at 2 bar|30 psi and a temperature of 134 °C for 30 minutes. Before beginning these tests and after 25 autoclaving cycles, the integrity of the MaxiCaps®, MidiCaps® and Capsules is verified through diffusion and bubble point testing, as well as the water flow rates.

In order to demonstrate that the Sartopore® 2 MaxiCaps®, MidiCaps® and Capsules have good thermal stability, multiple Capsules from three different manufacturing lots were tested under the following procedure:

1. The new MaxiCaps®, MidiCaps® and Capsules are wetted with water.

2. The filters are then integrity tested by bubble point and diffusion test. The flow rates were also recorded for these MaxiCaps®, MidiCaps® and Capsules.

3. The MaxiCaps®, MidiCaps® and Capsules are then autoclaved sterilized 25 times.

4. After the 25 cycles, the MaxiCaps®, MidiCaps® and Capsules are integrity tested by bubble point and diffusion tests. The flow rates were also recorded.

5. The MaxiCaps®, MidiCaps® and Capsules are then Bacteria Challenge tested to determine if the autoclaving cycles had any effect on the bacteria retention properties of the Sartopore® 2 MaxiCaps®, MidiCaps® and Capsules.

| 85

Lot Number Flow Rate prior to Flow Rate Autoclaving after 25 Autoclaving Cycle [L/min] [L/min]

008881 39.0 40.4

008981 41.5 40.6

010181 36.0 37.0

Lot Number008881 008981 010181

0

10

20

30

40

50

Flow Rate [L/min]

Flow Rate [L/min]

Prior to Autoclaving

After 25 Autoclaving Cycles

7.2.1 Effects on Water Flow Rates

Test ProcedureSartopore® 2 MaxiCaps® and MidiCaps® each from three different lot numbers were installed and wetted. The flow rate was measured at a differential pressure of 0.5 bar|7 psi. The following table contains the average values for the Capsules tested.Flow rate values have been standardized at 20 °C.

7.2.1.1 MaxiCaps® 0.2 µm

86 |

Lot Number Flow Rate prior to Flow Rate Flow Rate Autoclaving after 1 Autoclaving Cycle after 25 Autoclaving Cycles [L/min] [L/min] [L/min]

402243 17.1 16.1 17.9

402343 17.5 16.3 17.6

414243 16.9 16.1 17.2

Flow Rate [L/min]


After 1 Autoclaving Cycle


Lot Number402243 402343 414243

0

5

10

15

20

Flow Rate [L/min]

7.2.1.2 MidiCaps® 0.2 µm

| 87

7.2.2 Effects on Diffusion Values

Test ProcedureSartopore® 2 MaxiCaps® and MidiCaps® each from three different production lots were wetted. A Diffusion Test utilizing the following parameters was conducted utilizing an automated integrity test system, the Sartocheck® 4:

Test Pressure: 2.5 bar|36 psiStabilization Time: 5 minutesTest Time: 5 minutes

The following results are the averages for the Capsules tested.


Lot Number Flow Rate prior to Flow Rate Autoclaving after 25 Autoclaving Cycles [L/min] [L/min]

008881 13.8 12.2

008981 13.8 11.5

010181 12.4 11.4

Lot Number008881 008981 010181

0

3

6

9

12

15

Diffusion [mL/min]

Diffusion



88 |

Lot Number Diffusion prior Diffusion Diffusion to Autoclaving after 1 Autoclaving Cycle after 25 Autoclaving Cycles [mL/min] [mL/min] [mL/min]

402243 4.0 4.3 3.8

402343 4.1 3.4 4.0

414243 4.6 4.1 4.3

Diffusion




Lot Number402243 402343 414243

0

1

2

3

4

5

Diffusion [mL/min]


| 89

7.2.3 Effects on Bubble Point Values

Test ProcedureAfter diffusion testing, the same Sartopore® 2 MaxiCaps® and MidiCaps® are then tested by the Bubble Point Test, utilizing the Sartocheck® 4 automated integrity test system. The following results are then averages for the elements tested.


Lot Number Bubble Point Bubble Point prior to Autoclaving after 25 Autoclaving Cycles [bar|psi] [bar|psi]

008881 3.9|57 4.0|58

008981 4.0|58 4.2|60.9

010181 4.0|58 4.2|60.9

Lot Number008881 008981 010181

0

1

2

3

4

5

Bubble Point [bar]

Bubble Point



90 |

Bubble Point




Lot Number402243 402343 414243

0

1

2

3

4

5

Bubble Point [bar]


Lot Number Bubble Point Bubble Point Bubble Point prior to Autoclaving after 1 Autoclaving Cycle after 25 Autoclaving Cycles [bar|psi] [bar|psi] [bar|psi]

402243 4.1|59.4 4.0|58 4.4| 63.8

402343 4.1|59.4 4.3|62.3 4.4| 63.8

414243 4.2|60.9 4.3|62.3 4.5| 65.2

| 91

7.2.4 Bacteria Challenge Test Values

Test ProcedureWith the Sartopore® 2 MaxiCaps® and MidiCaps® previously mentioned, a Bacterial Challenge Test was performed. This test was conducted according to the HIMA-Document Number 3, Vol. 4, April 1982 ”The Microbiological Evaluation of Filters for Sterilizing Liquids”|ASTM F838-05 Guideline. The following bacteria concentra-tions are averages for the elements tested.


Lot Number Bioburden Bacteria Challenge Test [CFU] after 25 Steam Autoclaving Cycles




Lot Number Bioburden Bacteria Challenge Test [CFU] after 25 Steam Autoclaving Cycles





ConclusionThe results indicate that the integrity and bacterial retentive properties of the Sartopore® 2 MaxiCaps® and MidiCaps® are not effected by 25 autoclaving cycles at 134 °C for 30 minutes (12.5 hours total).

NoteThe service life of the filter elements is determined by process conditions, for instance the particle load of the solution being filtered. The service life can also be influ-enced by the steaming conditions. Different qualities of steam and steam process conditions may lead to variations in the service life of the filters when filters are in-line steam sterilized repeatedly. Additional service life can affected by product residues which are not removed completely from the membrane by flushing.

92 |

8. Testing According to USP

Test PurposeThe tests for extractable substances and particle release of Sartopore® 2 filter elements are performed in dynamic extraction mode. This methodology provides the best representative of actual filtration applications determining levels of extractable substances and particles present in varying filtrate volumes. The samples for all tests are taken after 1, 2.5, 5 and 10 liters flush volume for Standard Filter Cartridges and MaxiCaps® and after 0.5, 2 and 5 liters flush volume for Mini Cartridges.

According to the specifications given in section ”Sterile Water for Injection” of the current USP, filtrate samples of Sartopore® 2 filter elements are analyzed for Particulate Matter, Oxidizable Substances, pH and conductivity, Ammonia, Sulfate and Chloride. The tests are performed according to the descriptions given in the current USP. The test results obtained are compared to the relevant USP specifications and for Particulate Matter also to the specifications of the British Pharmacopoeia (BP).

The following filter types were used for extractables and particle release testing of Sartopore® 2 filters, representing the individual filter types available:

Standard Filter Cartridges: 5442507H1 5442558K1

Mini Filter Cartridges: 5441507H9 5441558K9

MaxiCaps®: 5441307H1--SS 5441358K1--SS

MidiCaps®: 5445307H9--SS 5445358K9--SO

Capsules 150: 5441307H4--SS 5441358K4--SS

Capsules 300: 5441307H5--OO 5441358K5--OO

| 93

Test Set-up

Water flow

Regulating Valve Pressure

Gauge

Particle Filtration


Particle Sensor System

Overflow

Collection Vessel

Balance

Sampling Vessel

8.1 Particle Content of the Filtrate

PurposeIn general, the particle release from the filters should be minimized. For parenteral solutions, the require-ments are define in the USP Monographs, which set maximum limits for particle content based on defined particle sizes. The particle release of Sartopore® 2 filter elements should lie well below the limits set forth in the current USP for ”Large Volume Parenterals for Single Dose Infusion“.

LimitsFrom the current USP, the following limits have been set as a maximum number of particles per mL of product (in this case, large volume injections for single dose infusion):

25 particles/mL $ 10 µm 3 particles/mL $ 25 µm

Test ProcedureTwo Standard Filter Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® from different production lots were being tested. As a wetting and flushing medium, deionized water (DI water) is used during the testing. An integrity test is performed to assure that only integral filters are used for this testing. In order to generate particle-free water, the water is first filtered through two 0.2 µm membrane Filter Cartridges. This water is used to flush the filter housing and all contact surface to remove surfac e particles prior to testing. The Filter Cartridges are then installed in the pre-rinsed system.

94 |

After attachment of the collection vessel that was also pre-rinsed with the filtered water, the inlet valve is opened and the water is filtered through the test Cartridges. The samples are taken after 1, 2.5, 5 and 10 liters for Standard Filter Cartridges, MidiCaps® 0.2 µm and MaxiCaps®, after 0.5, 2.5 and 5 liters for Mini Cartridges and after 150, 300, 500 and 750 mL for MidiCaps® 0.1 µm and Capsules 150|300 for analysis. The balance is used to determine gravimetrically when a sample should be taken. Particle analysis of the samples is conducted utilizing a particle sensor system. This system consists of a Pacific Scientific Hiac Royco sampler (Model 3000 SOS, serial No. 93023007), in which a particle sensor (Model HRLD 150, serial No. 9208-012) is installed to analyse the filtrate in accordance with the current USP requirements. The system also incorporates a particle counter (Model 8000, serial No. 91078805). The particle sensor system is calibrated twice a year in line with USP Standards.

A sampling vessel is placed into the sampler. The sample medium is drawn in through a glass bulb and a sample volume of 25 mL/min is set exactly on the sampler. The particle count begins automatically when the sampler is started. The average particle value is calculated from a total of six measurements, 25 mL each.

Summary of ResultsIn order to have an overview of the particle content of filtrates of the tested filters the following table contains the average values for the test performed. These averages are for the three diffe rent production lots previously noted.


Standard Cartridges 10"

Particle Size Particle Count Particle Count Particle Count Particle Count Limits [µm] per mL per mL per mL per mL according after 1 L Flush after 2.5 L Flush after 5 L Flush after 10 L Flush to USP

$ 10 0 0 0 0 25

$ 25 0 0 0 0 3

Standard Cartridges 5"


$ 10 0 0 0 0 25

$ 25 0 0 0 0 3

| 95

Mini Cartridges


$ 10 1 0 0 0 25

$ 25 0 0 0 0 3

MaxiCaps®


$ 10 2 5 5 2 25

$ 25 0 0 0 0 3

MidiCaps®


$ 10 0 0 0 0 25

$ 25 0 0 0 0 3

Capsules 150

Particle Size Particle Count Particle Count Particle Count Particle Count Limits [µm] per mL per mL per mL per mL according after 150 mL Flush after 300 mL Flush after 500 mL Flush after 750 mL Flush to USP

$ 10 3 1.5 1 1 25

$ 25 1 0 0 0 3

Capsules 300


$ 10 4 1 0 0 25

$ 25 0 0 0 0 3

96 |96 |

Standard Cartridges 10" Particle Size Particle Count Particle Count Particle Count Particle Count Limits [µm] per mL per mL per mL per mL according after 1 L Flush after 2.5 L Flush after 5 L Flush after 10 L Flush to USP

$ 10 2 1 0 0 25

$ 25 0 0 0 0 3

Standard Cartridge 5"


$ 10 0 0 0 0 25

$ 25 0 0 0 0 3

Mini Cartridges


$ 10 0 0 0 0 25

$ 25 0 0 0 0 3


| 97 | 97

ConclusionThe tables above show that for Sartopore® 2 filter elements the requirements of the current USP for particle content are met in the first liter of rinse volume. This shows that the initial filtrate conforms to these standards, as it is not techni-cally feasible to test the first mL of solution filtered. Accordingly, the Sartopore® 2 filter elements produce a filtrate that conforms with the current USP for particle content.

MaxiCaps®


$ 10 0 0 0 0 25

$ 25 0 0 0 0 3

MidiCaps®


$ 10 0 0 0 0 25

$ 25 0 0 0 0 3

Capsules 150


$ 10 3 2 1 1 25

$ 25 1 0 0 0 3

Capsules 300


$ 10 1 0.5 0.5 0.5 25

$ 25 0 0 0 0 3

98 |98 |

8.2 Determination of Oxidizable Substances of the Filtrate Test ProcedureThree Sartopore® 2 Standard Filter Cartridges, Mini Cartridges, MaxiCaps® from three different production lots were wetted (by soaking) and autoclaved. After installation into the filter housings the filter elements were flushed with Water for Injection and 100 mL samples were taken after 1, 2.5, 5 and 10 L flush volumes for Standard Filter Cartridges and MaxiCaps®; 0.5, 2 and 5 liters for Mini Cartridges and 0.5, 1 and 1.5 flush volumes for Capsules.

As described in the current USP to the 100 mL samples 10 mL of 2 N sulfuric acid were added and heated to boiling.

Than 0.2 mL of 0.1 N potassium permanganate were added and the solution was boiled for 5 minutes. If a precipitate forms, it is cooled to room temperature. If the precipitate remains its color after cooling to room temperature, the test sample and respectively the tested filter element meets the USP specifica-tions for oxidizable substances.

Standard Cartridges 10", 0.2 µm

Blank passed

Lot Test Results Test Results Test Results Test Results Number after after after after 1 L Flush 2.5 L Flush 5 L Flush 10 L Flush

920641 passed passed passed passed





Blank passed






Blank passed






Blank passed





| 99 | 99

Mini Cartridges, 0.2 µm

Blank passed

Lot Test Results Test Results Test Results Test Results Number after after after after 1 L Rinse 2.5 L Rinse 5 L Rinse 10 L Rinse






Blank passed





MaxiCaps®, 0.1 µm

Blank passed





MaxiCaps®, 0.2 µm

Blank passed

Lot Test Results Test Results Test Results Test Results Number after after after after 1 L Rinse 2.5 L Rinse 5 L Rinse 10 L Rinse




8.2.3 MaxiCaps®

100 |100 |

MidiCaps®, 0.1 µm

Blank passed





MidiCaps®, 0.2 µm

Blank passed





8.2.4 MidiCaps®

| 101 | 101

Capsules 150, 0.1 µm

Blank passed

Lot Test Results Test Results Test Results Number after after after 0.5 L Flush 1 L Flush 1.5 L Flush

024881 passed passed passed




Blank passed






Blank passed






Blank passed





8.2.5 Capsules

ConclusionThe Sartopore® 2 Standard Filter Cartridges, Mini Cartridges, MaxiCaps®, MidiCaps® and Capsules produced filtrates that, when measured by this method, were below the requirements set by the current USP Limits for Oxidizable Substances for ”Sterile Water for Injection”.

102 |102 |

8.3 Determination of pH Values and Conductivity of the Filtrate

Test ProcedureThree Sartopore® 2 Standard Filter Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® from three different production lots were wetted (by soaking) and autoclaved. After installation into the filter housings the filter elements were flushed with Water for Injection and samples were taken after 1, 2.5, 5 and 10 L flush volumes for Standard Filter Cartridges, MaxiCaps®, and MidiCaps®, 0.5, 2 and 5 liters for Mini Cartridges and after 0.5, 1 and 1.5 liters for Capsules.

Conductivity and pH value of the samples were measured using appropriate calibrated pH meters and conductivity meters according to the USP regulations.

Test LimitsThe following table lists the limits for pH and conductivity given by the current USP in conjunction with “Sterile Purified Water” and the filters were tested in the specified pH range of 5 to 7.

The relationship between the pH value and the maximum allowable conductivity for “Sterile Water for Injection“ according to the current USP is:

pH Value Maximum Allowable Conductivity [µS/cm]

5 4.7

5.1 4.1

5.2 3.6

5.3 3.3

5.4 3.0

5.5 2.8

5.6 2.6

5.7 2.5

5.8–6.1 2.4

6.2 2.5

6.3 2.4

6.4 2.3

6.5 2.2

6.6 2.1

6.7 2.6

6.8 3.1

6.9 3.8

7.0 4.6

| 103


Blank pH 5.9

Lot pH pH pH pH Number after after after after 1 L Flush 2.5 L Flush 5 L Flush 10 L Flush

920641 6.0 5.9 6.0 6.0

920741 5.55 5.55 5.55 5.6

920841 5.65 5.55 5.65 5.55


Results for pH Values


Blank pH 5.9


009141 5.9 5.9 5.9 5.9

009241 5.8 5.8 5.7 5.9

009341 6.0 5.8 5.8 5.8


Blank pH 5.85


036002 5.85 5.85 5.85 5.85

036102 5.85 5.85 5.85 5.85

036302 5.8 5.8 5.8 5.8


Blank pH 5.8


313742 5.8 5.8 5.8 5.8

314442 5.75 5.75 5.8 5.8

314642 5.8 5.8 5.8 5.8

Note Due to the interrelationship between the pH value determi-nation and the measurement of the conductivity, results for both tests must be viewed together.

104 |


Blank 0.7 µS/cm

Lot Conductivity Conductivity Conductivity Conductivity Number after after after after 1 L Flush 2.5 L Flush 5 L Flush 10 L Flush [µS/cm] [µS/cm] [µS/cm] [µS/cm]

920641 0.81 0.79 0.82 0.78

920741 0.87 0.87 0.62 0.62

920841 0.67 0.66 0.62 0.57

Results for Conductivity


Blank 0.9 µS/cm


009141 0.9 1.0 1.0 0.9

009241 0.9 0.9 0.9 0.9

009341 1.0 0.9 0.9 0.9


Blank 0.64 µS/cm


036002 0.71 0.67 0.68 0.67

036102 0.71 0.74 0.71 0.74

036302 0.72 0.72 0.71 0.72


Blank 0.52 µS/cm


313742 0.52 0.52 0.52 0.52

314442 0.79 0.76 0.56 0.52

314642 0.78 0.72 0.63 0.63

| 105 | 105


Blank pH 5.6

Lot pH after pH after pH after pH after Number 1 L Flush 2.5 L Flush 5 L Flush 10 L Flush

932941 5.6 5.6 5.6 5.6

932641 5.8 5.8 5.8 5.7

930541 5.6 5.6 5.6 5.7




Blank pH 5.85


011441 5.9 5.85 5.85 5.85

011541 5.95 5.85 5.85 5.85

011741 5.85 5.85 5.85 5.85


Blank 0.6 [µS/cm]

Lot Conductivity Conductivity Conductivity Conductivity Number after after after after 1 L Flush 2 L Flush 5 L Flush 10 L Flush [µS/cm] [µS/cm] [µS/cm] [µS/cm]

932941 0.7 0.7 0.6 0.7

932641 0.6 0.5 0.6 0.6

930541 0.5 0.6 0.6 0.6



Blank 0.6 [µS/cm]


011441 0.9 0.5 0.6 0.9

011541 0.8 0.6 0.6 0.9

011741 0.8 0.6 0.6 0.9

106 |106 |

MaxiCaps®, 0.2 µm

Blank pH 5.8


008881 6.3 6.0 5.9 5.8

008981 6.2 5.9 5.8 5.8

010181 6.2 5.8 5.8 5.8

8.3.3 MaxiCaps®


MaxiCaps®, 0.2 µm

Blank 0.6 [µS/cm]

Lot Conductivity Conductivity Conductivity Conductivity Number after after after after 1 L Flush 2 L Flush 5 L Flush 10 L Flush [µS/cm] [µS/cm] [µS/cm] [µS/cm]

008881 1.7 0.7 0.7 0.7

008981 1.6 0.7 0.6 0.6

010181 1.8 0.8 0.6 0.6


MaxiCaps®, 0.1 µm

Blank pH 5.9


009081 6.4 6.1 6.0 6.0

009081 6.4 6.1 6.0 5.9

009081 6.4 6.1 5.9 5.9

MaxiCaps®, 0.1 µm

Blank 0.7 [µS/cm]


009081 2.2 0.8 0.7 0.6

009081 2.9 1.0 0.8 0.6

009081 2.5 0.9 0.8 0.7

| 107 | 107

MidiCaps®, 0.2 µm

Blank pH 6.0


402243 6.0 6.0 6.0 6.0

402343 5.9 5.95 6.0 6.0

414343 5.9 5.9 6.0 6.0

MidiCaps®, 0.1 µm

Blank pH 6.0


403943 5.95 5.85 5.85 5.85

403943 5.85 5.85 5.9 5.9

404143 5.85 5.9 5.9 5.95

8.3.4 MidiCaps®


MidiCaps®, 0.2 µm

Blank 0.75 µS/cm


402243 0.7 0.7 0.7 0.7

402343 0.75 0.7 0.7 0.7

414343 0.7 0.7 0.7 0.7

MidiCaps®, 0.1 µm

Blank 0.75 µS/cm


403943 0.62 0.61 0.61 0.61

403943 0.63 0.61 0.62 0.62

404143 0.58 0.55 0.62 0.52


108 |108 |


Blank pH 0.6

Lot pH pH pH Number after after after 0.5 L Flush 1 L Flush 1.5 L Flush

100243 5.55 5.55 5.55

100343 5.55 5.55 5.55

100443 5.55 5.55 5.55


Blank pH 0.6


024881 5.75 5.75 5.75

024881 5.75 5.75 5.75

024981 5.75 5.75 5.75


Blank pH 0.6


919281 5.9 5.9 5.9

920881 5.85 5.85 5.85

920981 5.8 5.8 5.8


Blank pH 0.6


005781 5.9 5.9 5.9

005881 5.85 5.85 5.85

005981 5.8 5.8 5.8

8.3.5 Capsules


| 109 | 109

ConclusionBoth parameters, pH and pH dependent conductivity of the filtrate, when filtering with the Sartopore® 2 Standard Filter Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® are well below the limit requirements of the current USP.


Blank 0.75 µS/cm

Lot Conductivity Conductivity Conductivity Number after after after 0.5 L Flush 1 L Flush 1.5 L Flush [µS/cm] [µS/cm] [µS/cm]

100243 0.76 0.84 0.77

100343 0.77 0.85 0.76

100443 0.75 0.77 0.76


Blank 0.75 µS/cm


919281 0.83 0.69 0.71

920881 0.74 0.71 0.73

920981 0.8 0.74 0.7


Blank 0.75 µS/cm


005781 0.83 0.69 0.71

005881 0.74 0.71 0.73

005981 0.8 0.74 0.7


Blank 0.75 µS/cm


024881 0.78 0.51 0.5

024881 0.72 0.74 0.68

024981 0.66 0.52 0.53


110 |

8.4 Determination of Chloride, Sulfate and Ammonia in the Filtrate

8.4.1 Determination of Chloride

Test ProcedureThree Sartopore® 2 Standard Filter Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® from three different production lots were wetted (by soaking) and autoclaved. After installation into the filter housings the filter elements were flushed with Water for Injection and 20 mL samples were taken after 1, 2.5, 5 and 10 L flush volumes for Standard Filter Cartridges, MaxiCaps®, and MidiCaps®, 0.5, 2 and 5 liters for Mini Cartridges and 0.5, 1 and 1.5 liters for Capsules.

To the 20 mL samples 5 drops of nitric acid and 1 mL of silver nitrate are added and gently mixed. If the turbidity formed within 10 minutes is below the control reagent consisting of 20 mL of high purity water containing 10 µg of Chloride the test is passed.


Blank passed





8.4.1.1 Standard Cartridges

Test Results for Chloride


Blank passed






Blank passed






Blank passed





| 111

Blank passed





8.4.1.2 Mini Cartridges




Blank passed





Blank passed





8.4.1.3 MaxiCaps®


MaxiCaps®, 0.2 µm

MaxiCaps®, 0.1 µm

Blank passed




112 |112 |

MidiCaps®, 0.1 µm

Blank passed





Blank passed





8.4.1.4 MidiCaps®


MidiCaps®, 0.2 µm

| 113 | 113

Blank passed






Blank passed





8.4.1.5 Capsules




Blank passed






Blank passed





ConclusionSartopore® 2 Standard Filter Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® produced filtrates that, when measured by this method, were below the requirements set by the current USP Limits for Chloride for “Sterile Water for Injection”.

114 |114 |

8.4.2 Determination of Sulfate

Test ProcedureThree Sartopore® 2 Standard Filter Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® from three different production lots were wetted (by soaking) and autoclaved. After installation into the filter housings the filter elements were flushed with Water for Injection and 100 mL samples were taken after 1, 2.5, 5 and 10 L flush volumes for Standard Filter Cartridges, MaxiCaps® and Midicaps, 0.5, 2 and 5 liters for Mini Cartridges and 0.5, 1 and 1.5 liters for Capsules.

To the 100 mL samples 1 mL of barium chloride is added. If no turbidity forms the test is passed.


Blank passed






Test Results for Sulfate


Blank passed






Blank passed






Blank passed





| 115 | 115


Blank passed







MaxiCaps®, 0.2 µm

Blank passed





8.4.2.3 MaxiCaps®



Blank passed





MaxiCaps®, 0.1 µm

Blank passed





116 |

8.4.2.4 MidiCaps®


MidiCaps®, 0.2 µm

Blank passed





MidiCaps®, 0.1 µm

Blank passed





| 117

ConclusionSartopore® 2 Standard Filter Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® produced filtrates that, when measured by this method, were below the requirements set by the current USP Limits for Sulfate for “Sterile Water for Injection”.

8.4.2.5 Capsules



Blank passed






Blank passed






Blank passed






Blank passed





118 |118 |

8.4.3 Determination of Ammonia

Test ProcedureThree Sartopore® 2 Standard Filter Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® from three different production lots were wetted (by soaking) and autoclaved. After installation into the filter housings the filter elements were flushed with Water for Injection and 100 mL samples were taken after 1, 2.5, 5 and 10 L flush volumes for Standard Filter Cartridges, MaxiCaps® and MidiCaps®, 0.5, 2 and 5 liters for Mini Cartridges and 0.5, 1 and 1.5 liters for Capsules.

To 100 mL of each of the recircu-lation samples and sample blank, added 2 mL of ammonium oxalate (2 %). A sample passed the test, if no turbidity was produced.


Blank passed






Test Results for Ammonia


Blank passed






Blank passed






Blank passed





| 119 | 119


Blank passed







MaxiCaps®, 0.2 µm

Blank passed





8.4.3.3 MaxiCaps®



Blank passed





MaxiCaps®, 0.1 µm

Blank passed




120 |120 |

MaxiCaps®, 0.2 µm

Blank passed





8.4.3.4 MidiCaps®


MaxiCaps®, 0.1 µm

Blank passed





| 121 | 121

ConclusionSartopore® 2 Standard Filter Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® produced filtrates that, when measured by this method, were below the requirements set by the current USP Limits for Ammonia for ”Sterile Water for Injection”.

8.4.3.5 Capsules



Blank passed






Blank passed






Blank passed






Blank passed





122 |

8.5 Biocompatibility

PurposeThese tests are to determine that all components used in the manufacture of Sartopore® 2 Filter Cartridges, MaxiCaps®, MidiCaps® and Capsules are biosafe and meet or exceed the requirements for the current USP Class VI–121 °C Plastics Tests.

Test Method and ResultsSartopore® 2 Filter Cartridges, MaxiCaps®, MidiCaps® and Capsules were supplied to an independent testing facility for evaluation under the require ments of the current USP Class VI Plastics Tests, including the following tests:

Intracutaneous test Systemic injection test Implantation test (7 days)

The complete test report is available upon request.

ResultThe following certificate was released as a result of the testing of Sartopore® 2 Filter Cartridges, MaxiCaps®, MidiCaps® and Capsules. All material used in the construction of the Sartopore® 2 filter elements meet or exceed the requirements of the USP Class VI–121 °C Plastics Tests.

| 123

Certificate Standard Cartridges

124 |

Certificate MaxiCaps® Housings

| 125

Certificate MidiCaps®

126 |126 |

9. Endotoxin Testing

PurposeThe goal of these tests is to determine whether the amount of endotoxins released in the effluent of a Sartopore® 2 filter element meets the requirements of EP and USP monographs for "Sterile Water for Injection" (0.25 EU/mL).

Test MethodThe Filter Cartridges are shaken in a defined, smallest possible volume of endotoxin free water. Sartopore® 2 MaxiCaps®, MidiCaps® and Capsules from a variety of production lots are placed into glass vessels and filled with the required amount of endotoxin free water. In contrast to Filter Cartridges Capsules are filled from both sides of the plastic housing with the required amount of endotoxin free water.

Then the vessels or Capsules are placed on a shaker in order to free any endotoxins that may be present. They are shaken for 60 minutes with 100 rpm. Samples are then taken and evaluated with the LAL gel clot test with a sensitivity of 0.06 EU|mL.

Along with these samples a positive control containing 0.06 EU of endotoxin per mL is incubated as well as the endotoxin free water itself as a negative control. A clot indicates the presence of endotoxins. No clot indicates absence of endotoxins (i.e. < 0.06 EU/mL).

Results:

Standard Cartridges


Lot Number LAL Test Results

920641 passed

920741 passed

920841 passed



009141 passed

009241 passed

009341 passed

Mini Cartridges



932941 passed

932641 passed

930541 passed



011441 passed

011541 passed

011741 passed

MidiCaps®

MidiCaps®, 0.2 µm


402243 passed

402343 passed

414343 passed

MidiCaps®, 0.1 µm


403943 passed

403943 passed

404143 passed

ConclusionAll Sartopore® 2 Standard Cartridges, Mini Cartridges, MaxiCaps® and MidiCaps® tested, under the conditions of the extraction test described above, gave results below 0.18 EU/mL bacterial endotoxin.

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