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Disinfectant Trigger Sprays

Process Overview and Validation Support Guide

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Contents

1. Manufacturing Process Overview 2. Process Flow Chart 3. Product Codes 4. Component Specifications 5. Finished Goods Specifications 6. Materials Safety Data Sheets 7. Process Qualification 8. Example Product Certificates 9. Broth Fill Testing 10. TSE Statement 11. Efficacy and Mode of Action 12. ISO 9001:2008 Registration Certificate

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Manufacturing Process Overview

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Micronclean: Compliance 100® Disinfectant Trigger Sprays Process Overview & Validation Support Guide

Introduction Micronclean manufacture a range of sterile cleanroom disinfectant trigger spray products at our Skegness facility. Below is an overview of the manufacturing process including key quality assurance and qualification measures. Raw Materials Bottles and trigger sprays are purchased from approved suppliers. These raw materials, together with the necessary packaging materials, are stocked within a dedicated storage area in Skegness. Bottles are blow-moulded HDPE, with bayonet-type neck. Trigger sprays have a bayonet-style fastening, with a multidirectional pre-compression configuration. The trigger spray incorporates a hydrophobic ePTFE membrane, preventing liquid and microbial penetration while allowing gas exchange in both directions to equalise pressure. Alcohol (70% IPA / 30% USP WFI and 70% Denatured Ethanol / 30% USP WFI) is supplied by approved suppliers to the agreed specifications shown in Table 1 and Table 2. Table 1 – IPA

Parameter Specification

Alcohol type and concentration 70% USP Isopropyl Alcohol Diluent water type and concentration 30% USP WFI Filtration level 0.2µm Filtered Specific gravity tolerance 0.872 to 0.883 (at 20°C) Endotoxin (IPA/WFI) <0.25 EU/ml Non-volatile residues Does not exceed 0.01% w/v Acidity Complies with USP monograph Organoleptic Clear, colourless liquid with spirituous odour Blending and dosing standard GMP

Table 2 – Denatured Ethanol

Parameter Specification

Alcohol type and concentration 70% Trade Specific Denatured Alcohol 7 Diluent water type and concentration 30% USP WFI Filtration level 0.2µm Filtered Specific gravity tolerance 0.880 to 0.890 (at 20°C) Endotoxin (Denatured Ethanol/WFI) <0.25 EU/ml Non-volatile residues Does not exceed 0.01% w/v Acidity Complies with USP monograph Organoleptic Clear, colourless liquid with spirituous odour Blending and dosing standard GMP

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Prior to use conformance checks are carried out on the alcohol. Filling & Packaging The bottle filling area comprises a unidirectional airflow cabinet validated to BS EN ISO 14644-1 Class 5 (in use). The background environment is an ISO 14644-1 Class 7 (in use) cleanroom. Both areas are monitored microbiologically to agreed levels. Bottles are passed into the LAF cabinet and filled with alcohol. Each bottle is then sealed in outer polythene bags before being placed into a cardboard carton designed to meet all applicable UK safety legislation. Gamma Irradiation Packaged bottles are transported to Isotron at Bradford who gamma irradiate the product, providing a minimum radiation dose of 25kGy. Following irradiation the product is then returned to the Compliance 100® Warehouse at our Louth site prior to final release. Finished Goods Storage & Release All finished goods are stored in Micronclean’s dedicated Compliance 100® Warehouse at our Louth site. Goods are checked to ensure that gamma irradiation has occurred and batch paperwork has been reconciled prior to release. Batch Traceability All bottles are labelled with a batch number. This number provides traceability to:

• Bottle Lot

• Trigger Spray Lot

• Alcohol Lot

• Packaging Lot

• Gamma Irradiation Lot

Each box of trigger sprays has an electronic Certificate of Conformity referencing the appropriate batch number and is available on request. Quality System All Micronclean processes operate under BS EN ISO 9001:2008 registered systems. June 2013

Phil Brennan Technical Director

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Process Flow Chart

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Bottle/Trigger Spray Moulding

Alcohol Manufacture

Pouch/Carton Manufacture

Incoming Goods QC Approval

ISO 14644 Class 4 / EU GMP Class B Decontamination

Alcohol dosing under ISO Class 5

unidirectional air flow

Final Packing and Batch Documentation

Sterilisation Validated at 25 kGy

Final QC Approval and Certificate of

Compliance

Distribution by Micronclean’s own Transport Fleet

Approved Gamma Irradiation Sub-contractors

Micronclean Logistics

Micronclean • Multi-site back-up

capability

• BS EN ISO 9001:2008

• GMP

• Validated processes

Raw Materials Supply

Compliance 100® Trigger Sprays Process Flow

Secondary packaging in an ISO Class 7 (in use)

cleanroom

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Product Codes

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Description Reference

Code Disinfectant

Bottle Size (ml)

Primary Packaging

Secondary Packaging

Bottles per Case

ZSAL64499 70% IPA 30 % USP WFI

500

Natural HDPE bottle with tamper proof trigger spray

Double bagged in polythene pouches

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ZSAL64950 70% USP IPA 30 % USP WFI

950

Natural HDPE bottle with tamper proof trigger spray

Double bagged in polythene pouches

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ZSAL65499

70% USP Denatured Alcohol 30 % USP WFI

500

Natural HDPE bottle with tamper proof trigger spray

Double bagged in polythene pouches

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Sterile disinfectant trigger spray for use in cleanroom environments

ZSAL65950

70% USP Denatured Alcohol 30 % USP WFI

950

Natural HDPE bottle with tamper proof trigger spray

Double bagged in polythene pouches

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August 2011

Phil Brennan Technical Director

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Component Specifications

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500ml Bottle - Specification

Size: 500ml ‘Sumo’ Bottle Colour: Natural Material: HDPE Neck Fastening: TS2 Push-Fit

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950ml Bottle - Specification

Size: 950ml ‘Sumo’ Bottle Colour: Natural Material: HDPE Neck Fastening: TS2 Push-Fit

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Trigger Spray - Specification

Description: Trigger spray with ePTFE membrane at venting valve Material: PP/LDPE/Stainless Steel

Colour: Shroud – Blue Lever – Blue Nozzle – Blue Nozzle Type: Spray with On/Off settings Fastening: TS2 Push-Fit Dip Tube Length: 180mm Other Features: Multi-direction valve device

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Vent Filter - Specification

Description: Oleophobic ePTFE membrane at venting valve Support Material: Non-woven polyethylene / polypropylene

Pore Size: 3µm (maximum pore size)

Performance Characteristics:

Test Method Description QA Test Units

Airflow (Gurley Method) 219 seconds

Thickness 201 mils

Water Entry Pressure 584 psi

Oil Rating 713 n/a

Bond 709 psi

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Denatured Ethanol - Specification

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Denatured Ethanol – Microbiological Data

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Isopropyl Alcohol - Specification

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Isopropyl Alcohol – Microbiological Data

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Finished Goods Specifications

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IPA 70/30 500ml TRIGGER SPRAY SPECIFICATION

Product Code : ZSAL64499

Product Description : Sterile cleanroom trigger spray containing 70% isopropyl alcohol

/ 30% water for injection

Pack Size : 24 x 500ml bottles per carton

Technical Specification

Packaging : Natural HDPE Bottle with tamper-proof trigger spray

Bottle Contents : 500ml

Alcohol : 70% USP IPA / 30% USP WFI

Decontamination : Product filled in an ISO 14644-1 Class 5 (in use) unidirectional

flow cabinet, secondary packaging in an ISO 14644-1 Class 7 (in use) cleanroom

Sterilisation : Validated sterilisation by gamma irradiation (minimum 25 kGy)

Shelf-Life : Two years from manufacturing date

Quality Assurance : Product manufactured under BS EN ISO 9001:2008 registered systems

Issue no 1 30/06/10

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IPA 70/30 950ml TRIGGER SPRAY SPECIFICATION

Product Code : ZSAL64950

Product Description : Sterile cleanroom trigger spray containing 70% isopropyl alcohol / 30% water for injection

Pack Size : 15 x 950ml bottles per carton

Technical Specification

Packaging : Natural HDPE Bottle with tamper-proof trigger spray

Bottle Contents : 950ml

Alcohol : 70% USP IPA / 30% USP WFI

Decontamination : Product filled in an ISO 14644-1 Class 5 (in use) unidirectional

flow cabinet, secondary packaging in an ISO 14644-1 Class 7 (in

use) cleanroom

Sterilisation : Validated sterilisation by gamma irradiation (minimum

25 kGy)

Shelf-Life : Two years from manufacturing date

Quality Assurance : Product manufactured under BS EN ISO 9001:2008

registered systems

Issue no 1 30/08/11

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IMS 70/30 500ml TRIGGER SPRAY SPECIFICATION

Product Code : ZSAL65499

Product Description : Sterile cleanroom trigger spray containing 70% denatured ethanol / 30% water for injection

Pack Size : 24 x 500ml bottles per carton

Technical Specification

Packaging : Natural HDPE Bottle with tamper-proof trigger spray

Bottle Contents : 500ml

Alcohol : 70% USP Denatured Ethanol / 30% USP WFI

Decontamination : Product filled in an ISO 14644-1 Class 5 (in use) unidirectional

flow cabinet, secondary packaging in an ISO 14644-1 Class 7 (in use) cleanroom

Sterilisation : Validated sterilisation by gamma irradiation (minimum 25 kGy)

Shelf-Life : Two years from manufacturing date

Quality Assurance : Product manufactured under BS EN ISO 9001:2008

registered systems

Issue no 1 30/06/10

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IMS 70/30 950ml TRIGGER SPRAY SPECIFICATION

Product Code : ZSAL65950

Product Description : Sterile cleanroom trigger spray containing 70% denatured ethanol / 30% water for injection

Pack Size : 15 x 950ml bottles per carton

Technical Specification

Packaging : Natural HDPE Bottle with tamper-proof trigger spray

Bottle Contents : 950ml

Alcohol : 70% USP Denatured Ethanol / 30% USP WFI

Decontamination : Product filled in an ISO 14644-1 Class 5 (in use) unidirectional

flow cabinet, secondary packaging in an ISO 14644-1 Class 7 (in use) cleanroom

Sterilisation : Validated sterilisation by gamma irradiation (minimum 25 kGy)

Shelf-Life : Two years from manufacturing date

Quality Assurance : Product manufactured under BS EN ISO 9001:2008

registered systems

Issue no 1 30/08/11

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Materials Safety Data Sheets

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Process Qualification

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Irradiation Dosimeter Validation – 500ml

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Sterility Testing Protocol

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Example Product Certificates

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Certificate of Conformity

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Certificate of Irradiation

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Certificate of Sterility

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Broth Fill Testing

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Broth Fill Testing of the Micronclean Compliance 100® Trigger Spray System

Test Report

Testing is to be carried out to challenge the filter within the trigger spray head of Micronclean’s Compliance 100® Disinfectant Trigger Spray. The filter is incorporated in the trigger spray head in order to maintain the sterile environment within the bottle. To equalise pressure within the bottle when the trigger is pulled air is allowed back into the bottle through the filter. The filter has a pore size sufficient to filter out microorganisms and particulate from the air passing through it. In order to challenge this system a broth fill test has been arranged. This will involve filling sample trigger sprays with a microbiological growth medium, irradiating the bottles, and then operating the bottles’ triggers in an uncontrolled environment over time. At certain time intervals bottles will be incubated, and after incubation the growth medium will be assessed for microbiological growth to find out if the sterile environment has been maintained within the bottle. Test Method Fifteen bottles and compatible trigger spray heads were obtained. The integral filter was removed from three of the trigger spray heads; these were labelled ‘Positive Control’. The other trigger spray heads were labelled A to L. The bottles were each filled with 450ml of Tryptone Soya Broth and assembled. The filled bottles were then sent for gamma irradiation (minimum 25kGy radiation dose). On return from irradiation the bottles were stored in the laboratory – an uncontrolled environment. The trigger spray of each bottle was operated 5 times every 24 hours. After certain time intervals bottles were transferred to a 30°C incubator for a minimum of 48 hours as detailed in Table 1.

RD+T Department

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Table 1

Sample ID Test Point Date for Transfer to 30°C

Incubator

A Day 1 26-04-10

Positive Control 1 Week 03-05-10

B 1 Week 03-05-10

Positive Control 2 Weeks 10-05-10

C 2 Weeks 10-05-10

D 2 Weeks 10-05-10

E 3 Weeks 17-05-10

F 3 Weeks 17-05-10

G 4 Weeks 24-05-10

H 4 Weeks 24-05-10

I 6 Weeks 07-06-10

J 6 Weeks 07-06-10

Positive Control 12 Weeks 19-07-10

K 12 Weeks 19-07-10

L 12 Weeks 19-07-10

After at least 48 hours incubation the bottles were agitated and then opened. The broth in each bottle was checked for evidence of microbiological growth (turbidity); the results were recorded. Results The results of the broth fill test are shown in Table 2. TSB Batch No. 10040151

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Table 2

Sample ID Test Point Microbiological Growth

(Y/N)

A Day 1 N

Positive Control 1 Week Y

B 1 Week N

Positive Control 2 Weeks Y

C 2 Weeks N

D 2 Weeks N

E 3 Weeks N

F 3 Weeks N

G 4 Weeks N

H 4 Weeks N

I 6 Weeks N

J 6 Weeks N

Positive Control 12 Weeks Y

K 12 Weeks N

L 12 Weeks N

Growth promotion testing was carried out on broth showing no growth after incubation within bottles C and D (2 Weeks) and K and L (12 Weeks). The broth in each bottle was inoculated with an environmental isolate and incubated for a further 24 hours at 30°C. The broth from each of the 4 bottles appeared turbid after incubation showing that the broth was still able to support microbial growth. This testing shows that the trigger spray system maintains the integrity of the product through use. Even in this test, where the bottles were filled with a microbiological growth medium and operated in an environment uncontrolled in terms of contamination, the bottles’ contents showed no signs of microbial ingress after 12 weeks of use. The three Positive Control bottles, which the filters were removed from, each showed evidence of microbial ingress into the bottles through use. This shows that the filter has a pore size sufficient to filter out microorganisms from the air. 27-07-10 Greg Cochran R+D Manager

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TSE Statement

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Compliance 100® Trigger Sprays

Statement to confirm that the above product is not derived from animal origin This document confirms that the Compliance 100® Trigger Sprays produced by Micronclean Skegness, UK are not produced from any materials derived from animal origin. Additionally no part of the decontamination, filling, packing or sterilisation process, directly involved with the manufacture of the Compliance 100® Trigger Sprays, utilises any products derived from animal origin. 30th June 2010 Greg Cochran Research and Development Manager

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Efficacy and Mode of Action

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The Efficacy and Mode of Action of Alcohols Efficacy Alcohols, like other disinfectants, are tested using two types of method – suspension testing and surface testing. These tests are covered by the following standards: BS EN 1276:1997 Chemical Disinfectants and Antiseptics – Quantitative suspension test for the evaluation

of bactericidal activity of chemical disinfectants and antiseptics used in food, industrial, domestic and institutional areas.

A range of standard bacterial organisms are used for this test; these are Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Enterococcus hirae. The pass criterion for the test is that the disinfectant or antiseptic must cause a 5-log reduction in the number of viable bacterial cells present in a suspended culture. BS EN 1650:1998 Chemical Disinfectants and Antiseptics – Quantitative suspension test for the evaluation

of fungicidal activity of chemical disinfectants and antiseptics used in food, industrial, domestic and institutional areas.

Standard fungal organisms are used for this test; these are Candida albicans and Aspergillus niger. The pass criterion for the test is that the disinfectant or antiseptic must cause a 4-log reduction in the number of viable fungal cells present in a suspended culture. BS EN 13697:2001 Chemical Disinfectants and Antiseptics – Quantitative non-porous surface test for the

evaluation of bactericidal/fungicidal activity of chemical disinfectants and antiseptics used in food, industrial, domestic and institutional areas.

A range of standard bacterial and fungal organisms are used for this test; these are Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Enterococcus hirae, Candida albicans and Aspergillus niger. A suspension of the test organism is inoculated onto a stainless steel surface and dried. A sample of the disinfectant or antiseptic is applied to the dried inoculum for a given exposure time. The pass criterion for the test is that the disinfectant or antiseptic must cause a 5-log reduction in the number of viable bacterial cells, or 4-log reduction in the number of viable fungal cells, present on the surface. 70% aqueous solutions of isopropyl alcohol and ethanol that meet the requirements of the European Pharmacopoeia have been shown to give satisfactory test results in all of the above tests. Micronclean use alcohol solutions that meet the requirements of the European Pharmacopoeia.

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Mode of Action Alcohols, such as isopropyl alcohol and ethanol, have a number of effects on microorganisms that give them their antimicrobial action. Predominantly alcohols are antimicrobial as they cause coagulation and denaturation of proteins. Other known modes of action associated with this effect on proteins include disruption of cytoplasmic integrity, lysis of cells, and various interferences with cell metabolism (Ali, et al. 2001). Alcohols have optimal antimicrobial effect at concentrations of between 50% and 80% in aqueous solution. Their antimicrobial effects are much reduced in very low or very high concentration aqueous solutions. Alcohols are effective against a range of microorganisms including vegetative bacteria, fungus, yeast and viruses. Alcohols are not effective against microbial spores or non-lipid containing viruses. References Ali, Y. Dolan, M.J. Fendler, E.J. & Larson, E.L., 2001. Alcohols. In Block, S.S. 5th ed. Disinfection, Sterilization and Preservation. Philadelphia: Lippincott Williams & Wilkins, 2001, Chapter 12, pp.229 – 253. 30th June 2010 Greg Cochran Research and Development Manager

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ISO 9001:2008 Registration Certificate

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