New approaches to cleaning validation

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New approaches to cleaning validation

• Louise Lennard
• European Allergen Controls Project Manager
• HJ Heinz UK
• The Food Safety Centre Allergen Resource Bureau
  Conference
• Food Allergens Issues and Opportunities for the
  Food Manufacturer
• 16th February 2006
• Sydney, Australia

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Overview

• Allergen risk assessment
• Identifying cleaning requirements
• Cleaning validation
• Validation / verification

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Due diligence

• Allergen Policy
• Allergen Risk Assessment manufacturing sites,
  suppliers, co-packers
• Allergen questionnaires
• Product reformulation
• GMPs
• Training technical, product development,
  production, marketing, legal, production
  planning, consumer support, suppliers, co-packers
• Cleaning validation
• End product testing

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Heinz Europe approach to cleaning validation

• Conduct and document Allergen Risk Assessment
• List all allergens at supplier site /
  manufacturing site
• All mandatory GMPs that must be in place
• Allergen training appropriate to job role
• Identify where cleaning validation required
• Cleaning validation process

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Extract Heinz Europe Allergen Safety Assessment
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Essential components of cleaning program

• Change-over matrix include enough time
• Documented procedures
• Individual equipment cleaning procedure
• Time
• Method
• temperature
• Chemical concentration etc
• Validation eg. Visual, ATP, Coomassie

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Purpose of cleaning validation

• To ensure that the (current) documented method of
  cleaning is sufficient to remove allergen
  residues
• If allergen is detectable
• Need to revise current cleaning method
• If allergen is not detectable
• current cleaning technique is sufficient
• Then control the cleaning implementation
  ensure always followed, always documented
  (verify)

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Methods for cleaning validation

• We have evaluated these tests for our purpose
• Visual
• Bioluminescence testing
• PCR
• ELISA
• Non-specific protein tests

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ATP bioluminescence

• Testing for the presence of ATP
• A positive result is an indication of inadequate
  cleaning
• A negative result (not detectable) does not
  necessarily mean the surface / equipment is
  allergen-free

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PCR tests

• Not specifically for protein (allergen)
• Not cost effective
• Can have false results
• Time consuming (10 days)

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ELISA based tests

• Specific for a single protein
• false positives and false negatives
• Not all proteins have a specific kit
• Not cost effective
• Some kits are rapid, others lab based

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Bradford Coomassie test

• Non-specific protein test
• Advantages
• Most allergens are proteins
• Quicker than PCR / Std ELISA
• Cheaper than PCR / ELISA
• Easy to use???

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Pierce Coomassie Plus test

• Based on the Bradford colorimetric method
• Coomassie Brilliant Blue G-250 dye (CBBG)
  specifically binds to proteins at arginine,
  tryptophan, tyrosine, histidine and phenylalanine
  residues
• The free dye in solution is in the cationic form,
  absorbance maximum at 465 nm (red)
• Coomassie binds to residues in the anionic form,
  absorbance maximum at 595 nm (blue) measure in
  a a spectrophotometer
• Manufacturer of kit claims LOD 1ppm protein
• http//www.piercenet.com/resources/browse.cfm?fldI
  D45E20F41-58C0-48FF-8DE1-384477718DB1
• Bradford, M. M. (1976) Anal. Biochem. 72, 248

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Initial feasibility study

• a garlic sauce product was manufactured
• Rinse water samples were taken during the
  Cleaning-in-place (CIP) after production
• 1. Pre-rinse
• 2. Pre-rinse
• 3. Pre-rinse
• 4. Cleaning liquid
• 5. Post-rinse

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Protein Detection on Surfaces using Coomassie
Method

• For equipment or utensils which are cleaned
  manually
• For these experiments, metal plates were
  contaminated with a bovine serum albumin solution
  and dried with air in a sterile hood
• The plates then contained amounts of protein
  from 0 to 25 µg. The plates were swabbed using
  standard microbiological swabs. The swabs were
  put in sterile water and vortexed for 10 seconds.
  after which 1 ml of the water was assayed for
  proteins in the Coomassie test.

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Protein Detection on Surfaces using Coomassie
Method

• Results
• Protein was detected to a level of approx. 2- 5
  µg/mL (2-5ppm)
• Swabbing methods in general are not as precise
  as the liquid sample method.
• Factors that affect the reproducibility of the
  swab method are differences in the area swabbed
  and the swab technique used.

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Problems encountered with Pierce Coomassie Plus
kit as purchased

• Problems with method that accompanied the kit
• Blanks and lowest standard 2.5ppm both gave
  results lt2.5ppm
• Needed to improve the method to give larger
  differences in absorbance values for the standard
  curve, particularly between zero and the lowest
  standard

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Suggested modifications to method accompanying
Pierce Coomassie Plus kit

• Sample preparation centrifuge samples
• Use of disposable semi micro cuvettes
  increasing the path length from 5mm to 10mm
  (increasing sensitivity)
• Analyse all samples, standards and controls in
  duplicate
• Sandwich order of analysis standards,
  controls, samples, controls, standards
• Accounts for drift of absorbance over time

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Suggested modifications to method accompanying
Pierce Coomassie Plus kit

• Control materials - inclusion of appropriate
  control standard eg. skim milk powder
• Dedicated laboratory area for allergen testing to
  reduce possibility of cross-contamination
• Skilled technicians
• Limit of Detection improved method to quote
  results as lt1ppm protein in rinse water
• Conclusion valid method to test cleaning
  efficacy

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Wet cleaning validation
Consider May Contain labelling
Is wet cleaning used?
no
yes
Use Coomassie Protein test method to validate
cleaning method
Are protein residues detectable (gt1ppm) after
cleaning?
yes
Modify cleaning method
no
Cleaning method adequate
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Sample cleaning validation matrix
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Validation and verification

• Validation
• Check that the cleaning method is adequate for
  purpose
• Remove allergens

• Verification
• Check that the validated cleaning method is being
  implemented
• Records signatures of those responsible
• Training
• Recorded corrective action
• Before start-up checks eg. CCP

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Suggested cleaning check list eg. CCP

• Documented cleaning method has been followed - ?
• Final rinse water samples visually clear, pH in
  normal range ?
• Visual inspection of equipment no visible
  residue (include photos, hard-to-reach areas) ?
• Line approved for use signed ?

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Where we are now

• Heinz European manufacturing sites using
  Coomassie to validate CIP wet cleaning
• As a result of the cleaning validation project
• Confirmed many existing cleaning protocols
  suitable for removing protein
• Continuous improvement of cleaning methods for
  products
• Developing in-house swabbing method for manually
  cleaned equipment

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Thankyou

• Thanks to Gern Huijberts (Holland), Brian Walsh
  (UK), Andrea Gorga (Italy), Sinead MacGuinness
  (Ireland) for their input and hard work.

Louise Lennard European Allergen Controls Project
Manager HJ Heinz Company UK Hayes Park Middlesex
UK UB4 8AL Louise.lennard_at_uk.hjheinz.com Ph 44
20 8848 2283
This information is intended as a guide only.
Whilst the information it contains is believed to
be correct, it is not a substitute for specific
advice. Heinz can take no responsibility for the
actions taken based on information in this
presentation.