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Guidelines for Investigating & Assessing Product

Endotoxin Failures

Richard Fitzpatrick & Aoife Brady

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Content

• Summary of Irish Pharmaceutical/Medical Device Industry

• Background on Endotoxin

– What are endotoxins

– Where endotoxins are found

– Physiological effect of endotoxin on the body

• Regulatory requirements for endotoxin testing of medical devices

– AMI-ST72, USP 85, USP 161

• Implications of endotoxin Failures

– Patient Perspective

– Business Perspective

• Guidelines for Investigating Product Endotoxin Failures

– 5 Step guide for conducting product endotoxin investigations

– Examples of Product Endotoxin Root Causes

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• Over 120 pharma companies base in Ireland

• Nine of the top 10 largest pharma companies base

in Ireland

• Pharma sector worth 50.8 billion

• Employs over 25,000 people directly,

100,000 people indirectly

• Over 180 medical device companies base in Ireland

• Eleven of the of the top fifteen largest medical device

companies base in Ireland

• €8 billion industry supplying cardiovascular,

orthopaedic & diagnostic devices

• Employs over 27,000 people directly

Employs over 100,000 people indirectly

Summary of Irish Pharmaceutical/Medical Device Industry

Synergy Lotus

Wallstent

Cardiology

Peripheral

Intervention

Mustang

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Summary on Bacterial Endotoxins

• Endotoxins are components of the cell wall of Gram negative bacteria

• Specifically endotoxins are the lipopolysaccharides in the outer membrane of Gram negative bacteria

• The primary source of Gram negative bacteria for medical devices is water to which the product is exposed

• Upon entering the body endotoxins can induce septic shock and can be fatal

• Endotoxins are not destroyed by sterilization therefore control & prevention of endotoxin contamination is central to advoiding endotoxin issues

• Estimated that 1 x105 Gram negative bacteria equivalent to 1 EU

To exceed 20 EU in excess of 2 x107 Gram negative

bacteria would have to contaminate the device

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Physiological effect of endotoxin exposure

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Implications of product endotoxin failures

• Patient Perspective

• Upon entering the body endotoxins

can induce…..

– Septic shock

– Fever

– Organ Failure

– Death

• Business Perspective

• Endotoxin failures including

inadvertent release of contaminated

product for human use or inadequate

investigations can lead to …..

– Product Recall

– Back Orders

– Lost of earnings from scrappage of

impacted batches

– Penalties from competent authorities

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Examples of FDA 483s for Product Endotoxin Issues

Case Study 1

Company: Sanquin: Holland

Plasma Products

44 Batches Rejected

Reason

• Insufficient Investigation

• Investigation concluded alteration in cleaning process was source of endotoxin

• All tests on equip were negative for both endotoxin and bioburden

• Investigation failed to link a leak as the possible source of the endotoxin contamination

• No additional investigation for subsequent batches that had endotoxin alert levels

Case Study 2

Company: Sanofi Pasteur: France

Vaccine Products

13 batches rejected

Reason

• 13 batches failed over a 4 year period and no root cause identified

• A component was identified as the root cause but was only used in 4 of the impacted batches

• The impacted component was continually used in subsequent batches

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Regulatory Requirements for endotoxin testing

• All companies have a

responsibility to access their

products for routine endotoxin

testing

• As per AMI ST72 Products

that have direct or indirect

contact with vascular,

neurological or ophthalmic tissue

require routine endotoxin testing

Interventional

Cardiology

Cardiac Rhythm

Management

Urology

Neuromodulation

Electrophysiology

Peripheral

Intervention.

Endoscopy.

Watchout: FDA update to its 510(k) Sterility Review Guidance document 2016

calls out for endotoxin testing for all implants

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• Validated method for product endotoxin testing: AMI-ST72, USP161, USP85

• Rational for endotoxin testing of product Pre vs Post sterilization: AMI-ST72, USP161

• Batch Definition: AMI-ST72

• Routine product endotoxin sampling plan: AMI-ST72

• Control/Link Batch Release with Product Endotoxin Results AMI-ST72

• “Conduct an adequate & timely investigation” AMI-ST72

What scale of investigation satisfies an adequate investigation ???

Regulatory Requirements for Endotoxin Sampling Plan

Batch Size # of samples

<30 2

30-100 3

>101 3% of lot, up to

maximum of 10

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Responding to Product Endotoxin Failures

“Production”

The batch needs

to be released today !!

“Product Release”

We are going into

back order !

Release the batch?

We havent even started

the investigation !!

Do we have an Investigation Plan…..????????????

HELP

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Investigative Endotoxin Testing to Identify

Root Cause

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Five Step Guide for Investigating Endotoxin Failures

1. Conduct Lab OOS and confirm endotoxin result

2. Conduct initial investigation

– Assemble investigation team

– Contain product

– Assess impact to subsequent batches

– Conduct a line walk of the production process

– Apply Investigation tools such as

• Is/Is Not – Helps define the scope of the problem

• Fish bone diagram – helps to identify root causes of the endotoxin failure

3.Conduct review of endotoxin controls

– Product Bioburden Trends

– Product Endotoxin Trends

– Cleanroom Environmental Trends

– Process solutions that come in contact with product

– Change of suppliers/production staff or manufacturing process

– People behaviour/GMP/cleaning

4. Conduct investigative endotoxin testing to identify root cause

5. Evaluate investigative results and determine product disposition strategy

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BET Assay Assessment Flow Chart

Assay Criteria met %CV standard (<10%)

Slope/R2 >0.98

Blank reaction<

Lowest standard

Sample criteria met

%CV of sample <20%

Spike recovery 50-200%

Endotoxin

result within

specification

Test Passes

Result OOs

Test Fails

OOS investigation

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Laboratory Error – Potential Causes

OOS Result

Negative Controls Ok

Correct procedure?

Training Records

Any unusual events

during testing?

Contamination in other test?

Glucans?

Technician Error ?

Change in product formulation?

Test equipment

Test reagents

Test consumables

Contamination during dilution

• Retest originals

• Retest new samples

Sample

• Correct sample?

• Sampled correctly

• Condition

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Five Step Guide for Investigating Endotoxin Failures

1. Conduct Lab OOS and confirm endotoxin result

2. Conduct initial investigation

– Assemble investigation team

– Contain product

– Assess impact to subsequent batches

– Conduct a line walk of the production process

– Apply Investigation tools such as

• Is/Is Not – Helps define the scope of the problem

• Fish bone diagram – helps to identify root causes of the endotoxin failure

3.Conduct review of endotoxin controls

– Product Bioburden Trends

– Product Endotoxin Trends

– Cleanroom Environmental Trends

– Process solutions that come in contact with product

– Change of suppliers/production staff or manufacturing process

– People behaviour/GMP/cleaning

4. Conduct investigative endotoxin testing to identify root cause

5. Evaluate investigative results and determine product disposition strategy

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Schematic representation of production batch where

all test units pass endotoxin testing and batch is released

• Batch consist of 60 lot

• Each lot consist of 20 units

• Batch period is defined as a weeks production Mon-Fri

• Batch consist of 1200 units estimated value 250K

• As per AMI ST72 ten units are required for endotoxin testing and batch release (batch>100 units)

• 2 units are randomly pulled on each day of production. Randomly selecting 2 units from each day of production improves the power of the sampling plan

• In this scenario all units passed endotoxin testing and batch can be released

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Schematic representation of production batch where two units selected

from Wednesday production failed endotoxin testing and batch is

contained

• Two units selected from

Wednesday’s production failed

endotoxin testing. Batch is

contained

• All other units selected throughout

the batch passed endotoxin

testing

• Initial assessment of results

suggests endotoxin failure is not

systemic throughout the batch

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Guidelines for conducting investigative endotoxin

testing to determine root cause

• Hypothetical situation: 2 units from lot 32 failed

batch release endotoxin testing

• Step1: Test 10 additional units for endotoxin from

the impacted lot to determine if the root cause of

the endotoxin failure is systemic within the lot

• Step 2: Test 10 additional units for endotoxin from

lots either side of the impacted lot ie lot 31 and lot

33 to determine if root cause is confined to lot 32

or is more systemic in nature

• Step 3: Test 2 units from each of the 60 lots to

determine if root cause is systemic throughout the

batch or is confined to a specific

process/component batch

• Note test results obtained from investigative

testing can not be used for batch release.

Pending investigation outcome a 2nd round of

batch release testing is required to enable batch

release

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Case Study 1. All units selected for investigative

endotoxin testing throughout the batch failed

• All units selected for endotoxin

testing failed

• Root cause for endotoxin testing

is systemic throughout the batch

• Manufacturing process is not in

control

• Impacted batch should be

scrapped and manufacturing

ceased until root cause is

identified

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Case Study 2. All units selected from a specific days

production failed endotoxin testing

• All units selected from a Wednesdays production failed endotoxin testing

• All other units selected from the weeks production passed endotoxin testing

• This indicates that root cause for the failure is confined to a specific process / component lot unique to Wednesday.

• Test 10 additional units derived from Monday, Tuesday, Thursday & Fridays production lots for batch release testing

• If the 2nd round of batch release testing passes all units manufactured on Monday, Tuesday, Thursday & Friday can be released. All units manufactured on Wednesday should be scrapped

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Case Study 3. All units selected for investigative

endotoxin testing throughout the batch passed

• All units selected for investigative endotoxin testing throughout the batch passed

• Consideration should be give to whether the two units that failed endotoxin testing in lot 32 are

– Atypical to lot 32

– Atypical to Wednesday production

– Atypical to the entire batch production

• Review all investigative results to establish if the batch can be released on the basis that the two failed units are atypical to the batch

• Conduct 2nd round of batch release testing to disposition batch

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Examples Product Endotoxin Root Causes

• Contaminated Coating soln

– Inproper storage of glassware following washing facilitated formation of Gram

negative biofilim

• Contaminated Component Supply by external vendor

– Water used in the manufacturing process highly contaminated with endotoxin

• Labels used to identify units for endotoxin testing

– Labels manufactured using recycled paper which was highly contaminated with

endotoxin

• Saliva

– Saliva contains high amounts of endotoxin …800 EU/ml

– Hand mouth contact absolutely avoided in cleanroom

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Conclusion

• Five Step Guide for Investigating Endotoxin Failures:

1. Conduct Lab OOS and confirm endotoxin result

2. Conduct initial investigation

3. Conduct review of endotoxin controls

4. Conduct investigative endotoxin testing to identify root cause

5. Evaluate investigative results and determine product disposition strategy

• The goal of this 5 step guide is to facilitate an informed, structured and

timely conclusion to the endotoxin OOS investigation and the fate of the

impacted batch

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Responding to Product Endotoxin failures

An informed & structured Investigation Plan

will facilitate a timely conclusion to the endotoxin

OOS investigation and the fate of the impacted batch

Where do I start ?!