Validation & Biomanufacturing

What is Validation
Validation – An Essential Part of GMPs!

Validation is the scientific study of a system

 To prove that the facility/system/equipment/method is consistently doing

what it is supposed to do (i.e., that the process is under control).

 We want to make decisions based on good science and not hunches and
assumptions!

 To determine the process variables and acceptable limits for these variables,
and to set-up appropriate in-process controls.

 Is it ok if the wash from a chromatography column is pH 6.8 vs. 7.0 ?

FDA definition of validation
“Validation is a process of demonstrating, through
documented evidence, that a process, procedure,
method, piece of equipment, or facility will
consistently produce a product or result that meets
predetermined specifications and quality
attributes.”
Quality Attributes Remember these?
Identity
 21 CFR 211.84 (d) at least one test shall be conducted to verify the identity of each component of a drug product.
 Chemical, biological, Immunological
 Raw materials, In-process intermediates, final products.
 Safety
 21 CFR 600.3 (p) safety as the relative freedom from harmful effect to persons affected, directly or indirectly, by a
product when prudently administered, taking into consideration the character of the product in relationship to the
condition of the recipient at the time.
 Activity of active ingredients
 Activity of the excipients or additives
 Activity of process related impurities
 Efficacy
 Effectiveness of the product in achieving its medicinal purpose (therapeutic, prophylactic, diagnostic). Gathered at
phase II and Phase III trials.
 Potency
 21 CFR 600.3 (s) specific ability or capacity of the product, as indicated by its appropriate laboratory tests or by
adequately controlled clinical data obtained through the administration of the product in the manner indicated to
effect the given result.
 Purity
 21 CFR 600.3 (r) relative freedom from extraneous matters in the finished product, whether or not harmful to the
recipient or deleterious to the product.
 Cleaning Procedures
 Stability
 21 CFR 211.137 (a) to assure that a drug product meets applicable standards of identity, quality, and purity at the
time of use; it shall bear an expiration date determined by stability testing. Drugs may use accelerated time studies,
biologics must use real time studies.
 Consistency
 The ability of the product and/or process to reliably possess specified quality attributes on an ongoing basis. 3
consecutive batches of product meeting predetermined specifications is accepted as proof that a process is
consistent. However, in NDA data from up to twenty batches may be submitted.
 Biomanufacturing

 Biomanufacturing is a complex process involving multiple unit

operations many of which are critical to insuring patient safety and
product efficacy
 Block Flow Diagram of a typical Production Process
Seed Production
Innoculum Fermentati Fermentati Harvest
on on

UPSTREAM

DOWN- Ultrafiltrati Ultrafiltrati

STREAM Chrom. 1 Chrom. Viral
on on
1 2 Filtration
1 2

VIRAL

NON-VIRAL
Ultrafiltrati Final
Chrom.
on Formulation/
3
3 Sterile Filtration

Sterile Fill
Historical Basis for Validation
 Assumptions concerning virus inactivation resulted in ten
deaths and 200 children becoming paralyzed, from a
supposedly “inactivated” polio vaccine.
 Assumptions about sterilization caused severe infections
among burn victims given supposedly sterile solutions.

 Validation eliminates assumptions and relies on

experimental proof!
 Quality by Design
 A central concept in quality is that quality can not be tested for.
Quality must be designed and built into the production process.
 Requires careful attention to raw material specifications, in
process material specifications, and final product specifications.
 Validation and Quality
 Validating the performance of unit operations, analytical
methods, and critical process points (sterilization, viral
inactivation, cleaning procedures) is essential in insuring that
the process generates a quality product.
Validation in Biomanufacturing

 Validation does not replace testing, but it does reduce the

testing burden for raw materials, in-process materials, and
final product
Validation in Biomanufacturing
 Validation itself is a process that evolves with the
product.
 Validation requirements for production of pre-clinical
material less stringent then for phase III clinical
material.
 Critical operations must be validated: For example:
raw materials, analytical methods, viral clearance,
sterilization, cleaning.
Validation in Biomanufacturing
 A fully validated process is “locked in”
 Any change outside of the validated space invalidates process
 Change must be evaluated for effect on patient safety and
product efficacy-Change control !

Validated Production Process

 Difficulty in Changing and Binding Authority

More
LAWS

Regulation
s

Guidance

Less

Scientific Content & Detail about Implementation

Less

LAWS

Regulation
s

Guidance
More
Regulatory requirement for validation

21 CFR 211 Subpart H- Holding and Distribution

 211.165 – Testing and release for distribution

 “Requires that the accuracy, sensitivity, specificity, and

reproducibility of test methods employed by the firm
shall be established and documented. Such validation
and documentation may be accomplished in accordance
with 21 CFR 211.194 (a)(2)”
Regulatory requirement for validation
 Sec. 211.113 Control of microbiological contamination.
 (a) Appropriate written procedures, designed to prevent
objectionable microorganisms in drug products not required to be
sterile, shall be established and followed.
 (b) Appropriate written procedures, designed to prevent
microbiological contamination of drug products purporting to be
sterile, shall be established and followed. Such procedures shall
include validation of any sterilization process.
What does “validation of any
sterilization process” mean ?
 What parameters are critical to sterilization?

 Temperatures, pressures, time, pore size (filtration), radiation

dosage, chemical concentration.

 Must demonstrate that your autoclave reaches the temperatures,

pressures, and times necessary for sterilization.
 Must demonstrate that items representing real world samples achieve
those conditions ( 20 ft of 1 ½ hose; a 20 L carboy; a 500 ml bottle).
 Must challenge with worse case scenario (may take place in pilot
plant if scalability demonstrated).
Regulatory guidance on validation
 Guideline on General Principals of Process Validation
http://www.fda.gov/cder/guidance/pv.htm
 Guidance for Industry: For the Submission Documentation for Sterilization
Process Validation in Applications for Human and Veterinary Drug
Products. CDER CVM November 1994.
www.fda.gov/CDER/GUIDANCE/cmc2.pdf
 Working Party on Control of Medicines and Inspections
 Final Version of Annex 15 to the EU Guide to Good Manufacturing Practice
 Title: Qualification and validation
 http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-en/v4an15.pdf
 ICH Q7a Section 12 on validation
 http://www.fda.gov/cder/meeting/ICH_Q7A/index.htm
 A WHO guide to good manufacturing practice (GMP) requirements. Part 2:
Validation
 Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997. World Health
Organization, Geneva.
Critical Operations in
Biomanufacturing
 Some operations are more critical than others.
 Viral filtration, sterilization, cleaning, analytical methods.

 These operations will require greater validation efforts then less

critical operations (media blending).
Testing
 Usually done by the Quality Control Laboratory

 CFR requires that quality unit be under independent

supervision and report directly to senior management
Quality Assurance
 Reviews records from quality control and production departments
 Verifies that all specifications and production operations met /
performed
 Investigations necessary for any deviations
 Root cause
 Affect on quality
 Corrective action (CAPA)
 Approves final release of product
Designing Quality into the Product
 Design of production process and specifications all contribute
to a quality product:
 Absence of contamination

 Clean rooms, closed systems, use of BSC for critical

operations.

 Purity
 Separation process (chromatography) designed to
remove potential contaminants
 Viral purification / inactivation
Validation Plan
 Organizations must define an approach towards
validation

 What is to be validated
 How is it to be validated
 Who is to validate it
 Who is to approve the validation
 When it must be revalidated
Validation
Examples of individual
systems subject to validation:

HVAC systems
Autoclaves
pH meters
Depyrogenation Ovens
Lyopholyzers
Centrifuges
Steam generators
Water systems
Compressed air systems
Vacuum systems
Validation Plan
 Regulatory agencies (FDA, EMEA, WHO, etc) identify minimum
components of validation.

 “Industry standards” (the c in cGMP) can increase validation

requirements.

 New & Novel processes / equipment require greater scrutiny then

established processes / equipment.

 Validation requirements increase as a product moves through

development (phase I, phase II, phase III).
Validation Plans

The Validation Master Plan

 A high level document that outlines the organizations

philosophical approach to validation and revalidation. The
master validation plan becomes a guideline by which individual
validation protocol are developed and implemented.

 May contain a flow chart or other diagram of the validation

process
A prospective validation study
IQ

Calibration

OQ

PQ protocol
approval

PQ protocol
execution

Data
Analysis

Validation
Report

Approve
Conclusions
 Developmental studies

 Experiments designed to explore and define the limits of the system to be

validated
 Sterilization developmental studies may focus on “worst case ” or
hard to sterilize items
 Cleaning developmental studies may focus on “worst case” or hard to
clean items
 Analytical methods may focus on defining the limits of the procedure
(range, recovery, etc)
 Developmental studies then used to develop validation protocols and
refine SOP’s
Validation Protocol
 Specific protocols (SOP’s) that provide detailed information on what is to
be validated.

 Validation Protocols consist of:

 A description of the process, equipment, or method to be validated.
 A description of the validation method.
 A description of the sampling procedure including the kind and
number of samples.
 Acceptance criteria for test results.
 Schedule or criteria for revalidation.
Example of a protocol for the
IQ component of validating a
pH meter

As with all other SOP’s this

document will contain an
Objective, scope, and responsibility
Section.
Validation Protocol

 Validation Protocols may consist of multiple SOP’s each

describing specific steps in the validation process
Critical Systems

 How critical is the system being validated to final product quality?

 Media blending systems for cell growth vs. final fill & finish
operations

 Demonstrating that the device which fills, labels, and caps the
final product will require more extensive validation then the
blenders used to prepare media for bioreactors.

 Validation of complex devices may take years!

Validation
 Proceeds in stages with new facilities / equipment.

 Planning for validation should start with the design

process.

 Leaving validation to the last minute is asking for

trouble.
Stages of Validation
 Starts with Design & Receipt:
 Does the equipment meet the needs (is the autoclave big
enough?)
 Do you have the manuals, spare parts, can you plug it in?
 Is it installed properly (drain lines, vents, etc)
 Does it work?
 Does the autoclave reach the necessary temp. and pressure?
 Can the autoclave sterilize your equipment (worse case
situation)?
 How does it work in the manufacturing process?
 Can it handle production quantities?
 Will failure compromise product quality?
 IQ, OQ, PQ ?
Installation Qualification (IQ)
A process used to document that the piece of equipment was supplied and
installed properly and that appropriate utilities, i.e., electrical, steam, gas,
etc. are available to operate the equipment according to the manufacturers
specifications.

Operational Qualification (OQ)

A process designed to supply the documented evidence that a piece of
equipment operates as it is intended through all anticipated operational
ranges.

Performance (Process) Qualification (PQ)

Verifies that a process / piece of equipment performs as it is intended to in
the manufacturing process and produces product (in process or final)
meeting predetermined specifications.
Typical information in an IQ protocol
 Name and description of equipment, including model numbers
 Identification, including model and serial numbers
 Location of the equipment
 Any utility requirements, i.e. electrical voltage, steam or water
pressure, etc.
 Any safety features of the equipment, including alarms, interlocks,
or relief valves.
 That all documentation, including manufacturers contact
information, spare parts inventory, operational manual, and
installation drawings are available on site.
Typical OQ Protocol Components
 Objective
 Responsibility
 Equipment required (Calibration verification & Traceability)
 SOP(s) used
 Equipment Identification
 Parameters measured (Specifications)
 Documentation
Validation
 Ideally validation takes place prior to actual production runs,
however in some cases validation may take place as product
is produced, or past production runs may be used to provide
validation data.

 Prospective Validation

 Concurrent Validation

 Retrospective Validation
Revalidation
 Is the initial validation of a piece of equipment the end?
 No!
 Periodic revalidation may be necessary depending on the criticality
of the equipment
 Changes need to be evaluated for their impact on validation
 Deviations from specifications may require revalidation
 Revalidation spelled out in Master Validation Plan
Change Control
 Must assess impact of changes on FDA compliance and validation
state.

 Change control is a formal process defined in company SOP on how

process/equipment changes are evaluated.

 Any change that takes place outside the change control process can
jeopardize product quality (patient safety).
References
 Pharmaceutical Manufacturers Association’s (Pharmaceutical Research and Manufacturers of America) Validation
Advisory Committee “Process Validation Concepts for Drug Products” Pharmaceutical Technology, September 1985 p 82.
 Bismuth, G. Cleaning Validation: A Practical Approach. CRC Press, 2000. ISBN 1574911082.
 Pharmaceutical Process Validation, 3rd Ed. Edited by Robert Nash and Alfred Wachter, Marcel Decker, 2003. ISBN
082470838-5
 Validation of Pharmaceutical Processes: Sterile Products. 1998. 2nd Edition. Edited by Frederick J. Carlton and James
Agalloco. Marcel Decker, 1998. ISBN 0824793846.
 Validation Standard Operating Procedures: A step by Step Guide for Achieving Compliance in the Pharmaceutical, Medical
Device, and Biotech Industries, Syed Imtiaz Haider, St. Lucie Press, 2002. ISBN 1574443313.
 Good Manufacturing Practices for Pharmaceuticals: A Plan for Total Quality Control From Manufacturer to Consumer,
Sidney J. Willig. Marcel Decker, 2000. ISBN 0824704258.
 Voss, J. Cleaning and Cleaning Validation: A Biotechnology Perspective. CRC Press, 1995. ISBN 0939459507.
 LeBlanc, D.A. 2000. Validated Cleaning Technologies for Pharmaceutical Manufacturing. CRC Press. ISBN 1574911163.
 Cloud, P. 1998. Pharmaceutical Equipment Validation: The Ultimate Qualification Guidebook. CRC Press. ISBN
1574910795.
 Juran, Quality Control Handbook, 4th Edition., McGraw-Hill, 1988.
 DeSain C, Sutton C. (1995). Process development that supports process validation. Pharmaceutical Technology 19 (Oct.):
130-136, 1995.
 Garcia T, Wilkinson S, Scott J. The development of a blend-sampling technique to assess the uniformity of a powder
mixture. Drug Development and Industrial Pharmacy 27(4): 297-307, 2001.
 Chaloner-Larsson, G., Anderson, R., Egan, A. 1997. A WHO guide to good manufacturing practice (GMP) requirements
Part 2: Validation . World Health Organization, Geneva. www.who.int/vaccines-documents/DocsPDF/www9666.pdf
Accessed on October 2nd, 2006.
 Brown, F. 1993. Review of accidents caused by incomplete inactivation of viruses. Dev. Biol. Stand. 81: 103-7
 Nathanson, N. and Langmuir, A.D. 1995. The Cutter incident. Poliomyelitis following formaldehyde-inactivated poliovirus
vaccination in the United States during the Spring of 1955. II. Relationship of poliomyelitis to Cutter vaccine. 1963. Am. J.
Epidemiol. 142:109-40.
Autoclave Validation
 IQ-
 Design specifications meet users needs
 Proper installation, utilities, manuals, spare parts
The DQ, IQ, OQ process
insures that this autoclave
will meet the needs of the
manufacturing group.
Sample Format for Installation
Qualification of an autoclave.
Courtesy of WHO. Chaloner-Larsson, G.,
Anderson, R., Egan, A. 1997. A WHO guide to good
manufacturing practice (GMP) requirements Part 2:
Validation .
World Health Organization, Geneva
. www.who.int/vaccines-documents/DocsPDF/www9666.pdf
Accessed on October 2nd, 2006.
Autoclave Validation
 OQ
 Does it operate properly
 Does it reach the specified temperature and
pressure
 Do timers work
 Does the operator interface panel work
 Are safety interlocks functional
Sample Format for Operational
Qualification of an autoclave.
Courtesy of WHO. Chaloner-Larsson, G.,
Anderson, R., Egan, A. 1997. A WHO guide to good
manufacturing practice (GMP) requirements Part 2: Validation .
World Health Organization, Geneva
. www.who.int/vaccines-documents/DocsPDF/www9666.pdf A
ccessed on October 2nd, 2006.
Calibration

Figure 1: Operational
qualification of an
autoclave requires the
calibration of instruments
against traceable
standards. This figure
shows the calibration of a
validator (out of view)
against the IRTD probe.
Monitoring Temperature
Figure 2: A validator, used in the
operational qualification of an
autoclave. The validator is
attached to individual
thermocouples by wires coming
from the rear of the instrument
(arrow). Tha validator has been
previously calibrated and the data
gathered from the thermocouples
will be logged on the laptop
computer. The software on the
computer is also subject to
validation requirements.
Figure 4. The validator is
attached to individual
thermocouples (TC) by thin
wires that pass through the
wall of the autoclave
through a specially
designed port (arrow). This
picture shows the back side
of the autoclave. The
validator is out of view at
the lower left.
Figure 5. The inside of the
autoclave showing the maze
of wiring connecting the
individual TC’s to the
validator. The port through
which the wires pass is
visible in the middle left of
the picture. The individual
TC’s will be placed in various
areas of the autoclave or
equipment being autoclaved
to generate a thermal map of
the interior of the autoclave.
Figure 3. Output from the
validator. The temperature at
each connected thermocouple is
displayed. Accumulated lethality
(F 0 ) may also be displayed.
Notice how some TC have failed
and will not record a
temperature. Accounting for TC
failure is necessary to keep from
having to repeat a study.
Figure 6. Proving that sufficient
lethality (F0) is achieved within the
nooks and crannies of
biomanufacturing equipment
requires the placement of TC’s in
hard to reach areas, in this case
deep within a piece of tubing.
Special gaskets with openings for
the TC are used to insert the TC
within pieces of equipment.
Developmental studies
 Experiments designed to explore and define the limits of the
system to be validated
 Sterilization developmental studies may focus on “worst
case ” or hard to sterilize items
 Cleaning developmental studies may focus on “worst case”
or hard to clean items
 Analytical methods may focus on defining the limits of the
procedure (range, recovery, etc)
 Developmental studies then used to develop validation protocols
and refine SOP’s
Developmental Studies

Developmental studies are

used to identify hard to
autoclave items and to
test if item preparation has
an effect on ability to be
sterilized
Developmental Studies

Every little nook and cranny

Needs to be assessed ! We
have to prove that the inside
of the pipe reaches sufficient
temperature, for a long
enough time to insure sterility!
 Developmental Studies

Does bagging make a difference?

Cleaning Validation

Validating the cleaning cycle

on a loaded dishwasher.
Notice the various pipes and
parts with narrow openings.
Identification of hard to clean
and easy to clean areas starts
with developmental studies
How do we validate a dishwasher ?
 How do we identify hard to clean & easy to clean items?
 How do we test to see if they are cleaned?
Going over the documentation
 Cleaning Validation

Sampling, documenting, and

verifying is a labor intensive
process

Why is cleaning considered a

critical process?
Sampling
Figure 8. Swabbing is
commonly used to sample the
surface of cleaned materials.
The swab is then placed in a
sample vial and sent to the
Quality Control lab for
analysis. Proper technique is
essential in order to evaluate
the effectiveness of cleaning
techniques. Even so,
swabbing results are typically
corrected for known
deficiencies in recovery.
Cleaning Validation
Figure 9. Analysis of rinse water
for residual cleaning agents or
process materials is an essential
component of cleaning validation.
Insuring that the sample is not
contaminated requires vigilance
and properly following the
relevant SOP’s.
Analytical methods validation
 Considered a critical step in the manufacturing process
 Requirements for validated analytical methods explicitly written into
the CFR’s

 211.165 – Testing and release for distribution

 “Requires that the accuracy, sensitivity, specificity, and reproducibility of
test methods employed by the firm shall be established and documented.
Such validation and documentation may be accomplished in accordance
with 21 CFR 211.194 (a)(2)”
Testing For Identity

 Requires the development of validated analytical methods that can

determine identity.
 Chemical Tests:
 Is the molecule chemically what it is supposed to be?
 Biological Activity Tests:
 Does the molecules have the biologic activity that it is supposed to
have?
 Immunogenic Tests:
 Is the molecule immunogenic (allergic)?
Identity
 21 CFR requires testing of raw materials:

 Raw materials quarantined until identity verified

 Raw materials must meet predetermined specifications
 Vendors (and alternates) specified in BLA (NDA)
Identity

 21 CFR requires testing of in-process materials:

 Product from bioreactor / fermentor
 Product from purification steps
 Waste products from above

Must meet specifications, if not - stop the

process to investigate take corrective action
Regulatory guidance on validation
 Guideline on General Principals of Process Validation
http://www.fda.gov/cder/guidance/pv.htm
 Guidance for Industry: For the Submission Documentation for Sterilization
Process Validation in Applications for Human and Veterinary Drug
Products. CDER CVM November 1994.
www.fda.gov/CDER/GUIDANCE/cmc2.pdf
 Working Party on Control of Medicines and Inspections
 Final Version of Annex 15 to the EU Guide to Good Manufacturing Practice
 Title: Qualification and validation
 http://pharmacos.eudra.org/F2/eudralex/vol-4/pdfs-en/v4an15.pdf
 ICH Q7a Section 12 on validation
 http://www.fda.gov/cder/meeting/ICH_Q7A/index.htm
 A WHO guide to good manufacturing practice (GMP) requirements. Part 2:
Validation
 Chaloner-Larsson, G., Anderson, R., and Egan, A. 1997. World Health
Organization, Geneva.
Validation Protocol
 Specific protocol based on developmental studies
 Protocol is written, reviewed and approved
 Protocol is executed
 Report written and approved
 System is validated
Analytical Methods Validation
 Being a critical component of production
process analytical methods must be validated
 Raw material testing
 In process materials
 Final product specifications
What must be demonstrated
 Selectivity (specificity)
 Accuracy
 Precision
 Linear Range
 Limit of detection (LOD)
 Limit of quantification (LOQ or LLOQ)
 Robustness
Validated methods
 USP NF (United States Pharmacopeia National Formulary) contains
“validated analytical methods”

 Use of a USP method does not eliminate the organizations

obligation to demonstrate that the method performs adequetly
Selectivity (specificity)

 Does the analytical method detect the component it is supposed

to detect?
 Cross reactivity in antibody based methods

 Demonstrate specificity by conducting analytical method on

materials that may mimic analyte of interest
 Looking for “false positives”
Accuracy
 Ability of analytical method to accurately determine the
presence and amount of the analyte of interest
 Typically done by analyzing a traceable standard
Recovery

 Can we recover all of the analyte from a complex matrix

 May reflect sample preparation problems
 Typical recovery studies done using “spiked” samples
 Final results may be corrected by the % recovery
 Swab samples typically corrected to reflect recovery
Linear Range
 Must define the linear range of a method
 Assay may have multiple linear ranges
Precision

 How much variability does the assay exhibit when analyzing

the same sample
 Typically demonstrated by analyzing multiple aliquots of a
homogenous sample
 Acceptance criteria will depend on the assay and the
material being assayed (2-20% RSD)
 Typically expressed as % RSD (relative standard deviation)
Limit of Detection (LOD)
 Lowest level at which method can detect analyte
 Results reported as less than LOD
 Based on signal to noise specification (10:1, 20:1)
Robustness studies

 How sensitive is the method to minor variations in method

 Pipetting variation
 Temperature fluctuation
 Reagent stability
 Etc.
Detailed robustness studies will be reflected in final SOP’s
Limit of Quantification (LOQ)
 Lowest level at which method can accurately quantify analyte
 Based on signal-to-noise ratio specification (10:1, 20:1)
and precision specification
 Precision and LOQ related
 Lower LOQ will typically result in lower precision
Bradford Assay
for total protein

Well known colorimetric assay that relies on the binding of

Commassie G-250 dye to the proteins in an acidic solution
 Dye binding proportional to number of positive charges in
protein
 Proteins >3000 dal not detected
 Simple, quick, wide range, few interfering agents
Bradford Assay
Disadvantages
 Incompatability with surfactants
 Staining of glass and quartz cuvettes
 Use disposable polystyrene cuvettes
 Or wash with strong detergents and methanol
Validating the Bradford
 Selectivity (specificity)
 Accuracy
 Recovery
 Precision
 Linear Range
 Limit of detection (LOD)
 Limit of quantification (LOQ or LLOQ)
 Robustness
Some Questions
 A valve used to transfer material from a holding tank to the purification suite
jam’s closed. You have a spare valve that is an identical model. Can you change
this valve with the spare and continue operations? What if the valve is from a
different manufacturer?

 You notice that your autoclave loading plan leaves room for additional material.
Realizing that increasing that amount of material in the autoclave will shorten
the turn around time for the production line you contemplate increasing the
amount of material loaded into the autoclave then specified by the loading plan.
What should you do? What will be required to implement this change?

 An SOP for calibration of a pH meter calls for a two point calibration at pH 4

and pH 7. You notice that a single point calibration at pH 7 produces the same
result from pH measurements of your buffer solutions and allows you to take a
longer break. Is it Ok to do the one point calibration when the SOP calls for a
two point calibration? How would you go about changing the SOP to allow for a
one point calibration?
 What documents would provide information concerning the make and
model of a particular valve used to regulate the transfer of material from a
holding tank to the purification suite?

 Your supervisor is concerned that the fermentation vessel is not providing

sufficient aeration of the culture to get optimal growth and suggests
installing a different kind of baffle in the vessel. How would you
demonstrate that this change has no effect on product quality?