Keith Bader & Kelly Scalva

Translating Laboratory-
Developed Visual Residue
Limits to Process Area
Applications | IVT
Keith Bader and Kelly Scalva

ABStrAct
While considerable attention has been given to the development of
visual residue limits (VRLs) in a laboratory setting, translating the
bench scale values to the assessment of process surfaces has not yet
been thoroughly assessed. However, knowledge of both the critical
parameters that impact the determination of VRLs and the influence of
those parameters on visual inspection can provide a framework for the
development of a robust visual inspection program. Development of
such a program first entails the determination of constraints imposed
by equipment geometries and facility lighting. VRLs can then be deter-
mined for post-productions residues of concern, which, of course, car-
ries its own specific challenges. Once VRLs have been determined, they
cannot be immediately applied without considering certain strategic
cleaning program approaches and potential sources of variability.
Many factors influence how visual inspection will be conducted in a
manufacturing facility. Among the most critical are inspection condi-
tions in the facility, the condition of existing equipment surfaces, and
the physical characteristics of post-production residues deposited on
product contact surfaces. While many in industry embrace the im-
portance of visible residue limits (VRLs), few have a clear pathway to
translate VRLs determined in the laboratory to the manufacturing floor.
The intent of this paper is to provide the background and informa-
tion that will allow industry to formulate a plan of attack to practically
integrate VRLs into the visual inspection program. To begin, the best
starting place is to determine the constraints imposed by the manufac-
turing floor.

EquipmEnt inSpEction conditionS

Conducting a visual residue limit (VRL) determination study in the
laboratory must be completed such that important process equipment
parameters from the manufacturing floor are captured. To wit, the first
step is characterization of the conditions on the manufacturing floor
where visual inspection is put into practice. For any residue, there are
three parameters that must be considered for proper translation and
laboratory analysis: viewing angle, distance, and light intensity at the
product contact surface. The most critical variable for visual inspection
is, of course, the room lighting, so it is important to understand and
characterize lighting in the facility to properly later simulate it at the
bench scale.
The interior task lighting generally conforms to standards set by a
professional organization such as the Illuminating Engineering Society
of North America (IESNA). In the past, “over-lighting” was the norm
with standard luminance regularly reaching 1200 lux (1). Lighting was

Special edition: Cleaning Validation 45

Keith Bader & Kelly Scalva
figure 1: Determination of Viewing Distance and Angles.
also primarily done with less efficient incandescent
and full-spectrum lamps that inherently provided
good color rendering. However, the advent of current
sustainable design, legislated energy conservation
requirements, and project engineers that target seem-
ingly non-critical lighting designs in value engineer-
ing exercises means that manufacturers can no longer
assume that room lighting conditions will be consis-
tent or even adequate for visual inspection. Specifi-
cally, current legislation limits energy use for lighting
to 1.1–2.0 w/ft, depending on the space function,
forcing less experienced lighting designers to select
options that may not provide equivalent performance
to historical lighting choices.
Indeed, Forsyth et al noted that lighting levels can
vary from 400 to 1200 lux in a facility; such a range
is representative of that found in most manufactur-
ing facilities (2). Such variability is often unavoidable
as facility and process layout can change over time
to meet manufacturing needs, leading to shadows or
reflections unaccounted for in the original design.
Accordingly, it is recommended that the lighting be
assessed and recorded through the use of commonly
available light meters to acquire custom-tailored
ranges for developmental VRL studies.
The most convenient way of recording these
numbers is to record them on a layout drawing of the
facility. Furthermore, it is useful to note the style of
46 Special edition: Cleaning Validation
light fixtures and whether the color temperature of
the lamp closely replicates sunlight. Lighting at the
product contact surfaces within process vessels is a
unique and involved set of measurements to acquire,
as the distances and angles from which operators
view the product contact surface is dependent upon
available viewing ports and the differences in tank
configuration, shown in Figure 1. Measurement of
the distances and angles is most reliably performed in
the field using a laser distance meter and clinometers;
however, this information can also be determined
from mechanical drawings of the equipment.
As process equipment is surveyed, it is advisable
to develop a baseline record of known blemishes,
marks, imperfections, and visual obstructions to
ensure that they can be distinguished from post-pro-
duction residues. Establishing this baseline for each
process tank and training inspectors on the location
and appearance of these fingerprint imperfections
are critical to prevent the development of a viewing
bias that may impact the care used when inspecting
process equipment surfaces.
Characterized and appropriately restricted view-
ing angles developed during the survey will allow for
better translation of the VRL from the laboratory to
the floor.
Conditions in areas where manually cleaned
equipment or small parts are inspected after they
are cleaned out of place are important in that many
pieces of manufacturing equipment used for closed
processing operations do not allow for reliance on
ambient lighting and require the specification of an
independent light source for visual inspection. Ac-
cordingly, it is important to determine the maximum
inspection distance based on the size and configura-
tion of vessels and other closed processing equip-
ment. Once this is determined, it can be used not
only as a selection criterion for a light source but also
as one of the constraints for conducting laboratory
VRL studies.
thE SElEction of A light SourcE for ViSuAl
inSpEction
Before conducting the laboratory study or send-
ing technicians into the field to inspect equipment,
it is important to specify a hand-held light source.
Specifying a hand-held or tank-mounted light source
carries some additional considerations. This requires
understanding some lighting design terminology and
heuristics. The hand-held light should be selected
with three primary characteristics taken into con-
sideration: color temperature, color rendering index
(CRI), and light output or luminous flux.
Color temperature, or chromaticity, refers to the
color of the light emitted from the light source. Colors
similar to the warm yellow orange of a candle flame

Source CRI Chromaticity (°K)
Incandescent 100 2800K
Halogen 100 2900K
Fluorescent 62–90 2700–6000K
Mercury 15 5700K
High Pressure Sodium 22 2000K
Metal Halide 65–70 2700–4000K
Pulse Start Metal Halide 65–75 3000–4000K
are assigned a temperature of 1800 Kelvin; up the
scale is the bluish light cast by metal halide and the
fluorescent light sources that are assigned a color
temperature of 4200 Kelvin (3). The ability of observ-
ers to accurately perceive colors is also somewhat de-
pendent on the chromaticity of the light source based
on the second characteristic noted above, CRI.
CRI is quantified on a scale that assigns the light
source a rating from 0 to 100, with the best rendition
of color achieved with a CRI of 100. For detail-orient-
ed physical inspection work and accurate evaluation
of residues on process equipment surfaces, the CRI
of a light source should, at a minimum, be at a value
of 65 or greater (4). The general relationship between
color temperature, CRI, and various light sources is
shown in the Table.
Finally, the light output of the source should be
considered. Many are also tempted to get the bright-
est light they can find; however, this can lead to
problems with detecting residues because excessive
glare can actually inhibit detection. The inspection
of process vessels is thus facilitated by selecting a
tank light or handheld light source that replicates
levels of lighting equivalent to the recommended for
ambient task lighting within the vessel. Most light
manufacturers, however, do not provide the luminous
intensity at various distances from the source. Hand-
held light output, when rated, is often quantified
using lumens out-of-the-front (OTF). On occasion, a
specific intensity may be provided in Lux (lumen/m 2)
for some lamps at a specific distance.
To convert intensity at a given distance to one
matching the distances determined for the produc-
tion equipment in the field, it is important to know
that light intensity follows an inverse square relation-
ship relative to distance, as shown in Figure 2. As
the distance from the light source increases, the light
must diffuse over a larger surface thereby reducing
the surface brightness in accordance with a “one over
d squared” relationship. Alternatively, an inspec-
tion light of appropriate intensity can be determined
empirically in the laboratory by using a light meter
at the appropriate distance. For the most part, an ac-
ceptable CRI and chromaticity are provided in hand-
held light sources by either incandescent or halogen
Keith Bader & Kelly Scalva
figure 2: Light Intensity vs. Distance.
lamps as shown in the Table. Furthermore, a flash-
light with an adjustable spot size can also be used to
vary the intensity and provide greater flexibility over
a range of distances.
lABorAtory Vrl dEtErminAtion StudiES
Translating the angle, distance, and lighting param-
eters from the onsite equipment to the confines of
the laboratory is best examined within a range of
values. Most commonly tested in the lab are three
angles (30°, 45°, 90°), two to three distances (clos-
est and farthest away), a minimum of two different
lighting intensities (standard room task lighting and
the intensity from a hand-held light source), and a
variety of materials of construction (MOC). Assum-
ing the viewing variables were tightly and accurately
measured at the manufacturing site, the only other
test variables to be investigated in the laboratory
are the process soils (in varying concentrations),
materials of construction, and the assessment opera-
tors. As the potential variability in technicians is
the most difficult parameter to control and replicate,
they are discussed in greater detail in a subsequent
section.
ExpErimEntAl dEtAilS And logiSticS
First, coupons composed of materials of construc-
tion representative of process equipment should be
obtained (see Figure 3) and cleaned thoroughly to
remove any possible residue on the coupons; such
as dust, mechanical cutting oils; or, if the coupons
are not new, residue from previous tests. Typically,
a cleaning regimen consistent with United States
Pharmacopeia (USP) <1051> for cleaning glassware
employed for total organic carbon analysis will effec-
tively remove materials that can potentially confound
results. Cleanliness can then be confirmed using the
American Society for Testing and Materials (ASTM)
water break free test (5, 6). Clean coupons are then
rigorously inspected for visual cleanliness, surface
imperfections, water spots, discoloration, or oxidation
before use in the VRL Study. Coupons that appear to
have any noticeable imperfections are rejected from
the study.
Special edition: Cleaning Validation 47
Keith Bader & Kelly Scalva
figure 3: Coupons of Representative MOCs
Following a visual inspection of the coupons to
ensure their cleanliness, the process soils of inter-
est are diluted into a range of concentrations. These
concentration ranges are then spiked with equiva-
lent volumes onto the center of the coupon surface
and allowed to dry. Coupons commonly are dried at
ambient conditions overnight to ensure dryness, but
they can be dried at elevated temperatures depending
on process conditions. A visual target range is then
determined to find the appropriate loading density
and loading solvent for use in VRL testing.
Purified water is the common solvent used to make
the residue dilutions; however, surface tension ef-
fects combined with some process soils can create a
very visible ring around the periphery of the original
spiking area. As water dries at the edges of the spiked
solutions, it concentrates the residue, making them
more visible and quantification of the mass per unit
surface area more difficult. This concentrates the
residue to a greater degree in the laboratory setting
than that found on process equipment. Thus, it is
sometimes nessary to create a dilution mixture that
48 Special edition: Cleaning Validation
figure 4: Determining the Visibility Inflection Point.
includes a mixture of residue, purified water, and
isopropyl alcohol (IPA) to reduce the solution surface
tension and more uniformly distribute the residue
over the spiking area as the liquids dry, thereby
preventing concentration around the outer perimeter
or center of the spiking area. The exact ratios of IPA
to water must be determined on a per soil basis as
well as the solubility because each residue is differ-
ent. Furthermore, chemical interactions or protein
denaturation that alters the visual characteristics of
the soil should also be considered.
Once drying effects have been effectively contend-
ed with, the threshold concentration of each process
soil must be determined before multiple technicians
are screened. Specifically, the approximate concentra-
tion range visibility inflection point, when a residue
goes from visible to not visible, can be determined, as
shown in Figure 4.
Creating the initial spiking solutions in order of
magnitude intervals works well to initially narrow
the range. For example, a serial dilution of 1 ml resi-
due solution in 10 ml solvent followed by 1 mL of the
second solution in 10 mL of water, and so on, allows
a simple way to determine the range of concentra-
tions to be used for the VRL intermediate precision
testing assessment.
To prepare the coupons for testing, a consistent
volume of spiking solution is then spiked onto cou-
pons over a reproducible surface area. The concentra-
tions applied to the coupons span the range of Vis-
ible, Slightly Visible, and Not Visible as determined
in the initial bulk VRL screening exercise described
above. At least three replicates of each concentration
per material of construction should be used to ensure
statistical significance within the results. Four opera-
tors with 20/20 or corrected 20/20 vision are then
selected to establish the VRL intermediate precision
testing at different viewing angles and distances un-
der ambient and additional hand-held lighting.
To better simulate visual inspection conditions
on the manufacturing floor, it is important that the
coupons are placed on a background made of visually
similar material. The visually similar material reduc-
es operator bias during the laboratory study so that
contrast from the viewing background will not be
an issue. Thus, when testing the viewing of different
materials of construction, the background for these
assessments must either be constructed of the same
material as the coupon or must be consistent with the

figure 5: VRL Determination Conditions Based on Manufacturing Area
Assessment.
appropriate viewing parameters employed for the pro-
cess equipment. For example, if the major material of
construction is 316L SS EP20Ra with minor materi-
als such as borosilicate glass (commonly site glasses)
and/or PTFE (primarily tested to simulate valves), the
minor materials should have viewing backgrounds
equivalent or comparable to the major material of
construction. For example, 316L, 316, and 304 stain-
less steel would be visually comparable materials. If
glass were the major material of construction, match-
ing the background could also be somewhat complex
since interactions between process equipments and
the manufacturing area floors or walls could lead to a
more difficult situation to emulate in the laboratory.
Other variables to consider are the impact of personal
protective equipment (PPE) worn in the manufactur-
ing space, such as goggles, safety glasses, laboratory
coats, coveralls, or scrubs. The spiked coupons are
then arranged in a viewing area with controlled light-
ing and background.
To control for variations in visual acuity, operators
were screened to ensure that they had either correct-
ed or uncorrected 20/20 vision. Those without 20/20
vision were excluded from the study. Screening was
conducted by first asking each operator if they had
had a professional eye exam within the last twelve
months. If so, the operator was asked to view two dif-
ferent Snellen eye charts from distances of 6 ft. and
10 ft., respectively. A passing result for this portion
of the test required that the operator could correctly
recite the line on the chart corresponding to 20/20
without reading any of the letters incorrectly. This
practice is recommended to qualify technicians both
in the laboratory and on the manufacturing floor.
When transferring the measured angles and distances
to the laboratory, keep in mind that technicians
Keith Bader & Kelly Scalva
in the lab will not have the geometric restrictions
imposed by ancillary equipment, viewing ports, and
manways, thereby necessitating artificial limitations
on the amount of movement they may instinctively
employ as they try to observe spotted coupons. A
representative arrangement of coupons is shown in
Figure 5.
rEcommEndAtionS for tESting
In any quality system, the integration of quality-
by-design principles starts with the education of all
personnel involved in the production of a therapeutic
substance. Specifically, when developing training ma-
terials for visual inspection of the process equipment,
operators should be informed of the importance of
the activity as an important orthogonal method for
the prevention cross contamination. This will help
prevent technicians from treating the activity as “just
another task” and rushing to complete it at the poten-
tial peril of patients receiving a particular therapeutic
compound.
In the laboratory setting, the study was most influ-
enced by the level of training of the visual assessment
operators. For example, operators who had experi-
ence cleaning coupons in the lab were much more
adept at dismissing imperfections on the surface of
coupons that may have otherwise confused those
who had never before conducted visual inspection or
VRL testing with residues. Not only is it important
for operators to know and understand why they are
completing a task, but it is just as important for the
operators to have the chance to study a dirty vessel
directly after a process run and at the end of the dirty
hold time in addition to having historic areas of con-
cern and vessel imperfections pointed out to them.
A well-educated operator trained to recognize the
appearance of process equipment at both its worst
and best relative to cleanliness is much more likely
to properly ascertain the presence of residues on
product contact areas. Specifically, operators should
also have the opportunity during training to visually
inspect the vessel directly after the cleaning cycle.
If possible, they should also be able to observe the
equipment in a clean, wet state and a completely dry
state since residue is typically better seen on dry pro-
cess surfaces. Knowledge of how the vessel appears
when the surfaces are covered in process residues
typically makes the cleanliness easier to assess if an
idea of the range of cleanliness is known.
Bias can also be avoided in the laboratory evalu-
ation by arranging the coupons both in a random
loading concentration order and in rows of, or greater
than, five coupons to prevent operators from memo-
rizing which coupons they identified as soiled under
a previous lighting, distance, and viewing angle
combination. Based on the authors’ experience, more
Special edition: Cleaning Validation 49
Keith Bader & Kelly Scalva
consistent results are also obtained if the operators
verbally relay whether the residue is visible rather
than be required to fill out protocol worksheets.
Overall, it was found that slight movement of the
flashlight was necessary for the operators to attempt
to see the residues; directing the light towards the
coupon led to excessive glare that hindered the ability
of the operator to view residue. Residues were typi-
cally noted when they were on the periphery of the
hand-held light source spot. It was also noted that
operators who viewed the surfaces longer generally
saw more residue than operators who quickly glanced
at the coupons. However, with the use of the flash-
light, operator-viewing duration increased by about
50%. Interestingly enough, visibility and detection
of dirty coupons decreased when operators used the
hand-held light over diffuse ambient lighting.
While the procedures described above may seem
fairly complete and extensive, there are still many re-
lated topical permutations warranting further inves-
tigation. For example, most studies, to date, focus on
the intact API; however, cleaning processes employed
for biopharmaceuticals often employ alkaline clean-
50 Special edition: Cleaning Validation
ing agents that degrade post-production residues. As
with any chemical change to a compound, the physi-
cal properties can change, including visibility of the
compound on process surfaces. Future studies will
investigate this and other similar considerations.
rEfErEncES
1. E. Teicholz, “LIGHTING” in Facility Design and Management
Handbook, (McGraw-Hill Professional, 2001), AccessEngineer-
ing.
2. R.J. Forsyth, V. Van Nostrand, and G. Martin, “Visible-Resi-
due Limit for Cleaning Validation and its Potential Applica-
tion in a Pharmaceutical Research Facility,” Pharmaceutical
Technology 28 (10), 58–72, 2004.
3. T. Croft, W.I. Summers, F.P. Hartwell, “PRINCIPLES AND
UNITS” in American Electricians’ Handbook, Fifteenth Edition,
(McGraw-Hill Professional, 15th ed., 2009 2002 1996 1992
1987 1981 1970 1961).
3. R.C. Rosaler, “Lighting” in Standard Handbook of Plant Engi-
neering, Third Edition (McGraw-Hill Professional, 3rd ed.,
2002 1995 1983)
4. USP29–NF24 <1051>, Cleaning Glass Apparatus, 2896