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AQ-02755-000, Rev. 0
EC Declaration of Conformity
according to EC Electromagnetic Compatibility Directive 89/336/EECand according to EC Low Voltage Directive 73/23/EEC
We herewith declare,
Labsphere, Inc.
P.O. Box 70, Shaker Street
North Sutton, NH 03260 USA
that the following product complies with the appropriate basic safety and health requirements of the EC Directive based on its
design and type, as brought into circulation by us. In case of alteration of the product, not agreed upon by us, this declaration will
lose its validity.
Product Description: UV-2000S Ultraviolet Transmittance Analyzer
Model Number: UV-2000S
Applicable EC Directives(s): EC Electromagnetic Compatibility 89/336/EEC
EC Low Voltage Directive 73/23/EEC
Standards of IMMUNITY IAW EN 55022:1998 Class B
IEC 801-2:1991 for immunity to ESD
IEC 801-3:1998 for immunity to radiated electromagnetic energy
IEC 801-4:1998 for conducted immunity to electrical fast transient/bursts
Standards of EMISSIONS IAW EN 50082-1:1997 EMC-generic for class residential, commercial and light industry
Standards of EMISSIONS IAW EN 50081-1:1992 EMC-generic for class residential, commercial and light industry
Standards of IMMUNITY IAW EN 55024:1998 Class B
Standards of SAFETY IAW BS EN 61010-1:1993
Authorized Signature:
Title of Signatory: Vice President of Engineering Date: April 22, 2008
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UV-2000S
Ultraviolet Transmittance Analyzer
Introduction ...............................................................................................................................1
Unpacking and Inspection ........................................................................................................3
Installation and Assembly .........................................................................................................4
Power Input ..................................................................................................................4
UV-2000S Installation .................................................................................................5
Installing the Sample Stage Assembly .........................................................................5
Description of the UV-2000S ....................................................................................................6
Spectrometers ..............................................................................................................6
Optical Chambers ........................................................................................................7
USB-to-RS232 Converter ............................................................................................9UV-2000S Power Source .............................................................................................9
Sample Stage Assembly ...............................................................................................9
Navigating Through the UV-2000 Software ..........................................................................10
Installing UV-2000 .....................................................................................................10
UV-2000 Study Documents .......................................................................................11
Recording Blank Scans ..............................................................................................11
Sunscreen Studies without a Photoprotection Method ..............................................13
Sunscreen Studies with the COLIPA 2007 Method ...................................................18
Sunscreen Studies with the Boots Star 2008 Method ................................................21
Blank and Sample Preparation ..............................................................................................26Preparation of the Blank Plates ..................................................................................26
Preparation of the Sample Plates ...............................................................................28
Alternative Sample Preparations ...............................................................................28
Operating Procedures .............................................................................................................30
Setting the Optics Head Height .................................................................................31
Operating the Sample Stage Assembly ......................................................................31
Recording a COLIPA Sunscreen Analysis ................................................................31
Recording a Boots Star Sunscreen Analysis ..............................................................33
Instrument Validation .................................................................................................34
Maintenance .............................................................................................................................35
Appendix A Specifications ......................................................................................................37
Appendix B Schematics ...........................................................................................................38
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Introduction
The UV-2000S Ultraviolet Transmittance Analyzer is the most recent and highly application specificultraviolet
spectroscopy productoffering from Labsphere. The function of the UV-2000S is to measure the transmittance of
ultraviolet (UV) radiation through sunscreen products and compute new internationally recognized effectiveness
characteristics of the product. The UV-2000S system supports the cosmetic manufacturing industry as a research and
development or quality control tool for sunscreen products.
In the past and today, many regulatory agencies such as the US Food and Drug Administration (USFDA) and the
European Cosmetic Toiletry and Perfumery Association (COLIPA) require in vivo testing on human subjects as a meansof validating the effectiveness of sunscreen products. The in vivo tests are costly, time-consuming, and may not be
practical for routine product development/evaluation. Hence, these agencies now recognize the importance ofin vitro
testing. New regulations and test methods have been developed for evaluating the broad spectrum protection (UVA and
UVB) provided by sunscreen products. The Labsphere UV-2000S Ultraviolet Transmittance Analyzer is designed to
evaluate sunscreen samples using these new internationally recognized in vitrobroad spectrummethodologies.
The sunscreen manufacturing industry and cosmetic trade associations have done much work to improve the methods of
in vitro analysis on their sun protection products. The most celebrated sunscreen test methods are published by the
European Cosmetic, Toiletry and Perfumery Association (COLIPA), headquartered in Brussels, and Boots in the U.K.
Many international regulatory agencies have adopted the COLIPA methods or Boots Star method in their entirety and
incorporated the procedures into their respective countries. Sunscreen manufacturers in other countries are developing
their own broad spectrum protectiontest procedures that utilize simple sunscreen protection factor (SPF) calculations
with modified parts of the Boots Star or COLIPA methods.
For our customers who use the Boots Star Rating System or COLIPA photoprotection methods, the UV-2000S system
provides a total solution to theirin vitro testing process. The UV-2000 software application guides the operator through
the Boots Star or COLIPA methods step by step, saving all sample scans, statistical data and decision making parameters.
UV-2000 records and displays spectral transmittance data and calculates the same characterization parameters utilized by
the traditional test methods. Finally, the UV-2000S provides a readily expandible platform to meet the evolving needs of
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the cosmetic manufacturing industry.
The physical appearance of the UV-2000S analyzer has changed dramatically, compared to the UV-1000S, but the new
product retains the reliability and performance capabilities of the UV analyzers in the past. These capabilities include the
following:
One-touch sample analysis, sample scans within five seconds
Automatic calculations of spectral transmittance, SPF, critical wavelength and UVA:UVB ratiosPerformance validation routine
New capabilities of the UV-2000S include the following:
Compact benchtop footprint
Wavelength accuracy to + 1 nm
Measurement area 0.79 cm2
Dynamic range extension up to 2.7 AU
AutoFlash capability
USB computer interface
Absorbance, SPF and UVAPF in accordance with the COLIPA In Vitro Guideline 2007
UVA:UVB Ratio in accordance with the new Boots Star Rating System (2008)Manually operated UV-2000S sample stage assembly
UV-2000 is the software application that accompanies the UV-2000S system. UV-2000 supports the Microsoft Windows
XP and Windows VISTA operating systems.
Figure 1. UV-2000S Ultraviolet Transmittance Analyzerwith the sample stage.
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Unpacking and Inspection
The UV-2000S was thoroughly inspected and calibrated before shipping and should be ready to operate after completing
the set-up instructions. All Labsphere instrumentation is packaged and shipped in reinforced shipping containers. A list of
components you should receive with your shipment is provided below. Carefully check each component after unpacking
for any damage that may have occurred during shipping. If there is any such damage, file a claim immediately with the
freight carrier and contact the Labsphere Customer Service Department at
(603) 927-4266.
After unpacking, allow 30 minutes for the equipment to reach room temperature before applying power. The materials
used to package the UV-2000S are custom-designed to protect the instrument during shipping. You may choose to return
your UV-2000S in the future for re-calibration or refurbishment. The shipping box and foam inserts should be retained.
Instructions for shipping the instrument back to Labsphere are provided in the maintenance chapter of this instruction
manual.
Standard Components:
UV-2000S Ultraviolet Transmittance Analyzer
USB cable
Power cableValidation kit
UV-2000 installation CDROM
Instruction manual CDROM
Sample stage assembly with transparent sample mask templates
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Installation and Assembly
Installation and assembly of the UV-2000S analyzer is easy. You should place the instrument on a flat desk top where the
operator has easy access to the optical head. The optical head on the UV-2000S instrument emits a flash train of ultraviolet
light during each scan sequence. When mounting the instrument, make sure the optical head is not situated at eye level.
Table 1. Minimum computer requirements for the UV-2000 software.
System Component Required Minimum
Processor 1.6 GHz
Operating
System
Windows Windows XP or VISTA
Display SVGA 800 x 600
Memory Available HD Space 100 Mb
RAM 256 Mb
Drives CD ROM 1
Ports USB 2.0 1
Power Input
The power input module, located on the rear panel of the UV-2000S, features a fuse cartridge and rocker switch. The same
size fuse is loaded into each side of the cartridge - the cartridge does not need to be rotated. Electrical input requirements
of the UV-2000S are written on the rear panel overlay - the internal power supply is completely universal at 100 - 240
VAC, 50/60 Hz.
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UV-2000S Installation
Assemble the UV-2000S Ultraviolet Transmittance Analyzer system as follows:
Figure 2. Making cable connections tothe UV-2000.
1. The minimum computer requirements for the UV-2000 softwareapplication are listed in Table 1. Load the UV-2000 install CDROMinto the computer disk drive and install the software on your
computer. The disk has an autorun feature and will install .NETFrameworks 2.0, if not already on your computer, and the UV-2000application. When the installation is complete a prompt to restart thecomputer may appear on the screen - select Yes at the prompt.Remove the install CDROM from the disk drive when the installationis complete.
2. Connect the power cord to the rear panel of the instrument and turnthe UV-2000S on.
3. Connect the USB cable between the instrument rear panel and anavailable USB port on your computer the Found New HardwareWizard will appear on the computer screen. Select No at theWindows update prompt and make the follow-on selection to allowthe wizard driver installation to proceed automatically.
4. Launch the UV-2000 application and run an instrument validation as described in the chapter entitled Operating
Procedures.
Installing the Sample Stage Assembly
The sample stage assembly is included with the Ultraviolet Transmittance
Analyzer. The UV-2000S and stage assembly
Figure 3. Installing the sample stage.
must lie on a flat surface for
proper system operation.
1. Place the stage assembly in front of the UV-2000S instrument. Slidethe stage half-way out and make sure the aluminum holding arm thatnormally engages the sample plate is positioned away from theinstrument.
2. Lift up the front end of the instrument, slide the stage assembly probeunderneath. Lower the instrument until the probe mates with the base
plate cutout. It may be necessary to jiggle the front of the instrumentside to side until the top surface of the stage is level with the samplearea base plate.
3. Load an empty sample plate and mask transparency into the stage andadjust the optics head position per the height adjustment procedureprovided in the Operating Procedures chapter later in this manual.Operate the stage assembly in all directions. Observe that the stageglides smoothly into the sample area without clipping the plastic ormetal surfaces in the sample area.
4. Examine the empty plate in the sample area, noting that the plate restsflat on the sample floor.
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Description of the UV-2000S
The UV-2000S measures thespectral transmittance of ultraviolet light through a sunscreen material and calculates certain
characteristic parameters of the sunscreen sample using internationally recognized statistical methods. The term
transmittance refers to the percentage of radiant flux transmitted through the sample, relative to the incident energy. The
termspectral transmittance refers to the transmission of light at a single wavelength. The UV-2000S instrument measures
spectral transmittance across the 250 - 450 nm wavelength spectrum using an integrating sphere and two spectrometer
instruments. The sample beam is generated inside the integrating sphere by a high energy ultraviolet pulsed flashlamp.
During the blank scan, light collected by the integrating sphere is directed downwards through a blank plate, void ofsunscreen product, into the lower chamber below. When using PMMA substrates, the PMMA plate is typically coated
with a small amount of glycerin (15 micro liters or less) during the blank scan.Ultraviolet radiation from the incident
beam that is not reflected or absorbed by the blank plate is collected in the lower chamber of the optics train and measured
by Spectrometer No. 2 as the 100% transmittance value. During the sample scan, ultraviolet radiation is collected from the
sample beam that is not reflected or absorbed by the combined medium consisting of the sample plate and the sunscreen
material. This radiation is measured by Spectrometer No. 2 and compared to the signal detected during the blank scan.
The transmittance of the measured sample is equal to the ratio of the sample beams collected from the blank and sample
scans. The light collected from the integrating sphere by Spectrometer No. 1 is used to correct variations of the sample
beam energy from one scan to the next. A block diagram of the UV-2000S system is provided in Appendix B.
Components inside the UV-2000S enclosure include a power supply, two spectrometers, two optics chambers, USB-to-
RS-232 converter and flashlamp power supply. The UV-2000S instrument is supported on four feet that are attached to the
bottom plate. The instrument must be placed on a flat surface for proper operation with the sample stage.
Spectrometers
The principal components inside the UV-2000S analyzer are the two diode array spectrometers. The spectral data
recorded from these spectrometers operate in tandem for sunscreen characterization. The spectrometers are identical
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except that they operate as master and slave during the scanning process so that data collection from the integrating sphere
and lower chamber occurs simultaneously. Spectrometer No. 2 is the master unit; Spectrometer No. 1 is the slave.
The fiber optic cable that feeds Spectrometer No. 2 is located in the instrument base underneath the sunscreen sample
plate. The fiber optic cable that feeds Spectrometer No. 1 collects radiation from the Spectralon integrating sphere. Both
spectrometers collect spectral transmittance data across the 250 - 450 nm wavelength spectrum during each blank and
sample scan. The 250 - 450 nm data from the sample scan is displayed on the UV-2000 main operating screen even though
SPF, UVAPF and UVA:UVB calculations are performed across the more limiting 290 - 400 nm spectrum. The processeddata collection from both spectrometers generates transmittance spectra according to the equation
T ( )S2
B2------
B1
S1------
= Equation No. 1
where S1 and S2 are the sample scan recordings for Spectrometers No. 1 and 2, and B1 and B2 are the blank scan
recordings. All components in Equation No. 1, of course, exist as arrays spanning the spectrum 250 - 450 nm, each
component including a dark scan array. When the dark scan data is included, Equation No. 1 can be written:
T ( )S2 SD2
B2 BD2-----------------------
B1 BD 1
S1 SD 1-----------------------
= Equation No. 2
where SD1 and BD2 are the dark recordings for Spectrometers No. 1 and 2 respectively taken during the sample and blank
scans.
The diode array in the spectrometer instrumentation is not thermoelectrically cooled. An automatic dark current
measurement is incorporated into the UV-2000 software immediately before each blank or sample scan measurement. The
flashlamp is extinguished for the length of the dark scan.
The wavelength calibration of the UV-2000 instrument is established by six calibration coefficients, three for eachspectrometer, used by UV-2000 to convert photodiode array pixel number to wavelength. A set of coefficients is unique to
a specific spectrometer instrument. Since spectral transmittance is a relative parameter, the wavelength spectrum is the
only data component that requires calibration. The three coefficients reside in spectrometer memory and are uploaded to
UV-2000 when the software is launched.
Optical Chambers
The UV-2000S optical components, shown in Figure 4, are housed in upper and lower optical chambers called the optics
head and input optics. The optics head includes the integrating sphere, flashlamp and Spectrometer No. 1 fiber optic
sensor. The input optical chamber includes a planar-convex lens, flat mirror and Spectrometer No. 2 fiber optic sensor.
The integrating sphere is constructed of Spectralon, Labspheres proprietary highly diffuse reflective material. The
flashlamp is mounted inside the integrating sphere and is powered by a special power supply inside the instrumentenclosure. The power supply generates a pulse train to the flashlamp for each blank or sample scan measurement. A fiber
optic cable embedded into the sphere wall samples the irradiance at the sphere wall, transmitting the sampled light to
Spectrometer No. 1. An exit port at the bottom of the sphere provides an outlet for the ultraviolet sample beam. All
components of the optics head are mounted on a vertical stage assembly that moves up and down to accommodate sample
loading.
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The length of the flashlamp pulse train for a particular sample scan is determined automatically by a special feature of the
UV-2000S. This feature, called autoflash, is useful for enhancing the UV-2000S throughput and thereby extending the
measurement capabilities of the instrument beyond 2.7 AU. During autoflash operation, UV-2000 evaluates the
transmission properties of the PMMA plate or other substrate during the blank scan and determines the optimal pulse train
for subsequent sample scans within the sample set.
The optics head can be raised or lowered by the rotating knobs on the right and left-hand side of the UV-2000S enclosure.
The right hand knob adjusts the height of the optics head temporarily when loading a sample plate. The left-hand knobadjusts the lower limit setting for the optics head and must be pressed inward to engage the worm gear during operation. If
the optics head does not appear to respond to left-hand knob operation initially after shipping, press the knob inward and
keep rotating in the up position until the height adjustment responds. A height adjustment procedure is provided in the
Operating Procedures chapter later in this manual.
Figure 4. Optics head and input optical chamber components.
The blank or sample plate fits inside the gap between the
sapphire window at the sphere exit port in Figure 4 and
the lens embedded in the instrument base. To load a blank
or sunscreen sample plate, raise the optics head using the
right-hand knob on the instrument enclosure, insert the
blank or prepared plate between the two chambers so it
lies flat on the bottom chamber surface, and lower the
upper chamber until it reaches the bottom stop.
The light beam that exits the integrating sphere penetrates
the blank or sample plate held between the two chambers
where it is either absorbed, reflected or transmitted. The
sample area of the UV-2000S is 0.79 cm2. This area is
equivalent to a 1 cm diameter circle and is identical to the
area sampled by the previous UV-1000 instrument. The
area is defined by the clear aperture of the PL-CX lens
optic in the instrument base located directly underneath
the sample plate. Radiation transmitted into the lower
chamber is focused onto the fiber optic ferrule and
transmitted to Spectrometer No. 2.
Do not insert abrasive objects into the UV-2000S.
The UV-2000S analyzer uses the blank or empty sample plate as the reference for 100% transmittance. The 100%
transmittance is measured and stored in computer memory during the blank scan. It is imperative that the optics head
configuration during the blank scan closely matches the configuration during the sample scan, or else the presence of
stray lightmay affect your scan results. The termstray light, in this case, refers to light incident on the diode arrays from
sources other than the flashlamp. To preclude the possibility of scan error, make sure the upper chamber is lowered
completely to the bottom limit when running either the blank or sample scan. Both incandescent and fluorescent lighting
emits light below the 450 nm upper limit of your UV-2000S, and these emissions can produce errors in your scanning.
You can minimize the chance of stray light error by moving external light sources away from the vicinity of the optics
head.
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USB-to-RS232 Converter
The two spectrometer instruments and flashlamp power supply inside the UV-2000S are serial RS-232 devices.
Consequently, a USB-to-RS-232 converter resides inside the UV-2000S enclosure so that all communications can be
issued through the single USB port. The UV-2000 install routine copies the driver for the converter into the appropriate
Windows directory during installation. The Found New Hardware Wizard configures the driver when the initial USB
cable connection is made. The USB converter is powered solely from the USB cable. If the USB cable is disconnected
inadvertently during system operation, the UV-2000 may freeze up or a warning message may appear on the screen. If the
If the application freezes, the user will need to re-boot the software and any unsaved data is lost.
During operation, UV-2000 initiates the scan routine when the user executes a blank or sample scan from the main
operating screen. The USB converter formats the USB commands to RS-232, sending separate scan execution signals to
the master and slave spectrometers. The flashlamp power supply fires a series of flashlamp pulses automatically upon
trigger from the master spectrometer.
UV-2000S Power Source
The internal power supply within the UV-2000 supplies +12 VDC to the flashlamp board and the spectrometer
instrumentation. The power entry module connects to the power supply over two leads, so the power supply board is
completely universal. A green LED mounted in the front, top, center of the UV-2000S enclosure indicates when the +5
VDC output circuity of the internal power supply is operating.
Sample Stage Assembly
The sample stage assembly is an economic X-Y stage for positioning
a sample plate at prescribed locations within the
Figure 5. Sample stage assembly with a 75 mm maskoverlay.
UV-2000S sample
area. The stage is standard on the UV-2000S and attaches to the front
of the instrument. The stage is manually operated, incorporating a
mask to ensure accurate and consistent sample test results.
When the stage is withdrawn the PMMA plate loaded in the sample
area can fall in the gap between the instrument and the vertical stage
rail. To prevent this occurrence, a cap screw illustrated in Figure 6
limits the travel of the stage. This cap screw can be removed andmoved to the location in Figure 6 to allow full withdrawal of the
sample stage when testing large plates.
Figure 6. Lengthening the vertical travel of the sample stage.
The number of sample locations on a sample plate typically is
specified by the international test methods. These same
international standards recommend that the same sample
locations should be tested before and after the irradiation
process. The sample stage assembly is very effective in helping
the user realize these requirements.
A kit of three different transparent mask overlays accompanies
the stage assembly for 75 x 75 mm, 70 x 70 mm and 50 x 50 mm
sample plates. Nine sample locations are inscribed on each
mask. The user can construct a customized mask and apply his/
her own test locations according to company policy.
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Navigating Through the UV-2000 Software
The UV-2000 software is designed around a method-oriented framework - that is, the user selects a study that conforms to
the specific sunscreen product and a prescribed photoprotection method. The UV-2000 application executes the sunscreen
study either under auspices of a photoprotection method or a stand-alone analysis. A sunscreen analysis can be performed
in either mode of operation. When performing a study analysis under method control, UV-2000 displays a scan template
that guides the user through the selected method routine. When operating without benefit of a method, the user is free to
proceed with his study in any direction without the constraints of a template.
Labsphere defines a scan as an array of processed spectral data extracted from the UV-2000S instrument. There are twotypes of scans, the blank scan and the sample scan. A blank or sample scan is collected at a specific location on a sample
plate, and normally, each plate is sampled at a pre-determined number of locations. A study is a collection of blank scans,
sample scans, calculations and decision making parameters usually performed on a number of sample plates.
Installing UV-2000
Do not connect the USB cable to your computer until the UV-2000 software is installed. Installation files for the UV-2000
software are provided on CDROM. The Setup.exe file will load all applicable UV-2000 files including NET Frameworks
2.0 if not already installed on your computer. The UV-2000 software package is in 32-bit configuration. The UV-2000
root directory stores all program executable files, reporting data and the electronic files holding your scan data.
When the installation is complete, a prompt to restart the computer may appear on the screen - select Yes at the prompt.
Once the software is installed and the computer is re-booted, connect a power cord to the UV-2000 instrument, turn it on,and connect the USB cable between the instrument and computer the Found New Hardware Wizard will appear on the
computer screen. Select No at the Windows update prompt and make the follow-on selection to allow the wizard driver
installation to proceed automatically.
UV-2000 searches for the UV-2000S analyzer when the application is launched. If the instrument is found, the text
"Connected" appears in the Device Status text box, "Ready" appears next to the progress bar and the Blank Scan Status
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text box is empty in the status bar at the bottom of the main operating screen. A previously saved study can be viewed
when there is no instrument connection, but data collection cannot proceed until a UV-2000S instrument is detected.
UV-2000 Study Documents
UV-2000 collects and saves sunscreen characterization data in a container called a study. A UV-2000 study includes all
the sample scan data and characterization parameters collected and computed for the sunscreen in vitro analysis. UV-2000generates and saves study documents in binary format in the default My Documents folder on the computer with the .uv2
filename extension. Saved study documents from previous UV-2000S data sessions can be opened in UV-2000 for
viewing or for follow-on sunscreen testing where the last session left off.
The user can choose the test rules utilized when recording data within a study. These rules are based either on an
internationally recognized sunscreen method or no method at all. When UV-2000 is launched, a generic study document
named Study 1 is established automatically by the software. The operator can create a new study similar to Study 1 by
clicking the New Study selection in the File Menu. A study established automatically or through the File Menu does not
include a method, but utilizes the erythema action spectrum (CIE 1987) with the traditional standard sun options popular
on the older generation Labsphere UV-1000S (COLIPA 1994 (rounded average of spectra at Albuquerque, New Mexico
and 40 degrees North Latitude) or Melbourne, Australia), or the new spectral irradiance identified as "SSR" within the
COLIPA Method. There are no test rules when operating within a generic or traditional study and there are no software
prompts. If the user creates a study document through the Method... selection in the Study Menu, the study is based on
one of the methods selected in the Select Method Dialog Box. At present, UV-2000 offers two method options: COLIPA
Guideline (2007) orBoots Star (2008 Revision). UV-2000 applies strict test rules in a method-based study, displaying
modal prompts at each step of the test method routine.
Study filenames automatically assigned by UV-2000 are not
particularly descriptive and a document can be renamed by
Figure 7. Viewing open study documents in the Window Menu.
the
operator at any time during the UV-2000S data session. To
rename a study document, clickSave Study As from the File
Menu and enter a more appropriate filename. To open a
previously saved study document, select Open Study from the
File Menu. A study can be opened for read-only purposes or for a
continued data collection session. Leave the checkbox at the
bottom of the Windows save screen unchecked for a continued session and select the appropriate filename. When the
previously saved file is opened, the opened document becomes active, the filename of the opened study appears in theWindow Menu and subsequent operator actions are applied to the now active study, starting where the most recent data
session concluded. There is no limit to the number of study documents open at a time, but only one study can be active at
a time. To activate a study already opened in computer memory, click the corresponding study filename in the Window
Menu.
Study and data export files are stored in the My Documents directory in comma-separated values format with the .cvs
filename extension. These files in can be viewed using Microsoft Excel or other spreadsheet application. The format of
the exported depends on the photoprotection method of the study. To export a study, activate the applicable study from the
Window menu and clickExport Study from the File Menu. To export a set of scans, highlight the set in the Scan Sets
Window and clickExport Set.
Recording Blank ScansThe blank scan in a sunscreen analysis constitutes the 100% transmittance reference data used to convert the spectrometer
pixel data into units of transmittance. Blank scan data is recorded in units of counts and saved as part of the study
document when the operator clicks Save Study orSave Study As in the File Menu. The blank scan is a critical part of the
sunscreen analysis and the indiscriminate application of blank scan data may result in erroneous test results. Since the
PMMA plate is prepared completely separate from the sample plates, there typically is no reason to employ more than one
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blank scan per study unless the sample plates used in the study are drawn from more than one manufacturing lot. When
collecting sunscreen data within a COLIPA or Boots Star method, UV-2000 saves a single blank scan in the study
document that is utilized for the entire analysis. When operating without a sunscreen photoprotection method, UV-2000
makes provision for separate blank scans for each scan set in the study. The data held by an individualblank scan is
applied to every sample scan in the scan set - always! In addition to the individual blank scans, UV-2000 may save astudy
defaultblank scan. If recorded, the default blank scan remains in ready reserve to be copied to an individual set blank scan
array at the operators discretion.
Blank scan assignments are pre-programmed from the Scan
Options Dialog Box, when operating without a photoprotection
method, and modified by the operator during the analysis routine.
The sequence of blank scan execution and new set creation
depends on the scan options imposed. To invoke the Scan Options
Dialog Box, click the Scan Options selection from the Instrument
Menu. Scan options can be changed any time when operating
without a photoprotection method. Edits made in the Scan Options
Dialog Box take affect immediately when applied to the active
study and persist until the operator either exits UV-2000 or changes
the scan options at a later time. The Scan Options Dialog Box
Figure 8. Scan Options Dialog Box.
is
captured in Figure 8. The upper half of the dialog box controls UV-
2000 assignment of blank scan data. The lower half controls the
length of each blank and sample scan. Blank scan application is
controlled by a sequence of software messages issued by UV-2000
that may or may not appear on the screen during sunscreen
analysis. If the user selects Copy study default blank scan, no
prompts appear and the blank scan array for each set automatically
is assigned the data held as the default. If the radio button labeled
Prompt for blank scan is selected, UV-2000 issues blank scan
prompts to the operator during the course of the analysis.
Figure 9. Message appearing when theoperator first collects a blank scan.
When the Copy study default blank scan setting of Figure 8 is in effect for a new
study, the operator must execute a default blank scan immediately and the
information message in Figure 9 appears when the blank scan is complete. The
default blank scan data is copied to the blank scan for the individual setautomatically each time a new set is created. If the operator changes his mind and
performs a new blank scan, the option message in Figure 10 appears on the screen
immediately after blank scan execution. If he clicks Yes, the new blank scan data
is copied to the default study and the currently active set blank scans. If he selects
No, only the study default blank scan data is filled.
Figure 10. Option message that appearsfollowing a blank scan collected within ascan set.
When the Prompt for blank scan radio button in Figure 8 is selected, the
operator can elect to collect a blank scan immediately after creating a study, or
he can take the blank scan after generating the first scan set. If he executes a
blank scan first, the information message in Figure 9 appears after the scan
data is collected just as before. Since there is no scan set in the study, the blank
scan can only be applied as the default and the operator must select OKto
continue the analysis. If the operator creates a new study first, followed by theblank scan, the message in Figure 10 appears when the scan is complete. If he/
she selects the Yes button in Figure 10, the prompt in Figure 11 appears
immediately following the creation of all subsequent scan sets.
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So, what does all this mean to the operator? The operator should choose the blank
scan settings initially before embarking on the sunscreen analysis.
Figure 11. Message that appears when theoperator creates a new scan set.
The Take Scan
selection is never active in the Instrument Menu and no sample scanning ever can
occur until a blank scan is collected and the initial scan set created. If the operator
chooses the Copy study default blank scan option, a single array of blank scan
data is used for the analysis and there are no prompts - but the operator can
always change his/her mind and run a blank scan for an individual scan set. If the
operator chooses Prompt for blank scan, the software issues prompts at each
new set - but he/she can always change his mind and apply the default blank scan.
The newest feature incorporated into the UV-2000S and associated hardware is autoflash. This feature extends the range
capability of UV-2000S absorption measurement beyond 2.7 AU to support SPF product analysis well beyond SPF 50.
This feat is accomplished automatically by recording the blank data in three phases. Phase 1 is the recording of the dark
scan data with the flashlamp extinguished. Phase 2 is the trial scan where the flashlamp operates for a pre-determined
number of pulses - this sequence can last up to 25 seconds. UV-2000 evaluates the number of counts recorded by
Spectrometer No. 2 during the trial scan and adjusts the pulse train for subsequent blank and sample scans accordingly.
Phase 3 of blank scan execution is the full pulse train blank scan that constitutes 100% transmittance.
The lower half of the dialog box in Figure 8 controls the number of data sets collected when the operator clicks Take
Blank Scan orTake Scan from the Instrument Menu. The UV-2000S scan averaging feature is useful for two reasons.
First of all, the feature can be used to effectively extend the spectrometer integration time. The spectrometer integrationtime for each blank or sample scan is set automatically by theAutoFlash feature during blank scan execution. The
operator can effectively increase the instrument signal-to-noise ratio, without potentially saturating the spectrometer
arrays, by making Blank Scan orSample Scan entries greater than one. Secondly, the averaging feature may be useful in
recording the averaged blank scan at more than one location on the blank plate. Be careful - no prompt is displayed
between pulsed intervals when the scan averaging feature is used! UV-2000 always records only one set of averaged scan
data following each scan execution, no matter what scan averaging entries are applied.
When a study document is saved, the study default and individual blank scans are saved along with the sample scans in
the study document. The presence of multiple blank scans within a scan set or study can create considerable confusion
during subsequent viewing or data collection sessions. The operator can avoid this predicament by strategically renaming
each set and sample scan after generation.
The default settings in the Scan Options Dialog Box can be changed so the operator does not need to visit the dialog box
each time the application is opened. To change scan options, make the appropriate selections in Figure 8 and clickSave asapplication defaults. The new scan parameters are saved to a separate electronic file on your computer.
Sunscreen Studies without a Photoprotection Method
A new study identified in the Window Menu as Study 1 is created automatically when the UV-2000 application is
launched. Study 1 in its original format exists as a generic study in computer memory without the direction provided by a
photoprotection method. Sunscreen characteristics generated without a photoprotection method are calculated and
displayed in the same manner as the original Labsphere UV-1000S UV analyzer. These characteristics include the
following:
Spectral transmittance T(), absorbance A(), optical density OD(), and monochromatic protectionfactor mPF() data across the 290 - 450 nm wavelength spectrum
SPF calculated with user-selected solar spectral irradiance data
UVA:UVB Ratio
Critical wavelength cStatistical reporting of SPF and UVA and UVB test characteristics
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Generic test characteristics generated by UV-2000 are reported in the Data window on the left-hand side of the main
screen. There are no software prompts when collecting sunscreen data in this mode, and the operator should be familiar
with a typical test sequence as follows:
1. Create a new or select an existing study.
2. Create a new or select an existing scan set.
3. Record a blank scan.
4. Collect a sample scan.
5. Rename the sample scan curve.
Some of this test sequence is programmed into the UV-2000 application as described in the previous paragraphs. A
sample scan cannot be collected, for example, unless a scan set is selected and blank scan resides in computer memory for
the selected set. To create a new generic study, select New Study from the File Menu; to select an existing study already
in computer memory, select the study filename from the Window Menu. To record blank scan data, load the blank plate
into the UV-2000 sample area and select Take Blank Scan from the Instrument Menu. To select a scan set for your
anticipated scan data, click on the appropriate scan name in the Scan Sets Window. Finally, the sample scan is executed by
replacing the blank plate in the UV-2000 with a sample plate and selecting Take Scan.
Figure 12. Collecting sunscreen data in a study without a photoprotection method.
UV-2000 executes a blank scan by first collecting an initial dark scan across the wavelength spectrum followed by the
trial scan and data blank scan described in the previous section. The entire blank scan routine is performed automatically
when the operator clicks the Take Blank Scan selection in the Instrument Menu. Following blank scan execution, UV-
2000 subtracts the dark scan from the data scan and stores the blank scan data for future use. The operator executes a
sample scan by clicking the Take Scan button on the toolbar or by selecting Take Scan from the Instrument Menu. UV-
2000 records the sample scan in much the same manner as the blank scan and applies the blank and sample scan data to
the equations described in the previous chapter. Spectral curves are displayed on the main operating screen in one of the
formats selected from the Scan Display Options Dialog Box. SPF and critical wavelength calculations are displayed in the
data windows on the left-hand side of the main operating screen.
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Traditional Sunscreen Characteristics
Spectral scan data can be displayed on the graph in Figure 12 in units of transmittance, absorbance, optical density or
monochromatic protection factor. The operator can change the scale of the ordinate axis by selecting Graph and Display
Options from the Graph and Display Menu and clicking the appropriate entry in the Display Units select box. A brief
description of the UV-2000 calculations is provided here.
Transmittance T(). Spectral transmittance data is measured by the UV-2000S according to EquationsNo. 1 or 2 described previously in the last chapter. The data is displayed in units of percent on the spec-tral chart and in table format across the full 290 - 450 nm wavelength spectrum or a portion thereof.
Absorbance A(). Absorbance values for each wavelength are calculated from the recorded spectraltransmittance data according to the relationship
A T( )log=
where T() is expressed as a fraction. The data is displayed absorbance units (AU) on the spectral chartand in table format across the full 290 - 450 nm wavelength spectrum or a portion thereof.
Optical Density OD(). Optical density calculations are identical to spectral absorbance describedabove.
Monochromatic Protection Factor mPF(). The monochromatic protection factor is the spectralquantity equal to the inverse of transmittance:
mPF1
T------=
where T() is expressed as a fraction.
The spectral data from a sample scan is displayed by a table in the top left-hand data window of the main operating screen.
The order of display can be changed by clicking the header of each column. The spectral units of display can be changed
through the Display Options Dialog Box.
UV-2000 calculates the traditional SPF and critical wavelength sunscreen characteristics and displays the parameters for
the active scan at the top left-hand corner of the data window. The two parameters listed here are calculated using thetraditional definitions which may be different than similar parameters defined in the photoprotection methods. The
statistical data at the bottom left-hand corner displays the mean, standard deviation and coefficient of variation for the two
parameters calculated across the active scan set. The user can activate any scan and the corresponding scan set by clicking
on the scan name in the Scan Sets Window.
Sunscreen Protection Factor (SPF). SPF is a measure of the ultraviolet solar energy required to pro-
duce sunburn on skin treated with sunscreen, relative to the amount of solar energy required to produce
sunburn on unprotected skin. As the SPF rating of a sunscreen product increases, the sunburn protection
increases. Most sunscreen manufacturers use the in vitro method similar to the UV light test performed
by the UV-2000S to accelerate new product development, decrease in vivo product test cycles and cost
and to verify lot-to-lot manufacturing consistency. UV-2000 calculates the SPF characteristic according
to the ratio
SPF
ES d
290
400
EST d
290
400
---------------------------------= ,
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where E() is the erythema action spectrum, S() is the solar spectral irradiance, T() is the spectraltransmittance of the sample with the integral is calculate across the 290 - 400 nm wavelength limits.
Critical Wavelength (c). The critical wavelength is defined across the 290 - 400 nm spectrum by thefollowing relation:
c Mi n ( ),=
such that ' satisfies the relationship:
A
290=
A
290=
400
-------------------------- 0.9
where A() is the absorbance at wavelength .
UV-2000 displays the computed SPF and critical wavelength in the data window for the scan selected in Scan Sets
Window. The mean statistics are calculated for the entire set that applies to the scan or set selected in the Scan Sets
Window:
SPFdMeanSPF i( )
n----------------
i 1=
n
= LambdadCriticaldMean c i( )n
-----------
i 1=
n
=
The standard deviation statistics are calculated for the entire set that applies to the selected scan or set in the Scan SetsWindow:
SPFdSTD SPF i( ) SPFdMean( )2
n 1----------------------------------------------------------
i 1=
n
= LambdadCriticaldSTD c i( ) LambdadCriticaldMean( )2
n 1-----------------------------------------------------------------------------------------
i 1=
n
=
The coefficient of variation terms are defined for each set as:
COVSTD
Mean--------------100= %.
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Editing the Spectral Graph Format
A sample scan is always subordinate to a particular sample scan set. The scan sets
created in the currently active study are displayed in the Scan Sets Window on the
right-hand side of the main operating screen. To create a new set, click on the
Instrument Menu and select New Set - the newly created set will appear in the Scan
Sets Window and the graph will go blank. Spectral scan curves are displayed on the
graph and the scan names are listed in the Scan Sets Window automatically following
every sample scan execution. The scan curves are color-coded in the legend to the
right of the
Figure 13. Editing a scan set or indi-vidual scan within a set when operat-ing outside a photoprotection method.
graph. The operator cannot replace a scan already visible in the window,
but he can delete or rename a scan set or individual scan in a set by right-clicking the
name and making the appropriate selection in the popup menu. The title of the graph
and the scan label in the graph legend is updated each time a scan set or scan is edited.
There are no limits to the number of scans in a scan set or the number of scan sets in a
generic study. All scans that appear on the graph and the Scan Sets Window, for a
particular set, contribute to the statistical parameters reported in the statistical data
window in the lower left-hand corner of the main operating screen.
The operator can change the vertical axis scale and units of display on the spectral graph at any time without altering the
actual scan data or test analysis. To edit the spectral graph, clickGraph and Display Options from the Graph and
Display Menu.
Modifying the Sunscreen Characteristic Calculations
The solar spectral irradiance data accompanying your application are used to calculate the SPF characteristic. This data
can be viewed by clicking Solar Irradiance from the Study Menu. The erythema action spectrum (CIE-1987) is
embedded in the software and cannot be changed. Three different sets of solar spectral irradiance data are provided for the
SPF calculation when operating without a photoprotection method: Albuquerque NM*, Melbourne AU and COLIPA
2007. The solar irradiance data selected may impact the SPF test results significantly. Solar irradiance selections are not
saved in the study document. Therefore, it is suggested that the spectrum utilized for a study be denoted in the study
comments.The most recent solar irradiance data selection is stored in a separate electronic file on your computer by
clicking Save as application defaults for retrieval by the application during the next UV-2000S data session.
Reporting the Generic Study Test Results
The test results from a generic study can be displayed in report format or written to a separate file for subsequent analysis
by a spreadsheet application. Product information can be entered into the study by invoking the Study Information Dialog
Box from the Study Menu. To view a test report of the most recently updated sunscreen analysis, activate the applicable
study from the Window menu and clickReport in the Study Menu.
Data export files are stored in the My Documents directory in comma-separated values format with the .cvs filename
extension. These files in can be viewed using Microsoft Excel or some other spreadsheet application. To export a
complete generic study, activate the applicable study from the Window menu and clickExport Study from the File Menu.
To export a complete set of scans in a scan set, highlight the set in the Scan Sets Window and clickExport Set.
* For clarification, the "standard sun" spectrum referred to within this document as "Albuquerque, NM" is actually the rounded aver-age of 35 degrees north (Albuquerque) and 40 degrees North, as reported by COLIPA, SPF Test Method, May 1994. As notedwithin the COLIPA SPF Test Method, what is important is that the "standard sun" reference spectrum represents a realistic maximumsolar spectrum; defined as one typically encountered under conditions of a cloudless sky under high incidence at a fairly low altitude.Both these well published spectra, 35N and 40N, satisfy these requirements.
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Sunscreen Studies with the COLIPA 2007 Method
Sunscreen studies generated within a photoprotection method are calculated and displayed in a manner much different
from the generic studies. UV-2000 reports the following sunscreen characteristics when operating within a COLIPA 2007
study:
Spectral transmittance T(), absorbance A(), optical density OD(), and monochromatic protection
factor mPF() data across the 290 - 450 nm wavelength spectrumSPFin vitro calculated across the 290 - 400 nm wavelength spectrum for each sample plate
UVAPF0 calculated across the 320 - 400 nm wavelength spectrum for each sample plate prior to irradi-
ation
UVAPF calculated across the 320 - 400 nm wavelength spectrum for each sample plate post-irradiation
SPFin vivo: UVAPF ratio
Statistical reporting of the UVAPF test characteristic
COLIPA Method Characteristics
The sunscreen characteristics displayed under the COLIPA study method are more numerous and calculated differently
than their generic study counterparts:
Sunscreen Protection Factor (SPF) and Critical Wavelength (c). UV-2000 calculates the SPF andccharacteristics from a single scan in the same manner as the calculation in the non-method studyanalyses. The operator can view the SPF and c parameters in the Data Window on the left-hand side ofthe main screen. The statistical parameters for the scan set SPF and c characteristics are displayed inthe bottom left-hand corner.
In vitro SPF (SPFin vitro). The SPFin vitro characteristic is described in the COLIPA 2000 guidelines as
the ratio
SPFin dvitro
E ( )I ( ) d
290
400
E ( )I ( )10A0 ( ) d
290
400
---------------------------------------------------------= ,
where E() is the erythema action spectrum, I() is the COLIPA specified solar spectral irradiance, T()is the spectral transmittance of the sample, A() is the mean monochromatic absorbance of the testproduct layer before UV exposure, and the integration is performed over the 290 - 400 nm wavelength
spectrum.
UVAPF0. UVAPF0 is the pre-irradiation UVA Protection Factor calculated individually for each plate.
First, the SPFin vitro sunscreen characteristic described above is adjusted to the in vivo SPF value deter-
mined for the same sunscreen product, to determine the coefficient of adjustment, C. UV-2000 calcu-
lates the value of the Coefficient of Adjustment C automatically and applies the coefficient to the ratio
UVAPF0
E ( )I ( ) d
320
400
E ( )I ( )10A0 ( )C d
320
400
------------------------------------------------------------= .
The COLIPA Method recommends that C falls within a range between 0.8 and 1.2. UVAPF0 is deter-
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mined before irradiating the sample and is displayed in the Plate Data Table under the heading "UVAPF
Pre-irradiation".
UVAPF. The UVAPF characteristic is computed in the same manner as the UVAPF0 calculation above,
except that the calculation is made after the irradiance is applied to the sample plate. The value of the
coefficient C is the same in both calculations. UV-2000 displays the UVAPF characteristic for each
plate in the Plate Data Table under the heading "UVAPF".
Irradiation Dose. The irradiation dose is calculated for each sample plate after the pre-irradiation scansare complete. The dose parameter is equal to the UVAPF0 parameter for a plate multiplied by the value
1.2 J/cm2.
Mean UVAPF. UV-2000 computes the average of the included plate UVAPF values and reports this
characteristic at the bottom right-hand corner of the COLIPA Method Window as the "UVAPF Mean".
The characteristic does not appear on the screen until sufficient number of sample plates have been
tested.
SPF:UVAPF Ratio. The final sunscreen characteristic under the COLIPA method is the SPF:UVAPF
Ratio which is the in vivo SPF or SPFlabelparameter divided by the UVAPF mean.
Conducting a COLIPA Study
Studies incorporating one of the photoprotection methods inherent to UV-2000 are accessed through the Study Menu. Tocreate a COLIPA 2007 method study, click the Method... selection in the Study Menu, select the COLIPA Guideline
(2007) radio button from the Select Method Dialog Box and enter the appropriate parameters in the COLIPA Method
Options Dialog Box. When the COLIPA study is created, the default name of the study appears in Window Menu, a scan
template appears in the Scan Sets Window and the appearance of the main operating screen is changed to reflect the
screen shot in Figure 14.
Figure 14. Sunscreen characteristic peculiar to the COLIPA 2007 guidelines are reported inthe main operating screen modified by the COLIPA method characteristics display.
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When operating within the COLIPA method, UV-2000 displays prompts to the operator during the sunscreen analysis.
These prompts are visible in the upper left-hand corner of the COLIPA Method Window in Figure 14 under the heading
"Instructions". The software issues prompts to perform blank and sample scans, load plates into the UV-2000 sample area
and make sample rejection decisions based on software generated statistics. UV-2000 displays the next prompt
automatically after each step is accomplished. The operator can step through the prompted COLIPA routine or return to
previous steps in the process to correct an obvious mistake.
A scan template appears in the Scan Sets Window of the main screen immediately after a COLIPA method study iscreated. The template conforms to the number of plates and sample locations entered previously in the COLIPA Method
Dialog Box. Before generating a new COLIPA method, the operator should be sure to select the correct scan option.When
editing the COLIPA Method Dialog Box, the operator should be sure to enter the correct test parameters - once a COLIPA
study is created, the scan options and scan template cannot be changed. When performing a sunscreen analysis under a
COLIPA study, UV-2000 collects scan data for each sample plate starting with "Plate 1 Pre-irradiation" at the top of the
Scan Sets Window. The sample scans collected within the Plate 1 set are compared to the sample scans under the "Plate 1
Post-irradiation" set later in the analysis. A green check mark appears automatically in the checkbox for each programmed
scan when the scan is complete. The software prompts the operator to execute sample scans sequentially until all pre-
irradiation data is collected.
Test characteristics
generated by UV-2000 under
COLIPA guidelines arereported on the COLIPA
Method Window in Figure
15 that
Figure 15. Calculations displayed in the COLIPA Method Window.
appears when a
COLIPA method study is
created or a previously saved
COLIPA study is opened.
Most data collection
operations are accessed from
this window. The New Set,
Take Blank Scan, and Take Scan selections in the Instrument Menu are disabled when collecting data within a COLIPA
study - to record COLIPA study scan at the software prompt, click the Scan Execution button directly underneath the
prompt.
The structure of the scan template and identification of each scan in a COLIPA study cannot be changed, however, the
user can correct mistakes made during scan execution. To replace an incorrectly collected scan, load the appropriate
sample plate into the UV-2000S instrument and click the scan location number in the scan template - the scan highlights
while the instructions and scan execution button in the COLIPA Method Window changes to reflect the scan template
selection. Click the Scan Execution button to record the scan.
UV-2000 updates the Plate Data Table in Figure 15 automatically when each sample plate is complete. The pre-irradiation
updates include the SPF Mean, "C" coefficient UVAPF0, Irradiation Dose (D) and calculated irradiation exposure time
values. Exposure time calculations are based on the irradiation entry selected in the Exposure Lamp Irradiance selectin
box. The pre-irradiation and post-irradiation scans up for all locations are complete in Figure 15. During study execution
the sunscreen characteristics do not appear until for a plate until all scans have in the set are collected. The Coefficient of
Adjustment is calculated automatically by the software, for each sample plate, using an iterative routine and displayed in
the table under the heading "C Coeff". The same values of "C" are used for the post-irradiation computations as are used
in the pre-irradiation calculations. If the coefficient for a particular plate is not within the 0.8 to 1.2 range, the text "out of
range" appears in the "C Coeff" column and no COLIPA characteristics for the plate are displayed.
During the course of the sunscreen analysis, UV-2000 automatically tracks the scanning progress of each sample plate,
both pre-irradiation and post-irradiation. When scanning is complete on a sample plate, UV-2000 automatically
determines the legitimacy of the scan data. This determination is made by computing the coefficient of variation (COV) of
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absorbance values between the scans on the sample plate for each wavelength. If the COV exceeds 50%, the sample plate
is not legitimate and the checkbox for the plate under the "Include Data in Results" header automatically is left
unchecked. The operator can edit these checkboxes manually after each sample plate is complete and save the edits in the
study document. A checkbox cannot be selected unless the plate meets the 50% COV requirement and the Coefficient of
Adjustment is valid.
When sufficient post-irradiation data is collected, UV-2000 calculates the final COLIPA statistical characteristics for the
sunscreen analysis, including UVAPF Mean, the SPF/UVAPF Ration and UVA Balance. These statistics are based on thecheckboxes selected in the Plate Data Table of the COLIPA Method Window and are displayed in the bottom right-hand
corner. The statistics do not appear if the number of sample plates included in the results does not meet the COLIPA
requirements. The COLIPA COV requirement between plates is 20%, but the operator is free to include any individual
plate that meets the COV requirement discussed in the paragraph above.
The statistics displayed in the Statistics Window in Figure 16 reflect the same statistical analysis on the same traditional
sunscreen characteristics as the generic study format. Although saved in the COLIPA study document, statistics displayed
in this window do not apply to the COLIPA method - the statistics are displayed for information only.
Figure 16. Statistics Window for a COLIPAmethod study visible during the executionof the Plate 3 scan set from Figure 14.
The statistical parameters in Figure 16 are updated continuously after each scan is
executed from the COLIPA Method Window (if you remember, traditional SPF
statistics are based on the number of locations or scans - not plates). The value of
the SPF Mean characteristic in the Statistics Window may be significantly
different in magnitude from the COLIPA characteristics because the parameter
calculations are different and the traditional SPF analysis is not adjusted for the in
vivo test input.
The operator can change the solar irradiance data used in the SPF statistics in the
Statistics Window. The selected data does not appear in Figure 16 until the first
scan set is created. To change the irradiance data, click the button in Figure 16 or
select Solar Irradiance from the Study Menu and select either the Melbourne or
COLIPA data. The irradiance data set that appears here is used only for the
traditional SPF calculations and has no impact on COLIPA method characteristics.
The operator can apply the same spectral graph formatting under the COLIPA method as described previously with no
photoprotection method. Graph formatting does not affect the COLIPA test results.
Reporting the COLIPA Study Test Results
The test results from a COLIPA study can be displayed in report format or written to a separate file for subsequent
analysis by a spreadsheet application. Product information can be entered into the study by invoking the Study
Information Dialog Box from the Study Menu. To view a test report of the most recently updated COLIPA analysis,
activate the applicable study from the Window menu and clickReport in the Study Menu.
Data export files are stored in the My Documents directory in comma-separated values format with the .cvs filename
extension. These files in can be viewed using Microsoft Excel or some other spreadsheet application. To export a
complete COLIPA study, activate the applicable study from the Window menu and clickExport Study from the File
Menu. To export a complete set of scans on a sample plate, highlight the set in the Scan Sets Window and clickExport
Set.
Sunscreen Studies with the Boots Star 2008 Method
Sunscreen studies generated within a photoprotection method are calculated and displayed in a manner much different
from the traditional characteristics displayed without a method. The Boots Star method analyzes a sunscreen by
calculating the ratio of average sunscreen product absorbance for the UVA and UVB regions. This concept holds validity
because the effectiveness of sunscreens in the UVB spectrum is well documented by testing in the past. The Boots Star
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system uses the termspre-exposure andpost-exposure for the UV degradation analysis instead of the irradiance
terminology utilized by the COLIPA method. UV-2000 reports the following sunscreen characteristics when operating
within a Boots Star 2008 study:
Spectral transmittance T(), absorbance A(), optical density OD(), and monochromatic protectionfactor mPF() data across the 250 - 450 nm wavelength spectrum
SPFin vitro calculated across the 290 - 400 nm wavelength spectrum for each sample plate
UVA protection index calculated across the 320 - 400 nm wavelength spectrum for each sample plate
UVA protection index calculated across the 290 - 320 nm wavelength spectrum for each sample plate
UVA:UVB Ratio for each sample plate, pre-exposure and post-exposure
Star rating allocation for the analysis
Boots Star Method Characteristics
Sunscreen characteristics displayed within a Boots Star study method include some of the traditional parameters as well as
the Boots Star characteristics. The traditional parameters are calculated as described in earlier sections of this manual.
Sunscreen Protection Factor (SPF) and Critical Wavelength (c). UV-2000 calculates the SPF andccharacteristics from a single scan in the same manner as the calculation in the traditional study anal-yses. The operator can view the SPF and
cparameters in the Data Window on the left-hand side of the
main screen. The statistical parameters for the scan set SPF and c characteristics are displayed in thebottom left-hand corner.
UVA Protection Index. The UVA protection index (UVA) is a Boots Star characteristic defined as the
average absorbance calculated across the UVA spectrum:
UVA
A ( ) d
320
d
320
400
--------------------------= ,
where A() is the monochromatic absorbance averaged across the sample plate for each wavelength ofthe UVA spectrum. The integration is performed over the 320 - 400 nm spectrum. The UVA calculation
is made twice - once before the radiation exposure and once after exposure. UV-2000 does not actually
display the UVA values for each plate. Instead, the UVA:UVB Ratio for each plate is displayed in the
Boots Star Substrate Data Table when all scanning within the set is complete.
UVB Protection Index. The UVB protection index (UVB) is the average absorbance calculated across
the UVB spectrum:
,UV B
A ( ) d
290
d
290
320
--------------------------=
where A() is the monochromatic absorbance averaged across the sample plate for each wavelength ofthe UVB spectrum. The UVB wavelength spectrum is defined as 290 - 320 nm. The UVB calculation is
made twice - once before exposure and once after exposure. UV-2000 does not actually display the
UVB values for each plate. Instead, the UVA:UVB Ratio for each sample plate is displayed in the Boots
Star Substrate Data Table when all scanning within the set is complete.
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UVA:UVB Ratio. The UVA:UVB Ratio sunscreen characteristic is calculated for pre-exposure and
post-exposure conditions:
dUVA
UVB-------------=UVA:UVB Ratio
UV-2000 displays a UVA:UVB Ratio for each sample plate before and after the ultraviolet radiation
exposure.
The star assignments for each sample are based on the combined UVA:UVB Ratio parameters using the Boots Star criteria
in Table 2.
Table 2. Star Rating criteria, copied from "Measurement of UVA:UVB Ratios According to the Boots Star Rating Sys-tem (2008 Revision)" distributed by Boots (UK) Limited.
INITIAL Mean UVA:UVB RATIO
0.0 to 0.59 0.6 to 0.79 0.8 to 0.89 0.9 and over
POST
EXPOSURE
MeanUVA:UVB
RATIO
0.0 to 0.56 No Rating No Rating No Rating No Rating
0.57 to 0.75 No Rating *** *** ***
0.76 to 0.85 No Rating *** **** ****
0.86 and Over No Rating *** **** *****
UV-2000 evaluates the degradation of the Mean UVA:UVB characteristic from pre-exposure to post-exposure and assigns
the star rating automatically.
Conducting a Boots Star Study
Studies incorporating one of the photoprotection methods inherent to UV-2000 are accessed through the Study Menu. To
create a Boots Star method study, click the Method... selection in the Study Menu, select the Boots Star (2008 Revision)radio button from the Select Method Dialog Box and enter the appropriate parameters in the Boots Star Method Options
Dialog Box. When the Boots study is created, the default name of the study appears in Window Menu, a scan template
appears in the Scan Sets Window and the appearance of the main operating screen is changed to reflect the screen shot in
Figure 17.
When operating within the Boots Star method, UV-2000 displays prompts to the operator during the sunscreen analysis.
These prompts are visible in the upper left-hand corner of the Boots Star Method Window in Figure 17 under the heading
"Instructions". The software issues prompts to perform blank and sample scans, and load plates into the UV-2000 sample
area for each scan. UV-2000 displays the next prompt automatically after each step is accomplished. The operator can step
through the prompted Boots Star routine or return to previous steps in the process to correct an obvious mistake.
Sample plate rejection criteria under the Boots Star guidelines is not as strict as the rules under the COLIPA method. In
fact, except for comments regarding high standard deviation, the Boots guidelines stipulate no plate rejection
requirements. UV-2000 displays the highest standard deviation computed for each sample plate so the operator can applyuser policies to the scan data before completing the analysis.
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Figure 17. Sunscreen characteristic peculiar to the Boots Star guidelines are reported in the mainoperating screen that now includes the Boots Star Method Window.
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A scan template appears in the Scan Sets Window of the main screen immediately after a Boots Star method study is
created. The template conforms to the number of plates and sample locations entered in the Boots Star Options Dialog
Box. When editing the Boots Star Options Dialog Box, the operator should be sure to enter the correct test parameters -
once a Boots Star study is created, the scan template cannot be changed. When performing a sunscreen analysis under a
Boots Star study, UV-2000 collects scan data for each sample plate starting with Substrate No. 1 at the top of the Scan Sets
Window. A green check mark appears automatically in the checkbox for each programmed scan when the scan is
complete. The software prompts the operator to execute sample scans sequentially until all pre-exposure data is collected.Test characteristics generated by UV-
2000 under the Boots guidelines are
reported on the Boots Star Method
Window in Figure 18 that
Figure 18. Calculations displayed in the Boots Star Method Window.
appears when
a Boots Star method study is created or a
previously saved Boots study is opened.
Most data collection operations are
accessed from this window. The New
Set, Take Blank Scan, and Take Scan
selections in the Instrument Menu are
disabled when collecting data within a Boots study - to record a Boots Star scan at the software prompt, click the Scan
Execution button directly underneath the prompt.
The structure of the scan template for a Boots Star study cannot be changed, however, the user can correct mistakes made
during scan execution. To replace an incorrectly collected scan, load the appropriate sample plate into the UV-2000 and
click the scan location number in the scan template - the scan highlights while the instructions and scan execution button
in the Boots Star Method Window changes to reflect the scan template selection. Click the Scan Execution button to
record the scan.
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UV-2000 updates the Substrate Data Table in Figure 18 automatically when each sample plate is complete. The pre-
exposure updates include the highest standard deviation of transmittance, and UVA:UVB Ratio. Pre-exposure scans up
through Location No. 1 of Plate No. 3 are complete in Figure 18, for example, but the pre-exposure characteristics cannot
yet be displayed for Plates No. 3 and 4 because the scans have not yet been collected and the values can not yet be
computed.
During the course of the sunscreen analysis, UV-2000 automatically tracks the scanning progress of each sample plate,
both pre-exposure and post-exposure. Renaming the plates or scans in the Scan Sets Window does not impact thecorrelation between scans or sample plates. UV-2000 does not determine the legitimacy of the scan data when operating
under Boots Star guidelines. Instead, the standard deviation statistic provides the user a measure of consistency within the
analysis. This determination is made by computing the coefficient of variation (COV) of absorbance values between the
scans on the sample plate for each wavelength. When sufficient post-exposure data is collected, UV-2000 calculates the
final star ratings for the sunscreen analysis. These ratings are computed for each sample plate.
Figure 19. Statistics Window for a BootsStar method study displaying the traditional
sunscreen characteristics
UV-2000 calculates the and displays the traditional SPF and c sunscreencharacteristics during a Boots Star analysis as well as those stipulated by the
boots guidelines. The statistics displayed in the Statistics Window in Figure 19
reflect the same statistical analysis as when operating without a standard
method. Although saved in the Boots Star study document, statistics displayed
in this window do not apply to the Boots Star method and the statistics are
displayed for information only.
Reporting the Boots Star Test Results
The test results from a Boots Star study can be displayed in report format or
written to a separate file for subsequent analysis by a spreadsheet application.
Product information can be entered into the study by invoking the Study
Information Dialog Box from the Study Menu. To view a test report of the most
recently updated Boots analysis, activate the applicable study from the Window
menu and clickReport in the Study Menu.
Data export files are stored in the My Documents directory in comma-separated values format with the .cvs filename
extension. These files in can be viewed using Microsoft Excel or some other spreadsheet application. To export a
complete Boots study, activate the applicable study from the Window menu and clickExport Study from the File Menu.
To export a complete set of scans on a sample plate, highlight the set in the Scan Sets Window and clickExport Set.
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Blank and Sample Preparation
The in vitro characterization of a sunscreen can be performed by measuring the diffuse transmittance in the ultraviolet
spectrum of a carefully prepared sample. Preparation of the blank and sample plates is a critical issue when using the UV-
2000S, and there are two objectives sought in the preparation method. The first objective is to simulate the application
conditions used forin vivo testing - both the applied quantity of sunscreen product and substrate interaction. A test method
that accomplishes these conditions would produce a reliable in vitro SPF value that would positively predict the result of a
subsequent in vivo test. The second objective is consistency - the preparation method must generate reproducible results
on a sample-to-sample basis. It is difficult to devise blank and sample preparation routines that generates consistent test
results from sample to sample. In most cases, the consistency obtained depends on the sunscreen product and the
individual preparing the samples.
A blank scan is required for all sunscreen studies, whether or not a standard photoprotection method is utilized. When
operating with the confines of a photoprotection method, UV-2000 accommodates only one set of blank scan data for the
entire study so that errors made in plate preparation for the blank scan affect all sample plate analyses. There is no reason
to prepare more than one blank for these studies because the COLIPA and Boots Star substrates for the blank and sample
plates are now completely independent. The blank plates prepared with glycerin are not photostable and should be
scanned immediately at the beginning of a study. For sunscreen studies conducted without a standard photoprotection
method, the blank and sample plates may be one in the same, such that more than one blank scan may be useful for
generating independent plate analyses. UV-2000 accommodates these studies by permitting a separate blank scan for each
scan set.
Preparation of the Blank Plates
Labsphere has performed extensive testing on the UV-2000S using the COLIPA and Boots Star methods of sunscreen
analysis. The COLIPA method recommends a single side, roughenedpolymethylmethacrylate (PMMA) PlexiglasTM plate
as the optimum substrate for a blank or sunscreen sample. The plates are roughened on one side for simulating the skins
topography. The transmittance profile of the clean 2.5 mm thick PMMA plate, as measured by the UV-2000S, is a smooth
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curve starting from 52% and rising to approximately 71% at the high end of the spectrum. Three PMMA plate sizes fit the
UV-2000S stage assembly: 75 x 75 mm, 70 x 70 mm and 50 x 50 mm. The 50 x 50 mm holds nine sample locations and
the 75 x 75 mm plate is large enough to handle twelve sample locations easily. Nine position masks for each PMMA size
are included with the stage assembly. Plate selection depends only on the number of sample locations required for the
analysis. The same size PMMA plate should be used for blank and sample scans. Plates should be handled by the edges to
avoid damage or contamination of the surfaces.
The roughened PMMA or quartz plate to be used to capture a blank scanshould be prepared in a manner that produces thesame light scattering properties as the sunscreen. This preparation should include a thin coating of wetting agent. The
agent should be UV transparent and non-fluorescent. The most popular wetting agent as specified in the COLIPA and
Boots guidelines is glycerin,but other UV-transparent formulas can be used. Labsphere has produced good results with
both glycerin and ethyl alcohol.
Preparing a Glycerin Plate for a Blank Scan
This procedure provides guidelines for preparing a glycerin plate to be used subsequently for the blank scan. The
transmittance limits in Table 3 are provided in the COLIPA Guidelines, but may be useful for generic and Boots Star
studies as well.
1. Set the UV-2000S optics head to the correct height.
2. Remove the plastic tape from the bottom side of the plate. Wash both sides of the plate under clear, cold waterand blot the plate dry with a paper towel. Examine the plate for any foreign materials and discard the plate ifthey are evident.
3. Place the plate onto the analytical with the roughened side up and note the weight measurement.
4. Using a syringe or pipette, apply a number of small droplets of wetting agent onto the clean plate distributedevenly over the roughened surface until the total weight of the agent is 15 l for a 50 x 50 mm plate (34 l for a75 x 75 mm plate).
5. Remove the plate from the analytical balance and spread the agent across the entire surface of the plate evenlywith a finger and fingercot using light strokes as quickly as possible. Continue stroking the surface of the platein all directions until no puddles or areas of excess agent exist. Set the blank plate aside in a dark environmentfor at least 15 minutes.
Table 3. Transmittance limits for the wetted blank PMMA plate.
Wavelength (nm) Minimum
Transmittance (%)
Maximum
Transmittance (%)
290 60 70
300 69 79
320 81 91
6. Open a UV-2000S generic study. Enter the Study Menu, clickSubstrate Evaluation... and follow theinstructions on the screen. When the evaluation is complete, compare the transmittance results to Table 3.
7. Once the transmittance limits are achieved, proceed with the procedure for a COLIPA or Boots Star studydescribed in the Operating Procedures chapter in this manual.
Preparing an Ethyl Alcohol Plate for a Blank Scan
1. Set the UV-2000S optics head to the correct height.
2. Remove the plastic tape from the bottom side of the plate. Wash both sides of the plate under clear, cold waterand blot the plate dry with a paper towel. Examine the plate for any foreign materials and discard the plate ifthey are evident.
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3. Hold the PMMA plate over a paper towel with the roughened side up and dispense a few drops of alcohol to wetthe entire plate.
4. The blank scan must be collected on the plate immediately before the ethyl alcohol evaporates. Make sure thecorrect scan options are selected for your study and load the PMMA plate into the instrument with the wettedside up. Run a blank scan.
Preparation of the Sample PlatesSample prepara