EC Declaration of Conformity

according to EC Electromagnetic Compatibility Directive 89/336/EEC and 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: Model Number: UV-2000S Ultraviolet Transmittance Analyzer 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

AQ-02755-000, Rev. 0

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 ............................................................................................9 UV-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 ..............................................................................................26 Preparation 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

AQ-02755-000, Rev. 0

Introduction
The UV-2000S Ultraviolet Transmittance Analyzer is the most recent and highly application specific ultraviolet spectroscopy product offering 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 means of 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 of in 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 vitro broad spectrum methodologies. 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 protection test 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 their in 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 AQ-02755-000, Rev. 0 1

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 ratios Performance 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 Analyzer with the sample stage.

AQ-02755-000, Rev. 0

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 cable Validation kit UV-2000 installation CDROM Instruction manual CDROM Sample stage assembly with transparent sample mask templates AQ-02755-000, Rev. 0 3

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.
System Component Processor Operating System Display Memory Drives Ports Windows Required Minimum 1.6 GHz Windows XP or VISTA SVGA 800 x 600 100 Mb 256 Mb 1 1

Available HD Space RAM CD ROM USB 2.0

Table 1. Minimum computer requirements for the UV-2000 software.

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.

AQ-02755-000, Rev. 0

UV-2000S Installation
Assemble the UV-2000S Ultraviolet Transmittance Analyzer system as follows: 1. The minimum computer requirements for the UV-2000 software application are listed in Table 1. Load the UV-2000 install CDROM into the computer disk drive and install the software on your computer. The disk has an autorun feature and will install .NET Frameworks 2.0, if not already on your computer, and the UV-2000 application. When the installation is complete a prompt to restart the computer may appear on the screen - select Yes at the prompt. Remove the install CDROM from the disk drive when the installation is complete. Connect the power cord to the rear panel of the instrument and turn the UV-2000S on. Connect the USB cable between the instrument rear panel and an available USB port on your 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.

2. 3.
Figure 2. Making cable connections to the UV-2000.

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 must lie on a flat surface for proper system operation. 1. Place the stage assembly in front of the UV-2000S instrument. Slide the stage half-way out and make sure the aluminum holding arm that normally engages the sample plate is positioned away from the instrument. Lift up the front end of the instrument, slide the stage assembly probe underneath. Lower the instrument until the probe mates with the base plate cutout. It may be necessary to jiggle the front of the instrument side to side until the top surface of the stage is level with the sample area base plate. Load an empty sample plate and mask transparency into the stage and adjust the optics head position per the height adjustment procedure provided in the Operating Procedures chapter later in this manual. Operate the stage assembly in all directions. Observe that the stage glides smoothly into the sample area without clipping the plastic or metal surfaces in the sample area. Examine the empty plate in the sample area, noting that the plate rests flat on the sample floor.
Figure 3. Installing the sample stage.

2.

3.

4.

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Description of the UV-2000S

The UV-2000S measures the spectral 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 term spectral 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 of sunscreen 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-toRS-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 AQ-02755-000, Rev. 0 6

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 processed data collection from both spectrometers generates transmittance spectra according to the equation

S2 B1 T ( ) = ----- ----- B 2 S 1

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:

S 2 SD 2 B 1 BD 1 T ( ) = ---------------------- ---------------------- B 2 BD 2 S 1 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 each spectrometer, 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 instrument enclosure. 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. AQ-02755-000, Rev. 0 7

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 knob adjusts 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. 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.

Figure 4. Optics head and input optical chamber components.

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 light may affect your scan results. The term stray 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.

AQ-02755-000, Rev. 0

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 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 and moved to the location in Figure 6 to allow full withdrawal of the sample stage when testing large plates.

Figure 5. Sample stage assembly with a 75 mm mask overlay.

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.
Figure 6. Lengthening the vertical travel of the sample stage.

AQ-02755-000, Rev. 0

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 two types 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 AQ-02755-000, Rev. 0 10

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-2000 generates 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) or Boots 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 the operator at any time during the UV-2000S data session. To rename a study document, click Save 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 Figure 7. Viewing open study documents in the Window Menu. 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 the Window 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 click Export Study from the File Menu. To export a set of scans, highlight the set in the Scan Sets Window and click Export Set.

Recording Blank Scans

The 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 or Save 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 AQ-02755-000, Rev. 0 11

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 individual blank scan is applied to every sample scan in the scan set - always! In addition to the individual blank scans, UV-2000 may save a study default blank 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 is captured in Figure 8. The upper half of the dialog box controls UV2000 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 8. Scan Options Dialog Box.

Figure 9. Message appearing when the operator 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 set automatically 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.

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 OK to continue the analysis. If the operator creates a new study first, followed by the blank 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.

Figure 10. Option message that appears following a blank scan collected within a scan set.

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Figure 11. Message that appears when the operator creates a new scan set.

So, what does all this mean to the operator? The operator should choose the blank scan settings initially before embarking on the sunscreen analysis. 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 or Take 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 integration time for each blank or sample scan is set automatically by the AutoFlash 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 or Sample 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 click Save as application 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 protection factor mPF() data across the 290 - 450 nm wavelength spectrum SPF calculated with user-selected solar spectral irradiance data UVA:UVB Ratio Critical wavelength c Statistical 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. 2. 3. 4. 5. Create a new or select an existing study. Create a new or select an existing scan set. Record a blank scan. Collect a sample scan. 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, UV2000 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. UV2000 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. AQ-02755-000, Rev. 0 14

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 Equations No. 1 or 2 described previously in the last chapter. The data is displayed in units of percent on the spectral 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 spectral transmittance data according to the relationship A = log ( T ) where T() is expressed as a fraction. The data is displayed absorbance units (AU) on the spectral chart and 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 described above. Monochromatic Protection Factor mPF(). The monochromatic protection factor is the spectral quantity equal to the inverse of transmittance: 1mPF = ----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 the traditional 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 produce 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
400

E S d
SPF =
290 -------------------------------400

E S T d
290

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where E() is the erythema action spectrum, S() is the solar spectral irradiance, T() is the spectral transmittance 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 the following relation: c such that ' satisfies the relationship:

= Min ( ),

= 290

------------------------- 0.9 400

=
290

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:
n n

SPFdMean =

i=1

SPF ( i ) --------------n

LambdadCriticaldMean =

i=1

c ----------n

(i)

The standard deviation statistics are calculated for the entire set that applies to the selected scan or set in the Scan Sets Window:
n 2

SPFdSTD =

i=1

( SPF ( i ) SPFdMean ) --------------------------------------------------------n1

LambdadCriticaldSTD =

i=1

--------------------------------------------------------------------------------------- ( c LambdadCriticaldMean ) n1
(i)

The coefficient of variation terms are defined for each set as:
STD COV = ------------- 100 %. Mean

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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 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 Figure 13. Editing a scan set or individual scan within a set when operatgeneric study. All scans that appear on the graph and the Scan Sets Window, for a ing outside a photoprotection method. 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, click Graph 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 click Report 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 click Export 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 click Export Set.

* For clarification, the "standard sun" spectrum referred to within this document as "Albuquerque, NM" is actually the rounded average of 35 degrees north (Albuquerque) and 40 degrees North, as reported by COLIPA, SPF Test Method, May 1994. As noted within the COLIPA SPF Test Method, what is important is that the "standard sun" reference spectrum represents a realistic maximum solar 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 spectrum SPFin 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 irradiation 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 and c characteristics from a single scan in the same manner as the calculation in the non-method study analyses. The operator can view the SPF and c parameters 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 the bottom left-hand corner. In vitro SPF (SPFin vitro). The SPFin vitro characteristic is described in the COLIPA 2000 guidelines as the ratio
400

E ( )I ( ) d
A 0 ( )

290 SPF indvitro = -------------------------------------------------------, 400

E ( )I ( )10
290

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 test product 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 determined for the same sunscreen product, to determine the coefficient of adjustment, C. UV-2000 calculates the value of the Coefficient of Adjustment C automatically and applies the coefficient to the ratio
400

E ( )I ( ) d
320 UVAPF 0 = ------------------------------------------------------------. 400

E ( )I ( )10
320

A 0 ( )C

The COLIPA Method recommends that C falls within a range between 0.8 and 1.2. UVAPF0 is deterAQ-02755-000, Rev. 0 18

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 scans are 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 SPFlabel parameter 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. To create 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 in the 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 is created. 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 preirradiation data is collected. Test characteristics generated by UV-2000 under COLIPA guidelines are reported on the COLIPA Method Window in Figure 15 that appears when a COLIPA method study is created or a previously saved COLIPA study is opened. Most data collection operations are accessed from Figure 15. Calculations displayed in the COLIPA Method Window. 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 AQ-02755-000, Rev. 0 20

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 the checkboxes 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. 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 Figure 16. Statistics Window for a COLIPA select Solar Irradiance from the Study Menu and select either the Melbourne or method study visible during the execution COLIPA data. The irradiance data set that appears here is used only for the of the Plate 3 scan set from Figure 14. 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 click Report 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 click Export 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 click Export 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 AQ-02755-000, Rev. 0 21

system uses the terms pre-exposure and post-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 protection factor 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 and c characteristics from a single scan in the same manner as the calculation in the traditional study analyses. The operator can view the SPF and c parameters 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 the bottom 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:
400

A ( ) d
UVA =
320 ------------------------400

d
320

where A() is the monochromatic absorbance averaged across the sample plate for each wavelength of the 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:
320

A ( ) d
UVB =
290 ------------------------320

d
290

where A() is the monochromatic absorbance averaged across the sample plate for each wavelength of the 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. AQ-02755-000, Rev. 0 22

UVA:UVB Ratio. The UVA:UVB Ratio sunscreen characteristic is calculated for pre-exposure and post-exposure conditions:
UVA UVA:UVB Ratiod = -----------UVB

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.

INITIAL Mean UVA:UVB RATIO 0.0 to 0.59 POST EXPOSURE Mean UVA:UVB RATIO 0.0 to 0.56 0.57 to 0.75 0.76 to 0.85 0.86 and Over No Rating No Rating No Rating No Rating 0.6 to 0.79 No Rating *** *** *** 0.8 to 0.89 No Rating *** **** **** 0.9 and over No Rating *** **** *****

Table 2. Star Rating criteria, copied from "Measurement of UVA:UVB Ratios According to the Boots Star Rating System (2008 Revision)" distributed by Boots (UK) Limited.

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 apply user 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 main operating screen that now includes the Boots Star Method Window.

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 UV2000 under the Boots guidelines are reported on the Boots Star Method Window in Figure 18 that 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 Figure 18. Calculations displayed in the Boots Star Method Window. 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. AQ-02755-000, Rev. 0 24

UV-2000 updates the Substrate Data Table in Figure 18 automatically when each sample plate is complete. The preexposure 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 the correlation 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. UV-2000 calculates the and displays the traditional SPF and c sunscreen characteristics 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 click Report in the Study Menu.

Figure 19. Statistics Window for a Boots Star method study displaying the traditional sunscreen characteristics

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 click Export 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 click Export 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 UV2000S, and there are two objectives sought in the preparation method. The first objective is to simulate the application conditions used for in 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, roughened polymethylmethacrylate (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 AQ-02755-000, Rev. 0 26

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 scan should be prepared in a manner that produces the same 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. 2. 3. 4. 5. Set the UV-2000S optics head to the correct height. Remove the plastic tape from the bottom side of the plate. Wash both sides of the plate under clear, cold water and blot the plate dry with a paper towel. Examine the plate for any foreign materials and discard the plate if they are evident. Place the plate onto the analytical with the roughened side up and note the weight measurement. Using a syringe or pipette, apply a number of small droplets of wetting agent onto the clean plate distributed evenly over the roughened surface until the total weight of the agent is 15 l for a 50 x 50 mm plate (34 l for a 75 x 75 mm plate). Remove the plate from the analytical balance and spread the agent across the entire surface of the plate evenly with a finger and fingercot using light strokes as quickly as possible. Continue stroking the surface of the plate in all directions until no puddles or areas of excess agent exist. Set the blank plate aside in a dark environment for at least 15 minutes.

Wavelength (nm) 290 300 320

Minimum Transmittance (%) 60 69 81

Maximum Transmittance (%) 70 79 91

Table 3. Transmittance limits for the wetted blank PMMA plate.

6. 7.

Open a UV-2000S generic study. Enter the Study Menu, click Substrate Evaluation... and follow the instructions on the screen. When the evaluation is complete, compare the transmittance results to Table 3. Once the transmittance limits are achieved, proceed with the procedure for a COLIPA or Boots Star study described in the Operating Procedures chapter in this manual.

Preparing an Ethyl Alcohol Plate for a Blank Scan

1. 2. Set the UV-2000S optics head to the correct height. Remove the plastic tape from the bottom side of the plate. Wash both sides of the plate under clear, cold water and blot the plate dry with a paper towel. Examine the plate for any foreign materials and discard the plate if they are evident. 27

AQ-02755-000, Rev. 0

3. 4.

Hold the PMMA plate over a paper towel with the roughened side up and dispense a few drops of alcohol to wet the entire plate. The blank scan must be collected on the plate immediately before the ethyl alcohol evaporates. Make sure the correct scan options are selected for your study and load the PMMA plate into the instrument with the wetted side up. Run a blank scan.

Preparation of the Sample Plates

Sample preparation techniques stipulated by the COLIPA and Boots Star guidelines are very similar. The COLIPA procedure requires a sample application rate of 0.75 mg/cm2, spread by a "pre-saturated" fingertip. The Boots Star procedure prescribes a 1.0 mg/cm2 application rate but without the "pre-saturated" fingertip criteria. Consequently, only one standard sample procedure is provided here that satisfies both guidelines. 1. 2. 3. 4. 5. Calculate the weight of the sunscreen sample required by the guidelines of the analysis. The weight is based on the application rate of the method and the top surface area of the PMMA sample plate. Remove the plastic tape from the bottom side of the plate. Wash both sides of the plate under clear, cold water and blot the plate dry with a paper towel. Examine the plate for any foreign materials and discard the plate if they are evident. Place the plate on an analytical balance (capable of reading 0.0001 g recommended) with the roughened side facing up. Note the scale reading or zero the balance. Apply the product in a series of small droplets spread evenly across the plate with a syringe until the correct weight calculated in Step 1 is indicated. Remove the sample plate from the analytical balance and, using the "pre-saturated" fingertip as described in the two-phase COLIPA process or the gloved finger Boots Star technique, spread the product quickly and evenly across the entire surface of the plate. The spreading motion for distributing the sample product should be light stokes applied in the shortest amount of time. If conducting a COLIPA procedure, proceed to step 6, if Boots Star procedure, skip step 6 and proceed with step 7. Once the product is uniformly spread, rub the sample into the rough surface of the plate applying light pressure. This phase of application should take 20 to 30 seconds. When the sample is spread evenly with no gaps, smears or excess product buildup, immediately store the prepared sample plate in a dark area at room temperature. Allow the sample plate to dry for fifteen minutes.

6. 7.

Alternative Sample Preparations

Since the roughened PMMA plate technique is relatively new to the sunscreen industry, some manufacturers may still utilize the quartz plates for their traditional sampling. We offer two methods of preparation for the quartz sample plate these methods are the same procedures recommended and used successfully with the Labsphere UV-1000S instrument. Quartz plates should be handled by the edges to avoid damage or contamination of the surfaces.

Alternative Method 1 (Transpore Tape Substrate)

The main advantages of the Transpore tape are its low cost, ready availability, and ease of use. Another important advantage is that once prepared, the same blank plate can be used for all subsequent samples prepared from the same tape roll. There are a few disadvantages that need consideration when using Transpore tape: The tape will not absorb formulations that use alcohol or oil as a vehicle. Pore size can vary at the beginning and end of each roll. Therefore, it is recommended to discard the first two feet at the start and end of each roll. The pore size can vary from batch-to-batch it is advisable to screen at least 4 rolls to determine the suitability of a larger batch. Screening involves measuring a standard sunscreen formula of known SPF. The following procedure prepares the blank and sample quartz plates for sunscreen analysis. If the operator prepares one sample plate at a time, the timing for each plate will be staggered. AQ-02755-000, Rev. 0 28

1.

Examine the quartz plates for evidence of dust or other foreign materials. To clean a plate of dust, use clean high pressure air or nitrogen. If smudges or grease is visible on a plate, clean the plate surfaces with alcohol followed by a clean water. Rinse the plate with water until there is no evidence of residue or foreign material. Dry with a clean, soft, lint-free cloth. Cut a piece of Transpore tape for each plate being prepared and lay the tape pieces on a flat surface with the adhesive side up. Place a plate onto each piece of tape and then remove the overhanging edges of the tape with a sharp blade. Retain one of these prepared plates for the blank scan. Place the quartz sample plate on an analytical balance with the tape side up. Using a pipette, apply a number of small droplets of sunscreen product onto the clean plate distributed evenly over the tape surface until the application rate of the product by weight reaches Calculate the total weight of the product to achieve a 2.0 mg/cm2 application rate on the plate. Remove the plate from the balance and spread the sunscreen across the entire surface of the plate with a single finger and fingercot using light strokes as quickly as possible. Continue stroking the surface of the plate in all directions until no puddles or areas of excess sunscreen exist. Store the prepared sample plate in a dark location at ambient temperature for 15 minutes. Repeat Steps No. 3 through 6 for each sample plate.

2. 3. 4. 5. 6. 7.

Alternative Method 2 (Vitro-Skin Substrate)

Vitro-Skin, a registered trademark of IMS Inc. (www.ims-usa.com), is a synthetic skin substitute that has been widely used for in vitro analysis of sunscreens. Once hydrated, Vitro-Skin has a texture very similar to human epidermis. In addition, the hydrated material seems to help sunscreen emulsions break down in much the same way as human skin. Published data by the producer of Vitro-Skin indicates that the substrate gives excellent correlation with in vivo SPF measurements. The primary advantages of Vitro-Skin are its topographical similarity to human skin and the ability to break down emulsions. The disadvantages are: Relatively high cost per sample. Overall low UV transmittance, especially at low wavelengths. The need to hydrate the material before use and the relatively short lifetime of the sample plate. The following procedure prepares the blank and sample quartz plates for sunscreen analysis. 1. 2. 3. Cut a 1"x 2" section of Vitro-Skin for each sample plate prepared. Prepare a sufficient number of samples for each sunscreen to be tested. Hydrate the pieces using the method suggested by the manufacturer. The hydration procedure is accessed under the Testing Group Menu and the Protocol selection of the manufacturers website. Retain one of the plates for the blank scan. The blank slide can be used for all Vitro-Skin based measurements made from the same sheet for samples run during the same time period.

If the operator prepares one sample plate at a time, the timing for each plate will be staggered so that photostability is not an issue. The manufactures website provides a sample application procedure as well. 4. The Vitro-Skin is then placed rough side up on an analytical balance. Using a small micropipette or syringe with a fine blunt needle, deposit 12 l or other prescribed number of sample drops over the center area of the VitroSkin. It is our experience that approximately 2 mg /cm2 of the sample should be spread on the substrate, so you can adjust the number of sample drops to the surface area prepared. Care should be taken not to puncture the Vitro-Skin during the application process, nor should sunscreen be placed on the outer 5 mm edge which is used to hold the sample in the slide mount frame. Once the sunscreen sample has been accurately weighed, the VitroSkin sample is transferred, sunscreen coated side up, to a foam block-used to simulate the flexibility of human flesh--and gently rubbed in with a cot covered finger for 20 to 30 seconds. The rubbing motion should be circular at first, then slowly back and forth, as one would apply sunscreen to ones body. Once the sunscreen has been spread, the samples should be allowed to dry for 20 minutes to let the emulsion break down before beginning measurements. Failure to allow this dry-down period will result in inaccurate SPF measurements.

5.

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Operating Procedures
The UV-2000S supports in vitro testing using a methods framework. The user can select one of the embedded methods, either Boots Star or COLIPA, or he can collect sunscreen data using the traditional sunscreen characteristics of the past. Table 4 lists a brief comparison of the two current methods and the method proposed by the U.S. Federal Drug Administration. The 21 CFR method is not yet incorporated into the UV-2000 software.

Procedural Step Plate Material Sample Application Rate Minimum No. Sample Plates Minimum Readings per Plate Irradiation Sample Plate Rejection Analysis Rejection Characterization

COLIPA 2007 PMMA 0.75 mg/cm2 3 Total Area = 2.0 cm

2 2

BOOTS STAR 2008 Quartz or PMMA 1.0 mg/cm2 5 5, Total Area = 2.0 cm D = 17.5 J/cm None UVA:UVB Ratio Photostability degradation < 5% UVA:UVB Star Rating
2 2

21 CFR 352 Quartz 2.0 mg/cm2 5 12 D = (SPF)(200)(2/3) J/m2 COV > 10% of 5 nm T() values on a plate None UVA:UV Ratio & Star Rating

D = (UVAPF0)(1.2) J/cm

Absorbance COV > 50% Plate UVAPF COV > 20% UVAPF & SPF/UVAPF Ratio

Table 4. Summary of the approved in vitro test methods.

The procedures in this chapter provide guidelines for operating the UV-2000S instrument and performing the COLIPA and Boots Star analyses. No procedures for a generic study are provided.

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Setting the Optics Head Height

The UV-2000S optic head can be raised and lowered to accommodate loading and unloading a sample plate. The knobs on either side of the UV-2000 instrument adjust the height. The right-hand knob lifts the optics head up and down; the lefthand knob establishes the bottom limit of the upper chamber height above the sample plate. The left-hand knob is springloaded - the operator must push the left-hand knob inward to engage the worm gear inside. 1. 2. 3. 4. Engage and turn the left-hand knob counterclockwise to raise the lower limit of the optics head so there is plenty of clearance underneath. Load an empty PMMA plate into the sample area. Shine a flashlight into the sample area and carefully turn the left-hand knob clockwise until the optics head is positioned approximately two millimeters above the top surface of the plate. Disengage the left-hand knob and remove the plate from the sample area.

The distance between the optics head and sample plate should be maintained for the duration of all scans within a scan set.

Operating the Sample Stage Assembly

To install the stage assembly, lift up the front end of the instrument, slide the stage assembly probe underneath until it mates with the base plate cutout. It may be necessary to jiggle the front of the instrument side to side until the top surface of the stage is level with the sample area base plate. 1. 2. Select a transparent mask that corresponds to your sample plate format. Mount the mask into the stage assembly as shown in Figure 20 and twist the two brackets over the mask. Slide the stage half-way out from the sample area and load the blank or sample plate into the slide holder. Make sure the plate lies flat on the sample floor and lock the plate in place with the knurled knob as demonstrated in Figure 21. 3.
Figure 20. Installing the mask into the stage assembly.

Wearing either non-powdered latex gloves or finger cots, carefully slide the stage and sample plate into the sample area, taking care not to clip the edge of the sample floor or the bottom surface of the upper chamber optic. Position the stage assembly to the first mask position. Execute the scan at the software prompt or from the Instrument Menu, as appropriate, and move the stage to the next scan position. When all scans on the sample plate are complete, move the stage half-way out, loosen the knurled knob and remove the plate.

4. 5.
Figure 21. Loading a plate into the sample stage.

Depending on the number of locations programmed for a COLIPA or Boots Star study, the locations illustrated on the transparency masks may not correspond directly to the scan template visible in the UV-2000 software. If the number of sample locations is less than nine, the operator should devise a mapping method for identifying the actual sample location for each scan.

Recording a COLIPA Sunscreen Analysis

This procedure steps the operator through an entire sunscreen analysis from within a COLIPA study. Many of the tasks inherent to the procedure are time critical and the analysis may require the participation of more than one person. The COLIPA method requires at least three sample plates in the sunscreen analysis and three scan locations per plate. The three sample locations satisfy the minimum 2.0 cm2 sample area requirements of the COLIPA 2007 Guidelines. When entering the COLIPA method options for the study, be sure to account for potential sample rejections during the final steps AQ-02755-000, Rev. 0 31

of the analysis. 1. 2. 3. 4. 5. 6. 7. Power up the UV-2000S system and wait five minutes for spectrometer warm-up. Set the UV-2000S optics head to the correct height. Prepare the plate for recording the blank scan as described in the previous chapter. Open a COLIPA study from the Study Menu. Invoke the Study Information Dialog Box from the Study Menu and make the appropriate product and comment entries. Invoke the Scan Options Dialog Box and apply any necessary changes to scan options. Load the blank plate into the UV-2000S sample area. Record the blank scan data by clicking the Scan Execution button that now reads "Take blank scan". Save the study in the File Menu and remove the blank plate from the instrument sample area. Prepare the sample plates and label them according to the COLIPA method options selected in Step No. 5. An equilibration period of at least 15 minutes is required for each sample plate prior to collecting the sample scan. Typically, the sample plates are stored on a platform located in a dark area until the specified time period is elapsed. Load the first sample plate into the UV-2000S sample area or the sample stage, if applicable. Position the plate to the first sample location. Record the sample scan at this position by clicking the Scan Execution button that now reads "Scan location 1". Examine the scan curve and scan data for the plate just scanned. If the curve and data are acceptable, continue to the next step. Repeat the previous step for each programmed scan location on the sample plate, each time physically advancing the plate to the next sample location. When all sample scans are recorded satisfactorily for the plate, examine the sunscreen characteristic data displayed in the Statistics Window and the COLIPA Method Window. If the displayed results are satisfactory, click Save Study in the File Menu to save the recently collected data to the study document. Calculate the irradiation interval required for the sample plate based on the "Irradiation Dose" value displayed in the Plate Data Table. Remove the plate from the UV-2000S and place it on the plastic irradiation sheet. Record the start and anticipated stop times for the irradiation interval. Repeat Steps No. 8 through 11 for each plate on the scan template until all plates are complete. UV-2000 prompts the operator throughout the process in the "Instructions" heading in the COLIPA Method Window. The irradiation intervals for the sample plates will not necessarily elapse in the same order commenced, so the UV-2000 software prompts may not provide the optimum post-irradiation test sequence. Remove each sample plate from the irradiation sheet when the irradiation time interval is elapsed. Load the first sample plate off the irradiation sheet into the UV-2000S sample area or the sample stage, if applicable. Position the plate to the first sample location. Find the scan set for the sample plate in the post-irradiation listing of the scan template and left-click on the scan set name. Record the sample scan at this position by clicking the Scan Execution button that now reads "Scan location 1". Click again on the scan name for the scan just recorded and examine the scan curve and scan data. If the curve and data are acceptable, remove the sample plate from the UV-2000S and continue to the next step. Repeat the previous step for each programmed sample scan on the sample plate, each time physically advancing the plate to the next sample location. When all sample scans are recorded satisfactorily for the plate, examine the sunscreen characteristic data displayed in the Statistics Window and the COLIPA Method Window. If the displayed results are satisfactory, click Save Study in the File Menu to save the recently collected data to the study document. Repeat Steps No. 14 through 16 for each subsequent plate removed from the irradiation sheet until all plates are complete. When all sample plate scanning is complete, examine the checkbox for each plate under the "Include Data in Results" header. UV-2000 automatically includes a sample plate that meets the 50% COV absorbance criteria and leaves unchecked the checkbox for any plate that exceeds the 50% COV. A minimum of three qualified sample plates is required to proceed with the analysis and the COLIPA method instructs the user to measure additional sample plates if the minimum number of plates is not satisfied. Finally, examine the UVAPF COV value reported by UV-2000 for the analysis - the UVAPF COV between the sample plates should be no greater than 20%. When the sunscreen analysis is complete, make sure the study is saved and export the data or study to a .cvs file. 32

8.

9. 10. 11. 12. 13. 14.

15. 16. 17. 18.

19. 20.

AQ-02755-000, Rev. 0

Recording a Boots Star Sunscreen Analysis

This procedure steps the operator through an entire sunscreen analysis from within a Boots Star study. Many of the tasks inherent to the procedure are time critical and the analysis may require the participation of more than one person. The Boots Star method requires at least five sample plates in the sunscreen analysis and five scan locations per plate. 1. 2. 3. 4. 5. 6. Power up the UV-2000S system and wait five minutes for spectrometer warm-up. Set the UV-2000S optics head to the correct height. Prepare the plate for recording the blank scan as described in the previous chapter. Open a Boots Star study from the Study Menu. Invoke the Study Information Dialog Box from the Study Menu and make the appropriate product and comment entries. Invoke the Scan Options Dialog Box and apply any necessary changes to scan options. Load the blank plate into the UV-2000S sample area and position the plate to the first blank scan location. Lower the optics head down to the lower set limit. Record the blank scan data by clicking the Scan Execution button that now reads "Take blank scan" and advancing the blank plate to the next location at each prompt. Save the study in the File Menu and remove the blank plate from the instrument sample area. Prepare the sample plates and label them according to the Boots Star method options selected in Step No. 5. An equilibration period of at least 15 minutes is required for each sample plate prior to collecting the sample scan. Typically, the sample plates are stored on a platform located in a dark area until the specified time period is elapsed. Load the first sample plate into the UV-2000S sample area or the sample stage, if applicable. Position the plate to the first sample location. Record the sample scan at this position by clicking the Scan Execution button that now reads "Scan location 1". Examine the scan curve and scan data for the plate just scanned. If the curve and data are acceptable, continue to the next step. Repeat the previous step for each programmed scan location on the sample plate, each time physically advancing the plate to the next sample location. When all sample scans are recorded satisfactorily for the plate, examine the sunscreen characteristic data displayed in the Statistics Window and the Boots Star Method Window. If the displayed results are satisfactory, click Save Study in the File Menu to save the recently collected data to the study document. Remove the plate from the UV-2000S and place it on the plastic irradiation sheet. Calculate the irradiation interval required for the sample plate and expose the sample plate in accordance with the Boot Star guidelines. The current Boots Star approved exposure is 17.5 J/cm2. Record the start and anticipated stop times for the irradiation interval. Repeat Steps No. 8 through 11 for each plate on the scan template until all plates are complete. UV-2000 prompts the operator throughout the process in the "Instructions" heading in the Boots Star Method Window. Load the first sample plate off the UV exposure sheet into the UV-2000S sample area or the sample stage, if applicable. Position the plate to the first sample location. Record the sample scan at this position by clicking the Scan Execution button that now reads "Scan location 1". Click again on the scan name for the scan just recorded and examine the scan curve and scan data. If the curve and data are acceptable, remove the sample plate from the UV-2000S and continue to the next step. Repeat the previous step for each programmed location on the sample plate, each time physically advancing the plate to the next sample location. When all sample scans are recorded satisfactorily for the plate, examine the sunscreen characteristic data displayed in the Statistics Window and the Boots Star Method Window. If the displayed results are satisfactory, click Save Study in the File Menu to save the recently collected data to the study document. Repeat Steps No. 13 through 15 for each subsequent plate removed from the exposure sheet until all plates are complete. When all sample plate scanning is complete, examine the UVA:UVB ratios for each plate and choose the star rating according to organizational policy. When the sunscreen analysis is complete, make sure the study is saved and export the data or study to a .cvs file.

7.

8.

9. 10. 11.

12. 13.

14. 15. 16. 17. 18.

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Instrument Validation
The purpose of instrument validation is to ensure the instrument is functioning properly. The instrument validation routine tests the instrument for drift, detector response linearity, and wavelength accuracy. Should any test not pass, contact the Labsphere Customer Service Department at

(603) 927-4266. Instrument validation should be performed on a monthly basis and whenever instrument accuracy is suspect. Complete the steps that follow. You can cancel the validation from any dialog box in the procedure by clicking on Cancel. All data taken to that point will be lost if Cancel is selected. A validation kit accompanies the instrument that includes the six test filters required to complete the validation analysis. When loading a filter into the UV-2000S, the letter and transmittance value on the side of the optics holder should be upright with the flat side down. During the filter sample scans the lower part of the optics head should be protrude into the reamed cavity in the filter holder. 1. 2. 3. 4. 5. 6. Place two empty PMMA plates into the sample area and lower the optics head using the left-hand knob until the bottom surface of the optics head just touches the top PMMA plate. Lift the optics head with the right-hand knob and remove both plates. Select Instrument Validation from the Instrument Menu. A dialog box will appear on your screen asking for a number of wavelength and amplitude test parameters peculiar to your instrument. Enter the data requested. The test parameters are listed in the calibration certificate for the validation filter kit. The next message issued by the software is the blank scan prompt. The blank scan is recorded with the optics head at the low limit set in Step No. 1 and with no plate or filter loaded in the sample area. Execute the blank scan. The remaining steps in the validation routine include a series of sample scans. Follow the prompts issued by the software, loading a filter into the instrument each time. The final validation data collection is the 100% baseline scan. Upon completion of the routine, examine the validation results at the bottom of the screen. Any portion of the validation can be repeated by clicking a scan in the Scan Sets Window and following the prompt in the Validation Window. When no more scanning is required, click Save As from the File Menu and enter an appropriate filename that includes the instrument serial number.

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Maintenance
Do not attempt to disassemble the optics head and do not open the instrument enclosure - there are no user serviceable components inside.

DANGER: Some interior components operate on high voltage. Do not open the instrument enclosure.

Some routine maintenance the user can perform is as follows: 1. 2. 3. 4. Do not insert abrasive objects in the sample holder. To clean the optics holder, use isopropanol and a lint-free cloth to wipe down the optics on the upper and lower chamber. If the scan results generated by the instrument appear suspect or if the instrument fails the validation routine, obtain the validation files from the on your computer hard drive for the last six months and email the files to Labsphere for analysis. Check the number of flashes on the flashlamp periodically by clicking Device Information in the Instrument Menu. The manufacturers lifetime for the flashlamp is 108 shots. The instrument should be returned to Labsphere for flashlamp replacement and subsequent re-calibration before the existing flashlamp fails. The autoflash feature functions by performing a shortened transmittance scan on the sunscreen sample and adjusting the number of flashlamp flashes just prior to recording the full sample scan. One advantage of autoflash is that saturation of the spectrometer arrays cannot occur during normal operation. Nevertheless, the UV-2000 software monitors count levels from each spectrometer and issues a warning message should any pixel count reach saturation. If a saturation warning message appears during UV-2000S operation, contact your Labsphere representative or call us at the factory in the U. S. at

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(603) 927-4266.
5. The UV-2000S should be returned to Labsphere for re-calibration on a yearly basis. Items that should be returned include the validation filters and the UV-2000S instrument itself. Use the following packaging procedure when returning the instrument for re-calibration: a. b. Contact your Labsphere representative or call us at the factory in the U. S. You will receive a sales order or possibly an RMA number for your re-calibration. The original shipping box includes a re-enforced cardboard box with two foam inserts. Place the bottom foam insert into the box and set the UV-2000S instrument into the foam cutout as illustrated in Figure 22. Do not return the stage assembly unless it is defective. Place the top foam insert over the instrument so it fits over the instrument enclosure and surrounds the leftand right-hand operating knobs. Place the filter in their original plastic box and place the box into the smaller top cutout of the top foam insert as illustrated in Figure 23. Do not return any cabling and do not include any other loose objects in the box that may scratch the painted surface of the UV-2000S. Close the box and tape the edges. Print the sales order number or RMA number on the outside of the box. Ship the package to the address on your purchase order or as directed by your Labsphere sales representative.
Figure 22. Bottom shipping configuration in the original container.

c. d. e. f. g.

Figure 23. Top insert shipping configuration for the original container.

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Appendix A Specifications
Performance Specifications Spectral Range Wavelength Accuracy Dynamic Range (Absorbance) Data Interval Sample Size Scan Time Design Specifications Dimensions (w/o stage assembly) Dimensions (with stage assembly) Weight Integrating Sphere Sphere Diameter Flashlamp Lamp (Labsphere Part No. OC-03144-000) Power Supply Spectrometers Diode Array (512 Pixel) Integration Time Data Resolution Computer Interface Electrical Specifications Voltage Power (not including computer) * All system specifications are based on a wavelength range of 290-400 nm xenon Modified MVS-4000 OC-08023-100 (Master), OC-08023-200 (Slave) 250 - 450 nm 20 ms per flash 16-bit USB 2.0 Value 100 - 240 VAC, 50/60 Hz 20 Watts Value 250 - 450 nm* +1.0 nm 0 - 2.7 AU 1 nm 1 cm dia. < 5 sec. Value 11H x 12.6D x 12.3W in (27.9H 32.0D x 31.2W cm) 11H x 22.6D x 12.3W In (27.9H x 56.6D x 31.2W cm) 26 lbs. Spectralon 1.5 in.

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Appendix B Schematics

UV-2000S Instrument

UV-2000 Software

Flashlamp Power Supply Upper Chamber RS232/USB Converter

Trigger Module

sample(1) = Lsample(1) - Dsample(1)

32-bit Binary Data

Spectrometer No. 1

Data
(RS232)

(USB)

Blank(1) = LBlank(1) - DBlank(1)

32-bit Binary Data

Blank() = Blank(2) Blank(1)

16-bit Binary Data

sample() =

sample(2) sample(1)

T() = sample()

Data
(RS232)

100% Blank()

Spectrometer No. 2 (Master) Lower Chamber

Blank(2) = LBlank(2) - DBlank(2)

32-bit Binary Data

sample(2) = Lsample(2) - Dsample(2)

Trigger

32-bit Binary Data

LBlank(1) = Spectrometer No. 1 spectral flux during blank scan DBlank(1) = Spectrometer No. 1 dark current reading during blank scan Blank(1) = Corrected Spectrometer No. 1 spectral flux for blank scan LBlank(2) = Spectrometer No. 2 spectral flux during blank scan DBlank(2) = Spectrometer No. 2 dark current reading during blank scan Blank(2) = Corrected Spectrometer No. 2 spectral flux for blank scan

LSample(1) = Spectrometer No. 1 spectral flux during sample scan DSample(1) = Spectrometer No. 1 dark current reading during sample scan Sample(1) = Corrected Spectrometer No. 1 spectral flux for sample scan LSample(2) = Spectrometer No. 2 spectral flux during sample scan DSample(2) = Spectrometer No. 2 dark current reading during sample scan Sample(2) = Corrected Spectrometer No. 2 spectral flux for sample scan T() = Spectral transmittance of sample

Figure 24. Functional block diagram of the UV-2000S.

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