Spectrophotometric Standards Reference

Guide

NIST TraceableCustomizable

No Mixing or Diluting

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CHEMICALS ®

Ordering

ContentsTable of Contents

Continuous Improvement

In-Spec® UV Standards

In-Spec® Visible Standards

In-Spec® Patent Blue Standards

In-Spec® Amaranth Red Standards

In-Spec® BCA Purple Standards

In-Spec® Nano-Kit

Wavelength Standards

Stray Light Filters

Potassium Dichromate

Custom Standards

Amco Clear®Turbidity Standards

Application Notes

Troubleshooting Guide

Analytical & Lab Chemicals

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5-6

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Ordering InformationCustomer service representatives and technical support are available Monday through Friday from 7:30 am to 4:00 pm Eastern Time. Orders may be placed by phone, e-mail or fax.Every order receives an order confirmation that includes items ordered, pricing, shipping method and estimated shipping date.

GFS Chemicals, Inc.800 Kaderly Drive Columbus, OH 43228800-838-5567 extension 121 (U.S. and Canada) 740-881-5989 (Fax)001-740-881-5501 (International) in-spec@gfschemicals.com (e-mail)

www.in-spec.comwww.gfschemicals.com

Opening an AccountGFS Customer Service Representatives will be glad to help you open an account. For terms and conditions see Inside Back Cover.For complete Terms and Conditions, please visit:http://www.gfschemicals.com/statics/documents/aboutus/termsandconditions.html

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Continuous

Improvement

The In-Spec® product line is constantly growing and currently includes standards to check not just photometric accuracy but also wave-length, stray light, and resolution for a variety of instrumentation.

Some recent areas of focus have been short pathlength instruments and microplate readers used in many pharmaceutical and biotech laboratories.

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Features Benefits

GFS Chemicals has been manufacturing standards for over twenty years for unique customer applications. Many of our standards are specific to your industry.

SeTTINg The STaNDarD for SpeCTrophoToMeTrIC aNalySIS & verIfICaTIoN

Continuous improvement at GFS Chemicals means:• Development of new standards for the latest instruments. • R&D for new uses and applications for the In-Spec® product line.• Investment in a new laboratory to meet higher quality control standards for the demands of the most stringent industries.• Certification for ISO 17025 is underway.• Experienced technical support to troubleshoot and answer your specific questions.

Non-Toxic No disposal fees Green Alternative

Inexpensive Save cost compared to sealed cuvette alternatives

Easy to Use No wasted time with dilutions No special precautions

NIST Traceable Confidence in instrument readings Meets regulatory requirements

Live Technical Assistance Assistance for method development Help with troubleshooting

Customizable Made for your application Multiple pathlengths and wavelengths available

Manufactured by GFS In stock and ready to ship Trusted name for quality in the industry

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Selection Gui deHow to Use:Choose your wavelength of interest then go across the chart to find your closest target absorbance value. The top of the column will identify the item number and product name associated with that value.

Please note that the values in the table are representative values only and are to assist in picking out the appropriate standard for your application. The actual value of the lot you receive may vary. The data provided is representative of a 10mm pathlength cell.

In-Spec® Photometric Standards Selection Guide

In-Spec® UV Standards

In-Spec® UV Standards are de-signed to check the photometric accuracy of a traditional spectro-photometer from 250nm up to 400nm.

UV Standards - Item NumbersUV-1: 8301UV-2: 8302UV-3: 8303UV-4: 8304UV-5: 8305UV-6: 8306UV-7: 8307UV-8: 8308 UV-20: 8309UV-30: 8310

In-Spec® Visible Standards

The In-Spec® Visible Standards are designed to check the pho-tometric accuracy of a traditional spectrophotometer from 350nm up to 850nm.

Visible Standards-Item Numbers Vis 0.05: 8311 Vis 0.1: 8312 Vis 0.3: 8322 Vis 0.5: 8313 Vis 0.8: 8314 Vis 1.0: 8315 Vis 4.0: 8316

If you need a more precise value please see the custom products section on page 16.

Visible Standards

UV Standards

Available as sealed quartz cuvettes or bottles (60mL, 125mL, 250mL, 500mL, 1L)

Wave-length

vIS-0.05#8311

vIS-0.1#8312

vIS-0.3 #8322

vIS-0.5#8313

vIS-0.8#8314

vIS-1.0#8315

vIS-4.0#8316

400 0.122 0.260 0.724 1.254 1.882 2.436 3.437

450 0.082 0.177 0.500 0.871 1.323 1.780 3.326

500 0.059 0.126 0.360 0.628 0.957 1.298 3.238

550 0.043 0.093 0.270 0.470 0.719 0.975 3.127

600 0.034 0.072 0.212 0.368 0.566 0.767 2.917

650 0.027 0.056 0.169 0.294 0.450 0.611 2.524

700 0.022 0.045 0.137 0.239 0.365 0.497 2.111

750 0.018 0.037 0.111 0.195 0.299 0.406 1.740

800 0.017 0.030 0.092 0.161 0.247 0.334 1.440

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Wave-length

Uv-1#8301

Uv-2#8302

Uv-3#8303

Uv-4#8304

Uv-5#8305

Uv-6#8306

Uv-7#8307

Uv-8#8308

Uv-20#8309

Uv-30#8310

250 0.086 0.127 0.177 0.305 0.347 0.447 0.513 0.570 1.135 1.664

254 0.081 0.120 0.165 0.283 0.323 0.416 0.477 0.528 1.054 1.551

260 0.075 0.112 0.152 0.259 0.297 0.382 0.437 0.484 0.967 1.420

280 0.056 0.870 0.112 0.188 0.220 0.283 0.322 0.355 0.715 1.058

300 0.039 0.060 0.079 0.131 0.155 0.201 0.230 0.253 0.508 0.754

350 0.011 0.030 0.042 0.072 0.086 0.110 0.127 0.139 0.284 0.420

400 0.007 0.018 0.026 0.045 0.054 0.069 0.080 0.088 0.179 0.266

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Standards

In-Spec® Amaranth Red Standard Item #8414

In-Spec® Patent Blue Standard Item # 8416

Sizes Available60m l

125ml_250ml500ml

1lSealed Cuvette

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The In-Spec® Amaranth Red standard was designed to complement the In-Spec® Patent Blue but can be used ex-clusively for a variety of other processes. When used together, the In-Spec® Patent Blue and Amaranth Red standards cover a wider range of wavelengths that are associated with applications in pharma-ceutical and biotech laboratories. This reference standard is similar to the In-Spec® Patent Blue standard in that it can be used on microplate readers, traditional spectrophotometers, and small path-length instruments. Popular wavelengths for this standard are 280nm and 320nm.

This popular product can be customized to match a variety of wavelength and concentration levels. This reference standard is designed for small volume instruments and microplate readers but is equally adept on a traditional spectropho-tometer. Typical applications include analyses involving DNA, RNA, nu-cleic acids, proteins, and enzymes. A commonly utilized wavelength for this reference standard is 260nm.

Sizes Available60ml125ml250ml500ml1lSealed Cuvette

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Standards

In-Spec® Nano-Kit Item # 8476

In-Spec® BCA Purple Standard Item # 8587

The newest addition to the In-Spec® product line came about due to demand to validate colorimeters and spectrophotometers used in BCA pro-tein assays. The wavelength of interest for this application is 562nm. This reference standard is equally customi-zable as its Patent Blue and Amaranth Red counterparts.

Sizes Available60ml125ml50ml500ml1lSealed Cuvette

The In-Spec® Nano-Kit was awarded for the innovation in solving the issue of validating short pathlength instruments without a toxic material. Prior to the kit’s introduction the only option was hazardous potassium dichromate solutions. The In-Spec® Nano-Kit consists of our popular Patent Blue and Amaranth Red reference standards with standard data at 260nm, 280nm and 320nm. The data is certified with a 1mm pathlength cu-vette eliminating the conversion from the 10mm pathlength reading which is typical with many other standards. The reference standards are guaranteed for 1 year from the date of shipment unlike many com-peting materials that are a one-time use product.

Nano-Kit

 

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Wavelength AccuracyWavelength Accuracy Standards

Didymium Oxide Wavelength Standard Item # 8378 Peaks at: 298, 329, 355, 445, 469, 482, 512, 522, 576, 732, 740, 795, 802, 866nm

Holmium Oxide Wavelength Standard Item # 8340 Peaks at: 241, 250, 278, 287, 334, 345, 361, 386, 416, 451, 468, 485, 537, 641 nm

Samarium Oxide Wavelength Standard Item # 8379 Peaks at: 236, 267, 280, 290, 305, 318, 332, 345, 401, 416, 442, 464, 479 nm

The accuracy of your instrument’s wavelength scale is crucial when obtaining absorbance readings on your sample. A slight change of less than 1nm can cause an instrument’s read-ings to shift leading to incorrect data. This puts the laboratory at risk of passing a product that should have failed quality control.

GFS Chemicals offers 3 different wave-length reference filters that are used to validate the wavelength scale. These materials will add confidence to your quality control process.

Listed below are the wavelength refer-ence filters available with associated peaks. These materials are available in sealed quartz cuvettes certified for 4 years or in HDPE bottles with a 1 year shelf life. Recertification is available for the sealed quartz cuvettes.

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FiltersStray radiant light is energy reaching the detector that is not originating from the instrument light source. This can come from a variety of sources and impacts the readings output by the instrument. Stray light can be a problem at any wave-length but is more prevalent in the UV range.

The use of a stray light filter can verify that an external light source is not effecting the results. There are five commonly used materials to check the UV range of an instrument. Each standard has a ‘cut-off’ wavelength and eliminates light trans-mission below that level. Listed below are the standards offered and their associ-ated ‘cut-off’ wavelength. These are available in sealed quartz cuvettes certified for 2 years or sealed HDPE bottles with a 1 year shelf life. Recertification is available.

Stray Light Filters

Potassium Iodide: 260nmItem #: 8380

Lithium Carbonate: 227nmItem #: 8382

Potassium Chloride: 200nmItem #: 8384

Sodium Iodide: 260nm Item #: 8381

Sodium Chloride: 205nm Item #: 8383

Traditionally, potassium dichromate was used to check the photometric accuracy of an instrument. GFS Chemicals Inc. offers 5 different concentration levels. Using 3 or more of these values will allow you to check the linearity of your instru-ment. These standards come sealed in 10mm quartz cuvettes or in bottles and are designed for use in the UV spectrum. The sealed standards are certified for 1 year from shipment and recertification service is available. Custom concentrations are available. Please see the custom section on page 16.

Potassium Dichromate

Catalog Numbers20mg/L Item #835040mg/L Item #8368 60mg/L ltem #8369 80mg/L Item #8348 100mg/L Item #8372 Background Solution Item # 8349

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You may have a unique application that requires verification of certain absorbance values or wavelengths that aren’t listed for our stock products.

Our experienced staff has the ability to work with you to develop a refer-ence standard that will validate your instrument for your application.

Please call technical support for assistance.

In order to facilitate custom standard development there are several key pieces of data that are required. With this information, technical support will be able to quickly assess your needs.

• Wavelength(s) of interest• Pathlength• Instrumentation make and model• Desired or sample absorbance range

Our problem-solving lab team is available Monday through Friday from 7:30 am to 4:00 pm EST.

Custom

Custom Standards

AMCO Clear® Turbidity Standards

Amco Clear® is an EPA approved primary standard for cali-bration of turbidity sensors. It is available in a wide range of values. It has a guaranteed one year shelf life. The convenient submicron polymer suspensions are non-toxic primary calibration standards approved by the US EPA for drinking water analysis for turbidity. Amco Clear® is “The Clear Choice in Turbidity.”

AMCO Clear® Advantages: • Safe, non-toxic and disposable• Stable—does not settle out of suspension• Requires no mixing or inverting• Easy-to-Use/ No dilutions or preparations• Accurate to 1% lot-to-lot• Guaranteed one year shelf life from date of shipment• Available in a wide range of values• NIST Traceable• Custom Standards Available

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Application NotesSpecific Uv absorbance (SUva) for Total organic Carbon (ToC) analysis

Specific UV Absorbance (SUVA) is an analysis of drinking water that uses UV absorbance at 254nm to identify dissolved organic carbon (DOC) levels. UV absorbance is easily determined using a spectropho-tometer. Previous studies established a relationship between SUVA and the levels of humic substances that are removed during enhanced coagulation and/or enhanced softening. Utilizing this relationship allows for a cheap and easy method to determine whether enhanced coagula-tion or softening is required based off of the exemption allowed in Stage 1 D/DBPR and Stage 2 D/DBPR. Removing the extra disinfection from the process can save a water district money when it comes to TOC removal.

Under Stage 1 D/DBPR, Standards Method 5910B was the only EPA approved method for SUVA determination. Stage 2 D/DBPR approved EPA Method 415.3 (Revision 1.1, 2005) for TOC, DOC and SUVA determination. This one method not only combines all three tests, but also provides increased quality control procedures to ensure testing ac-curacy, including calibration verification procedures for all instruments utilized in the method.

According to the EPA method, the verification of the spectrophotom-eter can be done with a commercially prepared check standard. This method specifically mentions the In-Spec® photometric standards, manufactured by GFS Chemicals, Inc, which were utilized by the EPA to validate their spectrophotometer when developing 415.3. The In-Spec® photometric standards are NIST traceable, non-toxic, have a one-year shelf life and require no preparation to use.

Data provided at 254nm for In-Spec UV Standards 1 through 30.

Standards for validation of Microplate Readers & Short Pathlength Instruments

With growing demand to for smaller sample analysis, microplate readers are becoming more common place in the biotech/pharmaceutical industry.These instruments are typically used in performing various tests in relation to the analysis of DNA/ RNA, protein and enzymes. Many of the options that are used to verify the accu-racy of these instruments are either expensive or limited in the scope that they can check.

GFS Chemicals has developed standards to validate the accuracy of instruments with equal to or less than 1mm pathlengths. The most popular application at this time is for users checking at 260 and 280nm.

This type of validation is where the In-Spec® Color standards work perfectly. The color standards are designed to work with the built in software programs that manipulate the data obtained in the plate and displays it as optical density. The color standards have many useful wavelengths, as exhibited on page 8 and 9. These can also be made in a wide range of concentrations to better match the values you see in your daily testing.

The color standards are available in a wide range of sizes and have a 1 year shelf life so you can keep them on the shelf and ready to test your instrument when the need arises. All you need to do is pipet an aliquot into the wells on your plate and run them on the reader. Once the data is obtained you then compare that to the values on the certificate of analysis to verify that the instrument is performing as it should.

GFS Chemicals Inc. also offers Holmium Oxide solution in individual bottles so that you may verify the wavelength scale on your scanning microplate reader. This provides a much cheaper alternative to sealed plates with Holmium Oxide or a sealed quartz cuvette.

By utilizing both the color standards and the Holmium Oxide solution you will be able to say with confidence that the data you obtained on your microplate reader is accurate based off NIST-traceable standards.

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Notesamaranth red and its use in the Wine Industry

The In-Spec® Amaranth Red standard is ideal for use in the wine industry for some common testing that is performed on colorimeters or spectrophotometers. Wavelengths commonly tested in wine samples for color intensity, hue, and total phe-nolic concentration analysis are 520nm, 420nm, & 280 nm.

The color standards can be used to validate your instrument so that you know that it is performing as it should or it can be used as an inter-lab check so that you know your instrumen-tation in one lab is matching that in another lab. Over time an instruments light source may dim and the values you obtain will start to drift.

Knowing when this is occurring allows you to perform the necessary maintenance before the issue impacts the quality of the wine that you are producing. Multiple concentrations are available so that you may check a broad range of absorbance values.

Guide

TroubleShooting Guide

If your reference standards are not reading within the tolerance levels stated on the certificate of analysis utilize the following guide prior to calling for techni-cal support. Having the answers to the following questions will help expedite the troubleshooting process.

1) When was the last time the lamp was changed?a. GFS replaces the lamps in our instrument every 6 months, but with normal use, changing the lamps every 2 years should be sufficient.b. Many older spectrophotometers do not have diagnostic capabilities to tell the user when lamp energy is depleted or how many hours they have been in use.

2) Are your cuvettes/cells clean, scratch-free and suitable for your range of use?a. All data provided has been obtained in a 10mm FUV quartz cuvette unless stated in the certificate of analysis.b. Do not allow SDVB suspensions to dry on or inside your cells as it is difficult to remove.c. Disposable plastic cuvettes are generally not designed for the UV range unless specified on the package.

3) Was the supplied background solution* used to provide a baseline that reads 0.0?a. GFS has found that the “baseline” function alone may not provide a 0.000 ab-sorbance when the background solution is scanned later. It may take a combina-tion of “Auto zero” and “Baseline” over the range of interest.b. See Certificate of Analysis for appropriate lot number of background solution.

4) How recently has your instrument had a full calibration that included wavelength scale certification and stray light detection?a. GFS has this performed twice yearly by the manufacturerb. GFS provides NIST traceable wavelength and stray light check standards as well.

5) What spectral bandwidth are you using on your instrument?a. All provided data is for a 1nm bandwidth unless otherwise specified. b. Higher bandwidths often provide lower resolution and affect the values ob-tained.

*Supplied with In-Spec UV and Visible standards

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GFS Chemicals - Innovative Chemical Technology for Innovative Applications

Chemists and technicians can connect to a single company with manufacturing resources in all key areas of chemistry. Customers rely on us for our problem solving ability. Our proven technologies, technical expertise, engineering excellence and attention to detail allow us to work hand-in-hand with our customers every step of the way – from startup to commercialization. GFS technology and products have kept us steps ahead of the indus-try’s ever- changing chemical needs.

GFS Chemicals began when Professor G. Freder-ick Smith started producing perchloric acid ina small factory in Columbus, Ohio. Since then, GFS has grown to become a leading producer ofspecialty chemicals with performance-proven tech-nologies for laboratory, analytical, scientific,research and development, pilot scale production, and manufacturing applications.

Project Customization

Our production environment gives us the flexibility to offer you innovative solutions for all of your chemical needs. GFS research and development can support you with the chemicals that your application requires. Using our proven technology to customize your product, we create customer-specific item numbers and SKUs to streamline and safeguard your process. Our formalized and repeatable ISO manufacturing ensures that you get the same exact material every time. We package according to your requirements in small packages through drums.If your application requires more stringent condi-tions, we have experience with Good Manufac-turing Practice (GMP) processes. We are a key supplier of critical materials for numerous Fortune 100, GMP manufacturers. We understand the needs of our GMP customers – including pharma-ceutical manufacturers.

Unmatched Product Quality

GFS is an ISO 9001-2008 certified company. Our quality control team guarantees product consis-tency from lot-to-lot. We guarantee our products to meet the quality standards that we publish, and ship a Certificate of Analysis and a Material Safety Data Sheet (MSDS) with every order.

Environmental and Community Stewardship

GFS was founded on a business philosophy of community stewardship and respect for our envi-ronmental resources. As a long-standing member

of the Society of Chemical Manufacturers & Affili-ates (SOCMA) and the Ohio Chemistry Technology Council, GFS is committed to conducting our business with integrity and foresight. In addition, GFS is committed to full implementation of the SOCMA® ChemStewards® environmental, health, safety and security management system.

Why Choose GFS Chemicals?

• 80+ years of chemical manufacturing and chem-istry experience supporting process andcommercial applications• Knowledgeable chemists and operators who have a complete understanding of themanufacturing process• Scale-up flexibility from small sample sizes to drums to metric tons, to ensure consistent qualityfrom R&D to pilot to final production• International expertise shipping products to 85+ countries means your products will arrive on time,meeting all regulatory compliance regulations• Technical support and resources every step of the way• Dedicated, responsive customer support personnel and points of contact• Competitive pricing and reliable production lead times• U.S.-based quality manufacturing to ensure material availability, consistency and delivery• Experienced staff who understand the complexi- ties and dependencies of process chemistry

Associations and AffiliationsGFS Chemicals has been actively involved in the scientific community for generations. Member-ships include: • American Chemical Society • Society of Chemical Manufacturers and Associates (SOCMA)• Tier II ChemSteward® Certified for Environmental Health and Safety• American Water Works Association and Stan- dard Methods for the Examination of Water and Wastewater• Water Environment Federation• Ohio Water Environment Association• ASTM International - Committees D19, E13, E15 and E27• American Council of Independent Laboratories- Associate member• Ohio Manufacturers Association• Chemical and Specialties Management Council • Ohio Chemistry Technology Council• U.S. Chamber of Commerce• Ohio Chamber of Commerce• National Federation of Independent Businesses• National Association of Manufacturers• Tech Columbus

Analytical and Laboratory Chemicals

GFS manufactures over 6,000 organic and inorganic analytical reagent chemicals and specialties in a lean manufacturing operation. We have most chemicals needed for laboratory, bench, analytical, quality control, materials characterization, scientific or research work.

Our ultra-high purity double-distilled Veritas® and ACS Superior™ brand acids and Watermark® brand reagents are used throughout the world. Our AMCO Clear® Water Analysis Division manufactures the highest water calibration standards available for water treatment plants around the globe.

• Watermark® pyridine-free Karl fischer reagents• Veritas® hplC Solvents• HPLC/GC/UV-Vis Solvents• Reagent Grade Solvents• Spectroscopy Products• Primary Standard Salts• Normality Solutions • Colors and Indicators

• Silver Compounds• Reagents for Water & Wastewater labs• Organic Halogen Reagent – OHR/Halogen Level Field TestKits

• Polyethylene Glycol (small quantities)

• Conductivity Standards and ph Buffers

• AA and ICP Standard Solutions• Anion Standard Solutions• Standard Solutions for Trace Metal analysis

• ACS Reagents• ASTM Type I Deionized water

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Are you sure your instrumentation is properly reading your samples? Do your standard operating procedures require a vali-dation standard? Over time small changes in your instrumenta-tion can affect the data you obtain. Our reference standards are a high quality, in-expensive way to add confidence to your measurements.

GFS Chemicals Inc offers a full line of spectrophotometer validation standards.• Wavelength reference standards• Photometric reference standards• Stray Light reference standards

Contact information:e-mail in-spec@gfschemicals.com

800-838-5567 extension 121 for technical assistance, pricing and availability

(M-F 7:30 am to 4:00 pm Eastern Standard Time)614-824-3299

740-881-5989 FAX

Visit www.in-spec.com for technical information or Order online at www.gfschemicals.com

Team GFS

CHEMICALS ®